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Issue 1 ， 2019
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Research on Guidance of Prevention and Treatment of Vascular Diseases by Yingwei Theory of Vessel-collateral Theory
Abstract：The blood-vein theory is inherited and developed. Combined with the related theory features of Qi and blood of traditional Chinese medicine(TCM), the progress results in two national 973 plan projects undertaken are learned from, the vessel-collateral theory guiding the prevention and treatment of vascular lesions is systematically built, the academic development of TCM collateral disease discipline is pushed from " treatment of symptoms of collateral disease" to study on " vessel-collateral theory" , a new stage in the history. The pathogenesis, basic pathogenesis, clinical syndrome and syndrome differentiation therapy of vessel-collateral lesions are systematically studied, Yingwei theory, the core content of vessel-collateral theory, is proposed, the syndrome differential diagnosis standard of " vessel-collateral-vascular system disease" is established, and the blood-vessel theory of TCM is improved. The development of Yingwei theory of vessel-collateral theory is deepened. The theory of Yingwei sympathia and Yingwei's " opening by collaterals, intersecting and generating" are proposed to guide the study on prevention and control of vascular lesions. With tertiary collaterals-microvessel as a breakthrough point of research, four types of microscopic pathological features of microvessels are proposed. It is confirmed that dredging collaterals and protecting tertiary collaterals-microvessel is the common key mechanism for the treatment of major diseases of heart, brain and kidney(sugar). It protects microvessels, improves blood flow perfusion, protects tissue cells, and improves the viscera function. Dredging collaterals and protecting microvascular endothelial cells is the core mechanism of treating tertiary collaterals-microvessel lesions. It is proposed that protection of microvascular endothelial cells is a new strategy to solve the problem of microvascular lesions. After more than ten years, four studies of evidence-based medicine have been completed to solve the clinical problem of acute myocardial infarction without reflow. New drugs are provided to the international medical problem of cardiac insufficiency with premature ventricular contraction. The gap of fast and slow treatment, integration and rhythm adjustment of sinus bradycardia with premature ventricular contraction is filled. The clinical treatment effect of chronic heart failure is significantly improved, a major breakthrough in the treatment of microvascular lesions by TCM is achieved, and it is fully recognized and highly evaluated by the medical profession at home and abroad. It promotes the specialized subject platform construction and society development under the guidance of Yingwei theory of vessel-collateral theory, as well as the clinical application of syndrome differentiation diagnosis and treatment method and drugs of vessel-collateral lesions, and has significant social and economic benefits.Keywords：blood-vein theory;vessel-collateral theory;Yingwei theory;vessel-collateral-vascular system disease;Yingwei sympathia;tertiary collaterals-microvessel;microvascular disease- 102
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Coronary Heart Disease Phlegm-blood Stasis-stagnation-deficiency Innovation Theory and Clinical Application Based on Complex System
Abstract：There exits a trend of treatment for coronary heart disease(CHD) in terms of Chinese medicine(CM) in history, from warming Yang and dispelling cold, moving Qi and dissolving phlegm to invigorating blood and dissolving stasis, whose time-space development accords to the theory of complex system.Targeting the contemporary population of CHD, phlegm-blood stasis-stagnation-deficiency theory is proposed to direct clinical application to further improve effectiveness.From the perspective of theory, blood stasis-stagnation is the basic pathogenesis of CHD that runs through the whole period in the advance of CHD, while phlegm has been changing the feature of CM Zheng of CHD population with the development of the society, and nourishing the body according to the type of deficiency might delay the process of growing older, postpone the occur of disease and hinder the advance of disease, which genuinely differs from modern medicine to render the superiority of CM to work out the challenges of CHD.As for the clinical application, the treatment options could consider invigorating blood, dissolving stasis and removing stagnation as the basic method, such as CHD formula 2, Xuefu Zhuyutang, Zhizi Chitang and Danggui Niantongtang could be used in varied period of CHD.Moreover, boosting the body by herbal medicine could regulate the routine of body, then to prevent CHD and fear down the advance of CHD, for instance, Codonopsis Radix, Astragali Radix, Liuwei Dihuang pills, Zuoguiyin and Youguiyin.Furthermore, balmiest herbs could not only dissolve phlegm, unblock the meridian and relieve pain, but also calm the mind and eliminate the anxiety.For example, Aquilariae Lignum Resinatum, Santali Albi Lignum and Moschus could be used handily in the treatment of CHD.Keywords：phlegm-blood stasis-stagnation-deficiency;coronary heart disease;complex system;innovation theory;clinical application;promoting blood circulation to remove blood stasis;nourishing and conditioning- 19
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Monograph
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Establishment of DNA Fingerprints and Genetic Similarities Analysis of
Dendrobium officinale Abstract：This study adopted the 4 pairs of simple sequence repeats(SSR)primers selected by National Resource Center for Chinese Meteria Medica (CACMS) to detect 29 populations of Dendrobium officinale samples from 9 main places of production; 5, 7, 4 and 3 polymorphism bands were amplified from these 4 pairs of SSR primers. The DNA identities of different populations were constructed by SSR. The 29 D. officinale populations could be divided into 4 classes. The clustering result was related to the places of production. Samples from Yunnan, Guizhou, Sichuan provinces were classified into one category, while samples from Anhui and Guangxi provinces were classified into another category. Samples from Guangdong Danxia, Zhejiang Yongkang, Zhejiang Leqing and Taining belonged to a category. PopGene (version 1.32) software was applied to calculate the genetic similarity of the 29 D. officinale populations. The similarities was between 0.403 4 and 1.0.Based on the genetic similarity, the genetic consistency included three classes, A, B and C. Samples with a similar geographical location and landform environment have higher genetic similarities, which indicate the same genetic background. This paper provides reference information to study the identification, selection and breeding of good varieties.Keywords：Dendrobium officinale;DNA molecular identities;simple sequence repeats(SSR);genetic similarities- 13
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
HPLC Characteristic Spectrum Optimization of Flavonoid Glycosides on
Dendrobium officinale and Characteristics Analysis of Different ProvenancesAbstract：Objective:To optimize the HPLC characteristic spectrum of flavonoid glycosides of Dendrobium officinale, and identify the common and specific components of different provenances.Method:Kromasil 100-5 C18 column was adopted, with tetrahydrofuran-acetonitrile-methanol (10∶22∶5)-0.05% phosphoric acid as mobile phase (gradient elution). The detection wavelength was 340 nm, the column temperature was 30 ℃, and the flow rate was 1.0 mL·min-1.Result:13 flavonoid characteristic peaks were marked in 27 batches of D. officinale, and 7 characteristic peaks of 6 flavonoid C-glycosides (vicenin Ⅱ, vicenin Ⅰ, schaftoside, isoschaftoside, violanthin and isoviolanthin) and one flavonoid O-glycosides (rutin) was identified. 7-11 characteristic peaks were detected in different batches of samples. Among them, vicenin Ⅱ was a relatively stable common peak in different source samples, and the characteristic peaks of rutin, schaftoside and isoschaftoside were quite different. According to the relative abundance of the characteristic peaks, the samples could be divided into three categories. Among them, the first category had 10 batches of samples, which mainly came from Danxia landforms of Guangdong, Jiangxi, Fujian and Zhejiang (Wuyi) Province (which called " Danxia landform species" ) and characterized by detection of obvious peak of rutin. The second category had 11 batches of samples, which mainly came from Yunnan and Guangxi Province (which included " Yunnan Guangnan species" and " Guangxi Tiepilan species" ) and characterized by detection of violanthin and isoviolanthin. And the third category had 6 batches of samples, which were mainly derived from Zhejiang Province (which called " native species from Zhejiang" ) and characterized by detection of different degrees of rutin peak, but it was difficult to detect the characteristic peaks of violanthin and isoviolanthin. HPLC characteristic chromatograms of D. officinale in bionics wild cultivation and greenhouse of " Danxia landform species" and " Guangxi Tiepilan species" were compared. The results showed that the characteristic peaks in D. officinale planted in greenhouse could be detected stably, which verified the reliability of the source in D. officinale.Conclusion:The analytical method has a better separation effect on flavonoids of D. officinale, with a good reproducibility. The commonness and specificities of flavonoid glycosides components of D. officinale from different categories have basically confirmed. This suggests that Vicenin Ⅱ is suitable to be a reference peak for characteristic chromatogram. Both the relative abundance of rutin and the detection or relative abundance of violanthin and isoviolanthin peaks could be used as a reference to judge the categories of D. officinale in " Danxia landform species" or " Tiepilan species from Yunnan, South Guangdong and Guangxi" or " native species from Zhejiang" .Keywords：Dendrobium officinale;characteristic chromatogram;flavonoid glycosides;Danxia landform species;Yunnan Guangnan species;Guangxi Tiepilan species;native species from Zhejiang- 11
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Isolation and Identification of Main Flavonoid Glycosides of
Dendrobium officinale from Danxia Species and Yunnan Guangnan SpeciesAbstract：Objective:To extract, isolate, purify and identify the structures of the flavonoid glycoside in Dendrobium officinale from two different origin places (Danxia species and Yunnan Guangnan species), and provide experimental reference for confirming the common flavonoid glycoside components in D. officinale.Method:①70% ethanol was applied to extract the total flavonoids in leaves of D. officinale from two different species. Organic solvents petroleum ether, acetic ether and water saturated n-butyl alcohol were used in turn to extract the crude extraction. Then AB-8 Macroporous resin, Sephadex LH-20 and ODS chromatographic column were applied to isolate and purify the water saturated n-butyl alcohol extraction fraction. The structures of flavonoid glycoside were identified by studying physicochemical property, applying modern spectroscopy method like HPLC, ESI-MSn, 1H-NMR, 13C-NMR, etc. ②HPLC characteristic spectrum technique was used to analyse and compare the common flavonoid glycoside components in Dendrobium officinale from different origin places (Danxia species, Yunnan Guangnan species, Guangxi Tiepilan species and Zhejiang native species).Result:Five flavonoid glycoside compounds were isolated from the crude extractions of the leaves of D. officinale from two different species, and they were identified as rutin, vicenin Ⅱ, viceninⅠ, violanthin and isoviolanthin. The characteristic spectrum of vicenin Ⅱ and viceninⅠwere detected in stems of D. officinale from four different origin places (Danxia species, Yunnan Guangnan species, Guangxi Tiepilan species and Zhejiang native species), and vicenin Ⅱ had a better separation degree in the characteristic spectrum. However, the characteristic spectrum of violanthin and isoviolanthin were more obvious in Yunnan Guangnan species and Guangxi Tiepilan species, while rutin was obvious in the Danxia species.Conclusion:Vicenin Ⅱis the common flavonoid glycosides component in D. officinale from different origin places (Danxia species, Yunnan Guangnan species, Guangxi Tiepilan species and Zhejiang native species), and can be used as the internal reference material for the characteristic spectrum of D. officinale.Keywords：Dendrobium officinale;flavonoid glycoside;rutin;vicenin Ⅱ;vicenin Ⅰ;volanthin;isoviolanthin- 8
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Content Comparison of Mannose, Glucose and Narinhenin in
Dendrobium officinale andDendrobium huoshanense Abstract：Objective:To optimize the pre-column derivation high performance liquid chromatography (HPLC) content determination method of D-mannose and D-glucose as well as the content determination method of narinhenin in Dendrobium officinale and D. huoshanense, and compare the contents of D-mannose, D-glucose and narinhenin between D. officinale and D. huoshanense.Method:A pre-column derivation HPLC method modified by Chinese Pharmacopoeia(Ch.P) 2015 was used to simultaneously determine the contents of D-mannose and D-glucose, with acetonitrile-0.02 mol·L-1 ammonium acetate solution as mobile phase for gradient elution. Kromasil 100-5 C18 was performed with the wavelength set at 250 nm, and the flow rate was 1 mL·min-1; column temperature was 30 ℃. HPLC content determination of narinhenin was performed on Kromasil 100-5 C18 with the acetonitrile-methanol-0.4% phosphoric acid solution as mobile phase for gradient elution, and the wavelength was set at 290 nm; the flow rate was 0.8 mL·min-1, and column temperature was 40 ℃.Result:D-mannose and D-glucose showed a good linear relationship within the range of 0.15-3.0 μg and 0.075-2.25 μg (r=0.999 9); and their average recoveries were 99.01% (RSD 2.1%) and 101.69% (RSD 2.0%) respectively. In addition, the other methodological researches such as repeatability and durability all met the requirements. The contents of D-mannose(Cm), D-glucose(Cg) and sum of them (Cm+ Cg) were 12.75%-36.40%, 2.93%-18.39% and 19.23%-54.58% in 43 batch of D. officinale. Almost all of the results except very few samples reached the D-mannose standard in Ch.P 2015, and the total content of D-mannose and D-glucose was also up to the total polysccharide standard in Ch.P. The correlation between content and origin was not significant. The contents of D-mannose(Cm), D-glucose(Cg) and sum of them (Cm+ Cg) were 14.33%-29.47%, 6.64%-15.20%, and 25.73%-44.37% in 12 batch of D. huoshanense. These contents and ratio of peak areas of D-mannose to D-glucose (Am/Ag) were within the scope of D. officinale's; in addition, their average contents were basically the same with those in D. officinale (about 33%). Next, naringenin showed a good linear relationship within the range of 0.020 8-0.832 0 μg (r=0.999 9), and its average recovery was 101.96% (RSD 1.8%). The content of naringenin was 0.053 2-0.122 4 mg·g-1 (average value of 0.081 0 mg·g-1) in 11 batch of D. officinale, slightly higher than 0.040 3-0.090 0 mg·g-1 (average value of 0.068 3 mg·g-1 ) in 7 batch of D. huoshanense. All of these results of narinfenin did not reach the content lower limit in Ch.P.Conclusion:The method used to determinate the content of D-mannose and D-glucose is reproducible, and their sum content is possible to substitute the total polysccaride determination (with higher errors) in D. officinale; monosaccharide content determination can be used for quantitative quality control of D. huoshanense. However, it could not distinguish D. officinale and D. huoshanense by determining the contents of polysccharide, D-glucose, D-mannose and narinhenin, and shall be combined with other specificity methods for further identification.Keywords：Dendrobium officinale;D. huoshanense;D-glucose;D-mannose;naringenin;determination;HPLC- 9
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Antioxidation Activities in Vitro of Vicenin Ⅱ Isolated from Dendrobii Officinalis Caulis and Effect on HepG2 Cells
Abstract：Objective:To study the antioxidation activities in vitro of a comment flavonoid component named vicenin Ⅱ(Apigenin 6, 8-di-C-glucoside)in Dendrobii Officinalis Caulis from different origin places and investigate its effects on apoptosis of HepG2 cells.Method:The antioxidation activities in vitro of vicenin Ⅱ (0.005-1 g·L-1)were evaluated by 2, 2-diphenyl-1-picrylhydrazyl (DPPH), salicylic acid and 2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid(ABTS)and copper ion reduction assays. Methye thiazolye telrazlium(MTT)assay was used to test the inhibitory effect of vicenin Ⅱ(12.5~100 μmol·L-1)on proliferation of 6 tumour cells in vitro. In subsequent apoptosis experiment, the concentration of vicenin Ⅱ was 75 μmol·L-1. The morphological changes of HepG2 cells were evaluated by Hoechst 33258 under fluorescence microscope; and the cell apoptosis rate was detected by flow cytometry with AnnexinV/PI apoptosis assay kit. The mRNA expressions of mitogen activated protein kinase (MAPK) pathway related apoptotic genes were detected by Real-time PCR assay.Result:The 1 g·L-1 vicenin Ⅱ showed 48.82% and 22.01% for DPPH scavenging rate and Cu2+ reduction rate respectively(P<0.01). 0.5 g·L-1 vicenin Ⅱ showed 86.88% for ABTS scavenging rate(P<0.01). vicenin Ⅱ could significantly inhibit the proliferation and increase the apoptosis rate on HepG2 cells; after treatment for 48 h with 75 μmol·L-1 Vicenin Ⅱ, the cells survival rate was 45.69%(P<0.01)and apoptotic rate was 14.57%(P<0.01). Meanwhile, the mRNA expression levels of B-cell lymphoma-2 (Bcl-2) related X protein (Bax)/Bcl-2, Caspase-8, p38, extracellular signal-regulated kinases (ERK), c-Jun N-terminal kinase (JNK), and nuclear transcription factor (NF)-κB were increased(P<0.01).Conclusion:The general flavone glycosides component vicenin Ⅱ of Dendrobii Officinalis Caulis from different origins has a certain antioxidation effect and significant inhibitory effect on proliferation, and could induce apoptosis on HepG2 cells probably by regulating the expression of related genes in MAPK pathway and Bax/Bcl-2.Keywords：Dendrobii Officinalis Caulis;flavonoid glycoside;vicenin Ⅱ;antioxidant;HepG2 cell;apoptosis- 12
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Topic on Germplasm Resources of Dendrobii Officinalis Caulis
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Progress in Research of Kangxianling Decoction in Delaying Renal Fibrosis
Abstract：Renal fibrosis is a common pathological change in the later stages of all kidney diseases. It is a multi-cytokine, multi-signal pathway, multi-factor driven chronic kidney disease. It includes renal interstitial fibrosis, tubular sclerosis and glomerular sclerosis, which eventually leads to chronic renal failure. From health through injury to loss of function, the disease process is closely related to the degree of deterioration of renal function and the prognosis of chronic kidney disease. There is no effective western medicine for the treatment of renal fibrosis. Professor HE Liqun from Shuguang Hospital affiliated to Shanghai University of Traditional Chinese Medicine has summarized the Kangxianling decoction through decades of long-term practical experience. It has the effects in strengthening the spleen and replenishing Qi, clearing away dampness and heat, promoting blood circulation and removing phlegm, and strengthening turbidity. It is composed of Salviae Miltiorrhizae Radix et Rhizoma 15 g, Persicae Semen 12 g, Angelicae Sinensis Radix 12 g, Achyranthis Bidentatae Radix 9 g, Rhei Radix et Rhizoma 15 g. Kangxianling can affect the synthesis and secretion of cytokines and inflammatory factors by expanding blood vessels, and can improve renal tubular fibrosis. It has a good multi-channel, multi-target and multi-directional protective effect on renal function. It also can delay the progress of chronic renal fibrosis by significantly alleviating such symptoms as fatigue and edema in patients with chronic renal fibrosis, and reducing serum creatinine, urea nitrogen and urine protein. In this paper, 5/6 nephrectomy, ischemia reperfusion injury, adriamycin-induced nephropathy, unilateral ureteral obstruction and other different modeling methods are listed. The mechanism of Kangxianling decoction in antagonizing renal fibrosis is discussed and summarized, which further provided new ideas and directions for future clinical and scientific research.Keywords：renal fibrosis;Kangxianling decoction;5/6 nephrectomy;ischemia reperfusion injury;adriamycin-induced nephropathy;unilateral ureteral obstruction;Taohong Siwutang- 13
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect of Kangxianling Decoction on Renal Fibrosis and ACE-AngⅡ-AT1R Axis in 5/6 Nephrectomized Rats
Abstract：Objective:To study the therapeutic effect of Kangxianling decoction on renal fibrosis induced by 5/6 nephrectomy, and angiotensin converting enzyme-angiotensin Ⅱ-angiotensin Ⅱ 1 receptor (ACE-AngⅡ-AT1R) axis.Method:Totally 50 SD rats were randomly divided into the following groups: control group (n=10), sham-operation group (n=10), 5/6 nephrectomized renal fibrosis model group (n=30). After two weeks, the rats in operation group were divided into the model group, Kangxianling group, and losartan potassium group, n=10 in each group. Rats in losartan potassium group were administered with losartan potassium by gastrogavage, and rats in Kangxianling group were administered with Kangxianling by gastrogavage. Equal volume of saline was administered to rats in the other groups. The rats were put to death after 16 weeks of consecutive medication, and serum creatinine(SCr), blood urea nitrogen(BUN), 24 h urine protein(24 h-Pro) were measured in each group. Hematoxylin-eosin(HE) staining was used to observe the pathological changes of kidney tissues, and the degree of renal fibrosis was observed by Masson staining. The expressions of ACE1 and AT1R were detected by immunohistochemistry. The protein expression levels of ACE1, AngⅡ and AT1R were determined by Western blot.Result:Compared with control and sham-operation groups, SCr, BUN and 24 h-Pro in model group were significantly increased (P<0.01). Compared with model group, SCr, BUN and 24 h-Pro levels in the Kangxianling group and the losartan potassium group were significantly lower, with statistically significant differences (P<0.05, P<0.01). HE and Masson staining showed that the degree of renal fibrosis was reduced in Kangxianling group and losartan potassium group compared with model group. Western blot showed that ACE1, AngⅡ and AT1R in Kangxianling group, and the losartan potassium were significantly lower than the model group, with statistically significant differences (P<0.01). Immunohistochemistry results showed that the levels of ACE1 and AT1R showed the same changes as Western blot.Conclusion:Kangxianling decoction can delay the progress of renal fibrosis in 5/6 nephrectomized rats, which is closely related to the inhibition of ACE-AngⅡ-AT1R axis activation .Keywords：5/6 nephrectomy;renal fibrosis;Kangxianling decoction;angiotensin converting enzyme-angiotensin Ⅱ-angiotensin Ⅱ 1 receptor (ACE-AngⅡ-AT1R)- 11
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect of Kangxianling Decoction on Renal Function and Kidney ECM Expression in Rats with Renal Fibrosis
Abstract：Objective:To investigate the effect of Kangxianling decoction on renal function and expression of extracellular matrix(ECM) in renal tissue of rats with renal fibrosis induced by 5/6 nephrectomy.Method:Totally 50 SD rats were randomly divided into control group (n=7), sham-operation group (n=7), operation group (n=36). The 5/6 nephrectomized renal fibrosis model was established in operation group. After successful modeling, rats were randomly divided into model group, Kangxianling group and losartan potassium group. The losartan potassium group and the Kangxianling group were given losartan potassium (33.3 g·kg-1) and Kangxianling decoction (21 g·kg-1) every day. The control group, sham-operation group and model group were all treated with equal volume of normal saline. Rats were put to death after 24 weeks of consecutive medication, and renal function, 24 h urine volume and 24 h-Pro quantitation of each group were measured. The expressions of collagen Ⅰ(ColⅠ) and ColⅢ in renal tissues were determined by immunohistochemistry and Western blot.Result:Compared with the normal group and sham-operation group, serum creatinine(SCr), blood urea nitrogen(BUN), 24 h urine volume, 24 h-Pro quantitation, renal tissue ColⅠ and ColⅢ expression levels were significantly increased in the model group (P<0.01). Compared with the model group, the levels of SCr, BUN, 24 h-Pro quantitation, protein expression levels of renal tissue ColⅠ and ColⅢ were significantly decreased in the Kangxianling group and the losartan potassium group (P<0.05, P<0.01).Conclusion:Kangxianling decoction can down-regulate the expressions of ColⅠ and ColⅢ in kidney tissue, there by inhibiting the accumulation of ECM, and exerting the effect in delaying renal fibrosis and protecting renal function.Keywords：Kangxianling decoction;extracellular matrix(ECM);renal fibrosis;collagen Ⅰ;collagen Ⅲ- 11
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect of Kangxianling Decoction on TGF-β1/Smad Signal Pathway in Renal Fibrosis Rats
Abstract：Objective:To study the therapeutic effect of Kangxianling Decoction on renal fibrosis induced by 5/6 nephrectomization in rats, and discuss its action mechanism.Method:Totally 50 SD rats were randomly divided into normal control group (n=10), sham-operation group (n=10), and 5/6 nephrectomized renal fibrosis model group (n=30). After successful modeling, rats were randomly divided into the model group, Kangxianling group, and Losartan Potassium group. Rats in the Losartan Potassium group were administered with Losartan Potassium by gastrogavage; rats in the Kangxianling group were administered with Kangxianling by gastrogavage. Equal volume of distilled water was administered to rats in the other groups. Rats were sacrificed after 8 weeks of consecutive medication, and then serum creatinine (SCr), urea nitrogen (BUN), 24 hour urine protein (24 h-Pro) were measured in each group. Hematoxylin-eosin (HE) staining was used to observe the renal pathological changes and the score of Kidney damage was made. Masson staining was used to observe the degree of renal fibrosis. Western blot and real-time fluorescent quantitative polymerase chain reaction(Real-time PCR)were used to detect the protein and mRNA expression levels of transforming growth factor (TGF-β1), Smad2, Smad3, and Smad7 in kidneys.Result:As compared with the normal control and sham-operation group, the renal tissue fibrosis, score of kidney damage, SCr, BUN and 24 h-Pro levels were higher in model group (P<0.01), with increased protein and mRNA expression levels of TGF-β1, Smad2, and Smad3 (P<0.01), and significantly reduced protein and mRNA expression levels of Smad7 in the model group (P<0.01). As compared with the model group, the renal tissue fibrosis, the score of kidney damage, SCr, BUN, 24 h-Pro level were significantly decreased (P<0.01), protein and mRNA expression levels of TGF-β1, Smad2, Smad3 were reduced (P<0.05), while protein and mRNA expression levels of Smad7 were elevated in the Losartan Potassium group and Kangxianling group (P<0.01), but lower than those of normal control group and sham-operation group.Conclusion:Kangxianling decoction could alleviate renal fibrosis by inhibiting TGF-β1/Smad signal pathway.Keywords：Kangxianling decoction;renal fibrosis;5/6 nephrectomy;transforming growth factor-β1 (TGF-β1);Smad2;Smad3;Smad7- 11
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Topic on Treatment of Renal Fibrosis with Kangxianling Formulas
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Mechanism of Liuwei Dihuangwan in Enhancing Hepatocarcinoma Suicide Gene Therapy from Cx32
Abstract：Objective:To investigate the effect of Liuwei Dihuangwan on connexin 32 (Cx32) in hepatoma cell line CBRH7919 and its gap junction intercellular communication (GJIC), and furthermore study its mechanism of enhancing the bystander killing effect of suicide gene therapy.Method:Liuwei Dihuangwan (32 g·kg·d-1) and the same volume of normal saline were given to the rats by intragastrical administration. Blood was taken to prepare the medicated serum of Liuwei Dihuangwan and blank control serum, respectively. The hepatoma cell line CBRH7919 were treated by control serum and medicated serum of Liuwei Dihuangwan in different concentrations. There were four groups in experiment: the blank control group (volume fraction of 10%), medicated serum high dose group of Liuwei Dihuangwan (the volume fraction of 10%), medicated serum middle dose group of Liuwei Dihuangwan (the volume fraction of 5%), and medicated serum low dose group of Liuwei Dihuangwan (the volume fraction of 2.5%). The expression levels of Cx32 protein and mRNA in hepatoma cell line CBRH7919 were detected by indirect immunofluorescence assay (ⅡA) and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) assay. The fluorescence redistribution after photobleaching (FRAP) method was used to detect the function of GJIC of hepatoma cell line CBRH7919.Result:① The indirect immunofluorescence assay (ⅡA) analysis indicated that as compared with the blank control group, the cx32 expression of CBRH7919 cells was up-regulated in a concentration-dependent manner in each dose group of the serum containing Liuwei Dihuangwan (P<0.05 in low dosage group; P<0.01 in middle dosage and high dosage group), especially on cell membrane.② The results of Real-time PCR assay indicated that as compared with the blank control group, the expression of Cx32 mRNA was increased in each dose group (P<0.05 in low dose group, P<0.01 in middle and high dose group) in a dose-effect relationship. ③ The result of fluorescence redistribution after photobleaching (FRAP) analysis indicated that the average fluorescence recovery rates of drug groups were significantly higher than that in blank control group, and also showed a certain concentration-dependent trend.Conclusion:The mechanism of medicated serum of Liuwei Dihuangwan in enhancing the bystander killing effect of suicide geneis related to gap junction. Liuwei Dihuangwan may enhance the function of GJIC by increasing the localization of cx32 on the cell membrane of CBRH7919 cells and increasing the expression of cx32 mRNA and protein to achieve the synergistic action.Keywords：hepatocarcinoma cell;Liuwei Dihuangwan;serum pharmacology;gene therapy;bystander killing effect;gap junction- 8
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect of Modified Lichongtang Combined with 5-Fluorouracil on Epithelial Mesenchymal Transition in H22 Tumor-bearing Mice
Abstract：Objective:To investigate the effect of modified Lichongtang combined with 5-fluorouracil (5-FU)on tumor epithelial-mesenchymal transition (EMT)in H22 tumor-bearing mice.Method:Mouse models of transplanted hepatoma were constructed. After tumor formation, they were randomly divided into 4 groups: blank group, 5-FU group(2.5 mg·kg-1 5-FU intraperitoneal injection), modified Lichongtang combined with 5-FU group (5-FU+ Chinese medicine group), and modified Lichongtang group (Chinese medicine group, 25 g·kg-1 gavage), n=10 in each group. The effect of modified Lichongtang combined with 5-FU on the tumor inhibition rate of subcutaneous transplanted tumor was observed. The gene expression levels of E-cadherin, N-cadherin, Snail, and Twist in transplanted tumor were observed by Real-time PCR. The protein expression levels of E-cadherin, N-cadherin, Snail, and Vimentin were detected by using Western blot.Result:The tumor inhibiting rate was 59.18%, 84.42%, and 10.39% respectively in 5-FU group, 5-FU+ Chinese medicine group, and Chinese medicine group. All of these can inhibit the growth of liver cancer transplantation tumor, and the tumor inhibiting rate of 5-FU+ Chinese medicine group was significantly higher than that in 5-FU group (P<0.01). Thymus index and spleen index in 5-FU+ Chinese medicine group were significantly higher than those in 5-FU group (P<0.05, P<0.01). Results of Real-time PCR indicated that, as compared with blank group, expression levels of E-cadherin were up-regulated, but expression levels of N-cadherin, Snail, and Twist were down-regulated in 5-FU group, 5-FU+ Chinese medicine group, and Chinese medicine group (P<0.05, P<0.01). As compared with the 5-FU group, expression levels of E-cadherin were up-regulated, but expression levels of N-cadherin, Snail, and Twist were down-regulated in the 5-FU+ Chinese medicine group (P<0.05, P<0.01). Results of Western blot showed that, as compared with blank group, expression levels of E-cadherin were up-regulated, but expression levels of N-cadherin, Snail, Vimentin were down-regulated in 5-FU group, 5-FU+ Chinese medicine group, and Chinese medicine group (P<0.05, P<0.01); as compared with 5-FU group, expression levels of E-cadherin were up-regulated, but expression levels of N-cadherin, Snail, and Vimentin were down-regulated in the 5-FU+ Chinese medicine group (P<0.05, P<0.01).Conclusion:Modified Lichongtang, 5-FU and their combination have inhibitory effect on the growth of transplanted tumors of hepatocarcinoma mice, and the combination of the two drugs can enhance the effect of chemotherapy and to some extent inhibit the toxicity of 5-FU. The mechanism may be related to the inhibition of liver cancer EMT.Keywords：modified Lichongtang;H22 hepatocarcinoma cells;tumor-bearing mice;subcutaneous transplanted tumor;epithelial-mesenchymal transition- 7
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Protective Effect of Renshen Sinitang and Its Active Ingredients on Myocardial Cell Injury Induced by Pentobarbital Sodium
Abstract：Objective:To explore the protective effect and mechanisms of Renshen Sinitang and its active ingredients on cardiomyocyte injury induced by pentobarbital sodium.Method:H9C2 cells were sub-cultured with ginsenoside Rb2 0.01, 0.1, 1 μmol·L-1, Re 0.01, 0.1, 1 μmol·L-1, isoliquiritigenin 20, 40, 80 μmol·L-1, glycyrrhetinic acid 10, 20, 40 μmol·L-1, Renshen Sinitang, 10, 100, 400 mg·L-1, for 4 h. After treatment with 0.1% of sodium pentobarbital for 30 min, cell viability, lactate dehydrogenase (LDH), lipid peroxide malondialdehyde (MDA), Na+ -K+ -adenosine triphosphate(ATP)ase, Ca2+ -ATPase activity, and real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) were used to detect the expressions of peroxisome proliferative activated receptor-1α (PGC-1α), B-cell lymphoma-2 associated X protein(Bax) and cysteine aspartate-specific protease-3(Caspase-3) mRNA.Result:Renshen Sinitang and its active ingredients have a protective effect on heart failure cell model. Compared with the normal group, the cell survival rate of the model group decreased significantly, while the LDH and MDA contents increased significantly, and the Na+ -K+ -ATPase activity increased. Ca2+ -ATPase activity was significantly decreased, PGC-1α mRNA expression was down-regulated, Bax and Caspase-3 mRNA expressions indicates the modeling(P<0.01) . Compared with the model group, each administration group showed a significantly increased cell viability, decreased LDH, MDA content, inhibited Na+ -K+ -ATPase activity, increased Ca2+ -ATPase activity, up-regulated PGC-1α mRNA expression, and inhibited Bax and Caspase-3 mRNA expression (P<0.05, P<0.01).Conclusion:Renshen Sinitang and its active ingredients have a significant protective effect on heart failure cell model, and its mechanisms of action are related to anti-oxidation, improvement of mitochondrial energy metabolism and inhibition of mitochondrial apoptosis pathway.Keywords：heart failure;Renshen Sinitang;malondialdehyde(MDA);Na+ -K+ -adenosine triphosphate(ATP)ase;Ca2+ -ATPse;B-cell lymphoma-2 associated X protein(Bax)- 7
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect of Shenghuitang on Learning Memory, Levels of IL-6, TNF-α and COX-2 in Hippocampus of Chronic Sleep Deprivation Mice
Abstract：Objective:To observe the effect of Shenghuitang on learning and memory and expressions of interleukin-6(IL-6), tumor necrosis factor-α(TNF-α) and cyclooxygenase-2(COX-2) in hippocampus of chronic sleep deprived mice, in order to explore the possible mechanism of Shenghuitang in improving learning and memory ability.Method:Mice were randomly divided into sleep deprivation group, blank group, melatonin group(7.8×10-4 g·kg-1·d-1), high, middle and low-dose Shenghuitang groups(54, 27, 13.5 g·kg-1·d-1). The model of chronic sleep deprivation in mice was established using the " multi-platform water environment method" . 28 d sleep deprivation and intragastric administration were provided. Morris water maze was used to detect the learning and memory ability of mice in each group. Real time-PCR was used to detect mRNA expressions of IL-6, TNF-α and COX-2 in the hippocampus of each group.Result:The results of Morris water maze test showed that compared with the blank group, the total time spent on finding the platform and the total swimming distance of the model group were significantly prolonged (P<0.01), while the number of crossing platforms and the target quadrant were significantly reduced (P<0.01). The time for the original platform was significantly increased (P<0.01). Compared with the model group, the total time spent on finding the platform and the total swimming distance decreased significantly in each drug-administered group (P<0.05, P<0.01) reduced, whereas the number of times for crossing the platform and the target quadrant increased significantly (P<0.05, P<0.01). The time for the first arrival of the original platform was significantly reduced (P<0.05, P<0.01). The results of RT-PCR showed that mRNA expressions of IL-6, TNF-α, and COX-2 were increased in the model group compared with the blank group. Compared with the model group, mRNA expressions of IL-6, TNF-α, and COX-2 were decreased in the treated group. COX-2 mRNA expression was down-regulated.Conclusion:Shenghuitang may improve the learning and memory ability of mice by decreasing mRNA expressions of IL-6, TNF-α and COX-2 in hippocampus.Keywords：Shenghuitang;sleep deprivation;water maze;inflammatory factors- 9
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect of Wendantang on PI3K, Akt and GSK3β of Hippocampus in Rats with Schizophrenia
Abstract：Objective:To research the effect of Wendantang on phosphatidylinositol-3-kinases(PI3K)/protein kinase B(Akt)/glycogen synthase kinase 3β(GSK3β) signaling pathway of hippocampus in rats with schizophrenia.Method:Sixty SD rats were randomly divided six groups: normal group (A), model group (B), Clozapine group (C), high-dosage group Wendantang (D), medium-dosage Wendantang group (E) and low-dosage Wendantang group (F), with 10 rats in each group. The rats of normal group and model group were given normal saline. The rats of Clozapine group were given 20 mg·kg-1 Clozapine. The rats of high-dosage, medium-dosage, low-dosage Wendantang groups were respectively given 40, 20, 10 g·kg-1 Wendantang, once a day, for 21 days.Two hours later after the last administration with Wendantang or saline, except for group A, groups B, C, D, E, F were intraperitoneally given MK-801 0.6 mg·kg-1 to establish the rat schizophrenia model. After modeling, the changes in stereotyped behavior of each group were observed and recorded. After three days, the rats were put to death, and the hippocampus tissue were tested. According to Sams Dodd and Hoffman's standard, the stereotyped behavior of rats was scored. The protein expressions of PI3K, Akt and GSK3β of hippocampus were determined by Western blot. The mRNA expressions of PI3K, Akt and GSK3β of hippocampus were tested by Real-time PCR.Result:Compared with A, the stereotyped behavior score of the B was significantly increased (P<0.01). The expressions of PI3K, Akt, GSK3β protein and mRNA of hippocampus were significantly decreased (P<0.01) in rats of the B. Compared with the B, Wendantang administration groups could reduce the stereotyped behavior score(P<0.05), increased significantly the expressions of PI3K, Akt, GSK3β protein and PI3K, Akt and GSK3β mRNA of hippocampus (P<0.05, P<0.01).Conclusion:Wendantang could regulate the PI3K/Akt/GSK3 signaling pathway by improving the stereotyped behavior and increasing the expressions of PI3K, Akt, GSK3β of hippocampus in rats with schizophrenia, so as to achieve the purpose of treating schizophrenia.Keywords：Wendantang;schizophrenia;phosphatidylinositol-3-kinases(PI3K);protein kinase B(Akt);glycogen synthase kinase 3β(GSK3β)- 9
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Clinical Observation on Modified Huqianwan in Treatment of Rheumatoid Arthritis Liver-kidney Yin Deficiency Syndrome
Abstract：Objective:To explore the clinical efficacy and safety of modified Huqianwan in treatment of rheumatoid arthritis (RA) liver-kidney Yin deficiency syndrome, and investigate its possible mechanism.Method:A total of 184 patients with RA liver-kidney Yin deficiency syndrome were randomly divided into Chinese medicine group (62 cases), western medicine group (57 cases) and integrated Chinese and western medicine group (65 cases) according to the digital table method. The patients in Chinese medicine group were treated with Huqianwan; the patients in western medicine group were treated with methotrexate tablets and leflunomide tablets; and the patients in integrated Chinese and western medicine group received Huqianwan+ methotrexate tablets and leflunomide tablets, with a treatment course of 12 weeks in all groups. The pain visual analog scale (VAS), swelling and tenderness scores of 28 joints (DAS28), average hands grip strength, morning stiffness time and liver-kidney Yin deficiency syndrome differentiation of traditional Chinese medicine (TCM) syndrome score were compared between groups before and after treatment. The changes of erythrocyte sedimentation rate (ESR), C reactive protein (CRP), immunoglobulin (Ig)G, tumor necrosis factor-alpha (TNF-α) and rheumatoid factor (RF) were detected in all groups after treatment. Clinical efficacy, and incidence of adverse reactions such as gastrointestinal response, liver injury, leukopenia, serum glutamate oxaloacetic aminotransferase (GOT) and platelet (PLT) level changes were compared between the groups, so as to investigate the efficiency and safety of the different medicines.Result:After 12 weeks of treatment, the total clinical effective rate was 79.0%, 80.7%, and 92.3% respectively in Chinese medicine group, western medicine group, and integrated Chinese and western medicine group; the integrated Chinese and western medicine group was significantly better than the Chinese medicine group and western medicine group (P<0.01), but there was no difference between the Chinese medicine group and western medicine group. Every treatment group can effectively improve liver and kidney Yin deficiency syndrome in RA patients (P<0.05), and the effect in integrated Chinese and western medicine group was superior to that in Chinese medicine group (P<0.05); the effect in Chinese medicine group was superior to that in western medicine group (P<0.05). The incidence of adverse reactions was Chinese medicine group (1.61%, 1/62)<integrated Chinese and western medicine group (7.69%, 5/65)<western medicine group (22.81%, 13/57).Conclusion:The efficacy in treating RA liver and kidney Yin deficiency syndrome shows no significant difference between modified Huqianwan and methotrexate tablets+ leflunomide tablets. In the treatment of RA liver and kidney Yin deficiency syndrome, Huqianwan has fewer adverse reactions. Huqianwan combined with methotrexate tablets+ leflunomide tablets is superior to that in methotrexate tablets+ leflunomide tablets in treatment of RA liver-kidney Yin deficiency syndrome.Keywords：modified Huqianwan;liver-kidney Yin deficiency;rheumatoid arthritis;erythrocyte sedimentation rate (ESR);C reactive protein(CRP);tumour necrosis factor-α(TNF-α);rheumatoid factor (RF)- 8
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Intervention Effect of Fuyuan Huoxuetang for Postoperative Syndrome of Partial Splenic Arterial Embolization in Patients with Hypersplenism of Blood Stasis Type Liver Cirrhosis
Abstract：Objective:To analyze the effect of Fuyuan Huoxuetang for the postoperative syndrome of partial splenic arterial embolization (PSE) in patients with hypersplenism of blood stasis type hepatic cirrhosis.Method:86 patients with hypersplenism of blood stasis type hepatic cirrhosis treated with PSE were divided into observation group (44 cases) and control group (42 cases). Patients in both groups were treated with routine liver protection, antiviral, postoperative prophylactic anti-infection and symptomatic supportive care. Patients in treatment group started to take the Fuyuan Huoxuetang on the first day after the operation of PSE. The degree of pain, duration of fever, abdominal distention, and biochemical index changes of the two groups were observed 7 days after surgery.Result:After postoperative treatment, both groups of patients showed significant increases in the levels of -glutamyl transpeptidase (GGT) and white blood cell(WBC)(P<0.05), and significant decreases in albumin (ALB) (P<0.05). After surgery, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were stable, with no significant changes and no signs of liver failure. The WBC level of patients in observation group was significantly higher than that of control group (P<0.05) after the treatment with oral Fuyuan Huoxuetang. As compared with control group, the pain score was significantly lower in observation group (P<0.05); the duration of fever was significantly shorter (P<0.05); the total effective rate of relieving abdominal distension constipation was higher (P<0.05), and there was no obvious adverse reaction during the whole treatment process, with good safety in observation group.Conclusion:Fuyuan Huoxuetang could effectively interfere with the postoperative syndrome of PSE in patients with hepatic cirrhosis combined with hypersplenism by reducing pain, shortening the duration of fever and relieving the degree of abdominal distension and constipation.Keywords：Fuyuan Huoxuetang;traditional Chinese medicine therapy;hypersplenism;partial splenic arterial embolization (PSE);postoperative syndrome- 7
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect of Jiawei Changfengyin with Retention Enema Therapy on Th1/Th2 in Patients with Acute Radiation Proctitis
Abstract：Objective:To discuss the efficacy of Jiawei Changfengyin with retention enema therapy for acute radiation proctitis and investigate its influence on regulating levels of helper T cell(Th)1/Th2.Method:One hundred and twenty-eight patients were randomly divided into control group and observation group by random number table. Patients in control group (64 cases) got montmorillonite powder (3.0 g), dexamethasone (10 mg) and normal saline (100 mL), with retention enema therapy. Patients in observation group (64 cases) got Jiawei Changfengyin(Changfengyin+ Xileisan+ pearl powder), with retention enema therapy. Both groups of patients received enema once every night, and the treatment course was 4 weeks in both groups. Before and after treatment, scores of main symptoms, proctoscopy, routine examination of stool+ occult blood, and KPS scores of quality of life were graded. Levels of Th1 cell factors [interleukin(IL)-1β, IL-8 and interferon-γ(IFN-γ)], Th2 cell factors (IL-4 and IL-10) were detected.Result:The total effective rate for comprehensive clinical efficacy was 95.31% in observation group, higher than 82.81% in control group (χ2=5.132, P<0.05). Scores of each symptom and the total scores in observation group were lower than those in control group (P<0.01). Degree of rectal mucosa injury in observation group was lighter than that in control group (Z=2.764, P<0.05). Score of rectal mucosal injury was lower than that in control group (P<0.01), but score of KSP was higher than that in control group(P<0.05). Levels of IL-1β, IL-8 and IFN-γ in observation group were lower than those in control group, while levels of IL-4 and IL-10 were higher than those in control group (P<0.01).Conclusion:Jiawei Changfengyin can relieve symptoms of acute radiation proctitis, promote healing of rectal mucosa, improve quality of life, and regulate Th1/Th2 cytokines, with good repairing effect for intestinal mucosa.Keywords：acute radiation proctitis;Changfengyin;Xileisan;pearl powder, retention enema;Th1/Th2 cytokines- 10
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Classic Prescriptions
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Interventional Effect and Mechanism of Extract of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma on Vascular Endothelial Cellular Senescence Induced by High Glucose and High Fat
Abstract：Objective:To explore the mechanism of extract of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma in delaying the senescence of vascular endothelial cells induced by high glucose and high fat.Method:The 40 mmol·L-1 glucose and 100 μmol·L-1 palmitate were used to induce endothelial cell senescence. The experiment was divided into control group, model group and low, medium and high-dose traditional Chinese medicine groups (50, 100, 200 mg·L-1). The intervention lasted for 48 h. Cell proliferation was detected by cell counting kit-8(CCK-8); cell senescence was detected by senescence β-galactosidase staining; p16 and p21 protein expression levels were detected by Western blot; p-H2A.X(Ser139)expression, mitochondria ROS(mtROS) production and changes in mitochondrial membrane potential(MMP) were detected by immunofluorescence.Result:Compared with the control group, in model group, the cell proliferation ability and the number of SA-β-gal blue-stained cells decreased(P<0.01), while the mtROS production and the expressions of p16, p21 and p-H2A.X(Ser139)increased(P<0.05, P<0.01). Compared with the model group, extract of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma can increase cell proliferation and MMP levels(P<0.01), and reduce the number of SA-β-gal blue-stained cells, the mtROS production, and expression levels of p16, p21 and p-H2A.X(Ser139)(P<0.05, P<0.01).Conclusion:The above results suggest that extract of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma delay of endothelial cellular senescence induced by high glucose and high fat, and its mechanism may be related to increasing mitochondrial membrane potential and reducing DNA damage accumulation caused by ROS production.Keywords：replenishing Qi and activating blood circulation medicine;extract of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma and Chuanxiong Rhizoma;vascular endothelium;cell senescence- 11
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Mechanism of Salidroside in Improving Glucose and Lipid Metabolism in Liver of Diabetic Rats
Abstract：Objective:To explore the mechanism of salidroside(SAL) in improving glucose and lipid metabolism in liver of diabetic rats.Method:According to blood glucose, 24 male ZDF(fa/fa) rats of 6-7 week old were randomly divided into model group, SAL-low group (0.05 g·kg-1), SAL-high group (0.1 g·kg-1), 8 rats in each group. Another 8 ZDF (fa/+ ) rats were selected as normal group. All the rats were administered continuously for 6 weeks. After experiment, fasting blood glucose(FBG), triglyceride(TG), total cholesterol(TC), free fatty acids(FFA), superoxide dismutase (SOD), malondialdehyde (MDA), catalase (CAT), fasting insulin(Fins), and homeostasis model assessment insulin resistance(HOMA-IR) were measured. Western blot was used to detect phosphorylated PKR-like ER kinase(p-PERK), NF-E2-relatedfactor2(Nrf2), heme oxidase-1(HO-1) protein expression levels in liver.Result:As compared with normal group, FBG, Fins, HOMA-IR, TC, TG, FFA, and MDA were increased significantly in model group (P <0.05, P<0.01), while SOD, CAT levels were significantly decreased (P<0.05, P<0.01); the protein expression levels of p-PERK, Nrf2, and HO-1 in control group were significantly decreased in model group(P<0.05, P<0.01). As compared with model group, FBG, Fins, HOMA-IR, TC, TG, FFA, and MDA levels were decreased significantly (P<0.05, P<0.01), while SOD and CAT levels were significantly increased in SAL low and high groups (P<0.05, P<0.01); and the protein expression levels of p-PERK, Nrf2, and HO-1 in SAL group were significantly increased(P<0.05, P<0.01).Conclusion:The Salidroside can improve metabolism of glucose and lipid, IR in diabetic rats, and the mechanism may be associated with inhabiting oxidative stress through the up-regulation of p-PERK, Nrf2, and HO-1 protein expression.Keywords：salidroside;oxidative stress;diabetes;PKR-like ER kinase/NF-E2-relatedfactor2/heme oxidase-1(HO-1)- 8
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect and Mechanism of Mori Folium on Insulin Resistance in 3T3-L1 Cells
Abstract：Objective:To observe effect of Mori Folium-containing serum on glucose consumption and cell activity of fat cell line 3T3-L1 insulin resistance (IR) model, in order to screen out the optimal concentration of drug-containing serum, detect effect of Mori Folium on the content of inflammatory factors, and explore the possible mechanism.Method:3T3-L1 preadipocytes in logarithmic growth phase were selected, and induced with 10 mg·L-1 insulin (Ins), 0.25 mmol·L-1 dexamethasone (DEX) and 0.5 mmol·L-1 3-isobutyl-methylxanthine(IBMX) for 48 h and then with 10 mg·L-1 Ins for 48 h. After the cells were differentiated into mature adipocytes, they were induced with 1 μmol·L-1 DEX for 96 h to establish IR model. Glucose content in the supernatant of cells was detected by glucose oxidase after serum containing Mori Folium cultured for 12, 24, 36, 72 h. Methyl-thiazdyl-tetrazolium(MTT) was used to detect the effect of serum containing Mori Folium on IR cells activity. The content of tumor necrosis factor-α(TNF-α) was determined by enzyme-linked immunosorbent assay (ELISA). Meanwhile, the effects of inflammatory factors on the expressions of insulin signaling pathway proteins insulin receptor (InsR), insulin receptor substrate (IRS), p-IRS1 and glucose transporter 4 (GLUT4) were determined by Western blot.Result:Serum containing Mori Folium could significantly increase the glucose consumption rate and cell activity of IR cells (P<0.01), reduce the content of TNF-α (P<0.01), and regulate the expression of insulin signaling pathway proteins (P<0.05, P<0.01).Conclusion:Mori Folium can significantly improve IR status of 3T3-L1 cells, and its mechanism may be related to inhibiting TNF-α and promoting the expressions of insulin signaling pathway proteins.Keywords：Mori Folium;serum containing drug;type 2 diabetes mellitus(T2DM);tumor necrosis factor-α(TNF-α);insulin signaling pathway- 7
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect of Yiqi Yangyin Zhuyu Recipe in Anti-inflammation Activities in Vitro on LPS-induced RAW 264.7 Cells
Abstract：Objective:To investigate the effect of Yiqi Yangyin Zhuyu recipe on expressions of inflammatory factor and transformation of classically activated macrophages(M1)/alternatively activated macrophages (M2) inflammatory phenotype in lipopolysaccharide (LPS)-induced RAW 264.7 cells.Method:Methyl-thiazdyl-tetrazolium(MTT) reduction assay was used to detect the effect of different concentrations of Yiqi Yangyin Zhuyu recipe on the proliferation of the cells. The release of nitric oxide was detected by the Griess method. Enzyme linked immunosorbent assay(ELISA)was used to detect the release of M1/M2 inflammatory cytokines in cell supernatant. The expressions of the pro-inflammatory factor genes of M1-macrophages and the anti-inflammatory factor genes of M2-macrophages were detected by Real-time PCR. The protein expression levels of tumor necrosis factor-α(TNF-α), interleukin(IL)-6, IL-1 , nitric oxide synthase(iNOS) were detected by Western blot.Result:Results of MTT showed that Yiqi Yangyin Zhuyu recipe with the concentration of 2.0 g·L-1 and below had no effect on the cell proliferation. Results of Griess indicated that compared with blank group, the release of nitric oxide of LPS-induced group was increased (P<0.01); compared with LPS-induced group, Yiqi Yangyin Zhuyu recipe group could reduce the release of NO (P<0.01). Results of ELISA showed that compared with blank group, the release of M1/M2 inflammatory cytokines of LPS-induced group were both up-regulated (P<0.01); compared with LPS-induced group, the secretion of M1-macrophages inflammatory cytokines was down-regulated in Yiqi Yangyin Zhuyu recipe group (P<0.01), but with no change in M2-macrophages. Results of Real-time PCR indicated that compared with blank group, the expressions of M1/M2 were both increased (P<0.01, P<0.05); compared with LPS-induced group, the expression levels of M1-macrophages were down-regulated in traditional Chinese medicine group (P<0.01); anti-inflammatory factor genes of M2-macrophages had no significant change. Results of Western blot showed that compared with blank group, expression levels of TNF-α, IL-6, IL-1β, iNOS were up-regulated (P<0.01); compared with LPS-induced group, the expressions of TNF-α, IL-6, IL-1β, iNOS were down-regulated in Yiqi Yangyin Zhuyu recipe group, especially at the concentration at 2.0 g·L-1 (P<0.01).Conclusion:Yiqi Yangyin Zhuyu recipe could effectively inhibit the inflammatory reaction induced by LPS. The anti-inflammatory mechanism of Yiqi Yangyin Zhuyu recipe may be related to inhibition of macrophages to M1 phenotype polarization, so as to play the role of regulating immune and reducing the release of inflammatory cytokines, like NO, TNF-α, IL-6, IL-1β.Keywords：atheroscberosis(AS);Yiqi Yangyin Zhuyu recipe;macrophage;classically activated macrophages (M1);alternatively activated macrophages (M2);lipopolysaccharide(LPS);Gualou Xiebai Baijiu Tang;Danggui Buxue Tang- 9
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect of Canna edulis Type 3 Resistant Starch in Reducing Body Weight, Serum Lipid and Acute Toxicity
Abstract：Objective:To evaluate effect of Canna edulis type 3 resistant starch(RS3) on weight loss and lipid reduction in obese hyperlipidemia mice and acute toxicity in mice.Method:KKAy mice were fed with high-fat diet for 20 weeks to establish a hyperlipidemia model and then randomly divided into model group, positive group (4 mg·kg-1), high-dose resistant starch group and low-dose resistant starch group (2, 1 g·kg-1). Mice in normal group were fed with standard diet. The medication groups received corresponding drugs by gavage. Normal group and high-fat model group were given equal volume of deionized water. After 8 weeks, mice were put to death. The levels of total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in serum of mice were measured, and weigh fat mass, fat/body ratio, body fat rate and Lee's index were calculated accurately. The pathological changes of liver and adipose tissue were observed byhematoxylin-eosin (HE). The acute toxicity of RS3 to mice was evaluated by limit test. The mice were continuously observed for 14 days, and the toxicity of mice was recorded.Result:The indicators of high-dose RS3 group were significantly reduced, such as body weight, fat mass, body fat rate, fat/body ratio, Lee's index, and serum TC, TG, LDL-C, AST, ALT levels(P<0.05). Histomorphometric examination showed that the administration of RS3 starch could significantly improve the fatty lesions of liver tissue, and the liver-protecting effect was obvious, which could still inhibit the expansion of fat cells and reduce the accumulation of fat in mice.Among them, the high-dose group wsa better; After the maximum dose of 36 g·kg-1 was administered, no toxic reaction and death occurred in the animals.Conclusion:RS3-type Canna Edulis Resistant Starch has a good effect in reducing body weight and serum lipid, with a better effect in the high-dose group and no toxicity. And the commonly used clinical dose is safe and reliable.Keywords：Canna edulis type 3 resistant starch;obesity;hyperlipidemia;hypolipiclemtc effect;acute toxicity- 9
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Pharmacology
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Clinical Effect of Jinlida and Tongxinluo on Type 2 Diabetic Kidney Disease Under Vessel Collateral Theory
Abstract：Objective:To observe the clinical efficacy of Jinlida (JLD) granules and Tongxinluo (TXL) capsules on type 2 diabetic kidney disease (DKD) under the guidance of vessel collateral theory.Method:A total of 120 patients with type 2 DKD, were randomly divided into 2 groups: the normal control group (60 cases) and the treatment group (60 cases). The patients in normal control group were treated with dietary control and hyperglycemia control. Based on treatment in control group, patients in treatment group were additionally treated with JLD granules (1 bag, tid), and TXL capsules (4 capsules, tid). The treatment was lasted for 12 weeks. The clinical efficacy, traditional Chinese medicine(TCM) syndrome scores were observed and compared before and after treatment. At the same time, the levels of glucose metabolism indexes including fasting blood glucose (FBG), postprandial 2 h plasma glucose (2 hPG), glycosylated hemoglobin (HbA1c) and the insulin resistance (IR); the levels of lipid metabolism indexes including total cholesterol(TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and the high-density lipoprotein cholesterol (HDL-C); the levels of renal function indexes including urinary albumin excretion rate (UAER) and serum creatinine (SCr); as well as nailfold microcirculation were detected and compared.Result:①The total effective rate was 80.0% in treatment group, significantly higher than 61.67% in the normal control group (P<0.05). ②The improvement in TCM syndrome scores of the treatment group was better than that of the control group, and the difference was statistically significant (P<0.05). ③After treatment, the treatment group showed no significant difference with the control group in glucose metabolism indexes FBG, 2 hPG and HbA1c, but HOMA-IR improvement was better than that of control group (P<0.05). Lipid metabolism indexes TG, TC and LDL-C were significantly decreased (P<0.05), and HDL-C was increased in both groups (P<0.05). The improvement of renal function indexes SCr, UAER and urine Uβ2-MG in treatment group was significantly obvious than that in the control group (P<0.05). The nailfold microcirculation results showed that the microcirculation improvement in treatment group was better than that of control group (P<0.05).Conclusion:Tongxinluo combined with Jinlida can improve the clinical symptoms of patients with type 2 diabetic nephropathy and reduce urinary trace albumin, and its mechanism may be related to lowering glucose, regulating lipid metabolism and improving microcirculation.Keywords：vessel collateral theory;Jinlida;Tongxinluo;type 2 diabetic kidney disease (DKD)- 6
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect of Shugan Jieyu Capsules Combined with Repetitive Transcranial Magnetic Stimulation on Neuroendocrine Function During Perimenopause
Abstract：Objective:To discuss the clinical effect of Shugan Jieyu capsules combined with repetitivetranscranial magnetic stimulation (rTMS) on depression during perimenopause and neuroendocrine function.Method:One hundred and thirty-two patients were divided into control groupand observation groupby random number table. Patients (65 cases) in observation group got sertraline hydrochloride tablets, 50 mg/time, 2 times/days. Patients (67 cases) in control group got Shugan Jieyu capsules after breakfast and dinner, 2 grains/time, and treatment of rTMS, 20 minutes/time, 1 time/day. And one course of treatment was 5 days, there were a 2-day interval between two courses. And there were a total of 4 courses in the two groups. Both groups were continuously treated for8 weeks. Degree of depression was evaluated by Hamilton depression scale-17 (HAMD-17). Before and after treatment, scores of Hamilton anxiety scale (HAMA), quality index of sleep in Pittsburgh (PSQI), Kupperman and syndrome of kidney deficiency and stagnation of liver were graded. Levels of follicle stimulating hormone (FSH), Luteinizing hormone (LH), estradiol (E2), 5-serotonin (5-HT), dopamine (DA), norepinephrine (NE) and 5-hydroxyindolyl acetic acid (5-HIAA) were detected, and the safety was evaluated.Result:According to the variance analysis of repeated measurements, after treatment, score of HAMD-17 decreased (P<0.01). At the 8th day after treatment, score of HAMD-17of observation groupwas lower than that in control group (P<0.01). And scores of syndrome of kidney deficiency and stagnation of liver, HAMA, PSQI and Kupperman of observation group were lower than those in control group (P<0.01). And the total rate of depression in observation group was 95.52%, which was higher than 81.54% in control group (χ2=6.405, P<0.05). And the total rate of traditional Chinese medicine(TCM) syndromes in observation group was 92.54%, which was higher than 78.46% in control group (χ2=5.304, P<0.05). After treatment, levels of FSH and LH were lower than those in control group (P<0.01).Conclusion:Shugan Jieyu capsules combined with repetitivetranscranial magnetic stimulation can ameliorate symptoms of depression and anxiety, havea bettereffect than Sertraline hydrochloride tablets, and can regulate levels of sex hormone and monoamine neurotransmitter.Keywords：perimenopausal depressive disorder;syndrome of kidney deficiency and stagnation of liver;Shugan Jieyu capsules;repetitivetranscranial magnetic stimulation;sex hormone;monoamine neurotransmitter- 8
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Clinic
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Kinetic Analysis of Enzymatic Reaction of Active Components from Xinshao Formula in Liver Microsomes of Sham-Operated and Cerebral Ischemia-reperfusion Injury Rats
Abstract：Objective:To investigate and compare enzymatic kinetics of scutellarin, apigenin-7-O-glucronide and paeoniflorin from Xinshao fomula in liver microsomes of sham-operated rats and middle cerebral artery occlusion(MCAO) rats with focal cerebral ischemia-reperfusion injury.Method:Xinshao fomula were incubated respectively with liver microsomes of sham-operated rats and MCAO rats, UPLC-MS and substrate elimination method was employed, Michaelis constant(Km), maximum velocity of enzymatic reaction(Vmax) and intrinsic clearance(CLint) of these three components from Xinshao fomula in liver microsomes of sham-operated rats and MCAO rats were calculated, these parameters between different groups were evaluated by statistical analysis.Result:The Km values of scutellarin, apigenin-7-O-glucronide and paeoniflorin in liver microsomes of MCAO rats were (0.798±0.031), (0.213±0.017), (0.499±0.029) μmol·L-1, which were quite different to these in liver microsomes of sham-operated rats.Compared with the sham-operated group, Vmax and CLint values of scutellarin and paeoniflorin in liver microsomes of MCAO rats were significantly reduced(P<0.05, P<0.01), and Vmax of apigenin-7-O-glucronide in liver microsomes of MCAO rat was also significantly reduced(P<0.05).Conclusion:Metabolic rates of these three active components from Xinshao fomula in liver microsomes of MCAO rats with focal cerebral ischemia-reperfusion injury decrease with low elimination rate.Keywords：Xinshao fomula;cytochrome P450;enzymatic kinetics;ischemia-reperfusion injury;middle cerebral artery occlusion;puerarin- 8
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Drug Metabolism
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Investigation of Physicochemical Properties and Determination Methods of Liposomal Entrapment Efficiency of Dichroa Alkali Hydrochloride
Abstract：Objective:To study on the physical and chemical properties of dichroa alkali hydrochloride and to establish a method for the determination of entrapment efficiency of dichroa alkali hydrochloride liposomes.Method:HPLC was used to determine the content of dichroa alkali hydrochloride with mobile phase of acetonitrile-water-triethylamine-glacial acetic acid(9∶91∶0.35∶0.75) and detection wavelength at 265 nm.The oil-water partition coefficient of this compound in different pH range was measured by shake flask method.The stability of the dichroa alkali hydrochloride in phosphate buffer solution with different pH after sterilization at 125 ℃ for 30 min was investigated.Ammonium sulfate gradient method was used to prepare dichroa alkali hydrochloride liposomes.The microcolumn was prepared by dextran gel and cation exchange resin, respectively.Then the free drug and liposome were separated by centrifugation, the drug content was measured, and the encapsulation efficiency was calculated.The t-test was performed using SPSS 20.0 software, the differences between these two methods were compared.Result:In the pH 6-9, the oil-water partition coefficient of dichroa alkali hydrochloride increased with increasing of pH, which was between 0.016 and 1.44; the recovery rate of dichroa alkali hydrochloride after sterilization was 37.16%-57.91%.Between the dextran gel microcolumn centrifugation and the cation exchange resin microcolumn centrifugation, there was no significant difference in the entrapment efficiency of the liposomes.Conclusion:Dichroa alkali hydrochloride is suitable for preparation of liposomes.However, its stability is not ideal, so the experimental temperature should be strictly controlled in the preparation process.Dextran gel microcolumn centrifugation and cation exchange resin microcolumn centrifugation can be used to determine the entrapment efficiency of dichroa alkali hydrochloride liposomes, and the cation exchange resin microcolumn centrifugation is suggested after comparison.Keywords：dichroa alkali hydrochloride;liposomes;oil-water partition coefficient;stability;encapsulation efficiency;microcolumn centrifugation method;ammonium sulfate gradient method- 10
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect of Decocting Containers on Content of Sinapine and HPLC Specific Chromatograms of Sinapis Semen Decoction
Abstract：Objective:To compare the effect of four kinds of decocting containers on the content of sinapine and the HPLC specific chromatograms of Sinapis Semen decoction, so as to optimize decocting container for the development of classical formulas.Method:Selecting four kinds of decoction vessels, named traditional casserole, ceramic pot, round-bottom flask and stainless-steel pot as the research object, the content of sinapine in Sinapis Semen decoction and its HPLC specific chromatograms were used as indexes to investigate the influence of different decoction vessels on the decoction.Similarity evaluation of specific chromatograms was performed by the " Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine" (edition of 2004A).Result:The contents of sinapine in the decoction prepared by traditional casserole, ceramic pot, round-bottom flask and stainless-steel pot were 0.04%, 0.07%, 0.84% and 0.97%, respectively.Compared with specific chromatograms of the decoction prepared by traditional casserole, the similarities of specific chromatograms of the decoction prepared by ceramic pot, round-bottom flask and stainless-steel pot were 0.98, 0.82 and 0.68, respectively.Compared with specific chromatograms of the decoction prepared by ceramic pot, the similarities of specific chromatograms of the decoction prepared by round-bottom flask and stainless-steel pot were 0.79 and 0.62, respectively.Compared with specific chromatograms of the decoction prepared by round-bottom flask, the similarity of specific chromatograms of the decoction prepared by stainless-steel pot was 0.97.Conclusion:The content of sinapine and HPLC specific chromatograms of Sinapis Semen decoction obtained from different decocting containers are quite different.Keywords：Sinapis Semen;decocting containers;specific chromatograms;sinapine thiocyanate;similarity;decoction- 9
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Quality Evaluation of Andrographis Herba Based On Electronic-eye Technique
Abstract：Objective:Taking electronic-eye (visual analyzer) technique, based on the powder color of Andrographis Herba, to investigate the applicability of electronic-eye technique and evaluate the quality of Andrographis Herba with different commercial specifications.Method:HPLC was employed to determine contents of andrographolide, dehydroandrographolide, 14-deoxyandrographolide, neoandrographolide in 50 batches of Andrographis Herba with different commercial specifications(stems, leaves and aerial parts). Color of these samples were measured by electronic-eye technique.The data were analyzed by principal component analysis(PCA) and Pearson correlation analysis.The ability of electronic-eye to distinguish the different commercial specifications of Andrographis Herba was investigated and the correlation of chroma space system parameters (L*, a*, b*) with active components was investigated.Result:There was remarkable difference in contents of 4 diterpenoids in Andrographis Herba from different parts, their contents in leaves was the highest, followed by the aerial parts(mixture of stems and leaves), and their contents in stems was the lowest.The results of PCA was divided into two classes, namely the stem part, leaf and aerial parts, indicating that electronic-eye could be used to distinguish the quality of Andrographis Herba.The correlation results showed that there were significant negative correlation(P<0.01) between L*(lightness value) and the contents of andrographolide, dehydroandrographolide, 14-deoxyandrographolide, neoandrographolide and the total content of these 4 components.In addition, L* of samples that did not conform to the lower limit of determination in the 2015 edition of Chinese Pharmacopeia was≥69.5, and the L* of more than 90% of the samples in accordance with the requirements was<69.5.Conclusion:Electronic-eye technique provides a new method and idea for the quality evaluation of Andrographis Herba.Keywords：Andrographis Herba;electronic-eye;color values;diterpene lactones;correlation analysis;principal component analysis;content determination- 9
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Effect of PEG400 Concentration in Receiving Solution on Drug Transdermal Test in Vitro
Abstract：Objective:To investigate the effect of varying concentrations of polyethylene glycol(PEG)400 in receiving solution on in vitro transdermal test of drugs.Method:5-Fluorouracil(5-FU) was selected as a model drug, by preparing different concentrations of PEG400-phosphate buffer solution(PBS) as the receiving solution, the receiving chamber did not add drug, the excised rat skins were treated with various additives for 12 h, then replaced by PBS and added the saturated model drug into the donor compartment to determine the transdermal parameters of the drug.Meanwhile, scanning electron microscopy(SEM) was employed to monitor the effect of PEG400 with different concentration on the stratum corneum of rat skin.Result:The 10%, 15% and 40%PEG400-PBS groups had no significant effect on in vitro transdermal absorption parameters of the 5-FU.The steady transdermal rate and cumulative penetration rate of the drug in 20% and 30% PEG400-PBS groups were significantly higher than that in the PBS group(P<0.01, P<0.05). SEM indicated that wrinkle of the intact rat skin gradually disappeared and a number of flakes were desquamated from the skin when the concentration of PEG400 was above 20% in receiving solution.Meanwhile, 30% PEG400-PBS group and 40% PEG400-PBS group were extremely wrinkled.Conclusion:In the rat skin transdermal test, the concentration of PEG400 in receiving solution should be controlled below 20%.Keywords：receiving liquid;polyethylene glycol 400;in vitro transdermal test;scanning electron microscopy;phosphate buffer solution;5-fluorouracil- 8
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Pharmacy and Processing
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Analysis and Identification of Chemical Components in Trichosanthis Fructus by UPLC-LTQ-Orbitrap-MS
Abstract：Objective:To analysis and identify the chemical components in Trichosanthis Fructus by UPLC-LTQ-Orbitrap-MS.Method:Samples of Trichosanthis Fructus were extracted by ultrasonic with 70% methanol after smashing and sifting by 40 mesh sieve. Thermo Scientific™ Dionex™ UltiMate™ 3000 Rapid Separation LC system performed UPLC separations with Waters HSS T3-C18(2.1 mm×100 mm, 1.8 μm) column. The mobile phase was 0.1% formic acid water(A)-methanol(B)with a gradient elution. The volume flow was 0.3 mL·min-1. A Thermo Scientific™ LTQ-Orbitrap mass spectrometer equipped with a ESI probe was employed. The samples were respectively scanned in MS1 and MS2 mode of positive and negative ions. According to the chromatographic peak separation, mass signal intensity, and the number of molecular ions in MS1 model, the extraction condition, chromatogram and mass spectrum parameters were optimized. The chemical compounds were identified by the accurate mass measurement of molecular ions and fragment ion and comparation with reference substance.Result:91 chemical compositions in Trichosanthis Fructus were totally identified, including 14 amino acids, 5 monoterpenoids, 5 tetracyclic triterpenoids, 1 pentacyclic triterpene, 14 flavonoids, 17 organic acids, 3 polysaccharides, 7 nucleotides, 7 alkaloids and nitrogen compounds, 2 volatile components, 1 phytosterol, 5 other compositions.Conclusion:The established UPLC-LTQ-Orbitrap-MS method can be used to quickly analyze and identify the main chemical constituents of Trichosanthis Fructus. The chemical information concerning the constituents in Trichosanthis Fructus could be helpful to the quality control and further studies of Trichosanthis Fructus.Keywords：Trichosanthis Fructus;chemical composition;UPLC-LTQ-Orbitrap-MS- 10
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Isolation and Structural Elucidation of Chemical Constituents of Corydalis Rhizoma
Abstract：Objective:In order to clarify the characteristic chemical constituents and furnish applicable information to the basic research and quality control research related to the chemistry of traditional Chinese medicines for Corydalis Rhizoma, this paper investigated the chemical constituents of Corydalis Rhizoma extensively.Method:The dried-up and pulverized plant materials were extracted using 95% EtOH as solvent, the EtOH extract was fractionated using different solvents to afford the EtOAc-soluble and n-BuOH-soluble portion, respectively, among others. These two portions were subjected to procedures of isolation and purification on silica gel or ODS column chromatographies to afford monomers. 1D and 2D NMR and MS methods, along with comparison with the data of literatures, were used to identify the structures.Result:Twelve compounds, all belonging to alkaloids, were isolated and identified as d-corydaline(1), tetrahydrocoptisine(2), tetrahydropalmatine(3), tetrahydrocolumbamine(4), corybulbine(5), tetrahydrojatrorrhizine(6), dehydrocorydaline(7), dehydroglaucine(8), 8-oxodihydrocoptisine(9), protopine(10), taxilamine(11), and pontevedrine(12). Of these compounds, the structure of 12 was a revised structure which was assigned by combined examinations of their 1D and 2D NMR spectra and MS data.Conclusion:Compounds 6 and 11 were reported from Corydalis Rhizoma for the first time. The structure of pontevedrine was verified as 1, 2, 9, 10-tetramethoxy-6-methyl-4H-dibenzo[de, g]quinoline-4, 5(6H)-dione.Keywords：Corydalis Rhizoma;alkaloids;isolation and purification;structural elucidation;1, 2, 9, 10-tetramethoxy-6-methyl-4H-dibenzo[de, g]quinoline-4, 5(6H)-dione- 8
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Analysis of Zingiberaceae in E'mei Area Using ITS2 Sequences
Abstract：Objective:To evaluate the use of ITS2 sequences as DNA barcode to identify the Zingiberaceae medicinal plants from E'mei area.Method:The genomic DNAs were extracted from 43 Zingiberaceae medicinal plant samples from Sichuan E'mei area. The ITS2 sequences of these samples were amplified and bidirectionally sequenced by PCR. 40 ITS2 sequences were downloaded from the GenBank, and then the interspecific and intraspecific genetic distances were calculated and analyzed by using MEGA 6.0 to construct Neighbor-joining (NJ) tree; TAXON DNA software was also used to analyze intraspecific and interspecific variations and barcoding gaps. The differences in secondary structure of the ITS2 sequences were predicted and compared.Result:The minimum interspecific distance in Zingiberaceae samples was greater than the maximum intra specific distance, with obvious barcoding gap. The NJ tree showed that the samples were clustered into five different branches, Alpinia, Curcuma, Globba, Hedychium, and Zingiber respectively, and further cluster into sub-branches. Significant differences were also present in the secondary structures of ITS2 between different samples.Conclusion:ITS2 sequences as DNA barcode can be used to conduct accurate and rapid identification of the Zingiberaceae plants and clearly figure out the phylogenetic relationship among them, providing guidance for the study of the distribution of medicinal plants of this genus, as well as theoretical basis for the quality control, medication safety and rational development of Zingiberaceae medicinal plants in E'mei area.Keywords：Zingiberaceae;DNA bar code;ITS2;secondary structure;identification;resource utilization- 9
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Resource and Quality Evaluation
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Issues in Process of Inheritance and Innovation of Traditional Chinese Medicine Processing Discipline and Chinese Herbal Pieces Industry
Abstract：Based on the investigation for the status of inheritance, innovation and development of traditional Chinese medicine processing discipline and Chinese herbal pieces industry in recent years, several problems worth discussing were analyzed in this paper.The definition of decoction pieces of Chinese crude drugs, the contents and methods of inheritance and innovation were put forward.It is aimed at building a consensus and further promoting the development of disciplines and industries.Keywords：traditional Chinese medicine processing;Chinese herbal pieces industry;inheritance;innovation- 25
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09 -
Prevention and Treatment Effect of Traditional Chinese Medicine for Stroke Based on Regulating Gut Microbiome
Abstract：The gut microbiome is a unique ecosystem, known as the " forgotten organ" of the human body, and the " second genome" of human. Gut microbiome imbalance is associated with many central nervous system diseases such as Parkinson's disease, Alzheimer's disease, schizophrenia and disseminated sclerosis. Stroke is characterized by high morbidity, recurrence, mortality, and disability. Gut microbiome plays a key role in the occurrence and development of stroke, which can further affect the onset of stroke by affecting the body's absorption, metabolism, blood pressure, blood sugar, blood lipids and atherosclerotic plaque. In Chinese medicine, it is believed that the spleen and stomach qi and blood flow loss, the Yin and yang imbalance, the body's physiological dysfunction, metaplasia " wind, fire, phlegm, deficiency, silt" and other pathological products, can cause the occurrence of stroke. The spleen and stomach are mainly involved for decomposing food, transporting and transforming nutrients from foodstuff, and the gut microbiome affect the digestion and absorption of the diet. The function of gut microbiome in modern research is related to the dysfunction of the spleen and stomach in Chinese medicine. Therefore, adjusting the homeostasis of the gut microbiome can be used as a potential intervention target to prevent and treat ischemic stroke. Traditional Chinese medicine(TCM) intervention has already achieved good results in stroke, and whether it is related to the regulation effect of gut microbiome is worthy of further research in the future. At the same time, the research progress of active ingredients in TCM (berberine, baicalin, resveratrol, et al), single TCM prescriptions (Salviae Miltiorrhizae Radix Et Rhizoma, Rhodiolae Crenulatae Radix Et Rhizoma, et al) and compound TCM prescriptions (Buyang Huanwu Tang, Naoxintong capsule, Buzhong Yiqi Tang) in prevention and treatment of cerebral ischemia were reviewed in this study, providing new ways and ideas for the prevention of ischemic stroke and development of new therapies.Keywords：gut microbiome;ischemic stroke;bacterial-gut-brain axis;spleen and stomach;traditional Chinese medicine- 13
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<HTML><CITATION> <Bulk Citation>Published：2021-02-09
Academic Discussion
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