Abstract：Related records about discipline of diagnosis and treatment of exogenous febrile diseases by traditional Chinese medicine (TCM) were reviewed to summarize TCM knowledge about etiology, pathogenesis, progress and treatment of infectious epidemic diseases among exogenous febrile diseases. In consideration of onset time and major clinical manifestations of novel coronavirus pneumonia (NCP), this disease is considered as within the scope of pestilence in TCM, it is located in the lung, with etiology being poison of winter-warm syndrome, the viruses enter body via mouth and nose, and invade pulmonary collaterals. Based on the syndrome characteristics proposed in diagnosis and treatment schemes of various regions as well as the researches on medication rules, the present study points out the core pathogenesis of NCP is epidemic toxin invading the lung, pathogenic heat obstructing the lung and toxic heat blocking the lung. It also analyzes theoretical formula features of TCM Lianhua Qingwen and related clinical and experimental research results, in order to prove the clinical value of this drug in the outbreak of NCP.
Keywords：traditional Chinese medicine (TCM);severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2);novel coronavirus pneumonia;pestilence;poison of winter-warm syndrome;Lianhua Qingwen;epidemic theory
Abstract：ObjectiveTo analyze the clinical effect of traditional Chinese medicine Lianhua Qingwen in the treatment of coronavirus disease 2019 (COVID-19) and provide the basis for medication guides through a retrospective study in a cohort of COVID-19 confirmed patients.MethodA retrospective analysis of clinical records was conducted in COVID-19 confirmed patients at The Ninth Hospital of Wuhan and CR&WISCO General Hospital including the treatment group (21 patients, basic treatment in combination with Lianhua Qingwen granules, 1 packet/time, 3 times/day) and the control group (21 patients, basic treatment). Comparison between the two groups was made in terms of the disappearance rates of cardinal symptoms (fever, cough and weakness), duration of fever, and disappearance rates of other symptoms (muscle pain, expectoration, nasal obstruction, running nose, dry throat, pharyngalgia, shortness of breath, chest distress, dyspnea, dizziness, headache, nausea, vomiting, loss of appetite and diarrhea).ResultThe baseline data were similar between the two groups. When compared with the control group, patients in the treatment group had the higher clinical effect, including the disappearance rate of fever (85.7% vs 57.1%, χ2=4.200, P=0.040), the disappearance rate of cough (46.7% vs 5.6%, P=0.012), the disappearance rate expectoration (64.3% vs 9.1%, P=0.012), the disappearance rate of shortness of breath (77.8% vs 0, P=0.021), and the duration of fever [(4.6±3.2) d vs (6.1±3.1) d, P=0.218].ConclusionLianhua Qingwen can significantly relieve cardinal symptoms in COVID-19 confirmed patients by inhibiting fever and cough, reducing their duration, as well as improving individual symptoms. All these results provide preliminary clinical evidence for Lianhua Qingwen granules in the COVID-19 treatment.
Abstract：ObjectiveTo observe clinical efficacy of Lung-toxin dispelling formula No.1 treating patients of corona virus disease 2019(COVID-19) type severe/type extremely severe, and summarize experiences of diagnosis and treatment.MethodCollected and analyzed clinical informations of patients of COVID-19 type severe/type extremely severe, treated with Lung-toxin dispelling formula No.1, who were hospitalized in central hospital in Zhumadian and the first affiliated hospital of Henan university of traditional Chinese medicine from 31st January to 27th February.ResultAll patients had positive epidemiological history, major symptoms were fever, cough, tachypnea, weakness and sore heavy muscles, combined with bad appetite and diarrhea. The median age was 59, median time from onset to getting worse was 9 days, ground glass opacity, lamellar, nodular high density shadow were mostly displayed in both lungs， lesions progressedfaster. After treatment with Lung-toxin dispelling formula No.1 combined with western medicine, the median time of PCR-NAD-test from positive to negative was 16 days, the median hospitalization days were 20 days, all patients were cured and discharged.ConclusionLung-toxin dispelling formula No.1 had certain clinical efficiency in treating patients of COVID-19 type severe/type extremely severe, further large sample clinical verification is needed.
Keywords：corona virus disease 2019(COVID-19);type severe;type extremely severe;traditional Chinese medicine syndromes;Lung-toxin dispelling formula No.1
Abstract：ObjectiveTo observe the therapeutic effect of Kesuting syrups and Keqing capsules, which have the function of promoting lung and resolving phlegm, on a mouse model combining disease and syndrome of human coronavirus pneumonia with cold-dampness pestilence attacking lung.MethodThe therapeutic effects of Kesuting syrups (the doses of 22, 11 mL·kg-1) and Keqing capsules (the doses of 1.155, 0.577 5 g·kg-1) on this model were evaluated by the inflammatory changes of lung tissue, the expression of viral nucleic acid, the contents of inflammatory factors [interleukin (IL)-6, IL-10, tumor necrosis factor (TNF)-α and interferon-γ (IFN-γ)], and the percentages of immune cells in peripheral blood (CD4+ T cells, CD8+ T cells and B cells).ResultCompared with the model group, high- and low-dose groups of Keqing capsules and Kesuting syrups could significantly reduce the inflammatory damage in the lung tissues of mice, Keqing capsules could significantly increase the percentages of CD4+ T cells, CD8+ T cells and B cells in peripheral blood, Keqing capsules and Kesuting syrups could reduce the expression levels of IL-6, IL-10, TNF-α and IFN-γ, inhibit the viral load in lung tissue, as well as improve the pathogenic manifestations of lung tissue.ConclusionAs the first-line drugs for novel coronavirus pneumonia, Keqing capsules and Kesuting syrups have significant therapeutic effect on the mouse model combining disease and syndrome of human coronavirus pneumonia with cold-dampness pestilence attacking lung, and the mechanism may be related to regulating immune function and reducing cytokine storm.
Keywords：novel coronavirus pneumonia (COVID-19);method of promoting lung and resolving phlegm;mouse model combining disease and syndrome of human coronavirus pneumonia with cold-dampness pestilence attacking lung;Kesuting syrups;Keqing capsules;inflammatory factors;immune cells in peripheral blood
Abstract：In this paper, based on the supramolecular "Qi chromatography" of traditional Chinese medicine (TCM), combined with the characteristics of corona virus disease 2019 (COVID-19), the historical origin and characteristics of theoretical research on the treatment of epidemic diseases by TCM were analyzed retrospectively, and the key scientific problems for achieving its modernization were proposed, and the countermeasures were illustrated. In fact, the emergence and development history of TCM epidemic theory is a comprehensive and scientific understanding of biomolecular chemistry for epidemic caused by pathogen. From the exploration of the cause of abnormal Qi to the epidemic pathogenic factor, from the treatment based on syndrome differentiation by six meridians to by the defensive Qi and nutrient blood and the triple energizer, which is gradually close to the modern theoretical system of infectious diseases and immunology, all of which are stamped with brand of TCM supramolecular "Qi chromatography" theory. In the long history of biological evolution in nature, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) as epidemic pathogenic factor in TCM, human body and Chinese materia medica are biological supramolecules at a specific stage of evolution in nature, which are controlled by the "imprinting templates" of molecular society in gradual biomolecules, with the unchanged laws of mutual advancement, mutation, suitability and mutual restriction. Based on the analysis of the law of supramolecular action hiding in the prevention and treatment of epidemic diseases dealt with TCM on procession of principle-method-recipe-medicines, this paper discusses the key scientific problems of its modernization and puts forward the solutions. It can not only clarify the scientific principle of the prevention and treatment with TCM, develop the multi-component Chinese medicine, accelerate the control of the epidemic situation, but also promote the solution of original frontier key scientific problems such as the material basis of meridians, viscera and syndromes, and the compatibility law of the TCM prescription, in order to protect the existing medical resources and open up a new situation in the modernization of TCM research.
Keywords：epidemic theory of traditional Chinese medicine (TCM);severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2);corona virus disease 2019 (COVID-19);modernization of TCM;supramolecular;"Qi chromatography" theory;imprinting templates
Abstract：Moxa smoke plays an indispensable role in traditional Chinese medicine with the functions of dispelling poison, exorcizing evil and avoiding filth, dispelling cold, etc. It has been used to prevent pestilence for thousands of years. This paper reviewed the clinical application, mechanism and existing shortcomings of moxa smoke for preventing epidemic diseases, and found that moxa smoke can effectively inhibit a variety of pathogenic bacteria, fungi and viruses that cause infectious and epidemic diseases, and has good pharmacological effects of sterilization, antiviral and immunity enhancement, and has achieved remarkable effect in disease prevention, health care and plague prevention. It is suggested that attention should be paid to the air quality of moxibustion clinics in the future, it is urgent to establish a unified sanitary air quality standard system to create a good clinic environment. At the same time, the research on the clinical safe concentration range of moxa smoke should be strengthened to effectively avoid the occurrence of adverse reactions, give full play to the efficacy of moxa smoke in eliminating pathogenic factors and preventing epidemic, so as to provide the basis for the rational application and promotion of moxa smoke.
Abstract：ObjectiveTo observe the effect of modified Erchentang on the expression of CXC chemokine ligand (CXCL) 8-CXC chemotaxis factor receptor (CXCR) 1/2 genes in the lung tissue of rats with chronic obstructive pulmonary disease (COPD), in order to explore the anti-inflammatory molecular mechanism of Erchentang on COPD.MethodForty SD rats were randomly divided into normal group, model group, Jizhi syrup group and modified Erchentang group. COPD models in rats were prepared by cigarette smoke and dripping lipopolysaccharide (LPS) in the trachea. After modeling, normal and model groups were intragastrically given normal saline solution, Jizhi syrup group was given Jizhi syrup（10 g·kg-1），and modified Erchentang group was given intragastrically corresponding herbal drugs (10 g·kg-1) for 14 days. The levels of chemokines CXCL1, CXCL8 were detected by enzyme-linked immunosorbent assay in rat bronchoalveolar lavage fluid (BALF). The mRNA expressions of CXCL8, CXCR1 and CXCR2 were detected by quantitative real time PCR (Real-time PCR). Western blot was used to detect the levels of CXCL8, CXCR1 and CXCR2 protein, the pathological changes of lung tissues were observed by hematoxylin-eosin(HE) staining,and immunohistochemistry (IHC) method was used to detect the expressions of CXCL8, CXCR1 and CXCR2 protein in the lung tissue of all the groups.ResultThe levels of chemokines CXCL1, CXCL8 in rats BALF were increased significantly (P<0.01), the expressions of CXCL8，CXCR1 and CXCR2 mRNA and protein were increased significantly (P<0.05, P<0.01) in model group compared with normal group. Compared with model group, the expressions of CXCL8, CXCR1 and CXCR2 mRNA and protein were decreased significantly (P<0.05), and the levels of chemokines CXCL1, CXCL8 in rats BALF were decreased significantly (P<0.01) in modified Erchentang.ConclusionModified Erchentang has an anti-inflammatory effect on COPD. The mechanism may be related to inhibiting the expressions of CXCL8, CXCR1, CXCR2 mRNA and protein, and reducing the release of chemokines CXCL1, CXCL8.
Abstract：ObjectiveTo study the effect of Zuoguiwan on the differentiation of CD4-CD8-T cells in thymus during the development of kidney-deficient fetal rats and the application value of kidney and essence-tonifying method, in order to explore the theoretical connotation of "kidney being the origin of congenital constitution".MethodMale and female rats were paired at a ratio of 2∶1, and three stress methods were combined to construct the kidney-deficient model. Pregnant rats were divided into control group, model group, thymosin group (0.003 g·kg-1), and low-dose Zuoguiwan group and high-dose Zuoguiwan group (2,8 g·kg-1). Normal saline was given to the control group， model group was used for modeling and given normal saline by gavage, thymosin group was given thymosin capsule solution by gavage, and low-dose Zuoguiwan group and high-dose Zuoguiwan group were given Zuoguiwan suspension by gavage. Embryos that were 16 days, 19 days and born on the first day (E16, E19, P1) were selected to calculate fetal rat thymus index. Electron microscope was used to observe thymic epithelial cell ultrastructure of fetal rats. Enzyme-linked immunosorbent assay (ELISA) method was used to detect thymus thymosinβ4 (TMSβ4) and thymosinα1 (Tα1) content, fluid cytology detection of fetal rats on the surface of CD4, CD8, CD25, CD44 expression for identifying T cell type.ResultThe morphology of thymus epithelial cells in the model group was irregular, the nucleus was deformed, and the chromatin of the nucleus was significantly increased. The results of ELISA showed that the content of TMSβ4 and Tα1 in thymus increased in the model group and the low-dose and high-dose Zuoguiwan groups. Flow cytometry showed that, compared with the control group, CD4-CD8-T cells were significantly increased in the model group at E16, E19 and P1 stages (P<0.05), suggesting that T cells were blocked during the development from DN to DP. CD4-CD8-CD25-CD44-T (DN4) was significantly increased (P<0.05), suggesting that T cell development was inhibited during DN4-DP transition. Compared with the model group, CD4-CD8-T cells were significantly reduced in low and high-dose groups (P<0.05) at E16, E19 and P1 stages. At the same time, DN4 was significantly reduced (P<0.05), suggesting that Zuoguiwan could promote the development of T cells from DN4 stage to DP stage as thymosin.ConclusionZuoguiwan can improve the structure of thymus epithelial cells in rats with kidney deficiency, and induce the differentiation of T cells in thymus from DN4 stage to DP stage by increasing the secretion of TMSβ4 and Tα1 in thymus of embryonic rats, indicating that Zuoguiwan can promote the development and maturation of T cells and regulate the immune status of fetal rats with kidney deficiency by tonifying kidney and spermatogenesis.
Keywords：congenital kidney deficiency;Zuoguiwan;thymosinβ4 (TMSβ4);thymosinα1 (Tα1);double negative T cells
Abstract：ObjectiveTo investigate the effect of Saposhnikoviae Radix on Tongxie Yaofang in regulating water metabolism and 5-serotonin（5-HT） signal system in diarrhea type irritable bowel syndrome（IBS-D）rats.MethodThe 40 IBS-D SD rats were randomly divided into model group, Tongxie Yaofang wituout Saposhnikoviae Radix group (26 g·kg-1), Tongxie Yaofang decoction group (30 g·kg-1), Tongxie Yaofang with double Saposhnikoviae Radix group (34 g·kg-1),another 10 normal SD rats were selected as the normal group.Except for normal group, the rats in other groups were separated from their mothers and induced by acetic acid to establish D-IBS model. These rats in the each group were administered with corresponding drugs for 14 days. The diarrhea indexs and the water contents of the feces were observed and calculated. The Na+-K+-ATPase activities in the intestinal mucosa were detected by micro method. The contents of 5-HT were detected by enzyme linked immunosorbent assay (ELISA). The activities of monoamine oxidase A (MAO-A) were detected by chemiluminescence method. The expressions of aquaporin 4 (AQP4) in colon were detected by immunohistochemistry. Protein expression levels of HT receptor 3 (5-HT3R), HT receptor 4 (5-HT4R) and serotonin transporter (SERT) in hypothalamus and colon were detected by Western blot.ResultCompared with normal group, the fecal water contents, contents of 5-HT in hypothalamus and colon, activities of MAO-A, expressions of 5-HT3R were significantly increased in model group (P<0.01), and the activities of Na+-K+-ATPase, expressions of AQP4, 5-HT4R and SERT were reduced significantly (P<0.01). Compared with model group, the diarrhea indexs and fecal water contents, contents of 5-HT in hypothalamus and colon, activities of MAO-A, expressions of 5-HT3R were significantly decreased in each experimental group, and the activities of Na+-K+-ATPase, expressions of AQP4,5-HT4R and SERT were significantly increased(P<0.05，P<0.01),however, there was no significant difference in the expression of 5-HT3R protein in Tongxie Yaofang wituout Saposhnikoviae Radix group.Compared with Tongxie Yaofang wituout Saposhnikoviae Radix group,the diarrhea index, fecal water content, 5-HT content, MAO-A enzyme activity, and protein expression of 5-HT3R were significantly decreased in Tongxie Yaofang group and Tongxie Yaofang with double Saposhnikoviae Radix group(P<0.05，P<0.01), and the activity of Na+-K+-ATPase and protein expression of SERT were significantly increased(P<0.05，P<0.01).ConclusionSaposhnikoviae Radix can enhance the effects of Tongxie Yaofang on improving the water metabolism of rats with IBS-D and regulating the multi-target of 5-HT signaling system, further confirming the synergistic effect of Saposhnikoviae Radix.
Keywords：Saposhnikoviae Radix;Tongxie Yaofang;irritable bowel syndrome;water metabolism;5-serotonin（5-HT）signal system
Abstract：ObjectiveTo study the effect of the kidney-tonifying recipes on mouse syndrome and adrenal function induced by low-dose hydrocortisone reduction and discontinuation.MethodThe 48 male ICR mice were randomly divided into normal control group, hydrocortisone group, Liuwei Dihuangtang group and Guifu Dihuangtang group, with 12 rats in each group. Each group was given 1.32 mg·kg-1·d-1 hydrocortisone for 28 days, and half a dose for 7 days, and the administration was discontinued 14 days before modeling. In the reduction and discontinuation phase, 12.5 g·kg-1·d-1 Liuwei Dihuangtang and 13.5 g·kg-1·d-1 Guifu Dihuangtang were simultaneously administered to the corresponding group for 21 d. On the 28th and 49th day of the experiment, the characteristic information of mice was detected by the experimental methodology of syndrome differentiation and treatment in mice (holding power, body surface infrared temperature). The mice were sacrificed on the 50th day of the experiment, the spleen and thymus were separated, and the spleen and thymus index were calculated, serum corticosterone content is measured by enzyme-linked immunosorbent assay (ELISA), the total RNA of the adrenal gland was extracted, and the expressions of Star, cholesterol-side-chain cleavage enzyme (Cyp11a1), Cyp21a1, Cyp11b1, low-density lipoprotein receptor (Ldlr), scavenger receptor class B type I (Scarb1/SRB1), Hmgcr, acyl-CoA-cholesterol acyltransferase-1 (Acat1), hormone sensitive lipase (Lipe/HSL), insulin-inducible gene 1 (Insig1) and sterol regulatory element binding transcription factor 2 (Srebf2) were detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). Western blot was used to detect the expressions of LDLR, SRB1, CYP11A1 and cytochrome P450 family 21 subfamily A member 2 (CYP21A2) in adrenal gland.ResultCompared with the normal control group, the body weight and the body mean infrared temperature of the model group were significantly decreased after 28 days of continuous administration (P<0.01). After the administration of the drug, the mice in the model group showed a significant decrease in the grip strength (P<0.05), a significant increase in the thymus index (P<0.01), a significant decrease in the serum corticosterone content (P<0.05). Expressions of adrenal Cyp21a1 and Hmgcr genes decreased (P<0.05, P<0.01), Lipe and Acat1 gene expressions increased significantly (P<0.01), adrenal CYP11A1, SRBI protein expressions decreased significantly, and LDLR protein expression increased. Compared with the model group, the body weight of Guifu Dihuangtang group decreased significantly after 21 days of treatment with traditional Chinese medicine (P<0.01), the spleen index of Liuwei Dihuangtang group decreased significantly (P<0.01), expressions of Cyp11a1, Cyp21a1, Acat1 and Lipe genes in Guifu Dihuangtang group were significantly increased (P<0.05, P<0.01), expressions of Ldlr and Scarb1 genes were significantly decreased (P<0.05), and expressions of Ldlr, Acat1 and Lipe genes in Liuwei Dihuangtang group were significantly decreased (P<0.05, P<0.01), and expressions of CYP11A1 and SRBI proteins in Liuwei Dihuangtang and Guifu Dihuangtang increased.ConclusionGuifu Dihuangtang can promote the recovery of adrenal function in mice with drug-induced deficiency syndrome by correcting expression of adrenal cortex synthase.
Abstract：ObjectiveTo explore the effect of Ermiaosan（EMS） on the polarization of M1 by lipopolysaccharide（LPS）+interferon(IFN)-γ and M2 induced by interleukin(IL)-4+IL-13 in rat bone marrow-derived macrophages.MethodMacrophages from rat bone marrow were extracted in vitro, stimulated by macrophage colony stimulating factor(M-CSF), induced to macrophages (marked by F4/80), stimulated by LPS+IFN-γ and induced to polarize to M1，while stimulated by IL-4+IL-13 and induced to polarize to M2. After adding different concentrations of EMS (0.2，0.4，0.8 g·L-1), the phenotypes of M1 and M2 were detected by immunofluorescence, and the effect of EMS on M1(marked by CD68 and iNOS)/M2(marked by CD206 and Arginase) polarization of macrophages from rat bone marrow was detected.ResultCompared with control group, LPS + IFN-γ could increase the polarization of M1 (P<0.01)，while IL-4+IL-13 could increase the polarization of M2 (P<0.01); compared with LPS+IFN-γ/IL-4+IL-13 group, EMS (0.2，0.4，0.8 g·L-1) could inhibit the polarization of M1 induced by LPS+IFN -γ for 24 hours (P<0.05), but had no significant effect on polarization of M2 induced by IL-4+IL-13.ConclusionEMS can inhibit M1 polarization induced by LPS+IFN - γ, but has no effect on M2 polarization induced by IL-4+IL-13.
Abstract：ObjectiveTo study the effect of icariin on renin homologous protein A (RhoA)/Rho-related kinase (ROCK) pathway in rats with nephrotic syndrome (NS) and its protective mechanism.MethodTotally 54 clean-grade male SD rats were tested and randomly divided into normal group, model group, RhoA inhibitor group (Rhosin, 40 mg·kg-1·d-1) and three doses of icariin groups (low, medium and high corresponding dose， 30, 60, 120 mg·kg-1·d-1). Adriamycin hydrochloride 6.5 mg·kg-1 was given in tail vein of rats to induce NS model in rats. After the model was established, peritoneal administration was carried out. The normal group and the model group were given saline 2.5 mL·d-1, and the inhibitor group and all of dose groups were given corresponding doses of Rhosin and icariin for intervention. Total urinary protein (Alb), creatinine (Cre), total urinary protein/creatinine ratio (A/C) kit were detected in rats, ultrastructure of kidney was identified by transmission electron microscopy (TEM), and Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to detect the mRNA and proteins expressions of RhoA, ROCK1, ROCK2.ResultTEM showed that the basement membrane was intact and the foot process was regular in the normal group, in model group, basement membrane was damaged seriously, foot process disappeared, and fusion was serious, in the low-dose group, the basement membrane injury was alleviated, the number and density of foot process were improved, and the fusion was obvious， in the middle-dose group and the inhibitor group, the basement membrane thickening was alleviated, and the foot process was slightly fused, in the high-dose group, the basement membrane structure was more complete, and podocytes were longer and arranged tightly. Compared with the normal group, the levels of Alb, Cre and A/C in urine, and RhoA, ROCK1 and ROCK2 mRNA and protein expressions in kidney tissue of rats of the model group were significantly higher (P<0.05). Compared with model group, the levels of Alb, A/C in urine and RhoA, ROCK1, ROCK2 mRNA and protein expressions in kidney tissue in the inhibitor group and low, medium and high-dose groups, and Cre in urine in inhibitor group and high-dose group decreased significantly (P<0.05). Compared with the inhibitor group, the levels of Alb, Cre in urine and RhoA protein in kidney tissue in the high-dose group were significantly decreased (P<0.05), the levels of Alb, Cre, A/C in urine and RhoA, ROCK1, ROCK2 mRNA and protein expressions in kidney tissue of the low-dose group, and the levels of RhoA, ROCK1 and ROCK2 mRNA expressions in kidney tissue of the middle-dose group were significantly increased (P<0.05). Compared with the low-dose group, the levels of Alb, A/C in urine, and RhoA, ROCK1, ROCK2 mRNA and protein expressions in kidney tissue in the middle and high-dose groups, Cre in urine of the high-dose group were significantly decreased (P<0.05). Compared with the middle-dose group, the levels of Alb, Cre in urine, and RhoA, ROCK1, ROCK2 mRNA and protein expressions, ROCK2 mRNA expression in kidney tissue in the high-dose group were significantly decreased (P<0.05).ConclusionIcariin may protect glomerular endothelium and podocyte by affecting RhoA/ROCK pathway in the treatment of NS rats.
Keywords：icariin;nephrotic syndrome;rats;renin homologous protein A (RhoA);Rho-related kinase 1 (ROCK1);ROCK2
Abstract：ObjectiveTo investigate the pharmacodynamics effects of antiobesity, lipid-lowering and the regulations of serum bile acid profiles of Lidan Ruanjian prescription (LDRJ) in obesity rats induced by high-fat diet.MethodThe 42 rats were fed high-fat diet for 9 weeks to establish model of obese rats，24 rats were randomly divided into model group, high and low-dose LDRJ group (30,15 g·kg-1). Another 8 normal rats were selected as the normal group.The model group and normal group were given normal saline, and drug group was given the corresponding dose of drug for 4 weeks. Body weight, liver weight, white adipose tissue (WAT) weight were determined after administration medicine for 4 weeks. The bile flow of the rats was measured by bile duct intubation and fasting serum lipid levels of total cholesterol (TC), total triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) were detected by automatic biochemical analyzer. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) assay was used to test serum bile acid profile of each group rats.ResultCompared with the control group, the average body weight, liver weight, WAT weight of the model group were significantly increased (P<0.01), while the fasting serum TC, TG and LDL-C levels were elevated (P<0.05，P<0.01). The total bile secretion and bile flow at each test point within 2 h were decreased and the proportion of primary bile acids was decreased (P<0.05).The serum total bile acid content decreased significantly（P<0.01），levels of cholic acid (CA), deoxycholic acid (DCA), chenodeoxycholic acid (CDCA), hyodeoxycholic acid (HDCA), taurocholic acid (TCA), taurodeoxycholic acid (TDCA), taurochenodeoxycholic acid (TCDCA), taurohyodeoxycholic acid (THDCA) and glycodeoxycholic acid (GDCA) were significantly reduced (P<0.05，P<0.01). Compare with model group, body weight, liver weight in high and low-dose LDRJ groups reduced significantly(P<0.05，P<0.01). Fasting serum TC, TG and LDL-C levels were decreased in high-dose group(P<0.05，P<0.01), so did as TG levels in low-dose group(P<0.05). The bile flow rate increased significantly in high-dose group 1~1.5 h after administration (P<0.05). All dose treatment groups increased the proportion of primary bile acids (P<0.05) and changed the bile acid profile, especially elevated the bile acid levels of TCA, DCA, glycocholic acid (GCA), GDCA in high-dose LDRJ group (P<0.05，P<0.01), while TCA and TCDCA in low-dose group (P<0.05).ConclusionLDRJ has significant lipid-lowering and antiobesity effects and the mechanism might involve the increase of bile secretion, the stimulation of primary bile acid synthesis and the regulation of bile acid profile.
Keywords：diet-induced obesity;Lidan Ruanjian prescription;lipid-lowering and antiobesity;ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS);metabolic profile of bile acid
Abstract：ObjectiveTo study the protective effect of Tongxinluo capsule on cerebral ischemia-reperfusion injury, and explore the mechanism of Tongxinluo capsule in treating cerebral ischemia through network pharmacology.MethodThe C57BL/6 mouse middle cerebral artery occlusion(MCAO) model was established by improved suture method and divided into sham operation group, model group, low, medium and high-dose Tongxinluo groups (crude drug 1,2,4 g·kg-1, intragastric administration), Aspirin group (2.055 g·L-1, intraperitoneal injection). Then, neurological function score and 2,3,5-triphenyltetrazole chloride(TTC) staining method were used to determine the infarct size of mice at 24, 48, 72 h by cerebral ischemia-reperfusion. First, chemical constituents of Tongxinluo capsule were screened from the BATMAN-TCM database, and the targets were analyzed. Then, gene ontology (GO) enrichment and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis were performed, and traditional Chinese medicine(TCM)-active ingredient-target network was constructed. Finally, the multi-dimensional pharmacological mechanism of Tongxinluo capsule in the treatment of cerebral ischemia was predicted.ResultLonga score, HE staining and TTC staining all suggested that Tongxinluo capsule could alleviate brain injury in mice after cerebral ischemia and reperfusion, and the improvement degree of Tongxinluo capsule on brain injury was gradually enhanced with the increase of Tongxinluo capsule dose. A total of 132 active components and 240 intersection targets, including cyclic adenosine monophosphate(cAMP)-dependent protein kinase catalytic subunit alpha (PRKACA), adenylate cyclase 1(ADCY1), serine/threonine kinase 1 (Akt1), dopamine receptor D2 (DRD2) and discs large homolog 4 (DLG4) were screened from 12 TCM in Tongxinluo capsule. GO was enriched in cationic channel activity, ion gated channel activity, gate channel activity, neurotransmitter receptor activity, ion channel activity, etc. KEGG was enriched in cAMP signaling pathway, calcium signaling pathway, neuroactive ligand-receptor interaction, cyclic guanosine monophosphate(cGMP)/protein kinase G(PKG) signaling pathway and dopaminergic synaptic signaling pathway.ConclusionTongxinluo capsule can alleviate brain damage in mice with cerebral ischemia-reperfusion, and achieve brain protection through multiple targets and multiple links. Network pharmacology reveals effective components，targets and pathway of Tongxinluo capsule in the treatment of cerebral ischemia, which provides theoretical support for the mechanism of Tongxinluo capsule in the treatment of cerebral ischemia.
Abstract：ObjectiveTo investigate the anti-tumor efficacy, mechanism and safety of zeylenone on acute T lymphocytic leukemia.MethodIn vitro, Molt-4 cells were treated with various concentrations of zeylenone (0.2, 0.4, 0.8, 1.6, 3.2 μmol·L-1) for 48 h, and the cell viability was measured with cell counting kit-8 (CCK-8) assay. nonobese diabetic-severce combined immunodeficient mice（NOD/SCID） mice were randomly divided into six groups: normal group, model group, vincristine group (1 mg·kg-1), low-dose zeylenone group (12.5 mg·kg-1), medium-dose zeylenone group (25 mg·kg-1), high-dose zeylenone group (50 mg·kg-1). With the exception of normal group, mice were pre-irradiated with 60Co and inoculated subcutaneously with Molt-4 cells to establish the Molt-4 xenograft model. Then NOD/SCID mice were sacrificed after 13 days of administration. The tumor inhibition rates, relative tumor growth rates and organ indexes were calculated. Hematoxylin and eosin (HE) staining was used to observe the pathological changes of liver and spleen tissues in mice. The expressions of phosphorylation signal transducer and activator of transcription (p-STAT3), B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X protein (Bax) and cysteine aspartate-specific protease-3 (Caspase-3) were detected in tumor tissues by Western blot.ResultIn vitro, zeylenone had an obvious inhibitory effect on Molt-4 cells. IC50 values of zeylenone was 1.49 μmol·L-1. In vivo, compared with the model group, medium and high-dose zeylenone groups had significant tumor inhibition effects, with the inhibition rates of 50.24% and 60.75%, respectively (P<0.01). Additionally, liver and spleen injuries were slight in the above mentioned two groups compared with the vincristine group, indicating that zeylenone was safe. Western blot analysis showed that medium and high-dose zeylenone groups showed significant declines in proteins p-STAT3, Caspase-3 and Bcl-2, and marked increases in pro-apoptotic protein Bax compared with the model group (P<0.05, P<0.01).Conclusionzeylenone could obviously inhibit the proliferation and induce the apoptosis of Molt-4 cells; and its mechanism may be related to the down-regulation of p-STAT3, Caspase-3, Bcl-2 and the up-regulation of Bax expressions. In addition, zeylenone had less damage to liver and spleen, and was safer than vincristine.
Keywords：acute T lymphocytic leukemia;zeylenone;cell apoptosis;anti-tumor;Molt-4 cell
Abstract：ObjectiveTo study the regulatory level of Dima decoction on adenovirus E1B 19 kD interacting protein 3（BNIP3） protein and to understand the mechanism in intervening mitophagy and controlling inflammation in ulcerative colitis.MethodThe 60 SPF SD rats were randomly divided into normal control group, model control group, low, medium ang high dose Dima decoction（8.5，17.0，34.0 g·kg-1）group and mesalazine（3.0 g·kg-1） group，10 for each group. The model of UC rats was established by the method of environmental diet intervention +2,4,6-trinitro-Benzenesulfonicacid（TNBS）+ethanol，the administration group was administered for 14 days.Hematoxylin-eosin (HE) was used to detect pathological changes of colonic mucosal tissues in rats.Western blot and real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） were used to detect the expressions of BNIP3 protein and mRNA in intestinal epithelial cells.ResultHE staining results showed that a large number of inflammatory cell infiltration and non-cheesy granulation tissue were observed in the colon of rats in the model group, while the rats in each drug group showed different degrees of fibrous repair. Compared with normal group, the expression of BNIP3 protein and mRNA in model group were significantly increased (P<0.05). Compared with model group, the expression of BNIP3 protein and mRNA in low, medium and high-dose Dima decoction group and mesalazine group were significantly increased (P<0.01). Compared with low-dose group, the expression of BNIP3 protein and mRNA in medium and high-dose groups and mesalazine group were significantly increased (P<0.05).ConclusionDima decoction can increase the expression of BNIP3 in the intestinal epithelium of UC active rats, which may be one of the mechanisms of promoting mitochondrial autophagy against UC inflammation.
Keywords：Dima decoction;ulcerative colitis;adenovirus E1B 19 kD-interacting protein 3（BNIP3）;mitophagy;mechanism
Abstract：ObjectiveTo study the protective mechanism of oxymatrine on oxidative stress induced by high glucose in H9C2 cells.MethodH9C2 cardiomyocytes were cultured in groups and divided into normal group, high glucose (HG) group, low-dose oxymatrine (OMT) group (50 mg·L-1), high-dose OMT group (100 mg·L-1), positive drug vitamin E (VE) group (1×10-4 mol·L-1) and mannitol (M) wasotonic control group. Cell damage was detected by lactate dehydrogenase leakage, changes in cell superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were detected, intracellular reactive oxygen species (ROS) content and cellular mitochondria and functional integrity were detected by fluorescent probes, and Western blotting was used to detect the expressions of Bcl family proteins.ResultCompared with the normal group, the content of malondialdehyde and reactive oxygen species and the expression level of pro-apoptotic protein in the high glucose group were significantly increased (P<0.01), while the activity of superoxide dismutase and the expression levels of mitochondrial membrane potential (MMP) and anti-apoptotic protein were significantly decreased (P<0.01). Compared with the high glucose group, oxymatrine significantly reduced the leakage of lactate dehydrogenase, significantly inhibited the production of intracellular ROS (P<0.01), reduced the amount of malondialdehyde and down-regulated the expression of pro-apoptotic protein (P<0.05), increased the activity of superoxide dismutase, regulated MMP and improved the expression of anti-apoptotic protein (P<0.01).ConclusionOxymatrine can regulate oxidative stress by improving mitochondrial function, so as to inhibit the apoptosis of H9C2 cardiomyocytes induced by high glucose.
Abstract：ObjectiveTo observe the effect of polyphyllin Ⅰ on the expressions of forkhead box Q1（FOXQ1）and epithelial-mesenchymal transition (EMT)-related factors, in order to explore the possible mechanism of polyphyllin Ⅰ in inhibiting the metastasis of colon cancer.MethodAfter the treatment with 1.25，2.50 μmol·L-1 polyphyllin Ⅰ on HCT116 cells, Western blot and Real-time PCR were used to detect the expressions of FOXQ1，E-cadherin，Vimentin protein and mRNA.ResultCompared with the blank group, relative expressions of FOXQ1 protein and mRNA in low-concentration polyphyllin Ⅰ group were decreased, while relative expression of E-cadherin mRNA was increased, the differences were not statistically significant, and relative expressions of Vimentin protein and mRNA in low-concentration polyphyllin Ⅰ group were decreased (P<0.05)， and relative expression of E-cadherin protein in low-concentration polyphyllin Ⅰ group was increased (P<0.01). Compared with the blank group, relative expressions of FOXQ1, Vimentin protein and mRNA in high-concentration polyphyllin Ⅰ group were decreased，while relative expressions of E-cadherin protein and mRNA were increased (P<0.05, P<0.01). Compared with low-concentration polyphyllin Ⅰ group, relative expressions of Vimentin protein and mRNA in high-concentration polyphyllin Ⅰ group were decreased, but the difference was not statistically significant, relative expressions of E-cadherin protein and mRNA in high-concentration polyphyllin Ⅰ group were increased, whereas relative expressions of FOXQ1 protein and mRNA were decreased (P<0.05, P<0.01).ConclusionMechanism of polyphyllin Ⅰ inhibiting the metastasis of colon cancer may be related to the decrease of FOXQ1 and Vimentin expressions, and the up-regulation of E-cadherin.
Abstract：ObjectiveTo observe the clinical efficacy of modified Yihuo Qnggan Tuiyi Tang on herpes simplex keratitis with liver meridian and wind-heat，so as to explore its mechanism of action.MethodTotally 130 patients with herpes simplex keratitis with liver meridian and wind-heat were randomly divided into control group and observation group，with 65 cases in each group. The control group was treated with conventional western medicine，while the observation group was treated with modified Yihuo Qinggan Tuiyi Tang in addition to the therapy of the control group for 4 weeks. The clinical efficacy，traditional Chinese medicine（TCM） syndrome score，symptoms and signs，adverse reactions and recurrence rate of the two groups were compared. The levels of serum interferon gamma（IFN-γ），interleukin-6（IL-6），interleukin-8（IL-8） inflammatory factors，malondialdehyde (MDA)，antioxidant activity (TAC)，glutathione peroxidase（GSH-PX）of oxidative stress indexes were detected by enzyme linked immunosorbent assay （ELISA）. The levels of CD4+ and CD8+ were measured by flow cytometry. The recurrence rate was observed during 1-year follow-up visit.ResultThe total effective rate of observation group was better than that of control group (Z=2.161，P<0.05). After treatment，the scores of eye pain，shame，photophobia，tears，vision loss，redness and total eyelid swelling were significantly lower than those of control group (P<0.05). The total effective rate of TCM syndrome in the observation group was better than that of control group (Z=2.540，P<0.05). After treatment，the scores of pain，eyelid redness，conjunctival congestion，corneal punctate pannus，corneal pannus and corneal pannus depth of symptoms and signs in observation group were better than those of control group (P<0.05). The level of serum IFN-γ in observation group was higher than that of control group，and the levels of IL-6 and IL-8 were lower than those of control group (P<0.05). Levels of MDA in the observation group was higher than that of control group (P<0.05). The levels of CD4+and CD4+/CD8+ in observation group were higher than those of control group，and the levels of CD8+ were lower than that of control group (P<0.05). The recurrence rate in observation group was significantly lower than that in control group after withdrawal for 6 months and 12 months (P<0.05).ConclusionIn addition to the therapy of routine western medicine， modified Yihuo Qinggan Tuiyi Tang has a good clinical efficacy in treating herpes simplex keratitis with liver meridian and wind-heat，and can reduce the scores of TCM syndromes，alleviate symptoms and signs，and reduce the recurrence rate. Its mechanism may be related to reducing inflammation reaction，and improving oxidative stress index and immune function.
Keywords：herpes simplex keratitis;liver meridian and wind-heat;Yihuo Qinggan Tuiyi Tang;inflammatory factors;indicators of oxidative stress;immune function
Abstract：ObjectiveTo investigate the clinical efficacy of acupuncture combined with modified Shengyutang in treatment of cervical spondylotic radiculopathy with Qi-blood deficiency and its effect on T-lymphocyte subsets.MethodTotally 201 cases of cervical spondylotic radiculopathy with blood deficiency were randomly divided into 3 groups, with 67 cases in each group. Group A was treated with modified Shengyutang. Group B was treated with acupuncture. Group C was treated with modified Shengyutang combined with acupuncture. The effective rate of each group and the therapeutic effect of deficiency of Qi and blood were observed. Neck disabilitv index（NDI）, visual analogue score（VAS）, SF-36 mental component summary（SF-36MCS）and SF-36 physical component summary（SF-36PCS） were compared before and after treatment. Serum levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and calcitonin related peptide (CGRP) were measured before and after treatment. The levels of CD3+, CD4+, CD8+ and the ratio of CD4+/CD8+ were compared before and after treatment. The therapeutic safety indexes of each group were observed.ResultAfter treatment, the total clinical effective rate of group C was significantly higher than that of group A and B (P<0.05). NDI and VAS scores were significantly reduced (P<0.05). The scores of SF-36MCS and SF-36PCS were significantly increased (P<0.05). The score of Qi-blood deficiency syndrome was significantly lower (P<0.05). Serum levels of IL-1β, IL-6 and CGRP were significantly lower (P<0.05). The levels of CD3+, CD4+ and CD4+/CD8+ were significantly increased (P<0.05, P<0.01), whereas CD8+ was significantly decreased (P<0.05, P<0.01). No significant adverse reactions occurred in each group during the treatment.Conclusionacupuncture combined with Shengyutang has a significant clinical effect on cervical spondylosis with Qi-blood deficiency, and is worth popularizing.
Keywords：acupuncture;Shengyutang;cervical spondylotic radiculopathy;deficiency of Qi and blood;inflammatory factor;T-lymphocyte subsets;clinical observation
Abstract：ObjectiveTo explore the clinical efficacy of Chaihu Shugansan combined with saccharomyces boulardii in treating functional dyspepsia (FD) in children and the effect on serum cytokines and gastrointestinal hormones expression.MethodTotally 120 cases were divided into control group (60 cases) and observation group (60 cases)．The control group was treated with Duopanlitong Pian 0.3 mg·kg-1, po， tid， and Saccharomyces boulardii 0.25 g，po，bid. The observation group was treated with Chaihu Shugansan, po，qd， and saccharomyces boulardii 0.25 g，po，bid. Four weeks was a course of treatment．The changes of serum Gastric Hunger Hormone and cytokines, Hamilton depression scale (HAMD) score, symptoms improvement time and clinical effect in the two groups were observed. And the recurrence rate was observed after 6 months follow-up.ResultThe total effective rate was compared between control group (76.67%，46/60) and observation group (96.15%，57/60)，with significant differences (P<0.01). After treatment, serum tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in both groups were decreased, while serum levels of ghrelin of both group were increased. However，the decreasing range and the increasing range of observation group were statistically obvious than those of control group (P<0.01). HAMD score and disappearance time of clinical symptoms in observation group was significantly different from those in control group (P<0.01). In adverse reactions during treatment, the control group was inferior to the observation group (P<0.05). After the 6-month follow-up, the recurrence rate in observation group was lower than that in control group (P<0.01).ConclusionChaihu Shugansan combined with Saccharomyces boulardii was effective in treatment of FD in children, with a low recurrence rate and good compliance. The mechanism may be related to the effective regulation of brain-intestinal axis, balance of intestinal flora, inhibition of inflammatory reaction, regulation of cytokines and gastrointestinal hormone levels.
Abstract：ObjectiveTo discuss the efficacy of modified Qingzao Jiufei Tang combined with adjuvant chemotherapy on quality of life and survival period of patients with non-small cell lung cancer (NSCLC) and the moderation effect on immune function and cytokines.MethodOne hundred and twenty patients were randomly divided into control group (60 cases) and observation group (60 cases) by random number table. Patients in two group got paclitaxel and cisplatin chemotherapy.The control group took Yifei Qinghua granules orally,2 bags/time,3 times/day. In addition to the therapy of control group, patients in observation group were added with modified Qingzao Jiufei Tang from the first day after chemotherapy to the 12th week after chemotherapy, 1 dose/day. Six-month and one-year survival rates and survival period were recorded. Before and after treatment, lung cancer quality of life scale (FACT-L 4.0) and M.D. Anderson Symptom Inventory (MDASI-C) were scored. And levels of CD3+, CD4+, CD8+ and CD4+/CD8+ , natural killer cells (NK), cytokeratin 19 fragment 21-1 (CYFRA21-1), squamous cell carcinoma antigen (SCC-Ag), neuron-specific enolase (NSE), vascular endothelial growth factor (VEGF), nuclear transcription factor-κB (NF-κB), intercellular adhesion molecule 1 (ICAM-1), transforming growth factor (TGF)-β1 were all detected.ResultThe 6-month survival rate of in observation group was 78.33%(47/60), which was higher than 60.00%(36/60) in control group (χ2=4.728, P<0.05). The one-year survival rate in observation group was 48.33%(29/60), which was higher than 30.00%(18/60) in control group (χ2=4.232, P<0.05). And the survival period in observation group was 15 months, which was longer than 11 months in control group (P<0.01). And scores of FACT-L and various factors were all higher than those in control group (P<0.01). Scores of disease symptoms, gastrointestinal symptoms, fatigue symptoms and emotional symptoms were all lower than those in control group (P<0.01), and scores of effect of symptoms on daily life in six aspects, such as enjoyment of life, were all lower than those in control group (P<0.01). And levels of NK, CD3+, CD4+ and CD4+/CD8+ were higher than those in control group (P<0.05), while levels of CD8+, TGF-β1, NF-κB, ICAM-1, VEGF, CYFRA21-1, SCC-Ag and NSE were all lower than those in control group (P<0.01).ConclusionModified Qingzao Jiufei Tang combined with adjuvant chemotherapy can prolong the survival time, improve the survival rate, alleviate the effect of symptoms on daily life, improve the quality of life, inhibit the expression of tumor markers and cytokines, and stabilize the immune function of the body, with a positive clinical significance in improving the prognosis of NSCLC.
Keywords：non-small cell lung cancer;Qingzao Jiufei Tang;chemotherapy;survival period;survival rate;quality of life;immune function
Abstract：ObjectiveTo observe the effect of Jiajian Fuyuan Huoxuetang combined with electroacupuncture on neurological rehabilitation of patients with spinal cord injury, and to investigate its effect on neurotrophic and inflammatory factors.MethodOne hundred and twenty patients were randomly divided into control group (59 cases) and observation group (61 cases) by random number table. Both groups’ patients got monosialotetrahexosylganglioside by intravenous drip for 6 weeks, 200-400 mg/time, 1 time/day.Rat nerve growth factor for injection by intramuscular injection for 4 weeks, 20 μg/time, 1 time/day. Patients in control group additionally got electroacupuncture treatment,and tongluo huatan capsule,3 caps/time,3 time/days. The patients in observation group additionally got Jiajian Fuyuan Huoxuetang combined with electroacupuncture, 1 dose/day. The courses of treatment were 12 weeks in two groups. Scores of degree of spinal cord neurological impairment were graded by using impairment score of American Spinal Injury Association (ASIA). Lower extremity motor ability was discussed by using the walking index of spinal cord injuryⅡ (WISCⅠ Ⅱ) and 10 minutes’ walking time (10 MWT). Before and after treatment, functions of daily life were evaluated by using Barthel index (MBI). Bladder function was also discussed, and levels of brain-derived nerve growth factor in peripheral blood (BDNF), nerve growth factor (NGF), astrogenic calcium binding protein S-100 β (S-100 β), tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) were also detected.ResultAfter the treatment, scores of sensory and motor in ASIA scale in observation group were all higher than those in control group (P<0.01). Score of WISCⅠ Ⅱ in observation group was higher than that in control group, and 10 MWT was shorter than that in control group (P<0.01). Times of leakage of urine, urinary incontinence volume and residual urine volume were all less than those in control group, and bladder volume was more than that in control group (P<0.01). Functions of daily life in observation group were better than those in control group (Z=1.967, P<0.05), and the levels of BDNF and NGF were higher than those in control group, while levels of S-100 β, TNF-α and IL-1β were lower than those in control group (P<0.01).ConclusionOn the basis of routine comprehensive rehabilitation therapy of western medicine, Jiajian Fuyuan Huoxuetang combined with electroacupuncture can promote the recovery of nervous function, improve sensory and motor function, improve bladder function and daily living ability, and promote the expression of neurotrophic factors and inhibit inflammatory reaction in neurological rehabilitation for the patients with spinal cord injury.
Abstract：ObjectiveThe phase separation of Baihutang was carried out. The content of mangiferin，new mangiferin，calciumion，glycyrrhizin and ammonium glycyrrhizinate in the solution phase，the nano phase and the precipitated phase of Baihutang were measured，so as to define the effect of nanometer particles of Baihutang on the growth of active components，and explain the mechanisms of Baihutang in potent detoxification and heat removal.MethodThe phase separation of Baihutang was performed by high-speed centrifugation and dialysis. The contents of mangiferin，new mangiferin，glycyrrhizin and ammonium glycyrrhizinate in Baihutang were determined by HPLC. Chromatographic column Diamonsil C18 (4.6 mm×250 mm，5 μm) was adopted, with acetonitrile-25 mmol·L-1 potassium dihydrogen phosphate solution as the mobile phase, and eluted in a gradient mode. The detection wavelength was 257 nm，the column temperature was 30 ℃，and the flow rate was 1.0 mL·min-1. EDTA-2Na solution was used to calibrate the calcium concentration in different phase states of Baihutang.Result1 mL Baihutang nanoparticles contained 483.00 μg new mangiferin，1 068.88 μg mangiferin，219.93 μg glycyrrhizin and 187.10 μg ammonium glycyrrhizin，and the content of new mangiferin and mangiferin accounted for 89.4% and 89.9% respectively in 1 mL Baihutang. The new mangiferin and mangiferin in the nano phase were 230.0 and 23.3 times the true solution，and 8.5 and 14.4 times of the precipitation，respectively. The content of calcium ions in Baihutang in the nano phase was higher，accounting for 86.9% of Baihutang，and the content of calcium ions in Baihutang and Baihutang in the nano phase was higher than that in gypsum group.ConclusionThe content of main components in Baihutang in nanometer phase is significantly higher than that in other phases. The nanoparticles of Baihutang have a solubilizing effect on the main antipyretic components, such as mangiferin，mangiferin and calcium ions as well as the antitoxic components glycyrrhizic acid and glycyrrhizic acid. The mechanism of action of Baihutang is related to the formation of nanoparticles.
Abstract：ObjectiveTo analyze and compare the volatile oil components of pine needle of Pinus yunnanensis from different habitats.MethodThe volatile oils of pine needle of P. yunnanensis from 12 different habitats were extracted by supercritical CO2 extraction （SFE-CO2），the components were analyzed by gas chromatography-mass spectrometry （GC-MS），and relative percentage of each component was determined by peak area normalization method，and SPSS 20.0 software was used for principal component analysis.ResultThe 137 compounds were isolated from the volatile oil of pine needle of P. yunnanensis from 12 different producing areas，with 1 sample identified 57 species，2 samples identified 57 species，3 samples identified 54 species，4 samples identified 48 species，5 samples to identify 58 species，6 samples identified 60 species，7 samples identified 53 species，8 samples identified 55 species，9 samples identified 56 species，10 samples to identify 58 species，11 samples identified 54 species and 12 samples to identify 50 species，composed of hydrocarbon，acid，alcohol，ester，aldehyde，ketone，phenol，ether 8 compounds，including 11 common composition including alpha pinene，beta pinene，beta caryophyllene，7C，10C，16C-hexadecatrienoic acid and palmitic acid methyl ester，palmitic acid，phthalic acid dibutyl do acid，alcohol，stearic acid，mountain，wood tar acid.ConclusionIn this study，supercritical CO2 extraction （SFE-CO2） was used to extract the volatile oil of pine needle of P. yunnanensis from different habitats. It was found that the volatile oil of pine needle of P. yunnanensis was mainly composed of hydrocarbons，acids，esters and alcohols. And the content and species of volatile oil of pine needle of P. yunnanensis from different producing areas differ greatly which lay a foundation for the further development and utilization of pine needle of P. yunnanensis resources.
Keywords：pine needle of Pinus yunnanensis;volatile oil;supercritical CO2 extraction;gas chromatography-mass spectrometry
Abstract：ObjectiveTo accurately identify Bupleurum seeds by traditional morphological identification method combined with DNA barcoding technique.MethodA total of 41 seed samples on the market were collected and 75 ribosomal DNA internal transcribed spacer 2 (ITS2) sequences of 15 varieties were downloaded from the GenBank database as experimental materials. The seeds were measured and observed by stereomicroscope and vernier caliper, and their 1 000-grain weights were calculated. Genomic DNA was extracted from the seeds and used as a template, and ITS2 sequences were amplified using polymerase chain reaction (PCR) and bidirectional sequencing. Species identification was conducted based on BLAST method, neighbor-joining (NJ) phylogenetic tree method, Kimura two-parameter model (K2P) genetic distance method, and secondary structure of ITS2 sequence.ResultThere were slight differences in the length, width, cross-section, and 1 000-grain weight among Bupleurum seeds from different origins. The ITS2 sequences of B. chinense seeds had 2 intraspecific variable sites and 3 haplotypes, the maximum intraspecific genetic distance (0.009) was far smaller than the minimum interspecific genetic distance (0.032). B. chinense and B. scorzonerifolium in the NJ phylogenetic tree were clustered into independent branches with good monophyletic property. The secondary structure of ITS2 sequences could make up for the shortcomings of NJ tree in identifying variants. The collected 41 seeds included 30 B. chinense seeds, 3 B. scorzonerifolium seeds, 5 B. falcatum seeds, 2 B. marginatum var. stenophyllum seeds, and 1 B. smithii var. parvifolium seeds.ConclusionThe B. chinense seeds on the market have problems of diverse sources and chaotic origins. Based on the combination of ITS2 gentic barcoding and seed morphological identification, the Bupleurum seeds can be accurately identified, which provides scientific bases for establishing the quality standard of Bupleurum seeds, standardizing the cultivation of B. chinense, and solving the quality problems of B. chinense from the source, and provides a reference for the accurate identification of other medicinal plant seeds or seed medicinal materials.
Abstract：ObjectiveTo establish a characteristic spectrum to reflect the efficacy of Houpo Qiwutang.MethodBased on the correlation between the efficacy and the pharmacological action of each herb in prescription，the target substances of characteristic map were screened. The extraction solvent，detection wavelength and gradient of the active ingredients were optimized. Peak assignment was made by comparing individual drugs. Q-TOF was used to infer the molecular formula of each peak in the characteristic atlas，and the reference substance was identified by the reference substance. The reference substance was screened out according to the correlation of main efficacy and medicine.ResultThe characteristic spectrum of material standard of Houpu Qiwutang was established. Five of the seven herbal medicines were attributed. Nine characteristic peaks were selected and identified by Q-TOF as glycyrrhizin，including naringin，neohesperidin，ammonium glycyrrhizinate，rhein，honokiol，magnolol. According to the main efficacy of Houpo Qiwutang，neohesperidin was selected as reference substances. According to the separation of characteristic peaks and the retention time，the mark peak of the characteristic spectrum was determined.ConclusionThe characteristic spectrum of the material basis of Houpo Qiwutang was established by selecting the characteristic peaks and controlling the key components. This method not only reflects the situation of all the effective chemical components，but also focuses on the control of the key efficacy，so as to provide a theoretical basis for the subsequent development and quality control of Houpu Qiwu Tang.
Abstract：ObjectiveTo evaluate the efficacy and safety of traditional Chinese medicine (TCM) in the treatment of post stroke cognitive impairment (PSCI).MethodSeven databases, including CNKI, WanFang, VIP, CBM, PubMed, The Cochrane library and ClinicalTrials.gov, were electronically searched for relevant randomized controlled trials (RCTs) of TCM in the treatment of PSCI. The Cochrane risk of bias assessment tool was used to evaluate the methodological quality of the included studies, descriptive analysis was carried out on the included studies, and the Meta quantitative analysis was carried out with RevMan 5.3 software.ResultA total of 16 RCTs were included with 1 296 participants, and they were assigned to the intervention group (n=649) and the control group (n=647). The results showed that TCM combined with western medicine group and TCM group were better than western medicine group in improving the scores of Montreal Cognitive Assessment (MoCA), Mini-mental State Examination (MMSE), Barthel Index (BI), Activity of Daily Living (ADL), Chinese stroke scale (CSS) and National Institutes of Health stroke scale (NIHSS) of PSCI patients, and no serious adverse events were observed.ConclusionTCM has potential advantages in improving the cognitive function of patients with PSCI, and it also has certain efficacy in improving the daily living ability and neurological impairment symptoms, and no serious adverse events have been observed. Due to the low quality of methodology included in the studies, in order to provide reliable basis for clinical decision-making, high-quality of RCTs are still needed to study the efficacy and safety of TCM for PSCI.
Keywords：post stroke cognitive impairment (PSCI);traditional Chinese medicine;randomized controlled trial (RCT);Meta analysis;cognitive function level;activity of daily living;neurological impairment
Abstract：ObjectiveTo investigate the distribution of traditional Chinese medicine (TCM) syndromes in patients with coronary heart disease (CHD) in 6 months after interventional therapy, and to analyze relevant influencing factors.MethodThe clinical data of 1 000 patients with coronary heart disease in 6 months after interventional therapy, including the four diagnosis information of TCM, were collected, and the distribution of TCM syndromes and the influencing factors were analyzed.ResultAmong 48 kinds of information about the four diagnostic methods of TCM, chest pain was the most frequent (98.10%)， among 9 kinds of common TCM syndrome types, blood stasis was the most frequent (89.90%), and the others were heart-Qi deficiency syndrome, phlegm turbidity syndrome, cold-dampness syndrome, kidney-Qi deficiency syndrome, heart-Yin deficiency syndrome, kidney-Yin deficiency syndrome, Heart-Yang deficiency syndrome and kidney-Yang deficiency syndrome in turn. Among 6 common TCM syndrome types, Qi deficiency and blood stasis syndrome were the most frequent (35.40%), and the others were phlegm turbidity and blood stasis syndrome, cold congealing heart pulse syndrome, Qi-Yin deficiency syndrome, heart-kidney Yin deficiency syndrome and heart-kidney-Yang deficiency syndrome in turn. There was no significant difference in sex ratio among different syndrome types . Patients with heart-kidney Yang deficiency syndrome had no significant difference. Compared with the average age of other syndromes, there were significant differences. Common complications included hypertension, diabetes, cerebrovascular diseases and dyslipidemia, among which hypertension had the highest frequency, with significant differences from other diseases (P<0.05). Phlegm, turbidity and blood stasis were found in patients with hypertension. The risk of syndromes was higher (OR=3.29, 95% CI [2.11, 5.05]), while the risk of cold congealing heart pulse syndrome was lower (OR=0.56, 95% CI [0.32, 0.98]), the risk of Qi and Yin deficiency was higher (OR=2.88, 95% CI [2.01, 4.99]), whereas the risk of heart and kidney Yang deficiency was lower (OR=0.54, 95% CI [0.29, 0.95]) when complicated with cerebrovascular diseases. The risk of Qi deficiency and blood stasis was higher (OR=2.97, 95% CI [2.05, 5.28]), while the risk of heart and kidney Yang deficiency was lower (OR=0.54, 95% CI [0.29, 0.95])， the risk of phlegm turbidity and blood stasis was higher when complicated with dyslipidemia (OR=3.55, 95% CI [2.32, 5.29]), and the risk of heart and kidney Yang deficiency was lower (OR=0.54, 95% CI [0.29, 0.95]). The time distribution of the disease had obvious seasonal characteristics.ConclusionThe main distribution characteristics of TCM syndromes in 6 months after coronary heart disease intervention are basically the same as those in patients without intervention. The main TCM syndromes are Qi deficiency and blood stasis syndrome, phlegm turbidity and blood stasis syndrome, cold congealing heart pulse syndrome, Qi-Yin deficiency syndrome, heart-kidney Yin deficiency syndrome and heart-kidney-Yang deficiency syndrome. The distribution pattern may be related to age, complications and seasons.
Keywords：coronary heart disease;intervention;traditional Chinese medicine (TCM) syndromes;distribution;relevant factors
Abstract：Diabetes is one of the most common chronic non-communicable diseases. The incidence rate is on the increase year by year. As the disease progresses, it can cause damages to many tissues and organs, such as eyes, kidneys, nerves, heart and blood vessels, which seriously endangers human health. Traditional Chinese medicine (TCM) has outstanding clinical advantages in the prevention and treatment of diabetes, with distinctive characteristics of diagnosis and treatment. TCM has a long history of understanding diabetes. In recent years, researches on etiology, pathogenesis and treatment have gradually deepened and made certain progress. As early as in the Huangdi Neijing, there is a record of "If people’s lung, spleen viscera (stomach), liver, kidney are weak, they can easily get diabetes". This paper collected and summarized ancient literatures relating to diabetes. According to the basic construction principle of "integration-authenticity-innovation" in the theoretical framework of TCM，the causes of the disease include liver weakness and failure in dredging and regulation，emotional disorder, endogenous heat and dryness, heart weakness, heart Yin depletion, endogenous dryness and heat， spleen deficiency and failure in transportation and transformation， lung injury caused by dryness and heat, dysfunction of body fluid distribution, kidney Yin deficiency, astringent weakness, insufficient Qi transformation for maintaining body fluid metabolism balance. In general, Yin deficiency is the root cause, and dryness and heat are the appearance. A framework for the treatment of diabetes was built with five organs as spatial dimension. This paper summarizes the etiology and pathogenesis of diabetes caused by viscera deficiency, with the treatment of diabetes with the five internal organs as the basic framework. The main methods of treating diabetes are relieving Qi stagnancy in liver, regulating the mood，clearing the heart fire, Yin-enriching and dryness-moistening, reinforcing spleen Yang, invigorating spleen and removing dampness, clearing lung and moistening dryness, nourishing Yin, kidney Yin, warming kidney and promoting Yang. A clear and hierarchical theoretical framework was built as a bridge between the theory of TCM and clinical practice of TCM, in the expectation to provide new ideas for the study of ancient TCM literatures and the treatment of diabetes mellitus with TCM, and contribute to the inheritance and development of TCM theory.
Keywords：traditional Chinese medicine theory of visceral;diabetes;treatment framework;traditional Chinese medicine theory;diabetes
Abstract：Alpinia zerumbet is a commonly used drug for ethnic minorities in Guizhou province. It has the effects of warming and drying dampness，relieving pain and eliminating malaria，and treating chills，chest and abdomen，indigestion，vomiting and diarrhea，with a long history of nearly 200 years. The author reviewed Chinese and foreign literatures for the past 30 years，and reviewed the research progress of the chemical constituents and pharmacological effects of A. zerumbet in and abroad，in order to provide a theoretical basis for its medicinal value. With the development of instruments and technology，the chemical composition of A. zerumbet has attracted more and more attention. More than 200 compounds have been isolated and identified，including volatile oils，flavonoids，steroids，terpenoids and organic acids and many other chemical ingredients. Pharmacological studies have shown that A. zerumbet has many pharmacological activities，such as anti-oxidation，blood pressure lowering，antispasmodic analgesia and protection against endothelial cells. However，current studies on the chemical constituents only focus on volatile oil components，and only a few studies focus on other organic matter and polysaccharides and are not deep enough. In clinic，the use of A. zerumbet is more confusing. The roots，stems，leaves，flowers，fruits and seeds can be used in medicines，but the pharmacological effects of the various medicinal parts have not been clearly explained，which leads to confusion in clinical medication. In addition，the pharmacological mechanism is not clear，especially in the studies on traditional activities，such as analgesic，digestive and anti-ulcer. The studies only focus on the pharmacological activity，and with only a few studies on the mechanism of action. Besides，the existing studies are mainly in vitro activity experiments，and need further validation in clinical trials，so as to provide reference for further rational development and comprehensive utilization of medicinal resources.
Keywords：Alpinia zerumbet;Zingiberaceae;Miao medicine;essential oil;flavonoids;antioxidant;protection of endothelial cells
Abstract：Arbuscular mycorrhizal fungi （AMF） can establish mycorrhizal symbiosis system with most terrestrial plants，and play important roles in their growth and development. However，there is no systematic analysis and summarization of their roles in the growth, biosynthesis and accumulation of active substances of herbs，as well as stress-resistance mechanism. First，the main research methods of AMF were summarized in the paper，including the separation of AMF spores，morphological identification，chemical staining and molecular identification. The main morphological structures of some AMF were detailed in the table. In terms of growth promotion，AMF promoted the growth by prolonging mycelium，enhancing phosphatase secretion，organic acid，activation of soil and increasing absorption efficiency. In the aspect of biosynthesis and accumulation of flavonoids，terpenoids and other active substances in herbs，AMF improved the contents by regulating activities of signal substances and key enzymes involved in the metabolism of secondary products. In addition，AMF could alleviate a series of stresses caused by drought，heavy metal，high salt，high or low temperature by improving the activity of antioxidant enzymes，enhancing the ability of plants to scavenge free radicals，complexing toxic heavy metals，diluting high salt concentration，or inducing the expressions of key genes. Finally，the application prospects and in-depth study of AMF in the ecological planting of herbs were discussed, in order to provide reference for promoting relevant research.
Abstract：Spinal cord injury (SCI) is mostly caused by severe trauma,which often leads to motor,sensory,reflex and sphincter dysfunction from the injury plane.It is the main cause of paraplegia.With the development of transportation and industry in modern society,the incidence of SCI is increasing year by year,but there is still a lack of ideal treatment drugs.Therefore,finding effective,safe and reliable drug treatment for spinal cord injury is one of the hotspots in basic and clinical research.Traditional Chinese medicine（TCM）has the characteristics of cheapness and less side effects,and it has a long history to be used in clinical treatment of spinal cord injury. Recent studies have shown that some single TCM and their extracts can effectively alleviate or improve secondary spinal cord injury, such as local vascular dysfunction, inflammation, oxidative stress, cell apoptosis, axonal demyelination, glial scar formation, and confirm the scientific nature of their treatment of spinal cord injury.This paper summarized the mechanism of single TCM and its extracts in promoting the repair of spinal cord injury,including improving spinal cord edema,ischemia and hypoxia by regulating the expression of aquaporin-4（AQP-4）,hypoxia-inducing factor（HIF）-1α,and vascular endothelial growth factor（VEGF）,inhibiting inflammation by regulating tumor necrosis factor（TNF）-α,interleukin（IL）-1β,inhibiting anti-oxygen free radical damage and lipid peroxidation.Chemicals,inhibit neuronal apoptosis,promote nerve cell repair and regeneration,inhibit the formation of glial scars and other aspects to alleviate pathological damage of spinal cord and promote the recovery of nerve function. It shows that traditional Chinese medicine has unique advantages such as multi-target,multi-channel and multi- level.It has broad research prospects for treating SCI and improving prognosis.
Keywords：spinal cord injury;traditional Chinese medicine;mechanism of action;research progress