Abstract：Objective:To explore the effect of Wenjingtang on the levels of serum platelet activating factor (PAF) and β-endorphins in primary dysmenorrhea patients with cold stagnation and blood stasis syndrome.Method:The 120 cases of dysmenorrhea treated at Second Affiliated Hospital, Heilongjiang University of Chinese Medicine from March 2016 to December 2017 were selected as the observation subjects, and divided into control group and observation group according to the random number table method, with 60 cases in each group. The control group was treated with ibuprofen capsules, while the observation group was treated with Wenjingtang and ibuprofen capsules. Dysmenorrhea symptom scores, clinical efficacy, recurrence condition and serum levels of platelet activating factor and β-endorphins were compared before and after treatment.Result:After treatment, the dysmenorrhea symptom scores decreased significantly in both groups, and the scores of patients in observation group were lower than those in control group (P<0.05). After treatment, the total efficiency in observation group was 95%, which was significantly higher than 83.33% in control group (P<0.05). The level of serum PAF decreased significantly in two groups, and the serum PAF in observation group was significantly decreased compared with control group (P<0.05). The level of serum beta-endorphins in observation group was significantly higher than that of control group (P<0.05). After half a year, the recurrence rate of the observation group was 5%, which was significantly lower than 30% of control group (χ2=12.987, P<0.01).Conclusion:Wenjingtang contributes to the reduction of dysmenorrhea symptoms scores in primary dysmenorrhea patients with cold stagnation and blood stasis syndrome, and can improve the clinical efficacy, reduce the serum PAF level and increase the concentration of β-endorphins, reduce the recurrence rate, improve the quality of life of patients, and it is worth promoting.
Keywords：Wenjingtang;syndrome of cold congealing and blood stasis;primary dysmenorrhea;platelet activating factor
Abstract：Objective:To observe the effect of Shenling Baizhusan(SBS)on the mammalian target of rapamycin complex 1 (mTORC1)/signal transducers and activators of transcription 3 (STAT3) pathway in liver hepatocyte of nonalcoholic fatty liver disease(NAFLD)rats induced by high fat diet, in order to reveal the mechanism of SBS against rat NAFLD from the perspective of inflammation.Method:Totally 80 SD rats were randomly divided into 4 groups, normal control group, model group, high-dose SBP group(30 g·kg-1), and low-dose SBS group(10 g·kg-1), with 20 rats in each group. The rats of NAFLD model were established by being fed with high-fat diets for 8 weeks, and the treatment groups were fed with high or low dose of SBS respectively. After treatment for 8 weeks, blood and liver samples of rats were collected. Alanine aminotransferase (ALT), aspartate aminotransferase(AST), total cholesterol (TC), triglyceride(TG), high-density lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(LDL-C)levels in blood serum were detected with automatic biochemical analyzer. The liver tissues were observed by oil red O and hematoxylin-eosin (HE) staining. Hepatocytes were isolated by type Ⅳ collagenase perfusion in vitro. Tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-5 and IL-6 in hepatocytes were detected by enzyme-linked immunosorbent assay (ELISA), and the relevant gene and proteins expressions of mTORC1 and STAT3 in hepatocytes were detected by Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) and Western blot detection respectively.Result:Compared with the normal control group, the serum levels of TG, TC, AST, ALT and LDL-C were increased significantly, the levels of TNF-α, IL-1β, IL-5 and IL-6 in hepatocytes were increased significantly, and the expression levels of mTORC1, STAT3 mRNA and proteins in hepatocytes were increased significantly(P<0.01). Compared with the model group, the hepatic lipid accumulation of the medicine intervention group was relieved significantly, the serum levels of AST, ALT, TG and LDL-C were decreased significantly, the expression levels of TNF-α, IL-1β, IL-5 and IL-6 of hepatocytes were decreased significantly, and the expressions of mTORC1, STAT3 mRNA and proteins in hepatocytes were decreased significantly(P<0.05, P<0.01). In the high-dose SBS group, the effects in improving the lipid accumulation and inhibiting the inflammatory reaction were better than those of the low-dose SBS group, and the expressions of mTORC1 and STAT3 genes and proteins in hepatocytes were significantly decreased (P<0.05, P<0.01).Conclusion:SBS can improve the fat metabolism disorder and reduce liver lipid accumulation and inflammatory reaction in NAFLD rats induced by high-fat diet. The mechanism may be correlated with the inhibition of mTORC1/STAT3 pathway relating to genes and protein expression in hepatocytes.
Keywords：Shenling Baizhusan;nonalcoholic fatty liver disease;mammalian target of rapamycin complex 1 (mTORC1)/signal transducers and activators of transcription 3 (STAT3) pathway
Abstract：Objective:To explore the effect of Zuoguiwan on the bone mineral density (BMD) and the expressions of Ca2+ transport-associated protein in ovariectomized rats.Method:The 48 female SD rats were randomly divided into six groups: normal group, model group, sham operation group, estrogen group(0.167 mg·kg-1) and low and high-dose Zuoguiwan groups(9.6, 38.4 g·kg-1), with 10 rats in each group. Except for the sham-operated group, the ovariectomized rats in the other groups received the bilateral ovariectomy. Therapeutic intervention was given in each group for 3 months after the establishment of the model. After 12 weeks, BMD was measured using dualenergy X-ray absorptiometry. Tartrated presistant acid phosphatse(TRACP) and serum calcium were detected by biochemical kits.Protein expression in Ca2+ transport (Bone tissue) was detected by Western blot.Result:Compared with the normal group, the serum calcium of the model group was decreased(P<0.01). Compared with the normal group, BMD of the model group was decreased (P<0.01). The serum calcium of rats in high-dose group and western medicine group was higher than that of model group(P<0.01). BMD in model group was lower than that of Zuoguiwan groups and estrogen group(P<0.05). There was no significant difference in TRACP among the groups. Nilestriol and Zuoguiwan can down-regulate the expressions of TRPV5, NCX1, CaBP-D28K and PMCA1b in bone tissue of castrated rats(P<0.05, P<0.01).Conclusion:Zuoguiwan can down-regulate the expressions of Ca2+ transport-associated proteins (Bone tissues) in rat osteoclasts, with an efficacy on osteoporosis.
Abstract：Objective:To study the effect of warming and heat-clearing method (Wenyang Jiedu Huayu decoction) on the expressions of Forkhead box P3 (FoxP3), Retinoic acid-related orphan receptor gamma t (ROR-γt) in colon tissue of mice with acute-on-chronic liver failure (ACLF), in order to explore the possible regulatory mechanism on intestinal endotoxemia (IETM) in liver failure mice.Method:The 130 SD rats were randomly divided into normal group (10 rats) and model group (120 rats). The ACLF mice model was established through the subcutaneous injection with bovine serum albumin and the intraperitoneal injection with D-galactosamine(D-Gal) and lipopolysaccharide (LPS). The model mice were randomly divided into model group, heat-clearing group (Yinchenhao decoction, 6.68 g·kg-1), warming group (Yinchen Zhufu decoction, 7.09 g·kg-1) and warming and heat-clearing group (Wenyang Jiedu Huayu decoction, 19.53 g·kg-1). The normal group and the model group were given distilled water by gastric lavage, while the other groups were given equal volume of corresponding Chinese herbal medicines for a week. The value of each index at 1, 12 and 24 h was measured. The ratio of Treg/Th17 cell in peripheral blood were detected and calculated by flow cytometry. Real-time fluorescence quantitative PCR (Real-time PCR) was used to detect the expressions of FoxP3 and ROR-γt in colon tissues of mice at different time points. In situ hybridization and immunohistochemistry were used to observe the expressions of FoxP3 and ROR-γt genes and proteins.Result:Compared with normal group, the ratio of Treg/Th17 in the model group decreased significantly at each time point (P<0.01). Compared with the model group, the Treg/Th17 ratio increased only in the warming and heat-clearing method group (P<0.05). Compared with normal group, the expression of ROR-γt in the model group was significantly higher (P<0.01), and the expression of ROR-γt in the model group was higher than FoxP3.Compared with the model group, the expressions of FoxP3 and ROR-γt mRNA in the heat-clearing group and the warming group decreased at each time point (P<0.05), and the expressions of FoxP3 and ROR-γt in the warming and heat-clearing method group decreased significantly (P<0.01). The expressions of FoxP3 and ROR-γt mRNA in warming and heat-clearing group decreased compared with those in the warming group and heat-clearing group (P<0.05).Conclusion:The mechanism of the warming and heat-clearing method on IETM in liver failure may be related to the regulation of FoxP3 and ROR-γt expressions.
Keywords：warming and heat-clearing method;acute-on-chronic liver failure;intestinal endotoxemia;Forkhead box P3 (FoxP3);retinoic acid-related orphan receptor-gamma t (ROR-γt)
Abstract：Objective:To investigate the relationship between Toll-like receptor 4 (TLR4) mRNA and its downstream inflammatory factor-tumor necrosis factor-α (TNF-α) and hepatocyte apoptosis in mice with intestinal endotoxemia (IETM) of hepatic failure, and explore the regulatory mechanism of Wenyang Jiedu Huayu granule on endotoxin-induced hepatocyte apoptosis.Method:The 85 SPF male SD rats were randomly divided into normal group, model group, TLR4 monoclonal antibody group and Wenyang Jiedu Huayu granule group. D-galactosamine (D-Gal) intraperitoneal injection was performed to establish the IETM model of hepatic failure. The TLR4 monoclonal antibody group and the Wenyang Jiedu Huayu granule group were given Wenyang Jiedu Huayu granule solution by gavage 5 days before the modeling. The normal group and the model group were given isovolumetric distilled water. Each group was given by gavage until sacrifice. Rats in each group were randomly sacrificed at 24, 48, 72 h, respectively, and samples were collected. The levels of serum levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected at 24, 48, 72 h. The pathological changes of liver tissue were observed by hematoxylin and eosin(HE) staining. The expression of TLR4 in liver tissue was detected by real-time fluorescence quantitative PCR (Real-time PCR). The expression of TNF-α in liver tissue was detected by enzyme-linked immunosorbent assay (ELISA). The apoptotic rate of hepatocytes was detected by flow cytometry.Result:Compared with the normal group, ALT and AST were increased in model group, while the pathological injury degree of liver tissue was significantly increased. The expressions of TLR4 mRNA and TNF-α were increased (P<0.05, P<0.01), whereas the apoptosis rate of liver cells was significantly increased (P<0.05, P<0.01). Compared with the model group, ALT and AST were decreased in Wenyang Jiedu Huayu granule group (P<0.01), and the degree of pathological injury of liver tissue was significantly reduced (P<0.05). The expressions of TLR4 mRNA and TNF-α were significantly decreased (P<0.01), and the apoptosis rate of liver cells was also reduced (P<0.01).Conclusion:TLR4 mRNA and TNF-α are positively correlated with hepatocyte apoptosis in liver failure. Wenyang Jiedu Huayu granule can improve liver function, alleviate liver cell injury and reduce liver cell apoptosis in IETM mice with hepatic failure. The mechanism may be related to its ability to down-regulate the expression of liver TLR4 mRNA, inhibit the release of TNF-α, and reduce the rate of hepatocyte apoptosis.
Abstract：Objective:To detect the expression levels of peripheral blood Treg/Th17 cells and related cytokines in patients with Hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF) with different traditional Chinese medicine (TCM) syndrome " Yanghuang-Yinyanghuang-Yinhuang" , in order to explore the cellular immunological characteristics of different TCM syndromes of liver failure.Method:The 32 cases of patients with HBV-ACLF in early, middle and late stages in line with the " Yanghuang-Yinyanghuang-Yinhuang" TCM syndrome grouping were selected. Flow cytometry was used to detect the frequency expression of Treg/Th17 cells in peripheral blood. The expression levels of interleukin-10(IL-10), transforming growth factor-β(TGF-β), interleukin-17A(IL-17A), tumor necrosis factor-α(TNF-α) and interleukin-23(IL-23) were detected by cytometric bead array (CBA). The expressions of transcription factor forkhead box P3(FoxP3) and retinoid-related orphan nuclear receptor-γt(ROR-γt) mRNA were detected by Real-time PCR. The SPSS 20.0 software was applied in data statistics and processing to analyze the expression characteristics of Treg/Th17 cells and related cytokines in patients with different TCM syndrome types of HBV-ACLF.Result:The patients with HBV-ACLF Yanghuang syndrome were mainly distributed in the early stage of liver failure, those with Yinyanghuang syndrome were mainly distributed in the middle stage, and those with Yinhuang syndrome were distributed in the late stage. From Yanghuang syndrome, Yinyanghuang syndrome to Yinhuang syndrome, the frequency of Treg and Th17 cells gradually increased, and the differences among the groups were statistically significant (P<0.05). From Yanghuang syndrome, Yinyanghuang syndrome to Yinhuang syndrome, Treg cytokines IL-10, TGF-β gradually increased, and the differences among the groups were statistically significant (P<0.05). Th17 cytokines IL-17A, TNF-α, IL-23 gradually increased, of which IL-17A were differences between Yanghuang syndrome and the Yinyanghuang syndrome, as well as Yanghuang syndrome and Yinhuang syndrome (P<0.05). From Yanghuang syndrome, Yinyanghuang syndrome to Yinhuang syndrome, the expression of FoxP3 was gradually decreased, while that of ROR-γt was gradually increased, and the differences among the groups were statistically significant (P<0.01).Conclusion:There is a certain correlation between the different course of early, middle and late stages of HBV-ACLF and the distribution of TCM syndromes. The frequency of Treg and Th17 cells and the correlation of IL-17A, TGF-β and IL-10 with TCM syndrome differentiation are related, suggesting that Treg and Th17 cells have a certain reference value for the diagnosis of patients with HBV-ACLF and the syndrome differentiation of TCM syndromes.
Keywords：Hepatitis B virus related acute-on-chronic liver failure(HBV-ACLF);Treg cells;Th17 cells;traditional Chinese medicine syndromes
Abstract：Objective:Through the retrospective study of Hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF) patients, the clinical features and the characteristics of traditional Chinese medicine (TCM) syndrome distribution of HBV-ACLF complicated with bacterial infection were preliminarily expounded, so as to provide the corresponding theoretical basis for the diagnosis and treatment of HBV-ACLF secondary bacterial infection.Method:A unified data collection form for patients with slow plus acute liver failure was designed by the retrospective epidemiological investigation method. The clinical data of 307 patients with HBV-ACLF who were hospitalized at the Hepatology Institute of the First Affiliated Hospital of Hunan University of Chinese Medicine were collected in strict accordance with the inclusion and exclusion standards, and the main symptoms and the TCM symptoms were screened. Then, according to whether or not there was a bacterial infection, the patients were divided into a combined bacterial infection group and a non-combined bacterial infection group. The distribution characteristics of the main symptoms and the TCM symptoms in two groups were compared, and the distribution characteristics of TCM symptoms of HBV-ACLF with bacterial infection were obtained.Result:Compared with patients with uncombined bacterial infection, patients with combined bacterial infection had significantly increased abdominal distension, puffiness, purple and dark tongue color, abdominal varicose tendons, subcutaneous stasis spots, and tooth marks on the side of the tongue. Compared with patients with combined bacterial infections, patients with uncombined bacterial infections mainly showed bright yellow head, yellow tongue coating, and significantly increased dry mouth (P<0.05, P<0.01). The main symptoms of patients with combined bacterial infections was the combination of syndrome (93.6%), and more spleen deficiency syndrome (81.6%), patients with uncombined bacterial infections mainly suffered from a single syndrome (53.5%), which was mostly damp-heat syndrome (51.9%), patients of concurrent syndrome with combined bacterial infections mostly suffered from false and real syndrome (94.4%), which was significantly higher than those with uncombined bacterial infections (58.9%).Conclusion:The patients with HBV-ACLF without bacterial infection mainly suffered from damp-heat symptoms and signs. With the combination of bacterial infection, the symptoms and signs of spleen deficiency and blood stasis gradually worsened. HBV-ACLF patients with bacterial infection mainly suffered from spleen deficiency and blood stasis, while patients with unincorporated bacterial infection mainly suffered from the single damp-heat symptoms and signs.
Keywords：Hepatitis B virus related acute-on-chronic liver failure;bacterial infection;traditional Chinese medicine syndrome
Abstract：Objective:To observe the effect of Erchentang, Chaihu Shugansan, Erchentang+ Chaihu Shugansan and metformin on insulin receptor substrate-1(IRS-1)and insulin receptor substrate-2(IRS-2)mRNA and protein expression in liver tissues of model rats with polycystic ovary syndrome(PCOS), and to explore the biological mechanism of relieving depression and resolving phlegm to improve insulin resistance of polycystic ovary syndrome.Method:A model of polycystic ovary syndrome was replicated in rats by subcutaneous injection of dehydroepiandrosterone(DHEA) under the neck and back.The control group was the normal group, The model rats were divided into model group, Erchentang group(9.135 g·kg-1), Chaihu Shugansan group(6.615 g·kg-1), Erchentang+ Chaihu Shugansan group (Hefang group, 9.135 g·kg-1+ 6.615 g·kg-1), and metformin group(0.158 mg·kg-1). The corresponding drugs were given by intragastric administration once a day for 4 weeks. The ovarian and liver tissues of rats were isolated, and serum testosterone(T)and estradiol(E2)hormone levels in each group were detected by enzyme-linked immunosorbent assay(ELISA), the histopathological changes of ovary in each group were detected by hematoxylin-eosin (HE) staining, IRS-1 and IRS-2 mRNA and protein expression levels in rat liver tissues were detected by quantitative real-time fluorescence PCR(Real-time PCR)and Western blot.Result:Compared with normal group, the serum T and E2 level of model group were significantly decreased (P<0.01), the histology of ovary showed a large number of saccate dilated follicles, thin granular cell layer and rare corpus luteum, IRS-1, IRS-2 mRNA and protein levels in liver tissues decreased significantly (P<0.01). Compared with model group, the serum T and E2 level of each group significantly decreased (P<0.01), histopathology of ovarian tissue showed that the number of saccular dilated follicles decreased, and the granulosa cell layer thickened, showing follicles of all levels, IRS-1, IRS-2 mRNA and protein expression were significantly increased in Erchentang group(P<0.05, P<0.01), IRS-1 mRNA and protein expression were significantly increased in Chaihu Shugansan group(P<0.01), IRS-2 mRNA and protein expression were on the rise, IRS-1 mRNA and protein expression and IRS-2 mRNA expression were significantly increased in the Hefang group (P<0.05), while IRS-2 mRNA was on the rise, IRS-1 protein and IRS-2 mRNA and protein expression were significantly increased in metformin group (P<0.01), while IRS-1 mRNA showed an upward trend.Conclusion:Erchentang improved the insulin resistance of PCOS model rats more significantly than Chaihu Shugansan, and its mechanism may be related to IRS-1, IRS-2 mRNA and protein expression levels of key signaling molecules regulating insulin signal transduction pathway.
Abstract：Objective:To study whether long-term administration of Gastrodiae Rhizoma powder can improve the learning and memory ability of APPswe/PSldE9 double transgenic (APP/PS1) Alzheimer' s disease(AD) model mice and delay the progress of AD whether these effects are related to the regulation of antioxidant stress pathway in Kelch-like epoxylopropylamine-related protein 1(Keap1)-nuclear factor E2 related factor 2 (Nrf2)/heme oxygenase(HO)-1, and further explore the neuroprotective mechanism of Gastrodiae Rhizoma powder and its role in the prevention and treatment of AD.Method:APP/PS1 double transgenic mice model, the mice consisted of five groups: normal, normal administration group, model group, Gastrodiae Rhizoma powder prevention group, Gastrodiae Rhizoma powder treatment group.The mice in the normal administration group and the Gastrodiae Rhizoma powder prevention group were given the same dose of Gastrodiae Rhizoma powder (1.5 g·kg-1) daily at the age of 8 weeks.The normal group and model group were given the same amount of normal saline at the same time, until 24 weeks old, Morris water maze was used to test the learning and memory ability of mice, and the treatment group was treated with Gastrodiae Rhizoma powder at 22 weeks old.The mice were given the same dose of Gastrodiae Rhizoma powder (1.5 g·kg-1) every day for 2 weeks.The number of crossing platform, escape latency and platform residence time of mice were detected by Morris water maze from 24 weeks old to 24 weeks old.RNA, Real-time PCR was extracted from mouse hippocampus to detect the mRNA level of Nrf2, HO-1, Keap1, and Western blot was used to detect the expression of Nrf2, HO-1, Keap1 protein in mouse hippocampus.Result:Compared with normal group, the water maze test showed that the learning and memory ability of model group was lower than that of the model group (P<0.01), and the learning and memory ability of Gastrodiae Rhizoma powder prevention group and Gastrodiae Rhizoma powder treatment group was significantly higher than that of model group (P<0.01). Compared with normal group, the levels of Nrf2, HO-1 and protein in the hippocampus in model group decreased in varying degrees (P<0.05). Compared with model group, Gastrodiae Rhizoma powder prevented Nrf2, in the hippocampus of mice in model group.The level of HO-1 in mRNA and protein increased in different degrees (P<0.05, P<0.01). Levels of Nrf2, HO-1 mRNA in Gastrodiae Rhizoma powder treatment group was significantly higher than that in Gastrodiae Rhizoma powder group (P<0.05). There was no significant difference in the expression of Nrf2, HO-1 protein.There was no significant difference in mRNA and protein levels of Keap1 among different groups.Conclusion:Morris water maze test and other results showed that Gastrodiae Rhizoma powder could improve the learning and memory ability of APP/PS1 mice, and it may enhance the expression of downstream antioxidant genes by regulating Keap1-Nrf2/HO-1 pathway.And then improve the learning and memory ability of APP/PS1 mice.
Keywords：Gastrodiae Rhizoma;APP/PS1 double transgenic mice;Alzheimer's disease;Kelch-like epoxylopropylamine-related protein 1(Keap1)-nuclear factor E2 related factor 2 (Nrf2)/heme oxygenase(HO-1) pathway
Abstract：Objective:To observe the effect of icariin on damaged neurons from the perspective of endoplasmic reticulum stress, in order to explore some mechanisms for repairing damaged neurons.Method:PC12 cells were induced by nerve growth factor (NGF) to differentiate into neurons, and the positive rate of microtubule associated protein-2 (MAP2) and neuron-specific enolase (NSE) expressions was determined by flow cytometry. The experiment was divided into 4 groups, blank control group: PC12 induced differentiation into neuronal cells, solvent control group: PC12 induced differentiation into neurons+ 0.1% dimethyl sulfoxide (DMSO), thapsigargin group: PC12 induced differentiation into nerves Yuan+ 2 μmol·L-1 thapsigargin, and icariin group: PC12 induced differentiation into neurons+ 2 μmol·L-1 thapsigargin+ 0.1 μmol·L-1 icariin. The proliferation of the cells was detected by using cell counting kit-8(CCK-8) method, the apoptosis of the cells was detected by flow cytometry, the protein expressions of CCAAT/enhace-binding protein homologous protein(CHOP) and glucoseregulated protein 78(Grp78) were detected by Western blot, and the mRNA expressions of CHOP and Grp78 were detected by real-time quantitative PCR (Real-time PCR).Result:Compared with the solvent control group, the thapsigargin group inhibited the proliferation of neuron-like PC12 cells induced by NGF, promoted apoptosis, and up-regulated the expressions of CHOP and Grp78 (P<0.05, P<0.01). Compared with the thapsigargin group, the icariin group can alleviate the inhibition of neurotrophic activity by thapsigargin, reduce neuronal apoptosis, and down-regulate the expressions of CHOP and Grp78 (P<0.05, P<0.01).Conclusion:Icariin can inhibit endoplasmic reticulum stress by down-regulating the expressions of CHOP and Grp78 and promote the repair of damaged neurons.
Abstract：Objective:To study the protective effect of astragalus polysaccharide (APS) on micronucleus and sister chromatid exchange (SCE) in human bone marrow mesenchyml stem cell (BMSCs) exposed to formaldehyde, in order to initially explore the potential mechanism.Method:BMSCs were cultured in vitro, cells were randomly divided into five groups: control group, formaldehyde group, and APS 40, 100, 400 mg·L-1 groups. BMSCs were infected with 120 μmol·L-1 formaldehyde, meanwhile, APS 40, 100, 400 mg·L-1 groups were co-cultured with 40, 100, 400 mg·L-1 APS. Cell morphology was observed by inverted phase contrast microscope, micronucleus were detected by micronucleus test, SCE was detected by SCE test, and mRNA and protein expressions of proliferating cell nuclear antigen (PCNA), xeroderma pigmentosum B, D, F, G (XPB, XPD, XPF, XPG) were detected by quantitative real-time fluorescence polymerase chain reaction(Real-time PCR)and Western blot.Result:Compared with control group, cell counts decreased, and cell morphology of BMSCs in formaldehyde group significantly changed, they were all recovered gradually in 40, 100, 400 mg·L-1 APS groups. Compared with control group, the micronucleus and SCE increased significantly (P<0.01), PCNA mRNA and protein expressions down-regulated significantly (P<0.05), while XPB, XPD, XPF, XPG mRNA and protein expressions up-regulated significantly (P<0.05, P<0.01). Compared with formaldehyde group, BMSCs were treated with APS at 40, 100, 400 mg·L-1, micronucleus and SCE decreased significantly (P<0.01), and mRNA and protein expressions of PCNA, XPB, XPD, XPF and XPG up-regulated significantly (P<0.05, P<0.01). Among them, the 100 mg·L-1 APS group had the most obvious effect.Conclusion:APS can protect formaldehyde-induced BMSCs micronucleus and SCE, especially 100 mg·L-1 APS has the most obvious effect. The mechanism may be associated with the up-regulation of expressions of PCNA, XPB, XPD, XPF and XPG in the nucleotide exicision repair pathway (NER), which promoted the damage repair.
Abstract：Objective:This study aims to explore the effect and mechanism of Yuehua capsule serum for autophagy of macrophages infected with multi-drug resistant mycobacterium tuberculosis.Method:The rats were undertaken intragastric gavage with Yuehua capsule by 3.02 g·kg-1 once a day which was produced through low temperature condensation drying method. After 7 days, blood of abdominal aorta of rats was collected to prepare Yuehua capsule serum. RAW264.7 andmultidrug resistant tuberculosis were cultured in vitro.According to cell counting kit-8(CCK-8), 10% drug-containing serum was considered as the effective concentration. The cultured cells were divided into four groups: model groups(10% fetal bovine serum). Yuehua capsule serum(10% Yuehua capsule serum). Autophagy inhibitor group+ 3-MA+ Yuehua capsule medicated serum(3-MA+ 10% Yuehua capsule serum). Rapamycin (Rap) positive control group(200 mg·L-1 Rap+ 10% Yuehua capsule serum). Except for the normal group, the cells of each group were cultured for 24 h and infected for 4 h according to cell-bacteria 1∶10.Testing index: observation of autophagosomes under transmission electron microscope, the test of expression of microtubule-associated protein light chain-3Ⅱ(LC-3Ⅱ), microtubule-associated protein LC 3-Ⅱ/microtubule-associated protein light chain 3-Ⅰ(LC3-Ⅰ) and Beclin-1 with Western blot, indirect immunofluorescence staining for LC3B, and mRNA of Beclin-1 as well as LC3 with real-time fluorescent quantitative polymerase chain reaction(Real-time PCR).Result:Compared with normal group, model group did not see autophagy body cells, cells in the LC-3 Ⅱ, LC-3 Ⅱ/LC-3 Ⅰ, Beclin-1 protein and LC3, Beclin-1 mRNA gene expression level had no significant change, the cells without fluorescent particles, spots, no fluorescence intensity.Compared with model group, Yuehua capsules serum group and Rap positive control group can be observed the formation of phage, mRNA andprotein expression levelof LC-3 Ⅱ, LC-3 Ⅱ/LC-3 Ⅰ, Beclin-1 and LC3, Beclin-1 were significantly increased (P<0.05). Autophagy inhibitor group+ 3-MA+ Yuehua capsule medicated serum did not see autophagy, the mRNA and protein expression level of LC-3 Ⅱ, LC-3Ⅱ/LC-3Ⅰ, Beclin-1 and LC3, Beclin-1 were no significantly increased.Conclusion:Yuehua capsule medicated serum could induce autophagy of macrophages of RAW264.7.The mechanism was probably accomplished through regulating the expression level of autophagy key protein LC3, autophagosome mature protein Beclin-1 and relevant gene, meanwhile the conversion of LC3-I to LC3-Ⅱ was accelerated.
Abstract：Objective:To established the model of chronic alcoholic liver injury in rats by long-term(8 weeks) alcoholic gavage, to study the effects of Tibetan medicine Lagotis brachystachys extracts on Toll-like receptor(TLR)2/myeloid differentiation factor 88(MyD88)/nuclear factor kappa B (NF-κB)and NOD like receptor protein 3(NALP3) signaling pathways and study preliminary the mechanism of action of chronic alcoholic liver injury.Method:Sixty male Sprague-Dawley rats were randomly divided into normal group, model group, bifendate positive drug group (0.1 g·kg-1) and L. brachystachys low, medium and high-dose groups (0.5, 1, 2 g·kg-1), the corresponding drugs were given at 10 mL·kg-1 in each morning, and the 56 degree Liquor was administered by the afternoon gradient alcoholic gavage method.After 8 weeks, the levels of serum aspartate transaminase (AST), serum alanineaminotransfease(ALT), serum total cholesterol(TC), triglyceride(TG), interleukin-1β(IL-1β), and the liver levels of L-glutathione(GSH)were measured. The expression of TLR2, MyD88, NF-κB and NALP3 protein in liver were detected by Western blot.Hematoxylin-eosin (HE) staining was used to observe the pathological changes of liver tissue.Result:Compared with normal group, the serum levels of AST, ALT, TC, TG and IL-1β in model group were significantly increased (P<0.05, P<0.01). Compared with model group, the serum AST, ALT, TC, TG and IL-1β levels were decreased in the various doses of L. brachystachys, and the high dose group was particularly effective (P<0.05, P<0.01). Compared with normal group, the GSH level in the liver homogenate of model group decreased significantly, and the difference was not statistically significant. The levels of TLR2, MyD88, NF-κB and NALP3 in the liver tissue of model group were significantly increased (P<0.05, P<0.01). The GSH levels in the liver and the protein expression of TLR2, MyD88, NF-κB and NALP3 were decreased in L. brachystachys group (P<0.05, P<0.01). The liver pathological section showed that L. brachystachys can improve the pathological changes of rat liver tissue.Conclusion:L. brachystachys can protect liver from alcohol-induced chronic liver injury in rats. The mechanism was related to TLR2/MyD88/NF-κB and NALP3 signaling pathway.
Keywords：Lagotis brachystachys;Toll-like receptor(TLR)2/myeloid differentiation factor 88(MyD88)/nuclear factor kappa B (NF-κB)signaling pathway;NOD like receptor protein 3(NALP3);chronic alcoholic liver injury;mechanism
Abstract：Objective:To observe the clinical efficacy of Dahuang Zhuyu decoction for oral administration and enema on severe acute pancreatitis with syndromes of blood stasis and toxin and its effect on serum inflammatory factors.Method:Sixty eight patients with severe acute pancreatitis with syndromes of blood stasis and toxin who were admitted in Henan Provincial People's Hospital from September 2017 to December 2018 were randomly divided into treatment group (34 cases) and control group (34 cases). The control group was treated with western medicine. The treatment group was treated with Dahuang Zhuyu decoction for oral administattion and enema in addition to the therapy of the control group. Both groups were treated for 7 days. Before and after treatment, abdominal pain, bloating, nausea and vomiting were scored separately, serum amylase (AMS), lipase (LPS), interleukin-6 (IL-6), interleukin-10 (IL-10) and tumor necrosis factor-α (TNF-α) were detected, and abdominal pain disappearance time, bloating disappearance time and exhaust recovery time were recorded.Result:After treatment, the abdominal pain score, abdominal distension score, nausea and vomiting score, serum AMS, LPS, IL-6 and TNF-α were lower than those before treatment (P<0.05), and those in treatment group was lower than those in control group (P<0.05). After treatment, IL-10 level in both groups was higher than those before treatment (P<0.05), and that in treatment group was higher than that in control group (P<0.05). The disappearance time of abdominal pain, the disappearance time of abdominal distension and the recovery time of exhaust were shorter in treatment group than in control group (P<0.05). The total effective rate of the treatment group was higher than that of the control group (P<0.05).Conclusion:Dahuang Zhuyu decoction for oral administration and enema for severe acute pancreatitis with syndromes of blood stasis and toxin can alleviate clinical symptoms, reduce amylase and lipase levels, inhibit the expression of pro-inflammatory factors, promote the expression of anti-inflammatory factors, restore intestinal function, and improve clinical efficacy.
Keywords：Dahuang Zhuyu decoction;severe acute pancreatitis;syndromes of blood stasis and toxin;clinical efficacy;inflammatory factors;Dahuang Mudan Tang;Gexia Zhuyutang
Abstract：Objective:To observe the clinical efficacy of modified Zhishi Daozhiwan on slow transit constipation (STC) and the effect on gastrointestinal hormones and intestinal flora.Method:One hundred and sixty patients were randomly divided into control group and observation group by random number table. Patients in control group got Maren Wan, 1-2 pills/time, 2 times/days, and mosapride citrate tablets, 1 piece/time, 3 times/days. Patients in observation group got modified Zhishi Daozhiwan, 1 dose/day. A course of treatment was 4 weeks. Before and after treatment, the main symptoms of constipation and patient assessment of constipation symptoms (PAC-SYM) were scored. And times of spontaneous complete bowel movement (SCBM) and colonic transmission test were recorded. And levels of intestinal flora and motilin (MTL), vasoactive intestinal peptide (VIP), substance P (SP) and gastrin (GAS) were detected. The 12-week follow-up of patients (SCBM≥3) was recorded, and the relapse of disease was calculated.Result:According to rank sum test analysis, the clinical efficacy in observation group was better than that in control group (Z=2.275, P<0.05). After treatment, scores of the main symptoms of constipation, PAC-SYM, and the total score of PAC-SYM were all lower than those in control group (P<0.01). At the second, third and fourth weeks after treatment, the times of SCBM were more than those in control group (P<0.01). Ratio of residual marker at 24, 48 and 72 h was lower than that in control group (P<0.01). Count of enterococcus and enterobacter were lower than those in control group (P<0.01), while counts of bifidobacterium and lactobacillus were higher than in control group (P<0.01). Levels of GAS, MTL and SP were higher than those in control group, whereas level of VIP was lower than that in control group (P<0.01). And relapse rate in observation group was 33.85%, which was lower than 57.69% in control group (χ2=6.653, P<0.05).Conclusion:Modified Zhishi Daozhiwan can alleviate constipation and other symptoms, increase the number of SCBM, regulate gastrointestinal hormones and intestinal flora, and improve colonic transit function, with a good clinical efficacy and low recurrence rate, so it is worth clinical application.
Abstract：Objective:To observe the efficacy of Wenfei Jianpi decoction on moderate to severe persistent allergic rhinitis (AR) with Lung Qi deficiency and cold syndrome and its effect on substance P (SP) in nasal secretions and peripheral blood proinflammatory factors.Method:One hundred and sixty-six patients were randomly divided into control group and observation group by random number table. Patients in control group got budesonide nasal spray, 2 times/days, and cetirizine hydrochloride tablets, 10 mg/days. And patients in observation group got Wenfei Jianpi decoction for oral, 1 dose/day. A course of treatment was 8 weeks. Before and after treatment, nasal obstruction, itching, sneezing, runny nose, itchy eyes, tears and nasal sign were scored, nasal airway resistance was detected, and rhinoconjunctivits quality of life questionnaire (RQLQ) were scored. Before and after treatment, levels of SP immunoglobulin E (IgE) in peripheral blood, interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were detected, and eosinophil (EOS) were calculated.Result:By rank sum test, the clinical efficacy in observation group was better than that in control group (Z=2.073, P<0.05). Before and after treatment, scores of total score of nasal symptoms, ocular symptoms and subjective symptoms and nasal sign score were all lower than those in control group (P<0.05). Except sleep and emotion, scores of other dimensions and the total scores of RQLQ were all lower than those in control group (P<0.05). EOS in observation group was less than that in control group (P<0.05). And nasal resistance declined in observation group (P<0.05), and levels of IgE, SP, IL-4, IL-5, IL-6, TNF-α and nasal resistance were lower than those in control group (P<0.05, P<0.01).Conclusion:Wenfei Jianpi decoction can be used to treat moderate to severe persistent allergic rhinitis by relieving subjective symptoms and signs, improving quality of life, inhibiting inflammatory factors and reducing allergic reaction.
Keywords：allergic rhinitis;Lung Qi deficiency and cold syndrome;Wenfei Jianpi decoction;inflammatory factors;substance P
Abstract：Objective:A multi-organ chip of intestine-liver-breast cancer was constructed based on microfluidic technology and used for pharmacokinetics-pharmacodynamics (PK-PD) study of drugs in vitro.Method:A multi-organ chip comprising a 4-layer polydimethylsiloxane (PDMS) substrate and a 2-layer poly(methyl methacrylate) (PMMA) cover was constructed by microfluidic technology. The connection between cells was investigated by staining the 21-day-grown human colon cancer cell line Caco-2 cell layer and the 3-day-grown human umbilical vein endothelial cell line HUVEC cell layer with CellTracker Red/Green and Hoechst, respectively. The transmission rates of 2 g·L-1 fluorescein sodium and 33.28 mg·L-1 propranolol acrossing the cell layer were employed to verify the function of the constructed intestinal module. The metabolic level of the liver module was investigated by comparing the inhibition rate of 125 μmol·L-1 cyclophosphamide against human breast cancer cell line MCF-7 cells treated with human hepatoma cell line HepG2 cells in a conventional well plate and chip liver module for 48 h. The secretion of albumin by HepG2 cells in the chip was detected to verify the synthesis function of hepatic module. Caco-2 cell layer, HUVEC cell layer, HepG2 cell layer, MCF-7 cell layer and dialysis membrane were assembled on the chip, the culture medium containing 55 mg·L-1 propranolol was injected into the upper channel of the chip for 4 h, and then changed into the normal culture solution. The mass concentration of propranolol in the lower circulating culture medium at each time point within 72 h was determined, and the drug-time curve was drawn. The culture medium containing 125 μmol·L-1 cyclophosphamide, 5 μmol·L-1 paclitaxel, 50 μmol·L-1 capecitabine was injected into the circulating fluid in the upper layer of the chip, in order to study the inhibition rates of the three anti-tumor drugs on the MCF-7 cell layer on the chip within 72 h, and the results were compared with those of the 96-well plate.Result:The constructed chip performed well. The Caco-2 and HUVEC cell layers were tightly connected. The transmission of fluorescein sodium and propranolol between the cell layers demonstrated that the constructed intestinal module had good absorption and transport function. The inhibition rate of MCF-7 by 125 μmol·L-1 cyclophosphamide after metabolism of HepG2 cells on the well plate was 22.12%, and the inhibition rate of MCF-7 by the unmetabolized cyclophosphamide was 1.84%. The inhibition rate of MCF-7 increased to 32.13%after injected 125 μmol·L-1 cyclophosphamide from the upper layer of the chip liver module, and the inhibition rate of MCF-7 after injection from the lower layer of the chip liver module was 7.23%. The mass concentration of propranolol on the chip changed with time, which was basically consistent with that in vivo. The inhibition rate of MCF-7 on the plate with 125 μmol·L-1 cyclophosphamide was lower than that on the chip, and the inhibition rates of MCF-7 on the plate with 5 μmol·L-1 paclitaxel and 50 μmol·L-1 capecitabine were higher than those on the chip.Conclusion:The constructed multi-organ chip of intestine-liver-breast cancer has the absorption and transport function of the intestine and the metabolic function of the liver. The chip can reflect the pharmacokinetic properties of propranolol in vivo, and can be used for pharmacodynamic studies of paclitaxel and capecitabine.
Abstract：Objective:To establish HPLC fingerprints of Aurantii Fructus and its processed products, and to quantitatively analyze the contents of four flavonoids in these products.Method:HPLC was employed with Inertsil ODS-3 C18 column (4.6 mm×250 mm, 5 μm), the mobile phase of acetonitrile-0.1%phosphoric acid aqueous solution for gradient elution, the detection wavelength of 283 nm, and the flow rate of 1.0 mL·min-1. HPLC fingerprints of raw products, stir-fried bran products and processing products of Aurantii Fructus were established. Similarity evaluation and cluster analysis were used to analyze the chromatographic data. At the same time, the contents of narirutin, naringin, hesperidin and neohesperidin were determined.Result:HPLC fingerprints of Aurantii Fructus and its processed products were established, taking naringin as the reference peak, 8, 15, 11 common peaks were demarcated for raw products, stir-fried bran products, processing products, respectively, the similarities of fingerprints were >0.95.Contents of the above four flavonoids in raw products were 0.574 7%, 5.986 3%, 0.302 2%and 3.574 7%, respectively. After processing, the contents of these four components in stir-fried bran products turned into 0.948 4%, 5.103 4%, 0.549 3%and 3.533 7%, their contents in processing products turned into 0.605 3%, 4.762 3%, 0.404 7%and 3.264 9%, respectively.Conclusion:The HPLC fingerprint of Aurantii Fructus changes significantly before and after processing. The contents of four flavonoids change to a certain extent before and after processing. The order of contents of narirutin and hesperidin in samples was stir-fried bran products>processing products>raw products, while the order of contents of naringin and neohesperidin was raw products>stir-fried bran products>processing products.
Keywords：Aurantii Fructus;Aurantii Fructus stir-fried with bran;processing products of Aurantii Fructus;fingerprint;cluster analysis;flavonoids;determination
Abstract：Objective:To establish a rapid evaluation method for Cinnamomi Cortex decoction pieces by near infrared spectroscopy.Method:The contents of coumarin, cinnamalol, cinnamic acid and cinnamaldehyde in 86 batches of Cinnamomi Cortex of different origins were determined by HPLC. And the NIR spectra of different batches of Cinnamomi Cortex were also collected. With NIR spectrum as independent variable and coumarin, cinnamalol, cinnamic acid and cinnamaldehyde as dependent variables, a quantitative analysis model of four components in cinnamon was established by partial least squares method.Result:The correlation coefficients (r) of coumarin, cinnamic alcohol, cinnamic acid and cinnamaldehyde near infrared quantitative analysis models were 0.952 8, 0.977 7, 0.961 9, 0.992 2, root mean square error of cross(RMSEC) were 0.012 2, 0.006 1, 0.004 3, 0.82 g·g-1, root mean square errorof cross-validation(RMSECV) were 0.015 8, 0.011 2, 0.002 0, 1.481 1 g·g-1, and root mean square error of prediction(RMSEP) were 0.017 8, 0.010 3, 0.010 3, 0.005 5, 1.63 g·g-1.Conclusion:The established NIR quantitative analysis model of four active ingredients in Cinnamomi Cortex slices has a good accuracy, and provides a basis for rapid evaluation of the quality of Cinnamomi Cortex slices.
Abstract：Objective:Camptosorus sibiricus is a kind of herbal medicine and famous folk medicine. However, the bioactivities or pharmacological effects of the C. sibiricus remain to be unclear. Therefore, it is necessary to make a systematic study on chemical constituents from C. sibiricus, so as to further study its potential medicinal value, and provide certain chemical basis and foundation for the comprehensive development and the search for pharmacological activity.Method:Various column chromatographic technologies, (silica gel, Sephadex LH-20 and ODS column chromatography) as well as HPLC were adopted to separate chemical constituents of C. sibiricus extract. The structure of the purified compounds was elucidated by nuclear magnetic resonance (NMR) and high-resolution electrospray ionization-mass spectrometry (HR-ESI-MS).Result:Totally 10 compounds have been isolated from water extract of C. sibiricus. By spectroscopic methods, they were elucidated as 7-dien-3-on-9-O-β-D-glucoside (1), bridelionoside F (2), (3R, 5S, 6S, 7E, 9S)-megastigm, an-7-ene-3, 5, 6, 9-tetrol 9-O-β-D-glucopyranoside (3), (6S, 7E, 9R)-roseoside (4), (3S, 5S, 6R, 9R)-3-hydroxy-5, 6-epoxy-β-ionol-9-O-β-glucopyranoside (5), 6, 9-dihydroxy-4, 7-megastigmadien-3-one (6), (3R, 6R, 7E, 9R)-3, 9-dihydroxy-4, elaphoside A (7), ptelatoside-A (8), n-butyl-a-β-D-fructofuranoside (9), dibutylphthalate(10)based on physical and chemical properties.Conclusion:All compounds were obtained from C. sibiricus for the first time. The discovery of these compounds further enriched the chemical constituents of C. sibiricus, and provided experimental and scientific basis for the comprehensive development and utilization of C. sibiricus.
Keywords：Camptosorus sibiricus;water extract;(7E)-9-hydroxymegastigma-4, 7-dien-3-on-9-O-β-D-glucoside;bridelionoside F
Abstract：Objective:To study the degradation of salvianolate lyophilized injection (SLI) and establish a stability-indicating analysis method.Method:UPLC-Q-TOF-MS/MS was used to conduct a qualitative study on the main components of SLI, and a stability-indicating analysis method was established for simultaneous determination of the original components of SLI and its degradation products. The stability of SLI were systematically assessed under physicochemical conditions of high temperature, oxidation, metal ions.Result:Totally 13 main active ingredients in SLI were identified, and a semi-quantitative analysis was performed. Under the conditions of high temperature, oxidation, light, trivalent ion and divalent ion, 6, 4, 3, 4 and 1 new degradation products were added respectively. The established stability-indicating analysis method can simultaneously determine the degradation products of the main components and their active components in SLI, with a good separation effect.Conclusion:According to the degradation mechanism of the main ingredients in SLI, macromolecular polyphenol acid compounds are degraded into small molecular compounds, such as tanshinol and protocatechu aldehyde by a series of reactions, like benzofuran open-loop, hydrolysis of ester bond and removal of DSS. The stability-indicating analysis method can be used for the stability quality control of traditional Chinese medicine Salvianolate Lyophilized Injection (SLI).
Abstract：Objective:Astragali Radix is an important medicinal and edible herb. To achieve standardized cultivation of Astragali Radix and improve cultivation results, Astragalus membranaceus var. mongholicus was cultivated with a row spacing of 30 cm and different plant spacing (8, 10, 12, 14, 16 cm) in a test base of Longxi County, Gansu Province.Method:The growth and development dynamics of green strains and the yield and quality of medicinal materials were measured, and the comprehensive evaluation of membership function was used to determine the rational transplanting density and explore the standard production technology of A. membranaceus var. mongholicus.Result:The transplanting topping had a significant effect on the growth and development of A. membranaceus var. mongholicus under the condition of the row spacing of 30 cm. With the increase of topping, the biomass of astragalus on the ground decreased, and the ratio of root to shoot increased, but the yield of medicinal material per unit area decreased. The appearance traits were improved as the transplant density decreased. Under the row spacing of 14 cm, the content of astragaloside and calycosin-7-glucoside were the highest. Under the row spacing of 8-16 cm, the content of root extract of Astragali Radix was better than the Chinese Pharmacopoeia standard. The comprehensive evaluation index of each plant spacing treatment was 14 cm>16 cm>10 cm>8 cm>12 cm in turn.Conclusion:Combined with the comprehensive evaluation of yield and economic benefit, the optimal transplanting plant spacing and row spacing were 30 cm×14 cm (with the density of 238 100 strains/hm2). Under this density, A. membranaceus var. mongholicus grows vigorously, with thick stems, big root crowns, a high yield and high quality.
Keywords：Astragalus membranaceus var. mongholicus;transplant density;growth and development;officinal yield and quality
Abstract：Objective:To determine the relationship between the characters and the main components of Chelidonii Herba pieces.Method:The main components of the samples were determined by HPLC, and the characters of Chelidonii Herba Pieces were evaluated by sensory evaluation. The correlation between the characters and components was calculated by correlation formula.Result:The content of 6 components, such as chelidonine, was determined by HPLC. Based on the result of sensory evaluation, there was a certain correlation between the characters and the components. Character descriptions that " some have visible white powder, some have white pubescence, and sometimes they have visible yellow florets" had a very low similarity with total alkali, indicating that these characters and total alkali content was not related. The similarity between the description of " hollow stem" had a high similarity with total alkali content, indicating that the amount of stem was related to the total alkali content. The character description of " more broken leaves" was negatively correlated with total alkaloids in the similarity, which indicated that the content of total alkaloids was less when there were more leaves(or more broken leaves), otherwise, the content of total alkaloids was relatively higher.Conclusion:The established HPLC method is simple and feasible. This study objectively quantifies the descriptions of Chelidonii Herba pieces characters, correlates them with the main components of Chelidonii Herba pieces, and then preliminarily judges the quality of Chelidonii Herba pieces according to the appearance of the characters, which provides a theoretical basis for the identification of Chelidonii Herba pieces in the market by experience, and ideas for the study of the characters of other traditional Chinese medicines.
Abstract：Objective:To study the effect of different packaging methods and storage conditions on the quality of Notopterygii Rhizoma et Radix pieces, in order to determine the optimal packaging method and suitable storage conditions for Notopterygii Rhizoma et Radix pieces.Method:Different packaged Notopterygii Rhizoma et Radix pieces were stored in different environments in a one-year long-term stability experiment. The appearance, water content, extract content and volatile oil content of Notopterygii Rhizoma et Radix pieces were regularly observed.Result:During the 1-year storage period, the Notopterygii Rhizoma et Radix pieces under different packaging and storage conditions showed different degrees of quality changes. Among them, the samples packed in polyethylene plastic bags and polyethylene aluminum foil composite bags were better preserved. The fluctuations in water content of the sample packed in polyethylene plastic bags were relatively low, and the RSD value of water content during the month was less than 11.5%. The extracts and volatile oil contents of each sample were reduced to different degree, but the samples packed in plastic sealed bags and protected from light had the smallest annual loss of extracts (1.27%), with the lowest monthly loss rate of volatile oil (0.08%).Conclusion:The quality of Notopterygii Rhizoma et Radix pieces can be well preserved in plastic sealed bags and storage in dark and cool conditions, which is suitable for the storage of Notopterygii Rhizoma et Radix pieces.
Keywords：Notopterygii Rhizoma et Radix pieces;package;storage;water content;extract;volatile oil
Abstract：Objective:Bioinformatic analysis was used to compare the gene expression profile between asthma patients and healthy people, and the gene characteristics of asthma were preliminarily identified and the potential mechanism and drugs were revealed.Method:The GSE74986 gene expression profile was downloaded from the gene expression omnibus (GEO) and the differentially expressed genes (DEGs) were analyzed by GEO2R. Then the gene heat map of DEGs was made by Morpheus, and their gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) analysis were performed by DAVID 6.8. Moreover, the protein-protein interaction (PPI) network and hub genes were constructed by String 10.5. Finally, the significant modules were analyzed by MCODE in Cytoscape 3.6.1, small molecule drugs related to asthma were screened through Coremine Medical.Result:A total of 510 DEGs were screened, including 29 up-regulated genes and 481 down-regulated genes. DEGs were mainly involved in these biological processes and pathways, including chromatin silencing, transcriptional regulation of RNA polymerase Ⅱ promoter, protein transport, messenger RNA (mRNA) processing, RNA splicing, ubiquitin-mediated proteolysis, protein processing in the endoplasmic reticulum, RNA transport, and myeloid differentiation factor (MyD)-dependent Toll-like receptor signaling pathway, platelet activation, nucleotide binding oligomerization domain (NOD)-like receptor signaling pathway and so on. A total of 9 hub genes were obtained, including T-complex protein 1 subunit theta (CCT8), T-complex protein 1 subunit alpha (TCP1), 26S protease regulatory subunit S10B (PSMC6), heat shock protein 90 alpha (HSP90A)A1, cell cycle protein C (CCNC), HSP90AB1, 26S proteasome non-ATPase regulatory subunit 6 (PSMD6), ubiquitin-specific protease 14 (USP14) and eukaryotic translation initiation factor 4E (EIF4E). Two important modules were obtained. The genes in two modules mainly involved these biological process, such as splice, ubiquitin-mediated proteolysis, protein modification, RNA modification and so on. Some potential molecular drugs for the treatment of asthma, such as anisomycin and genistein, have been developed.Conclusion:DEGs and hub genes can contribute to understanding the molecular mechanism of asthma and providing potential therapeutic targets and drugs for the diagnosis and treatment of asthma.
Keywords：asthma;differentially expressed genes;hub genes;gene ontology;Kyoto encyclopedia of genes and genomes;protein-protein interaction;small molecule drugs
Abstract：The inheritance of traditional clinical value of famous classical formulae is an important direction for the development of traditional Chinese medicine industry.Compared with the previous research and development of new drugs, the management requirement of " material reference" was introduced into the famous classical formulae research, which is used as the reference of process optimization and quality control.The characteristics of compound preparation of famous classical formulae are also reflected in the core concept of " quality inheritance of classics" in the road of industrial development.How to implement the above requirements and concepts into product development and industrial production? There are many specific common problems to be solved in practical research.How to effectively establish the " material reference" of famous classical formulae of different dosage forms? How to use " material reference" to guide the process optimization of compound preparation of famous classical formulae? How to determine the daily dose of famous classical formulae? How to take effective measures in the selection of raw material to reduce quality fluctuation range? This paper discusses the key issues such as production process and quality evaluation from the following aspects.Firstly, the management regulations and research and development guidelines are analyzed, and the specific implementation methods are given.Then, the possible problems in the Requirements for Declaration Documents (Draft for Opinions) are pointed out, and relevant suggestions are given.Finally, based on the research experience of standard decoction and famous classical formulae in the laboratory, an example is given to provide reference for the development of compound preparation of famous classical formulae.
Abstract：Objective:To investigate the distribution status of medicinal plants in the wild areas of Russian Caucasus and Altai, and clarify the types and efficacy information of medicinal plants in the area, so as to dig deep into new resources and new functions of medicinal plants in the countries along the Belt and Road.Method:Medicinal plants in the wild were searched and collected to make waxy specimens, and sent back to the country to extract the total DNA of the leaves of the leaves. Internal Transcribed Spacer(ITS)sequence universal primers were used for Polymerase Chain Reaction (PCR)amplification. The PCR products were sent for the two-way sequencing, and the sequencing results are spliced by software according to National Center for Biotechnology Information(NCBI). The same ITS sequence of the highest similarity species obtained by database BLAST was analyzed by DNAman software to identify the ITS sequence of the species and the ITS sequence of the same species. The MEGA 7 software was used as the phylogenetic tree, and the Kimura-2 parameter genetic distance was used to construct the neighbor joining(NJ) phylogenetic tree by the neighbor-joining method. The confidence of each branch of the development tree was tested by the bootstrap test method. A total of 2 000 cycles were performed, and the results were identified based on the clustering results. On this basis, the key medicinal plants in the Russian Caucasus and Altay wild areas were summarized and analyzed.Result:After BLAST alignment in NCBI database, the ITS sequences of each specimen were clustered with the login sequences on the NCBI database, which were separated from the outer group. The species classification of the specimens to be identified was determined by combining the characteristics of the specimens. A total of 51 plants were identified from the specimens collected in the field, covering 44 genera of 17 families, and 29 plants had clear efficacy records. The National Drug List of the Russian Federation and the Chinese Pharmacopoeia were retrieved to summarize commonly used medicinal plants in China and conclude that 20 kinds of Chinese and Russian common medicinal materials have different medicinal effects in local areas. This study has a reference significance for expanding the scope and clinical experience of traditional Chinese medicines, and provides a basis for strengthened local species conservation, development and utilization of wild medicinal plant resources.
Keywords：medicinal plants;internal transcribed spacer sequence;DNA barcode;National Center for Biotechnology Information Database
Abstract：Objective:To deeply understand important scientific problems and key technical problems in the cultivation of Chuanxiong, the Chuanxiong cultivation situation of 247 households in 23 towns of 5 counties in Sichuan province were investigated, providing basic data for the standardized cultivation of Chuanxiong.Method:On the basis of reported literatures, a questionnaire survey was conducted on the farmers of cultivating Chuanxiong in main producing areas, field measurements were also preformed, and the cultivation status of Chuanxiong was comprehensively analyzed.Result:The proportion of plain-breeding mainly existed in Pengzhou and Shifang, where per farmer had a small scale of planting areas, was 65%, while the proportion of mountain-breeding mainly existed in Meishan, Qionglai and Dujiangyan, where per proprietor had a large scale of planting areas, was 92%. The planting density and yield of Chuanxiong varied greatly among individuals. The growth period of Chuanxiong in Meishan and Qionglai is about 30 days shorter than that in Pengzhou, Shifang and Dujiangyan. About half of farmers applied base fertilizer and top dressing in spring. The proportion of NPK compound fertilizer input was high (>90%). Chuangxiong has a few diseases and insect pests, the incidence of which in new production areas was obviously lower than that in old production areas. The application of herbicide was not standardized. 52% farmers used Chu cao ling No.1 which was made by agricultural material store owners. The labor cost accounted for the highest proportion (37%) in Chuanxiong cultivation.Conclusion:Now in Sichuan clonal propagated patterns of Chuanxiong were both mountain-breeding and plain-breeding. It is suggested to make clear the differences between mountain-breeding nodes and plain-breeding nodes, and breed high qualities of Chuanxiong nodes. The mechanization research and production of Chuanxiong should be promoted, realizing timely planting and harvesting. Farmers should be guided to increase the application of organic fertilizer, and meanwhile standardize the application of agrochemicals.
Abstract：Platycodonis Radix, which was first recorded in the Agriculture God's Canon of Materia Medica. It is a multi-functional drug with a wide range of applications. The processing of Platycodonis Radix has been recorded as early as in the Jin dynasty, and has a long history of processing. Today, in addition to the washing, cutting and stir-frying with honey, there have also been more than 20 kinds of processing methods, such as stir-frying with wine, stir-frying with bran, stir-frying with Lilii Bulbus juice and so on. The ancients believed that Platycodonis Radix could enhance the effect of diffusing the lung, promoting pharynx and relieving cough by processing. In terms of the chemical compositions in Platycodonis Radix, more than 100 compositions, like triterpenoid saponins, flavonoids, phenols, sterols, polysaccharides and polyacetylenes, have been isolated and identified from it. Among them, triterpenoid saponins are the essential compositions. In addition, Platycodonis Radix has the pharmacological effects of expectorant, antitussive, anti-inflammatory, anti-tumor, etc. The medicinal ingredients of Platycodonis Radix are mainly triterpenoid saponins and polysaccharides. Among them, triterpenoid saponins have diverse biological activities, which lead it to be one of the hotspots of current researches. Platycodonis Radix has a good role in promoting lung and removing phlegm. After being processed, its medicinal effects are enhanced. It is complex and diverse in compositions of Platycodonis Radix so that has rich pharmacological activities. On the basis of sorting out the literature, this paper discusses the processing history, chemical composition and pharmacological effect of Platycodonis Radix, in order to provide reference for the special processing and modern research of Platycodonis Radix. Furtherly, it provides a theoretical basis for the research of its processing mechanism and quality control.
Abstract：Emodin is a common Chinese medicine compound with anti-inflammatory, antibacterial, anti-oxidant and lipid-lowering effects. Modern studies have found that emodin activates adenylate-activated protein kinase (AMPK) signaling molecules and regulates transcriptional factors and biological functions of relevant pathways. Nonalcoholic fatty liver disease is a chronic liver disease with a high incidence in China. With the global prevalence of obesity and metabolic syndrome, the development of nonalcoholic fatty liver disease (NAFLD) is closely related to the expression of the metabolism-related signal molecule AMPK. AMPK is a key enzyme in glycolipid metabolism that can involve different stages of NAFLD development to non-alcoholic steatohepatitis (NASH) by regulating energy metabolism in the body. In recent years, many studies have suggested that the activation of AMPK signaling molecules is related to the function realization of emodin, and lipid synthesis, fatty acid oxidation, insulin sensitivity and mitochondrial function-related transcription factors affected by AMPK downstream signaling molecules and other biological effects can be interacted with each other. The detailed mechanism of action associated with AMPK activation provides new thought about the treatment of NAFLD by emodin. This paper mainly summarizes the research progress of emodin by participating in the various stages of NAFLD by AMPK-related signaling pathways through literature retrieval and comprehensive analysis. It lays a foundation for further research on the therapeutic effect and mechanism of emodin on NAFLD.
Keywords：emodin;adenylate-activated protein kinase (AMPK);adiponectin;nonalcoholic fatty liver disease
Abstract：Hypertension is one of the most common chronic diseases, also an important risk factor for a series of cardio-and cerebra-vascular diseases. Due to its polygenic, multi-factorial nature and heterogeneity, the underlying cause has not been fully elucidated, satisfied therapeutic effect hasn't been totally achieved either. Metabolomics is used to evaluate metabolic changes of organisms from a holistic perspective, associating with biological processes to reveal the whole situation of the body. In recent years, researchers have used metabolomics to study the pathogenesis of hypertension, potential biomarkers, effects of lifestyle interventions, and mechanisms of antihypertensive drugs. Targeted or untargeted ways are applied to study metabolites form blood, urine, or tissues of human or animals. Metabolic pathways of gut microflora, oxidative stress, fatty acids, and amino acids have drawn more attention, and the discovered metabolites may become potential biomarkers, further the diagnostic biomarkers and treatment targets. In addition, traditional Chinese medicine (TCM) is an integrated complex system in syndrome diagnosis and treatment, and metabolomics coincides well with the concepts of it. TCM researchers also use this method to study the biological basis of syndromes in hypertension and the mechanism of antihypertensive Chinese medicine. There are significant differences in the metabolites between different syndromes. TCM treatments can restore the metabolite disturbance caused by high pressure, which is probably one of the pharmacological pathways of antihypertensive Chinese medicine. Metabolomic studies in hypertension have achieved great progress, but there're still challenges in data analysis, integration with other metabolomic studies and other omic studies and causal relationship in further study.
Abstract：With the improvement of people's living standard and the change of dietary structure, the prevalence of gout has increased gradually with the increased intake of protein, sugar and fat. There has been a positive correlation between gout and age, and the age of onset decreased gradually. The inflammation induced by sodium urate crystal is the pathological basis of gout, which activates innate immunity, releases many kinds of inflammatory mediators, such as interleukin(IL)-1β, IL-6 and tumor necrosis factor(TNF)-α, and then causes inflammatory cascade reaction and acute attacks, such as joint redness, swelling and heat pain. There is a spontaneous remission mechanism in gout. For one thing, macrophages reduce the stimulation of monosodium urate(MSU) through phagocytosis of MSU crystals as foreign bodies, for another, differentiated and mature macrophages secrete anti-inflammatory factor transforming growth factor(TGF)-β1, inhibit the expression of inflammatory factors and promote spontaneous relief of acute gout attack. In addition to the activation mechanism of intracellular signaling molecules associated with inflammatory response, the inflammatory mechanism of gout also involves complement activation, cell activation and other pathways. The complications caused by gout, such as cardiovascular system damage and joint destruction, are seriously harmful to human health. At present, western drugs, such as allopurinol and febuxostat, exert an effect in inhibiting xanthine oxidase. Benzimarone has effect in reducing renal absorption of uric acid and promoting uric acid excretion by inhibiting uric acid transporter 1(URAT1) and glucose transporter 9(GLUT9). Even Lesinurad and other medicines in current studies are based on the inhibition of uric acid re-absorption, but with adverse reactions that limit the clinical application. The treatment of gout with traditional Chinese medicine(TCM) has multi-target characteristics, with advantages in reducing uric acid, resisting inflammation and improving joint function and a high safety. It has been gradually popularized and applied in clinical treatment of gout. Therefore, it is a promising research direction to treat gout with TCM and western medicine based on the pathomechanism of gout.
Keywords：gout;traditional Chinese medicine;pathomechanism;interleukin(IL);uric acid transporter 1(URAT1);glucose transporter 9 (GLUT9)
Abstract：The study of traditional Chinese medicine(TCM) syndrome model started late but developed rapidly. In recent years, with the deepening of the study of TCM diseases and prescriptions, to develop the modernization of TCM, the study of TCM syndrome model has been required to be more specific and went deeper. Qi deficiency and blood stasis syndrome, as a common clinical syndrome type, often occurs in cardiovascular and cerebrovascular diseases, digestive system diseases, chronic kidney disease, cancer and other diseases. With the increase in the aging of population in China, the research on Qi deficiency and blood stasis syndrome has become a hot topic in the field of TCM due to the physical characteristics of the elderly people with multiple Qi deficiency and blood stasis. The animal models of Qi deficiency and blood stasis syndrome established in recent years were classified into pathological model, etiological model and combined disease and syndrome model. By observing the macroscopical representation, limb behavior and hemorheology of experimental animals in line with the theory of TCM, this paper evaluates the method of modeling, and provides ideas and reference for the method of modeling for Qi-deficiency and blood-stasis syndrome. The research on the model of Qi-deficiency and blood-stasis syndrome should follow the thought of cause-pulse-syndrome-treatment, namely etiology-external manifestation-pathological mechanism-syndrome differentiation and treatment, and combine the study on the TCM syndrome model with modern science, so as to standardize the development of modeling methods and build relatively improved animal models of Qi deficiency and blood stasis, and provide model support for exploring the pathogenesis of Qi deficiency and blood stasis syndrome and new treatment ideas in the future.
Keywords：Qi deficiency and blood stasis syndrome;animal model;multi-factor combination;syndrome;evaluation modeling method
Abstract：Psychiatric diseases represented by depression have gradually become one of the major health problems of people in the fast-paced, high-pressure society. Severe cases can cause suicides, huge harm or disaster to families and the society. Although modern medicine has made great progress in the field of anti-depressant drug therapy, depression still cannot be cured. At the same time, traditional Chinese medicines(TCM) with a definite curative effect, few adverse reactions, and mild efficacy have received increasing attention. TCM valerianae Jatanmansi Rhizoma et Radix has been widely used to alleviate sleep disorder, and its root extract is popularly known as valerian and used as a mild sedative for a long time in European. Tagara takes Valerianae Jatamansi Rhizoma et Radix as the key ingredient for treatment of depression-type insomnia, and is available abroad. It is reported that iridoid, sesquiterpenes, flavonoids or extract from Valerianae Jatamansi Rhizoma et Radix has a superior anti-depression activity in both animal and clinical trials, and the mechanism is mainly related to the regulation of neurotransmitters in the brain, the improvement of the function of the hypothalamic-pituitary-adrenal (HPA) axis, the resistance of free radicals and inflammation, and the neuroprotective effect. However, there is still lack of report on the anti-depression system and in-depth research of Valerianae Jatamansi Rhizoma et Radix. Therefore, it is urgently necessary to systematically collect and summarize the anti-depressant activity and explain the relevant mechanisms, so as to provide reference for the further development of Valerianae Jatamansi Rhizoma et Radix medicinal resources.
Keywords：depression;Valerianae Jatamansi Rhizoma et Radix;mechanism
Abstract：Xiao Chengqitang, derived from Treatise on Febrile Diseases, is composed of Rhei Radix et Rhizoma, Magnoliae Officinalis Cortex and Aurantii Fructus Immaturus. It is often used to treat Yangming Fushi light syndrome. This formula was included in the Catalogue of Ancient Classical Prescription (The First Batch), which has a high clinical application value. Current researches on the chemical composition, pharmacological effect and clinical application of Xiao Chengqitang have been carried out, but the history of Xiao Chengqitang is unclear and the dose conversation method is vague. It has great difficulties in clinical rational applications of Xiao Chengqitang and the research and development of its compound preparations. In this article, the literature reports on Xiao Chengqitang were summarized from the aspects of historical evolution, literature researching of Chinese herbs, dosage conversation method, modern research in case to clarify the historical context of Xiao Chengqitang and to provide basis for the clinical application and modern scientific research of this formula.
Keywords：Xiao Chengqitang;famous classical formulas;dose conversation;historical evolution;origin of medicinal materials;processing of decoction pieces;clinical application