Abstract：ObjectiveTo study the therapeutic effect of Jinchai Kangbingdu capsule based on human coronavirus pneumonia with 'Hanshi Yidu Xifei' syndrome model, in order to provide experimental basis for evaluating its effect in preventing and treating coronavirus infection.MethodThe 48 Balb/c mice were randomly divided into normal group, virus infection group, cold-dampness group, cold-dampness epidemic toxin lung syndrome model group, and high and low-dose Jinchai Kangbingdu capsule groups (1.76, 0.88 g·kg-1·d-1). A cold-dampness stimulation combined with human coronavirus 229E infection was used to imitate human coronavirus pneumonia with 'Hanshi Yidu Xifei' syndrome model. Behavioral characteristics, lung index, viral load, and lung tissue pathological changes in Balb/c mice were observed to evaluate the therapeutic effect of Jinchai Kangbingdu capsules. The contents of interleukin-6（IL-6），IL-10，tumor necrosis factor-α（TNF-α），interferon-γ（IFN-γ） in lung tissue and motilin（MTL），gastrin（GAS） in serum were detected by enzyme-linked immunosorbent assay(ELISA), and the contents of CD4+ T cells, CD8 + T cells, and B cells in peripheral blood were detected by flow cytometry.ResultCompared with the cold-dampness epidemic toxin lung syndrome model group, Jinchai Kangbingdu capsule can increase the activity and response ability of 'Hanshi Yidu Xifei' syndrome model mice, and change the skin and stool status of mice. High and low-dose Jinchai Kangbingdu capsule groups can significantly reduce the lung index (P<0.01), while significantly increased the content of CD4+ T cells, CD8+ T cells, and B cells (P<0.05, P<0.01). Low-dose Jinchai Kangbingdu capsule group could significantly decrease the MTL content in serum and the levels of IL-6, IL-10, TNF-α, IFN-γ in lung tissue (P<0.01), whereas alleviate the pathological damage of lung tissue.ConclusionJinchai Kangbingdu capsule showed a therapeutic effect on human coronavirus pneumonia with 'Hanshi Yidu Xifei' syndrome model, and can improve the behavioral characterization and gastrointestinal index level of cold-dampness syndrome, while reduce lung index and viral load in lung tissue. The mechanism may be related to the decrease of the content of inflammatory factors and the increase of the number of lymphocytes.
Keywords：Jinchai Kangbingdu capsule;coronavirus;'Hanshi Yidu Xifei' syndrome;inflammatory factor;combination of disease and syndrome
Abstract：ObjectiveTo study the clinical medication regulation for the prevention and treatment of new coronavirus pneumonia with traditional Chinese medicine（TCM），especially prescriptions，on the basis of "treatment in accordance with seasonal conditions".MethodThe guidelines and suggestions on novel coronavirus pneumonia formulated and published by national，provincial and municipal governments and experts before February 17，2020, were retrieved and summarized. Data was recorded in stages according to "prevention period，clinical observation period，pre-clinical period，middle clinical period，late clinical period and recovery period". The frequency analysis of TCM and its efficacy，prescription and proprietary Chinese medicine was carried out，and then the factor analysis and cluster analysis of TCM were carried out to obtain rational drug combinations.ResultTotally 172 suggestions on combined structure of drugs，50 suggestions on prescriptions and 31 suggestions on proprietary Chinese medicine were involved in the study，and provided by 24 official agencies and 24 medical experts， 147 kinds of herbs，44 prescriptions and 16 proprietary Chinese medicines were collected. No matter for the drug category or for the specific drug selection，the law of drug use in different stages was not the same. Huoxiang Zhengqi capsule，Shufeng Jiedu capsule，Lianhua Qingwen capsule and Jinhua Qinggan capsule can be used in the observation period. Xuanbai Chengqitang can be used in the middle of the disease. Angong Niuhuangwan，Suhexiangwan or Zixuedan, and Angong Niuhuangwan can be selected in the middle of the disease. And Qingfei Paidu decoction can be used in all stages of the disease.ConclusionDoctors need to adjust their prescriptions along with the progress of the disease，because the law of medication in each stage of the new coronavirus pneumonia is different.
Keywords：novel coronavirus pneumonia(COVID-19);different time;compatibility pattern of herbs;literature research
Abstract：Since the outbreak of corona virus disease-2019 (COVID-19), the timely intervention of Chinese medicine has played an important role， it can significantly improve the clinical symptoms of patients, alleviate fever, cough, shorten the course of disease and reduce the conversion rate of mild disease to severe disease， moreover, it has obvious advantages in improving survival rate, prognosis and quality of life in patients. In order to make better use of the therapeutic effect of Chinese medicine and overcome the epidemic situation as soon as possible, various provinces, municipalities and autonomous regions have introduced local Chinese medicine treatment schemes in accordance with the geography, climate and epidemic situation. In this article, we have analyzed the similarities and differences among treatment schemes from the perspective of three factors, and it is believed that the onset season is the same for this disease, and the differences between treatment schemes mainly depend on geographical and climatic factors. The results showed that in seven regions of China, the early symptoms in warm and humid regions such as Central China, South China, East China and Southwest China were characterized by "wet, warm, hot, and toxic", and treatment schemes mainly adopted the recipes with clearing heat and detoxifying, strengthening spleen and removing dampness functions. The early symptoms in cold regions such as Northeast China, North China, and Northwest China were mostly characterized by "cold, wet, wind, and toxic", and the therapeutic recipes emphasized on the effects of expelling wind and cold, removing toxicity and dampness. Among them, the national plan with the Hubei epidemic situation as an important reference has gradually evolved from a single syndrome attribute of each period to both cold and hot syndromes with multiple treatment methods. With the development of the disease, epidemic toxin and toxic heat in the lungs are the main characteristics in the medium term, when the treatment mainly focuses on purging heat and removing toxins. In severe cases and advanced stage, internal block and outward desertion are the main symptoms, when the treatment mainly focuses on block dredging and relieving desertion. In the recovery period, deficiency of lung and spleen Qi, and deficiency of Qi and Yin are the main symptoms, when the treatment is mainly used to invigorate the spleen, tonify the lungs, and nourish Qi and Yin. The physical factors are mostly reflected in the prevention program, so they are rarely mentioned in the treatment program. The Chinese medicine treatment schemes of COVID-19 in different regions show obvious differences. It is of great significance to analyze these differences and sum up the corresponding laws for the differential prevention and treatment of infectious diseases like COVID-19.
Keywords：corona virus disease-2019 (COVID-19);treatment in accordance with three categories of etiologic factors;Chinese medicine;treatment scheme;Chinese medicine plan
Abstract：The outbreak of corona virus disease-2019 (COVID-19) in Wuhan, China, in December 2019, is highly contagious and epidemic, and belongs to the category of "plague" in traditional Chinese medicine, which seriously threatens human life and health. In the face of this sudden public health event, based on the theory of Wenyilun, this paper analyzes the onset characteristics, transmission, and syndrome differentiation-based treatment of COVID-19 in clinical practice. According to the characteristics of this epidemic, the pathogeny of COVID-19 is "epidemic pathogenic factors", and the pathogenesis is that the epidemic suppresses the dampness. Its transmission follows the regularity of "nine transmissions between exterior and interior" recorded in Wenyilun, which can be divided into sequential transmission and reverse transmission. The course of the disease has certain regularity and stages. In the early stage, the pathogen hides in the pleurodiaphragmatic interspace, and the epidemic suppresses the dampness， the treatment shall reach the pleurodiaphragmatic interspace, clear the dampness and turbidness, diffuse the lungs and dredging the lung meridian. In the middle stage, the pathogen can be transmitted to the exterior， the treatment shall be pungent in flavor and cool in property and could relieve the exterior syndrome, so as to remove the pathogen by shivering and sweating. If the dampness toxin enters the interior and be transmitted into the heat, and caused damage to the lung meridian, obstruction of Qi in the lungs, interior excess and heat accumulation, the treatment shall dredge bowel and purge heat, and expel the pathogen out. Any delay in treatment, mistreatment or excessive pathogen can lead to pathogenic invasion into the pericardium, and cause delirium, dysphoria, reversal cold of limbs, barely palpable pulse and other critical manifestations of internal blocking causing external depletion. The treatment shall focus on both attack and supplement, open blocking and relieve depletion. In the recovery period, due to the residual pathogen and the Yin and Qi impairment caused by fever, the treatment shall prevent the recurrence of the residual pathogen, nourish Yin and blood, and restore the vital energy. During the treatment, WU You-ke put forward three principles of "removing the pathogen as the priority, and then avoiding false sweating, and caring for the spleen and stomach". The theory of Wenyilun has a systematic understanding of the pathogeny, pathogenesis, and syndrome differentiation-based treatment of "the epidemic", and provides valuable experience for fight against plagues in later generations, which is also of great significance for the prevention and treatment of COVID-19.
Keywords：Wenyilun;corona virus disease-2019 (COVID-19);syndrome differentiation-based treatment;plague;nine transmissions between exterior and interior
Abstract：Some patients with corona virus disease-2019 (COVID-19) experienced a severe exacerbation of the disease due to the occurrence of inflammatory storm during the development of the disease. They are complicated with acute respiratory distress, multiple organ dysfunction syndrome and other serious complications, with a poor prognosis and a high mortality. For the inflammation storm, western medicine mostly adopts glucocorticoids, nutritional support, artificial ventilation assistance and other measures at present. The development of artificial liver, blood purification therapy, extracorporeal membrane oxygenation and other technologies have also reduced the mortality of patients to some extent. However, due to the high requirements for equipment, the measures have not yet been widely carried out. From the perspective of traditional Chinese medicine(TCM), the basic pathogenesis of COVID-19 is epidemic toxin invasion, lung and spleen being affected by pathogens, damaging vital Qi, and pathological properties involving dampness, heat, poison, stasis and deficiency. At the stage of inflammation storm, the pathogens are abundant, while the vital Qi is deficient， and the pathogens occlude lung, and disturb the heart and mind, and blood stasis and toxicity are combined with Qi-Yin deficiency. In severe cases, even both Yin and Yang exhaustion occurs. At present, a number of studies have shown that a variety of Chinese herbal medicines have multi-target immunomodulatory effect on viral pneumonia and cytokine storm. TCM participates in whole process of the occurrence and development of inflammation storm, mainly eliminating pathogens in early stage, controlling inflammation and blocking occurrence of inflammation storm， eliminating pathogens and strengthening the body resistance to eliminate the pathological products of the inflammation storm, and promoting the dissipation and absorption of inflammation in the middle stage， and saving lives in late stage by benefiting Qi and relieving depletion, and restoring Yang and rescuing from collapse. On the basis of pathophysiological mechanism of COVID-19 inflammation storm and the theory of TCM syndrome differentiation and treatment, this paper summarized the pharmacological studies on intervention on inflammatory storm with relevant Chinese herbal medicine, Chinese medicine prescriptions and Chinese medicine preparations, and discussed the intervention measures of TCM in different development stages of inflammatory storm, in expectation of providing the guidance for clinical treatment.
Keywords：corona virus disease-2019;inflammatory storm;cytokines;traditional Chinese medicine（TCM）;dialectical treatment
Abstract：The outbreak of corona virus disease-2019 (COVID-19) not only endangers the public life and health, but also poses threats to the public mental health. Under the atmosphere of tension and fear, the public often produces adverse emotional reactions to psychological stress, and even has extreme behaviors, which seriously affects physical and mental health. Traditional Chinese medicine （TCM） emotional therapy is based on the TCM consciousness theory in adherence to the holistic view of "integration of mind and body". It adopts safe and effective emotional conditioning methods to regulate mood, eliminate bad emotions, maintain mental health, and enhance the body's immunity to achieve the effect of treating or preventing physical and mental diseases. It has differentiated applications in many medical fields, such as post-traumatic stress disorder, generalized anxiety disorder, cancer and hospice, with good clinical efficacy. By analyzing the theoretical origin, research status, application range of TCM sentiment therapy, as well as the current public psychological state of the COVID-19 epidemic, we innovatively proposed TCM sentiment therapies, such as calm emotion method, diversion of attention method, suggestive induction method, obedient mind method, music delight method, TCM refreshing method and auricular pressing method, and comprehensively applied them in the public psychological adjustment during the outbreak, with the aim to provide rational and effective psychological guidance, help the public to correctly understand various bad emotions during the outbreak, enhance their abilities to cope with it and adjust psychological problems, maintain a good mentality, and pass the epidemic period smoothly. At the same time, in order to make better use of the advantages of TCM sentiment therapy and benefit the public, it is advocated to widely apply TCM sentiment therapy in similar public health events and public psychological care in the daily life, for the purpose of preventing before disease outset and preventing progress of disease, and effectively protecting the physical and mental health of the people.
Keywords：corona virus disease-2019 (COVID-19);traditional Chinese medicine （TCM） emotional therapy;psychological stress;physical and psychological health;psychological adjustment
Abstract：ObjectiveTo investigate the protective effect of Wendantang-containing serum on astrocytes in glutamate environment and its effect on phosphatidylinositol 3 kinase (PI3K)/protein kinase B (Akt)/glycogen synthetase kinase 3β (GSK3β) signal pathway.MethodTotally 60 rats were randomly divided into 5 groups, normal group (n=20), clozapine group, and high, medium and low-dose Wendantang groups, with 10 rats in each group. Normal group was given 20 mL·kg-1 normal saline, clozapine group was given 20 mg·kg-1 clozapine, Wendantang groups were given 40, 20, 10 g·kg-1 Wendantang, once a day. Eight days later, the rats were killed, their blood was taken, serum was centrifuged, inactivated, filtrated, sterilized and filled in separate centrifugal tubes. The astrocytes were divided into normal group, model group, clozapine group, and high, medium and low-dose Wendantang groups. The normal group and the model group were cultured with normal serum, and the rest groups were cultured with corresponding drug containing serum. The other groups were treated with 10 mmol·L-1 glutamic acid for 24 hours, and then the apoptosis of astrocytes was detected by flow cytometry. Western blot and Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) were used to determine protein, and mRNA expressions of PI3K, Akt, GSK3β in astrocytes.ResultCompared with the normal group, apoptosis in model group increased significantly (P<0.01). Compared with the model group, apoptosis in Wendantang groups decreased significantly (P<0.01). Compared with the normal group, protein and phosphorylation expressions of PI3K, Akt, GSK3β in model groups decreased significantly (P<0.05, P<0.01). Compared with the model group, protein and phosphorylation expressions of PI3K, Akt, GSK3β in Wendantang groups increased (P<0.05, P<0.01). Compared with the normal group, expressions of PI3K, Akt and GSK3β mRNA decreased in model group(P<0.01). Compared with the model group, expressions of PI3K, Akt and GSK3β mRNA increased significantly in Wendantang groups group (P<0.05，P<0.01).ConclusionWendantang-containing serum can effectively increase expressions of PI3K, Akt and GSK3β, so as to regulate PI3K/Akt/GSK3β signal pathway and protect nerve cells.
Abstract：ObjectiveTo study the effect of Huangqi Guizhi Wuwutang on receptor of advanced glycation end products(AGEs)/advanced glycation endproducts (RAGE)/nuclear transcription factor-kappa B p65 (NF-κB p65) signaling pathway in the diabetic peripheral neuropathy rats through an animal modeling experiment, and discuss the mechanism of Huangqi Guizhi Wuwutang in alleviating diabetic peripheral neuropathy.MethodRat model of diabetic peripheral neuropathy was established by high-fat diet and intraperitoneal injection with streptozotocin (STZ). After successful modeling, Huangqi Guizhi Wuwutang intervention began in the fifth week. The patients in high-dose group (19.40 g∙kg-1∙d-1), middle-dose group (4.85 g∙kg-1∙d-1) and low-dose group (2.43 g∙kg-1∙d-1) were given by gavage continuously for 12 weeks. The western medicine control group was given 25 mg∙kg-1∙d-1 by gavage. After the experiment, serum interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA). Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) was used to detect RAGE and NF-κB p65 mRNA expressions in sciatic nerve tissue. The expressions of RAGE, NF-κB and phosphorylation(p)-NF-κB p65 proteins in sciatic nerve tissues were detected by Western blot (WB).ResultCompared with the normal group, serum IL-1β and TNF-α protein levels, RAGE mRNA and NF-κB p65 mRNA levels, RAGE protein, NF-κB p65 protein and p-NF-κB p65 protein levels were significantly increased in the model group (P<0.01), the ratio of p-NF-κB p65 to NF-κB p65 was increased, and the phosphorylation of NF-κB p65 was enhanced (P<0.01). After the intervention of Huangqi Guizhi Wuwutang, compared with the model group, serum IL-1β and TNF-α protein levels, RAGE and NF-κB p65 mRNA levels, RAGE protein, NF-κB p65 protein and p-NF-κB p65 protein levels were all decreased (P<0.01), the ratio of p-NF-κB p65 to NF-κB p65 was decreased in high-dose group (P<0.01). The effect was obvious with the increase of dose of astragalus cassia twig.ConclusionHuangqi Guizhi Wuwutang can alleviate diabetic peripheral neuropathy, and its mechanism may be related to blocking the expression of RAGE on tissue cell surface in AGEs/RAGE/NF-κB signaling pathway, inhibiting the activation of NF-κB and inducing TNF-α triggered oxidative stress and excessive inflammatory response, so as to avoid cell damage and dysfunction.
Abstract：ObjectivesTo investigate the therapeutic effect and mechanism of modified Fuzi Lizhongtang on ulcerative colitis （UC） model rats.MethodThe 72 male SD rats were randomly divided into normal group，model group，sulfasalazine group（0.5 g·kg-1），modified Fuzi Lizhongtang high，medium and low-dose group （23.62，11.81，5.91 g·kg-1）. These rats were used to replicate the UC rat model by 2，4，6-trinitrobenzene sulfonic acid （TNBS）-ethanol composite modeling and treated by gavage for 2 weeks. The general condition of rats in each group was observed. After anesthesia，blood was collected from abdominal aorta and colonic tissue was taken. Semi quantitative evaluation by the colon mucosa damage index （CMDI），the pathological changes of colonic tissue were observed by the hematoxylin and eosin （HE） staining. The contents of serum interleukin-4 （IL-4），IL-6，IL-10 and tumor necrosis factor-α （TNF-α） were detected by enzyme-linked immunosorbent assay （ELISA）. The expressions of mammalian target of rapamycin（mTOR） and phosphorylated ribosomal protein S6 kinase 1 （p-S6K1） in colonic mucosa were detected by immunohistochemistry （IHC） and Western blot.ResultCompared with normal group，the CMDI score of the model group rats was significantly increased （P<0.01）. The contents of IL-4 and IL-10 in serum were significantly decreased，the contents of IL-6 and TNF-α were significantly increased （P<0.01）. The expression levels of mTOR and p-S6K1 in colonic mucosa were up-regulated （P<0.01）. Compared with model group，the CMDI score of the modified Fuzi Lizhongtang high dose group was significantly decreased （P<0.05）. In modified Fuzi Lizhongtang high and medium dose group，the contents of IL-6 and TNF-α were significantly decreased （P<0.01） and the contents of IL-4 and IL-10 in serum were significantly increased （P<0.05，P<0.01）. In the modified Fuzi Lizhongtang high dose group，the expression level of mTOR and p-S6K1 protein was down-regulated significantly （P<0.05，P<0.01）.ConclusionModified Fuzi Lizhongtang high dose group can significantly reduce the congestion and edema，inflammatory cell infiltration，gland distortion，disorder of arrangement and other pathological manifestations of UC colon mucosa，and its mechanism may be related to its down-regulation of mTOR/p-S6K1 signal and the regulation of inflammatory factors secretion.
Keywords：modified Fuzi Lizhongtang;ulcerative colitis;inflammatory factors;mammalian target of rapamycin（mTOR）;phosphorylated ribosomal protein S6 kinase 1（p-S6K1）
Abstract：ObjectiveTo observe the effect of Danggui Buxuetang on lung histopathology and protein kinase D1 (PKD1), nuclear transcription factor-κB (NF-κB) and manganese superoxide dismutase (MnSOD)-mediated oxidative stress pathway in rats with pulmonary fibrosis induced by bleomycin, so as to explore the mechanism of intervention of pulmonary fibrosis.MethodThirty-two male SPF SD rats were randomly divided into sham operation group, model group, Danggui Buxuetang group and prednisone group, with 8 rats in each group. Except the sham operation group, the other groups were prepared through the intratracheal instillation with bleomycin. After modeling for 24 h, the rats of Danggui Buxuetang group were administered with Danggui Buxuetang (0.81 g·kg-1). The rats of prednisone group were given aqueous solution of prednisone (0.005 g·kg-1). The rats of sham operation group and model group were given the same volume of saline. After 14days of administration, blood was collected from the femoral artery, serum was separated, and the lungs were taken by thoracotomy. The pathological changes of rat lung tissues were observed by hematoxylin-eosin staining (HE) and Masson trichrome staining, and graded by Szapiel score and Ashcroft score at the same time. The content of serum malondialdehyde (MDA), and the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) were determined. Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to measure mRNA and protein expressions of PKD1, NF-κB, MnSOD.ResultCompared with the rats in sham operation group, the rats in model group had higher Szapiel scores and Ashcroft scores (P<0.05), higher serum MDA content , but lower SOD, CAT and GSH-Px activities(P<0.01), moreover, the rat lung tissues in model group had higher mRNA and protein expressions of PKD1, NF-κB and MnSOD (P<0.01) than those in sham operation group. Compared with the rats in model group, the Szapiel scores and Ashcroft scores of the rats in Danggui Buxuetang group were decreased significantly（P<0.05）. The serum MDA content was decreased significantly, and SOD, CAT, GSH-Px activities were increased, whereas mRNA and protein expressions of PKD1, NF-κB, MnSOD in the rat lung tissues were decreased（P<0.05，P<0.01）.ConclusionDanggui Buxuetang can reduce the degree of pulmonary fibrosis by regulating the anti-oxidation pathway of PKD1/NF-κB/MnSOD mitochondrial nucleus and improving the body's antioxidant capacity.
Abstract：ObjectiveTo observe the effect of Xiao Jianzhongtang on Adenylate-activated protein kinase/peroxidase proliferation-activated receptor coactivator 1-α （AMPK/PGC1-α） signaling pathway in skeletal muscle of exercise fatigue mice.MethodForty Kunming mice were randomly divided into normal group， model group， Buzhong Yiqitang group and Xiao Jianzhongtang group， with 10 mice in each group. The model group， Buzhong Yiqitang group and Xiao Jianzhongtang group were trained on the treadmill to establish a fatigue model， and the normal group did not apply any intervention. At the same time as the treadmill training， the model group was given the same amount of normal saline. Xiao Jianzhongtang was administered with 5 g·kg-1 of medicine， and Buzhong Yiqitang was administered with 2.8 g·kg-1 of medicine for 6 days. After the experiment， the weight of each group of mice and the time of running out of exhaustion were measured，the colorimetric method was used to detect the serum urea （UREA）， lactate dehydrogenase （LDH）， muscle glycogen （MG）， and skeletal muscle of each group of mice Na+-K+-ATPase， Ca2+-Mg2+-ATPase content， pathological changes of skeletal muscle of each group were observed by hematoxylin-eosin （HE） staining， Western blot was used to detect the protein expression of AMPK and PGC1-α in skeletal muscle of each group .ResultCompared with normal group， the body weight of model group significantly decreased （P<0.01）， and the contents of Na+-K+-ATPase， Ca2+-Mg2+-ATPase， LDH， and MG significantly decreased （P<0.05，P<0.01）. The content of UREA increased significantly （P<0.01）， and the expression of AMPK and PGC1-α protein increased significantly （P<0.01）. Compared with model group， the mice in the Xiao Jianzhongtang group had significantly increased body weight （P<0.05）， significantly increased the time spent on treadmill exhaustion（P<0.01）， Na+-K+-ATPase， Ca2+-Mg2+-ATPase， LDH， and MG. The content increased significantly（P<0.05， P<0.01）， the content of UREA decreased significantly （P<0.01）， and the expression of AMPK and PGC1-α protein increased significantly （P<0.01）.ConclusionXiao Jianzhongtang has an anti-exercise fatigue effect， which may be related to enhancing skeletal muscle AMPK/PGC1-α pathway，enhancing mitochondrial oxidative phosphorylation，reducing accumulation of metabolites，slowing down glycogen consumption and decomposition，and enhancing skeletal muscle energy synthesis.
Abstract：ObjectiveTo investigate the effect of different dose of Realgar compatible with Indigo Naturalis on the transitional constituents of Indigo Naturalis in rat serum based on the compatibility of Qinghuangsan.MethodIndigo Naturalis test solution, the drug-containing serum of three different proportions of Qinghuangsan (10 g of Indigo Naturalis compatible with 52.5, 105, 210 mg of Realgar for group A, B and C, respectively) and blank serum were detected by UPLC-Q-TOF-MS/MS, in combination with the chemical components identified in Indigo Naturalis test solution, the differences of transitional constituents of Indigo Naturalis in rat serum from the group A, B and C were analyzed. HL-60 cells (human leukemia cells) were treated with the three groups of Qinghuangsan drug-containing serum and the effect of drug-containing serum on the activity of HL-60 cells was detected by cell counting kit-8 (CCK-8) assay.ResultA total of 19, 22, 25 of transitional constituents were detected in Qinghuangsan drug-containing serum from group A, B and C, respectively. The three groups of drug-containing serum all contained 5 prototype components from Indigo Naturalis test solution, including tryptanthrin, indigo, indirubin, 2-aminobenzoic acid and N-phenyl-2-naphthylamine, respectively. The results of CCK-8 assay showed that Qinghuangsan drug-containing serum of group C had the strongest inhibitory effect on HL-60 cells.ConclusionAfter fixed Indigo Naturalis dose, with the increase of Realgar dose, the transitional constituents in rat serum increase and the inhibitory effect on HL-60 cells also gradually enhances, which indicates that Realgar may promote the absorption of active components in Indigo Naturali in vivo, thus enhance the efficacy, further explains the compatibility law and pharmacodynamic material basis of different proportions of Realgar and Indigo Naturalis.
Keywords：Indigo Naturalis;Realgar;Qinghuangsan;serum pharmacochemistry;transitional constituents;alkaloids;pharmacodynamic material basis
Abstract：ObjectiveTo preliminarily interpret the compatibility of Ginseng Radix et Rhizoma Rubra and Rhodiolae Crenulate Radix et Rhizoma in chemical and pharmacodynamic levels，and provide theoretical basis for its clinical application.MethodRapid resolution liquid chromatography coupled with quadrupole-time-of-flight tandem mass spectrometry（RRLC-Q-TOF-MS） was applied to identify and analyze the changes in chemical components of the Ginseng Radix et Rhizoma Rubra and Rhodiolae Crenulate Radix et Rhizoma before and after compatibility. The anti-fatigue activity before and after compatibility of Ginseng Radix et Rhizoma Rubra and Rhodiolae Crenulate Radix et Rhizoma was detected by weight-loading swimming experiment and determination of levels of serum urea，blood lactic acid and hepatic glycogen.ResultA total of 51 compounds were identified in mixture decoction of Ginseng Radix et Rhizoma Rubra and Rhodiolae Crenulate Radix et Rhizoma. Malonyl ginsenoside mRg1，mRb1，mRb2，mRb3 and mRd contents were significantly decreased，while ginsenoside Rb1，Rb2，Rb3，Rd，F2 and Rg3 contents were significantly increased in the compatibility mixture. According to pharmacodynamics study，as compared with those in the blank control group，swimming time of mice was significantly prolonged in all other groups （P<0.01），serum urea nitrogen（P<0.05，P<0.01） and lactic acid（P<0.05，P<0.001） levels of mice in the combined decoction and the single decoction groups were significantly lowered，while liver glycogen levels were significantly elevated（P<0.05，P<0.01）. The anti-fatigue ability of the combined decoction of Ginseng Radix et Rhizoma Rubra and Rhodiolae Crenulate Radix et Rhizoma was higher than that of the single decoctions.ConclusionIn this article, the effect enhancing mechanism of compatibility of Ginseng Radix et Rhizoma Rubra with Rhodiolae Crenulate Radix et Rhizoma was revealed based on the chemical changes, providing theoretical reference for the clinical application and development of products.
Keywords：Ginseng Radix et Rhizoma Rubra;Rhodiolae Crenulate Radix et Rhizomachemical;component;compatibility;anti-fatigue
Abstract：ObjectiveTo investigate the effect of celastrol on painful and the emotional of anxiety and depression comorbidity on neuropathic pain model animal and to explore its possible mechanism．MethodMice were randomly divided into sham group， model group， pregabalin group(25 mg·kg-1)， low， medium and high-dose celastrol groups (5，10，20 mg·kg-1). The mice model of neuropathic pain were established by the L5 spinal nerve ligation (SNL). After successful modeling， the treatment groups were given intragastric administration， the sham group and the model group were given the same volume of warm water.Mechanical pain were detected by Von Frey tests， anxiety and depression behaviors were separately detected by the open field and the tail tailing experiments， the pathological changes of microglial cells in hippocampus of mice in each group were observed by immunohistochemical staining (IHC). The inflammation of BV2 microglial cell made by 1 mg·L-1 lipopolysaccharide (LPS). Real-time quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression levels of tumor necrosis factor-α (TNF-α). The expression levels of TNF-α protein were detected by immunofluorescence（IF） staining.ResultCompared with sham group， significant change of mechanical pain thresholds， anxiety and depression were detected in the SNL mice (P<0.05，P<0.01)， the significant decreases of the body size of hippocampal microglia (P<0.05). Compared with SNL model group， 20 mg·kg-1 celastrol significantly increased the 50% paw withdraw threshold and the time of the open feld tests (P<0.05，P<0.01)，and decreased the time of the tail tailing experiments in the SNL mice (P<0.05)， and the cell body area of hippocampal microglia in SNL mice was reduced (P<0.05). Experiment in vitro show， compared with control group， the expression of TNF-α mRNA and protein expression in LPS-induced BV2 microglia increased significantly from 2-4 h (P<0.05，P<0.01). Compared with the LPS group， after 100 nmol·L-1 celastrol administration， LPS-induced microglia inflammatory factor TNF-α mRNA and TNF-α protein expression were significantly decreased (P<0.01).ConclusionCelastrol can relieve pain-emotion comorbidity on neuropathic pain model mice， and its mechanism may be related to the anti-inflammation in the central nerves system.
Abstract：ObjectiveTo observe the effect of Shuangshen Ningxin capsule in alleviating myocardial ischemia/reperfusion injury in rats by regulating mitochondrial adenosine triphosphate（ATP）-sensitive potassium channels.MethodA total of 56 adult male Sprague-Dawley rats were randomly divided into sham-operated control group (sham), model group (model), Shuangshen Ningxin group (SSNX, 90 mg·kg-1).Shuangshen Ningxin and mitochondrial ATP-sensitive potassium channel (MitoKATP) channel inhibitor group 5-hydroxyl-acid group (SSNX+5-HD, 5 mg·kg-1), with 14 rats in each group. Except the sham operation group, the other three groups received occlusion of left anterior descending coronary artery (LAD) for 45 min, and were sacrificed 3 h after reperfusion. Myocardial ischemia and infarct size were observed by TSC Evans blue staining, and myocardial tissue damage degree was observed by hematoxylin-eosin(HE) staining. The kit was used to measure serum lactate dehydrogenase (LDH), creatine kinase (CK) and creatine kinase isoenzyme (CK-MB). The ultrastructural changes of mitochondria and mitochondrial autophagy were observed under transmission electron microscope. The changes of mitochondrial membrane potential in cardiomyocytes were detected by fluorescent probe.ResultCompared with the sham group, myocardial infarct size and myocardial ischemic area percentage in the model group were significantly increased, myocardial tissue arrangement was disordered and loose, individual myocardial fibers were broken, cardiomyocytes were necrotic, and serum CK, CK-MB, LDH activities were significantly increased (P<0.01). Mitochondrial membrane potential was significantly decreased (P<0.01), and mitochondrial structure was destroyed by transmission electron microscopy. Compared with the model group, the myocardial tissue of the SSNX group was arranged orderly, and a small amount of cell edema was mildly degenerated. The percentage of myocardial infarct size and myocardial ischemic area was significantly decreased, serum CK, CK-MB, and LDH activities were significantly decreased (P<0.01), while mitochondrial membrane potential increased (P<0.01). Compared with the model group, the SSNX+5-HD group had mild myocardial tissue disorder and mild degeneration of cell edema in some areas, the percentage of myocardial infarct size and myocardial ischemic area was significantly reduced, serum CK, CK-MB, and LDH activities were significantly decreased (P<0.01), and mitochondrial membrane potential increased (P<0.01). Compared with SSNX group, SSNX+5-HD group had significant increase in serum CK, CK-MB and LDH activities (P<0.01), significant increase in the percentage of myocardial infarct size and myocardial ischemic area, and mitochondrial membrane potential Reduced (P<0.05).ConclusionSSNX protects rat myocardial ischemia-reperfusion injury by opening mitochondrial ATP-sensitive potassium channel.
Abstract：ObjectiveThe SD rat model of hyperplasia of mammary gland（HMG） and the ultrahigh-performance liquid chromatography and mass spectrometry (UHPLC-LTQ-Orbitrap MS) technology were used to explore the pharmacological material basis of Shuangjin Sanjie granules (SJSJG) for the treatment on HMG.MethodSD rat models of HMG were administered in groups, and the nipple height and the diameter were measured; the levels of estradiol (E2), progesterone (P) and prolactin (PRL) in serum were detected, pathological examination was conducted for the hyperplasia of breast tissue. Histochemical methods were used to detect the expressions of estrogen receptor α (ERα), androgen receptor (AR), progesterone receptor (PR), tumor necrosis factor-α (TNF-α) proteins. Finally, UHPLC-LTQ-Orbitrap MS technology was used to detect the main chemical constituents of SJSJG, and the pharmacodynamic substance basis was analyzed based on the pharmacological effect.ResultThe results of animal experiments showed that compared with the normal group, nipple height and diameter of the model group increased remarkably (P<0.01), serum E2 significantly increased (P<0.01). Pathological examination showed abnormal hyperplasia of breast tissue， expressions of ERα, AR, PR and TNF-α increased， compared with the model group, the nipple height and diameter of the SJSJG group decreased remarkably (P<0.01), serum E2 was decreased significantly (P<0.01)， pathological examination showed weakened abnormal hyperplasia of breast tissue, ERα, AR, PR and TNF-α protein expressions were significant decreased (P<0.01). The results of basic material study showed that 85 chemical components were identified from SJSJG, including 16 alkaloids, 7 flavonoids, 15 terpenes, 9 phenolic acid compounds, 3 coumarin compounds, 10 esters and lactone compounds, 7 fatty acids compounds, 4 amino acids compounds, and 14 other types of ingredients， among them, alkaloids and terpenoids chemical drug substances were closely related.ConclusionSJSJG can effectively improve the condition of breast hyperplasia, and its medicinal substance basis may include saikosaponin A, Saikosaponin D, verticinone, peimine.
Keywords：Shuangjin Sanjie granules;hyperplasia of mammary gland;pesticide effect;substance basis
Abstract：ObjectiveTo observe the effect of serum of kidney Yang deficiency rats on the expression of β-catenin，osteoprotegerin(OPG) and nuclear transcription factor-κB receptor activator ligand (RANKL) in the co-culture system and regulatory of icariin on it， and to explore the possible mechanism of inducing osteoporosis.MethodThe 16 male SD rats were randomly divided into blank group and model group， 8 rats in each group. 10 mL·kg-1 adenine was administrated to stomach to establish kidney yang deficiency model. Serum was separated and extracted after the model was established successfully. Isolation and culture of osteoblast（OB） and osteoclast（OC） in vitro， OB was observed and identified by alkaline phosphatase（ALP），alizarin red and Giemsa staining， OC was identified by tartrate resistant acid phosphatase（TRAP） staining， OB-OC co-culture system was established in transwell cell， icariin group（100 μmol·L-1）， blank group， icariin（100 μmol·L-1） + serum group， serum group and Dickkopf1（DKK-1） drug（100 μg·L-1） were set up in group ， 2 days after intervention of co-culture system， OC was counted， ALP and TRAP in supernatant were detected by microplate enzyme labeling， and the expression of OPG，β-catenin and RANKL in each group was detected by Western blot.ResultCompared with blank group， the ALP activity，β-catenin and OPG protein expression in serum group were significant reduction （P<0.05）， while the OC quantity， TRAP activity and RANKL protein expression were marked increase （P<0.05）. Compared with serum group， ALP activity of icariin group decreased significantly （P<0.01）， Compared with icariin group， ALP activity and OPG protein expression decreased （P<0.05）， trap activity and RANKL expression increased （P <0.05） in icariin + serum group.ConclusionThe serum of kidney Yang deficiency rats can induce the occurrence of osteoporosis， and the mechanism of action may be through inhibition of ALP activity， down regulating the expression of β-catenin and OPG protein， increasing the activity of TRAP and the expression of RANKL protein.
Keywords：tratidional Chinese medicine syndrome type;kidney Yang deficiency rat serum;induction;osteoporosis;osteoblasts-osteoclasts（OB-OC） co-culture system
Abstract：ObjectiveTo study the protective effect of Ficus pandurata extract on acute alcoholic liver injury based on pyroptosis mechanism.MethodThe 56 male C57BL/6 mice were randomly divided into normal control group， model control group， positive control group（60 mg·kg-1）， fresh medicine water extract group（48 g·kg-1）， dry drug water extract group（48 g·kg-1），dry drug 50% alcohol extract group（48 g·kg-1） and dry drug 95% alcohol extract group （48 g·kg-1）， 8 mice in each group.Positive control and different solvent extract groups of Ficus tenuifolia were intragastrically administrated for 18 days，once a day，while normal group and model group were given the same volume of pure water intragastrically. After 15 days of continuous gavage， mice received 50％ ethanol（12 mL·kg-1）intragastrically for 3 days to induce acute alcoholic liver injury model except for the normal control group. At 14 h after the last treatment，serum and liver samples were obtained，the serum content of alanine aminotransferase （ALT） and aspartate transaminase（AST） were determined， the histopathologic changes of the hepatic tissues were observed by hematoxylin ecosin（HE） staining.The content of malondialdehyde (MDA) in liver was determined by thiobarbituric acid (TBA) and the content of lactate dehydrogenase (LDH) was determined by microplate method. Western blot and TUNEL assay kit was used to detect the cell pyroptosis rate.ResultCompared with normal group， ALT， AST， MDA and LDH levels in the model group were significantly increased， liver index was significantly increased，TUNEL staining positive， inflammatory factors and pyroptosis related protein expressions were significantly increased （P<0.05）. Compared with model control group， the ALT，AST ，MDA and LDH of the drug intervention group decreased significantly （P<0.05）. The liver index decreased in different degrees， and the expression of inflammatory factors and pyroptosis related protein in the water extract treatment group decreased significantly （P<0.05）.ConclusionThe root extract of Ficus pandurata Hance has protective effect on acute alcoholic liver injury， and the mechanism of water extract might relate to inhibiting hepatocyte pyroptosis.
Abstract：ObjectiveTo observe the clinical efficacy and safety of modified Chaihu Jia Longgu Muli Tang in treating mild to moderate essential hypertension complicated with depression and liver-Yang hyperactivity syndrome.MethodTotally 121 mild to moderate hypertensive patients complicated with depression in line with the inclusive criteria were randomized into treatment group and control group. All of the enrolled patients in treatment group and control group were treated with conventional therapy. In treatment group, patients were given modified Chaihu Jia Longgu Muli Tang, one dose per day. The treatment course lasted for 4 weeks. Blood pressure, patient health questionnaire-9 (PHQ-9) score, score of traditional Chinese medicine syndrome, C-reactive protein (CRP), endothelial-dependent vasodilation, and adverse effect were observed in this study.ResultBoth systolic blood pressure and diastolic blood pressure were significantly lowered when compared to control group (P<0.05). PHQ-9 score was significantly improved in treatment group (P<0.05). The score of traditional Chinese medicine syndrome was significantly improved in treatment group compared to control group (P<0.05). CRP was significantly improved in treatment group compared with control group (P<0.05). Endothelial-dependent vasodilation was significantly improved in treatment group compared with control group (P<0.05). No severe adverse effect was observed in this research.ConclusionChaihu Jia Longgu Muli Tang has a creation clinical efficacy in the treatment of mild to moderate essential hypertension with depression. In addition to the effect in reducing both systolic and diastolic blood pressure, modified Chaihu Jia Longgu Muli Tang was also effective in improving depression, traditional Chinese medicine syndrome and endothelial-dependent vasodilation, and reducing the level of CRP with little adverse effect.
Abstract：ObjectiveTo observe the effect of Zikun decoction (ZKD) in treatment of Yin deficiency and blood dryness in delayed menorrhea due to decreasing ovarian reservation．MethodA total of 60 cases were randomly divided into two groups．The observation group was given ZKD orally，and the control group was treated with complex packing estradiol tablets/estradiol and dydrogestero for three months．The control group began to take medicine on the first day of menstruation for 28 days, while the observation group began to take medicine on the fifth day of menstruation, and stopped taking medicine on the first day of the next menstruation. Three menstrual cycles were followed up. The clinical syndrome scores, follicle-stimulating hormone (FSH), luteinizing hormone (LH), inhibin B (INHB), FSH/LH, Estradiol (E2), anti-müllerian hormone (AMH), and antral follicle count (AFC) scores of the patients before and after treatment were compared.ResultAfter treatment, there was no significant difference in clinical efficiency between observation group and control group, but the traditional Chinese medicine syndrome score of observation group was significantly reduced (P<0.05), observation group and control group had the same curative effect in improvement of FSH, FSH/LH, AMH and AFC (P<0.05), but ZKD was better than complex packing estradiol tablets/estradiol and dydrogestero in improvement of INHB.ConclusionZKD has a good treatment effect in the treatment of DOR. It can effectively improve patients' sex hormone levels, increase antral follicle count, improve the ovarian reserve function and promote menstruation.
Abstract：ObjectiveTo observe clinical effect of Tiaopi Huxin decoction to chronic heart failure (CHF) and to study mechanism of action of anti-inflammatory and anti ventricular remodeling.MethodOne hundred and forty patients were randomly divided into control group (70 cases) and observation group (70 cases) by random number table. And during the study, there were four patients for falling off and missing visit, two eliminated patients in control group, and 64 patients finished the study, and there were six patients for falling off and missing visit in observation group, so 64 patients finished the study. Two groups of patients got losartan potassium tablets, 50 mg/time, 1 time/day, metoprolol tablets, 200 mg/time, 1 time/day, spironolactone tablets, 20-40 mg/time, 1 time/day, control group Jingshe Yiqi granule 1 bag/time,3 times/day,flushed with boiling water, patients in observation group added Tiaopi Huxin decoction, 1 dose/day. The courses of treatment were 12 weeks. Before and after treatment, left ventricular ejection fraction (LVEF), left ventricular end diastolic dimension (LVEDd), leftventricular end-stolic diameter (LVEDs), stroke volume (SV), cardiac output (CO) and ratio of peak value of early diastolic velocity of mitral valve and peak velocity of late diastolic mitral valve (E/A) were recorded by echocardiography. And scores of Lee's heart failure, Qi deficiency and blood stasis, Minnesota Heart Failure Quality of life questionnaire (MLHFQ) and 6 minutes' walking test（6 MWT）were graded. And levels of N-terminal B-type natriuretic peptide (NT-proBNP), galactose lectin-3, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-17, transforming growth factor (TGF) - β and matrix metalloproteinase-9 (MMP-9) were detected.ResultAfter treatment, LVEDd and LVEDs were lower than that in control group (P<0.05). LVEF, SV, CO and E/A were higher than those in control group (P<0.05). And scores of Lee's heart failure, Qi deficiency blood stasis and MLHFQ were lower than those in control group (P<0.01). And 6 MWT was more than that in control group (P<0.01). And levels of NT-proBNP, galactose lectin 3, IL-17, IL-6, TNF-α, TGF-β and MMP-9 were lower than those in control group (P<0.01). Effect of cardiac function was better than that in control group (Z=2.191, P<0.05).ConclusionOn the basis of conventional western medicine treatment and according to theory of heart and spleen, Tiaopi Huxin decoction can improve the clinical symptoms and signs of heart failure, and it had effect of anti-inflammatory, and can improve ventricular remodeling, improve the exercise tolerance and quality of life of patients, improve the prognosis of heart failure, and its clinical effect is better than that of Western medicine alone.
Keywords：chronic heart failure;Qi deficiency and blood stasis syndrome;phlegm Yin syndrome;heart spleen related theory;tiaobihuxin recipe;Shipiyin;Baoyuantang;ventricular remodeling
Abstract：ObjectiveTo discuss the clinical efficacy of modified Zuoguiwan on postmenopausal osteoarthritis of knee (KOA) with deficiency of liver and kidney based on theory of syndrome differentiation and treatment of kidney, and its effect on endocrine hormone and cartilage metabolism.MethodOne hundred and forty patients of KOA were randomly divided into control group (70 cases) and observation group (70 cases) by random number table. Patients in two group got glucosamine hydrochloride capsule for 12 weeks, 1 capsule/time, 2 times/day, and those with obvious pain was added celecoxib capsules for 4 weeks, 0.2 g/time, 1 time/day. The control group took Kang Zengsheng pills orally. patients in observation group was also added with modified Zuoguiwan for 12 weeks, 1 dose/day. Before and after treatment, pain level during activity and rest by visual simulation of pain (VAS) were scored. And visual scale of osteoarthritis index (WOMAC) of Western Ontario and McMaster University, knee osteoarthritis severity index (ISOA), deficiency of liver and kidney, self-rating anxiety scale (SAS) and self-rating depression scale (SDS) were scored. And levels of serum estradiol (E2), follicle stimulating hormone (FSH), luteinizing hormone (LH), transforming growth factor-β (TGF-β), interleukin-1β (IL-1β), matrix metalloproteinase-3 (MMP-3) and tumor necrosis factor-α (TNF-α) were scored.ResultScores of VAS during activity and rest in observation group were lower than those in control group (P<0.01). The total score of WOMAC, scores of pain, stiffness, joint function, deficiency of liver and kidney, SAS and SDS were all lower than those in control group (P<0.01). Total score of ISOA, symptoms and signs, maximum walking distance, daily life were lower than those in control group (P<0.01). Levels of E2, FSH, LH, TNF-α, IL-1β and MMP-3 were lower than those in control group (P<0.01). Level of TGF-β was higher than that in control group (P<0.01). And total clinical effective rate in observation group was 89.23%(58/65), which was higher than 74.60%(47/63) in control group (χ2=4.793, P<0.05).ConclusionModified Zuoguiwan can obviously alleviate symptoms, improve joint function, regulate the level of endocrine hormone, relieve anxiety and depression, regulate the environment of cartilage metabolism, inhibit inflammatory reaction, and improve the patients' ability of daily life and clinical efficacy.
Keywords：knee osteoarthritis;menopause;deficiency of liver and kidney;tonifying kidney method;Zuoguiwan;endocrine hormone;cartilage metabolism
Abstract：ObjectiveTo establish a method for qualitative analysis of components in Perilla frutescens leaves and stalks by liquid chromatography-mass spectrometry (LC-MS)，so as to explore the substance basis of pharmacodynamics differences between P.frutescens leaves and stalks.MethodP. frutescens leaves and stalks were extracted by 80% methanol-water ultrasound. The samples were analyzed by UPLC-Q-Exactive-Orbitrap-MS comprehensively. Halo-C18 column (2.1 mm×100 mm，2.7 μm) was used for gradient elution with 0.05% formic acid aqueous-0.05% acetonitrile formate as mobile phase in positive and negative ion modes. The flow rate was 0.3 mL·min-1，the column temperature was 40 ℃，and the injection volume was 5 μL.ResultThe chemical compound in P. frutescens was deduced and identified based on the retention time of chromatography，and the exact molecular weight，excimer ion peaks，fragment ions and reference materials in Xcalibur software. The chemical composition of P. frutescens was identified by Mass Frontier 7.0 software. Totally 4 amino acids，7 phenylpropanoids，10 flavonoids，12 triterpenoids，7 organic acids，4 fatty acids，10 unknown compounds and 54 compounds were identified. Among them，6 triterpene acids, including glochidone, were identified in P. frutescens for the first time. The structures of five characteristic compounds were analyzed. There were 45 constituents in P.frutescens leaves and 32 constituents in P. frutescens stalks. They had 23 common constituents.ConclusionLC-MS can identify the components of P. frutescens rapidly and effectively. This study provides an important theoretical basis for the quality control of different parts of P. frutescens and the development and utilization of P. frutescens.
Abstract：ObjectiveTo develop a simple and accurate method for molecular authentication of Panax ginseng and P. quinquefolius.MethodThe mitochondrial cox Ⅱ sequences of P. ginseng and P. quinquefolius were amplified by polymerase chain reaction（PCR）with universal primers. PCR products of the two species were sequenced in both directions, and sequence alignments were conducted for intron length polymorphisms exploitation. Multiplex PCR was established for the identification of P. ginseng and P. quinquefolius with their specific primers，which were designed respectively based on their insertion sequences. And the limit of detection of the multiplex PCR was also determined.ResultThe insertion/deletion sequences were exploited in mitochondrial cox Ⅱ. Under the established multiplex PCR assay，P. ginseng generated a 729 bp specific band, while P. quinquefolius yielded a 141 bp specific amplicon，and the mixture of the two species yielded both 729 bp and 141 bp fragments. The established multiplex PCR assay could detect 0.1% of intentional adulteration of P. quinquefolius into P. ginseng, with down to 0.001 ng of genomic DNA.ConclusionThe established multiplex PCR assay can accurately identify P. ginseng and P. quinquefolius from different sources, without the optimization of reaction system and the introduction of additional mismatches，so as to provide a new molecular marker method for identifying botanical origin of P. ginseng and P. quinquefolius.
Abstract：ObjectiveAphanamixis grandifolia，a perennial herb of genus Aphanamixis （Meliaceae），has effects in soothing activating collaterals，dredging paisy，expelling wind-evil and removing wetness. This paper aimed to investigate terpenoids from the stems and leaves of A. grandifolia and reveal the effective substances.MethodTotally 22 kg leaves and twigs of A. grandifolia were extracted with 90% EtOH for three times by heating reflux. These extracts were decompressed and concentrated，and then dissolved in water. The solvent was successively extracted with petroleum ether and ethyl acetate. The chemical constituents from petroleum ether and ethyl acetate fractions were isolated by macroporous，silica gel，Sephadex LH-20 and ODS columns，and their chemical structures were determined through MS，NMR analysis（1H and 13C-NMR）and spectroscopic data from literatures，respectively.ResultTwelve compounds were obtained and identified as 16α-hydroxy-3-oxo-24-methyllanosta-7，9（11），24（31）-triene-21-oic acid （1），23（E）-cycloart-en-25-ethoxy-3-ol（2），23（Z）-9，19-cycloart-ene-3β，25-diol（3），23（E）-cycloart-en-3β，25-diol（4），labda-8，13-（E）-dien-15-ol（5），labda-7，13-（E）-dien-15-ol（6），vulgarol（7），（S，E）-6-[6-（5，5-dimethyl-4-oxo-4，5-dihydrofuran-3-yl]-2-methylhepta-1，6-dien-1-yl]-4-methyl-5，6-dihydro-2H-pyran-one（8），nemoralisin（9），nemoralisin C（10），1S，4R，5S，6R，7S，10S-1（5），6（7）-diepoxy-4-guaiol（11） and 1S，4S，5S，10R-4，10-guaianediol（12），respectively.ConclusionThe structures involves triterpenoids，diterpenoids and sesquiterpennoids，and eight of them （1-3，5-8 and 12） are obtained from A. grandifolia for the first time. Those compounds are also isolated from the geneus Aphanamixis for the first time.
Abstract：ObjectiveTo reveal the dynamic changes of flavonoids secondary metabolites and relevant genes expressions in the process of germination of tartary buckwheat seeds by investigating the content of catechins，epicatechins，rutin，and quercetin，and the expressions of their relevant genes in tartary buckwheat sprouts and seedlings，in order to provide scientific basis for the selection of high-quality, high-nutrition tartary buckwheat sprouts.MethodContents of catechin，epicatechin，rutin，and quercetin in tartary buckwheat sprouts and seedlings were detected by UPLC-ESI-QQQ-MS，and the expression levels of genes relating to flavonoids synthesis in tartary buckwheat sprouts and seedlings were detected by real-time quantitative PCR.ResultThere were differences between tartary buckwheat sprouts and seedlings in the relative contents of catechin，epicatechin，rutin and quercetin，as well as the expressions of relevant genes in the synthesis pathway, including FtPAL，FtC4H，Ft4CL，FtCHS，FtCHI，FtF3H，FtF3'H，FtFLS，FtDFR，FtLAR，FtANS，FtANR. The contents of flavonoids and the expressions of relevant genes in tartary buckwheat sprouts were higher than those in tartary buckwheat seedlings.ConclusionThe higher accumulation of secondary metabolites and flavonoids in tartary buckwheat sprouts may be related to tartary buckwheat seeds' resistance to the external environment in the initial growth stage of germination. From the perspective of application，there are more flavonoids in tartary buckwheat sprouts than in tartary buckwheat seedlings, indicating that tartary buckwheat sprouts have a higher nutritional value.
Abstract：ObjectiveBased on UPLC-Q-Orbitrap HRMS and network pharmacology, the material basis, processing principle and molecular mechanism of bile processed Coptidis Rhizoma (BPRC) for reducing excess fire of liver and gallbladder were elucidated.MethodThe chemical ingredients of BPRC were analyzed by UPLC-Q-Orbitrap HRMS. Chromatographic separation was achieved with 0.1% formic acid solution (A)-acetonitrile (B) as the mobile phase in gradient elution (0-20 min, 5%-80%B; 20-30 min, 80%-95%B; 30-30.1 min, 95%-5%B; 30.1-35 min, 95%-5%B). The flow rate was 0.2 mL·min-1, electrospray ionization (ESI) was applied and operated in positive and negative ion modes, the acquisition range was m/z 100-1 500. Based on the clinical manifestations and pathogenic factors of excess fire of liver and gallbladder, the potential effective ingredients, targets and functional characteristics of BPRC were predicted and analyzed by online database. Based on the characteristics of the new active ingredients after processing, the processing principle of BPRC was investigated by network pharmacology.ResultA total of 19 ingredients in BPRC were identified, six of which were newly added cholic acids after processing. It was determined that the alkaloids, including worenine, epiberberine, jatrorrhizine, coptisine, berberrubine, berberine, palmatine and cholic acids, including glycohyodeoxycholic acid, taurohyodeoxycholic acid, glycochenodeoxycholic acid, hyodeoxycholic acid and taurochenodeoxycholic acid, were identified as material basis of BPRC. A total of 66 targets of reducing excess fire of liver and gallbladder of BPRC were screened. There were 16 common targets and multiple same signaling pathways between cholic acids and alkaloids of BPRC, and many lesions of excess fire of liver and gallbladder were target organs of cholic acids. By acting on some targets, including albumin (ALB), Caspase-3 (CASP3), mitogen-activated protein kinase 14 (MAPK14), glucocorticoid receptor (NR3C1) and other targets and some signaling pathways, including interleukin (IL)-17, phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt), MAPK and other pathways, BPRC could reduce excess fire of liver and gallbladder.ConclusionBPRC has the characteristics of multi-component, multi-target and multi-pathway on reducing excess fire of liver and gallbladder. Bile and Coptidis Rhizoma have synergistic effect and bile can enhance the intensity of BPRC in lesions, which confirms the processing theory that the effect of BPRC on excess fire of liver and gallbladder enhance after being processed by bile.
Keywords：bile processed Coptidis Rhizoma;bile;alkaloids;cholic acids;excess fire of liver and gallbladder;network pharmacology;ultra-high performance liquid chromatography-quadrupole/electrostatic field orbitrap high-resolution mass spectrometry (UPLC-Q-Orbitrap HRMS)
Abstract：ObjectiveTo investigate the mechanism of Nardostachyos Radix et Rhizoma and Corydalis Rhizoma in treatment of atrial fibrillation by predicting targets and signaling pathways based on network pharmacology.MethodThe traditional Chinese medicine system platform (TCMSP) database was used to screen out active components of Nardostachyos Radix et Rhizoma and Corydalis Rhizoma,predict targets,and construct the active component-predicted target network.Through the Online Mendelian Inheritance in Man (OMIM),Therapeutic Target Database (TTD),and Genecards databases,potential target information of atrial fibrillation was retrieved.STRING 11.0 database was used to obtain the protein-protein interaction data of relevant targets,and the results were visualized by Cytoscape 3.7.1 software to construct protein-protein interaction network relating to atrial fibrillation.The predicted targets of Nardostachyos Radix et Rhizoma and Corydalis Rhizoma were mapped to the potential targets of atrial fibrillation.The intersection targets were the potential targets for the treatment of atrial fibrillation with Nardostachyos Radix et Rhizoma and Corydalis Rhizoma.Then,Visualization and Integrated Discovery (DAVID),a database for annotation,was used to analyze biological functions and pathways of the potential targets of Nardostachyos Radix et Rhizoma and Corydalis Rhizoma in the treatment of atrial fibrillation.Finally,Cytoscape3.7.1 software was utilized to construct active component-potential target-signal pathway network of Nardostachyos Radix et Rhizoma and Corydalis Rhizoma in treatment of atrial fibrillation.ResultTotally 51 active components of Nardostachyos Radix et Rhizoma and Corydalis Rhizoma were screened out,and 18 potential targets for the treatment of atrial fibrillation with Nardostachyos Radix et Rhizoma and Corydalis Rhizoma were predicted.The effect was mainly correlated with the regulation of interleukin-6 (IL-6),sodium channel protein type 5 subunit alpha (SCN5A),tumor necrosis factor (TNF),nitric-oxide synthase,endothelial (NOS3),potassium voltage-gated channel subfamily hmember 2 (KCNH2),collagen alpha-1(I) chain (COL1A1),retinoic acid receptor RXR-alpha (RXRA),tissue factor (F3),alpha-1B adrenergic receptor (ADRA1B) and other target proteins,cyclic guanosine monophosphate (cGMP)/cGMP-dependent protein kinase G (PKG) signaling pathway,phosphatidylinositol-3-kinases (PI3K)/protein kinase B (Akt) signaling pathway,transcriptional disorders in cancer,calcium signaling pathways,and adrenergic signals in cardiomyocytes.ConclusionNardostachyos Radix et Rhizoma and Corydalis Rhizoma treat atrial fibrillation based on multiple components,multiple targets and multiple channels,and provide a scientific basis for subsequent experimental studies for further explainning its mechanism of action.
Keywords：Nardostachyos Radix et Rhizoma;Corydalis Rhizoma;atrial fibrillation;network pharmacology;mechanism
Abstract：ObjectiveTo investigate the medication rules of traditional Chinese medicine (TCM) for the treatment of diabetic peripheral neuropathy (DPN).MethodThe literature published in China Knowledge Resource Integrated Database (CNKI), Wanfang Database, China Biomedical Literature Service System (SinoMed), VIP Database and PubMeb from 2008 to 2019 were retrieved by setting the topics of diabetic peripheral neuropathy and TCM. After screening, a database was established to analyze the medication rules (efficacy, frequency, flavor and meridian tropism, common couplet medicinals and core medicines) of TCM by frequency statistics, association rules and data statistical methods of constructing complex networks.ResultA total of 461 papers for treatment of DPN were included in this study, including 275 kinds of TCM and a total frequency of 6 361 times. Astragali Radix had the highest frequency. Among all kinds of medicinal materials, activating blood circulation and removing stasis was the most commonly used medicine, followed by Qi-invigorating medicine. Flavor of medicines was mainly sugariness and warm, and most of their meridian tropism was liver meridian. After the analysis by association rules, the couplet medicinals with the highest support was Astragali Radix-Angelicae Sinensis Radix. The core medicines obtained by complex network analysis were Astragali Radix, Angelicae Sinensis Radix, Chuanxiong Rhizoma, Spatholobi Caulis, Cinnamomi Ramulus, Carthami Flos, Pheretima, Paeoniae Radix Rubra, Salviae Miltiorrhizae Radix et Rhizoma and Persicae Semen.ConclusionThis study comprehensively analyzes the medication rules of TCM clinical treatment of DPN. The main treatment methods of TCM for DPN are invigorating Qi and blood, activating blood circulation and removing stasis, activating meridians to stop pain, which can provide guidance for the TCM clinical use and new Chinese medicines research and development of DPN.
Abstract：This paper, taking the formulation of national drug standards for traditional Chinese medicine (TCM) dispensing granules as a case study, explores the improvement of the formation mechanism of national drug standards, and promotes the reform of streamline administration, delegate powers, and improve regulation and services of national standards management, so as to release the vitality of the research and development of standards of drug manufacturers. After nearly two decades of pilot production of TCM dispensing granules, a large number of researches and discussions have been conducted on the formulation of unified standards of TCM dispensing granules from manufacturing enterprises to national standard administration departments, it was found that this work was difficult on the basis of the original drug standard formation mechanism. The authors tried to improve and innovate the formation mechanism of national drug standards, to provide methods and ideas for the formulation and unification of national standards for TCM dispensing granules, and to provide references for the formulation of other national drug standards.
Keywords：streamline administration, delegate powers, and improve regulation and services;traditional Chinese medicine dispensing granules(TCMDG);national drug standards;formation mechanism;pilot production;standard decoction;evaluation indexes
Abstract：The suitable production area can guarantee the quality of Dai medicine， but the research on the suitable ecological factors and suitable production area of plant Dai medicine is relatively scarce at present. In this study， gmpgis was used to analyze the ecological suitability of Dai medicine in China. For the first time， the range of ecological factors and main soil types in the main distribution areas of Dai medicine were extracted， such as annual average temperature of 8.6-23.7 ℃， annual average precipitation of 1212-1881 mm， annual average sunshine of 149.7-157.4 W·m-2， the main soil types are strong eluvial soil， alluvial soil， high activity strong acid soil， etc. Based on 179 sampling points， through the analysis of ecological similarity， the largest ecological similarity area of plant Dai medicine in the world was obtained， mainly including China， Brazil， the United States， Myanmar， Laos and other countries. In China， it is mainly concentrated in the West and south of Yunnan， including Xishuangbanna Dai Autonomous Prefecture， Dehong Dai Jingpo Autonomous Prefecture， Pu'er City， Baoshan City and Lincang City， which are suitable for cultivation. In addition， by analyzing the current situation of Dai medicine industry， this paper summarizes the problems such as the lack of talents， the decreasing of Dai medicine resources， the limitation of cultural heritage， and the weakness of scientific research， and puts forward strategies such as training high-quality talents of Dai medicine， the investigation and protection of Dai medicine resources， promoting the collection and arrangement of Dai medicine data and ancient books， and strengthening the basic and applied research of Dai medicine Enter the development of Dai medicine. This study provides a basis for guiding the rational layout， introduction and breeding of plant Dai medicine production base， and lays a foundation for the scientific and standardized production of high-quality Dai medicine.
Abstract：Holistic medicine is an interdisciplinary field that integrates all types of biological information (proteins, small molecules, tissues, organs, external environmental signals, etc.) to provide predictable and operational models for healthcare and disease treatment. Despite the global and integrative character of this discipline, scientific evidences to clarify the mechanism of holistic medicine is still lacking. Systems pharmacology, as a newly developed discipline based on the interdisciplinary and integration of classical pharmacology, computer technology, bioinformatics, and network pharmacology, has systematically studied the interaction between drugs and the human body, as well as its law and nature at the molecular, cellular, organ and network levels. Since its birth in 2011, this discipline has been widely used in the research of the promotion of holistic medicine, compound analysis, the development of new drugs, and interpretation of basic theories of traditional Chinese medicine (TCM). In this paper, in order to provide new methods and ideas for the modernization of holistic medicine, the authors introduce the research ideas of systems pharmacology of TCM in detail, and the mechanism of holistic medicine in the treatment of complex diseases is explained by sorting out the researches of systems pharmacology on multi-component, multi-target, multi-pathway, multi-function, compatibility of TCM, etc.
Keywords：systems pharmacology;holistic medicine;traditional Chinese medicine;network pharmacology;mechanism of action;compatibility;formulas
Abstract：Chuanxiong Chatiaosan was first recorded in Taiping Huimin Heji Jufang, which was made up of 8 herbs, including Chuanxiong Rhizoma, Menthae Haplocalycis Herba, Asari Radix et Rhizoma, Schizonepetae Herba, Saposhnikoviae Radix, Angelicae Dahuricae Radix, Notopterygii Rhizoma et Radix and Glycyrrhizae Radix et Rhizoma. This prescription mainly contains a variety of alkaloids, flavonoids, phenylpropanoids, volatile oils and other compounds, which play the biological activity of promoting blood circulation and relieving pain. Modern pharmacological studies have confirmed that Chuanxiong Chatiaosan can reduce blood viscosity, improve cerebral circulation, and has central analgesic effect to treat migraine effectively. However, the mechanism for treating migraine of this prescription is still unclear. The author elaborated the research status of Chuanxiong Chatiaosan from four aspects, including quality control method, chemical composition, preparation technology and pharmacological research, hoping to provide references for rational clinical application and explanation of pharmacological mechanism of this prescription.