Abstract：ObjectiveTo study the effect and mechanism of Zuoguiwan on the autophagy of cells in the skin during the development of congenital kidney-deficient mice， and to explore the application value of kidney tonifying and essence filling method in abnormal growth and development.Method36 rats with a 2∶1 ratio of male to female were paired， and the pregnant rats were stimulated by combined stress method to build a model of congenital kidney-deficient rats. According to different treatment methods， the pregnant rats were divided into the control group， the kidney-deficient group， and the Zuoguiwan low and high dose groups （2， 8 g·kg-1）. The control group was given normal saline without modeling. Kidney deficiency group was modeled before intragastric administration of normal saline. Zuoguiwan small dose and high dose groups received intragastric administration of Zuoguiwan suspension. 21 days after the birth of mice， the back skin was taken to observe the skin microstructure and morphology of the mice by HE staining and detect the thickness of dermis and epidermis. The gene and protein expression levels of Wingless3a and β-catenin in the skin of neonatal rats were measured by Western blot and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. Double immunofluorescence assay was used to detect the skin autophagy indicators Microtubule-associated protein light chain 3 （LC3）and Sequestosome 1 （SQSTM1-p62）.ResultIn the control group， the skin layers were clear and the structure was normal， in the kidney deficiency group， the collagen arrangement in the dermis was loose， the skin composition in Zuoguiwan low and high dose groups was basically normal. Compared with the control group， the epidermal layer of the kidney deficiency group was thickened and the dermis was thinned （P<0.05）. Compared with the kidney deficiency group， the thickness changes of epidermal layer and dermis layer were significantly restored in Zuoguiwan low and high dose groups （P<0.05）. Compared with the control group， the expression of Wnt3a and β-catenin protein and gene in the kidney deficiency group significantly decreased （P<0.05）. Compared with the kidney deficiency group， Wnt3a and β-catenin protein and gene expression increased in Zuoguiwan low and high dose groups （P<0.05）. At the same time， immunofluorescence assay showed that compared with the control group， the expression of LC3 was up-regulated and the expression of SQSTM1-p62 was down-regulated in the kidney deficiency group， while Zuoguiwan could reverse such abnormal expression （P<0.05）.ConclusionZuoguiwan can down-regulate the autophagy level of skin cells and improve the abnormal skin development of congenital renal deficiency by increasing the expression of Wnt3a and β-catenin protein and gene in the skin of neonatal rats.
Keywords：autophagy;Zuoguiwan;skin;development;kidney deficient;offspring rat
Abstract：ObjectiveTo establish ultra performance liquid chromatography （UPLC） fingerprint of Shengyutang and quantitative analysis method of 11 index components in this famous classical formula.MethodUPLC-diode array detector/evaporative light scattering detector （UPLC-PDA/ELSD） was used， two chromatographic conditions were established by different detectors according to the polarity of chemical components. Conditions of fingerprint 1 were as follows：ACQUITY UPLC HSS T3 column （2.1 mm×100 mm， 1.8 µm） with the mobile phase of acetonitrile （A）-0.6% formic acid solution （C） for gradient elution （0-4 min， 0-4%A； 4-8 min， 4%A； 8-9 min， 4%-8%A； 9-14 min， 8%-9%A； 14-21 min， 9%-15%A； 21-26 min， 15%-17%A； 26-30 min， 17%-20%A； 30-35 min， 20%-32%A； 35-40 min， 32%-40%A； 40-50 min， 40%-80%A； 50-55 min， 80%A）， the flow rate of 0.3 mL·min-1， PDA with detection wavelengths of 280 nm and 321 nm， the column temperature at 30 ℃. Conditions of fingerprint 2 were as follows：the CORTECS C18 column （3.0 mm×100 mm， 2.7 µm） with the mobile phase of acetonitrile （A）-water （D） for gradient elution （0-11 min， 19%A； 11-16 min， 19%-25%A； 16-34 min， 25%-28%A； 34-47 min， 28%-47%A； 47-60 min， 47%-80%A）， the flow rate of 0.4 mL·min-1， ELSD with drift tube temperature of 95 ℃， the carrier gas （air） flow rate of 2.0 L·min-1， and the column temperature at 30 ℃. UPLC-PDA/ELSD fingerprints of 15 batches of Shengyutang were established， and the similarity was evaluated by similarity evaluation system of chromatographic fingerprint of traditional Chinese medicine （2012 edition） issued by the Chinese Pharmacopoeia Commission， and the contents of eleven index components in this famous classical formula were determined.ResultThe similarities of UPLC-PDA/ELSD fingerprints of 15 batches of Shengyutang were >0.98 by comparing with the control fingerprint， 27 and 16 common peaks were identified in fingerprint 1， 2， respectively. It was tested and verified that the precision， repeatability， stability， linear relationship and other results of this method all met the requirements of the 2020 edition of Chinese Pharmacopoeia. The contents of chlorogenic acid， ferulic acid， calycosin glucoside， verbascoside， senkyunolide I， senkyunolide H， senkyunolide A， ginsenoside Rg1， ginsenoside Re， ginsenoside Rb1 and astragaloside A in 15 batches of Shengyutang were 0.063-0.193， 0.509-0.638， 0.160-0.318， 0.012-0.056， 0.394-0.519， 0.110-0.143， 0.031-0.097， 0.382-0.595， 0.292-0.505， 0.590-0.803， 0.142-0.367 mg·g-1， respectively.ConclusionThe established detection method meets the requirements of the 2020 edition of Chinese Pharmacopoeia， which can characterize the overall characteristics of chemical components in Shengyutang， and provide experimental basis for the quality standard research of this famous classical formula.
Abstract：ObjectiveTo explore the possible mechanism of Huangqintang in treating ulcerative colitis （UC）.MethodThe animal model of UC was induced by dextran sodium sulfate （DSS）．The experimental animals were divided into control group， model group，Huangqintang low dose （4.55 g·kg-1）， medium dose （9.1 g·kg-1）， and high dose（18.2 g·kg-1） groups. Intragastric administration was also given in the modeling process for 7 consecutive days. At the end of the 8th day， colon tissues were collected to measure colon length and mass， and calculate the colon mass index. Pathological changes were observed by hematoxylin-eosin （HE） staining. Serum iron content， superoxide dismutase （SOD）， glutathione （GSH）， catalase （CAT） and myeloperoxidase （MPO） were determined by biochemical assay. Western blot was used to detect the protein expression of glutathione peroxidase 4 （GSH-Px4）， long-chain acyl-CoA synthetase 4 （ACSL4） and ferritin heavy chain 1（FTH1）. The mRNA expression levels of tumor trotein 53 （P53） and solute carrier family 7 member 11 （SLC7A11） in colon tissues were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）.ResultThe experimental studies showed that compared with normal group， serum MPO and iron content， ACSL4 protein level and relative P53 mRNA expression in the model group significantly increased （P<0.05）， while serum SOD， CAT， GSH content， GSH-Px4， FTH1 relative protein expression level and relative SLC7A11 mRNA expression in the model group significantly decreased （P<0.01）. Compared with model group， serum MPO and iron content， ACSL4 protein level and relative P53 mRNA expression significantly decreased （P<0.05）， while serum SOD， CAT， GSH content， GSH-Px4， FTH1 relative protein expression level and relative SLC7A11 mRNA expression significantly increased （P<0.05） after the intervention of Huangqintang， and the effect was most significant in the high-dose group （P<0.05）. The results of general condition， colon length， colon mass index and HE staining showed that Huangqintang could relieve clinical symptoms and histopathological changes in UC mice.ConclusionThese results indicated that Huangqintang had therapeutic effect on ulcerative colitis mice， and its mechanism might be related to inhibiting the oxidative stress and ferroptosis.
Abstract：ObjectiveTo investigate whether there is inflammatory reaction and cell pyroptosis in impaired glucose tolerance （IGT） rats induced by high-fat diet and the intervention effect of Huanglian Wendantang.MethodHealthy male SD rats were fed with 45% fat content diet for 20 weeks to replicate the IGT model. The rats in line with the model establish criteria were randomly divided into 3 groups， with 10 rats in each group， another 10 rats were selected as the blank control group. Huanglian Wendantang group was given 7.8 g·kg-1·d-1 compound decoction of Huanglian Wendantang， and the positive control group was given 0.05 g·kg-1·d-1 aqueous solution of metformin hydrochloride， with the dose volume of 10 mL·kg-1·d-1. The blank group and the model group were given the same volume of distilled water. After continuous intragastric administration for 4 weeks， the body weight， body length and abdominal circumferences were measured， the Lee's obesity index was calculated， and the levels of fasting plasma glucose （FPG） and 2-hour plasma glucose （2 h PG） were detected in each group. Serum interleukin-6 （IL-6） content was detected by enzyme-linked immunosorbent assay （ELISA）. The mRNA and protein expressions of nuclear factor kappa B （NF-κB）， cysteinyl aspartate specific proteinase-1 （Caspase-1） and NOD-like receptor protein 3 （NLRP3） in liver tissues were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. Immunofluorescence was used to detect the expression of Caspase-1 and NLRP3 in rat liver. Hematoxylin and eosin （HE） staining was used to observe the morphology of liver tissue.ResultCompared with blank group， the body weight， abdominal circumference， body length and Lee's index in model group were significantly increased （P<0.05， P<0.01）， the levels of FPG and 2 h PG were significantly increased （P<0.05， P<0.01）， serum IL-6 content and NF-κB， Caspase-1 and NLRP3 gene and protein expressions in liver tissue were significantly increased （P<0.01）， immunofluorescence technique showed that Caspase-1 and NLRP3 expressions increased obviously in IGT rat model’s liver tissues， and inflammatory cells infiltration was observed obviously in HE stained rat liver tissue model. Compared with model group， Huanglian Wendantang and metformin hydrochloride groups could effectively reduce the body weight of IGT rats （P<0.01） and abdominal obesity， and correct the levels of FPG and 2 h PG （P<0.01）， while effectively reducing serum IL-6 content and gene and protein expressions of NF-κB， Caspase-1 and NLRP3 in liver tissues of IGT rats， and alleviating inflammatory cell infiltration.ConclusionInflammatory response exists in IGT model rats induced by high-fat diet. Huanglian Wendantang can obviously reduce its inflammatory response and alleviate liver cell pyroptosis.
Abstract：Wuzhuyutang is widely used in clinical practice nowadays due to its remarkable effect. It is the fifth prescription included in Ancient Classic Prescriptions Catalogue （the first batch）. Based on relevant medical literatures from the Eastern Han dynasty to the period of the Republic of China，combined with the methods of phiology and metrology，this paper systematically studies the origin，composition，evolution of prescription meaning，main indications，dosage of prescriptions，processing of various herbs，preparation method and administration method. According to the findings， Wuzhuyutang was created by ZHANG Zhongjing's Treatise on Febrile Diseases in the eastern Han dynasty. It was composed of Euodiae Fructus，Zingiberis Rhizoma Recens，Ginseng Radix et Rhizoma and Jujubae Fructus. In history，it had such synonyms as Zhuyutang，Yicujicuxinfang，Zhuyu Renshentang，Wuyutang，Sanwei Shenyutang，Shenyutang，Sishentang. It can be applied in internal medicine， gynecology，pediatrics，department of facial features and other miscellaneous diseases， which belong to Jueyin liver meridian and is manifested by upwardly reversed turbid Yin and invading other meridians， with cold vomiting （retching or salivation），abdominal pain，headache，dizziness，hand and foot cold as the main clinical symptoms. In terms of dosage，the commonly used dosage of Euodiae Fructus is 3.7-18.6 g in the previous dynasties. In clinical practice，if the main clinical manifestations are vomiting and salivation clearing，the dosage of about 3 g is recommended. Severe headache and abdominal pain can be mainly used，the dosage of about 10 g is recommended. The research and development of the formula can comply with the maximum dose of 5 g as stipulated in the 2015 edition of the Chinese Pharmacopoeia. The dosages of Ginseng Radix et Rhizoma and Zingiberis Rhizoma Recens can be calculated based on 1 liang equivalent to 3 g，and 1-5 pieces of Jujubae Fructus can be added. Zingiberis Rhizoma Recens and Ginseng Radix et Rhizoma should be processed according to the method of the Chinese Pharmacopoeia，reed should be removed，Zingiberis Rhizoma Recens should be washed，then peeled and sliced， and Jujubae Fructus should be dried in the sun，then cut and seeded before being used as medicine.
Abstract：ObjectiveTo study the effect of Aidi injection （AD） on the expression of cytochrome P450 isoenzyme 1A2，2E1，3A2，2C11（CYP1A2，2E1，3A2，2C11）mRNA and protein in rats with N-nitrosodiethylamine （DEN） chemically induced primary hepatocellular carcinoma（HCC）.MethodThree healthy SD male rats were randomly selected as the blank group， and the remaining rats were treated with DEN intermittently induced primary hepatocellular carcinoma rat model. After success of the model， the rats were randomly divided into model group and AD group， with 3 rats in each group. The rats in the blank group and model group were intraperitoneally injected with 10 mL·kg-1 saline， while those in the AD group were intraperitoneally injected with 10 mL·kg-1 AD once a day， a total of 14 d intervention. Real-time quantitative polymerase chain reaction（Real-time PCR） and Western blot were used to detect the mRNA and protein expressions of CYP1A2， CYP2E1， CYP3A2 and CYP2C11， respectively.ResultReal-time PCR results showed that after 14 days of drug administration， compared with the blank group， the mRNA expressions of CYP1A2， CYP2E1， CYP3A2 and CYP2C11 were all down-regulated in para-cancerous tissue （PCT） and cancerous tissue （CT） in model group， and there were significant differences （P<0.05，P<0.01）. Compared with the model group， the mRNA expressions of the four subtype enzyme were significantly down-regulated in PCT in the AD group（P<0.05，P<0.01）， while the mRNA expressions of the four subtype enzyme were significantly up-regulated in CT （P<0.05）， and the expression was down-regulated overall. Western blot results showed that compared with the blank group， the protein expressions of CYP1A2， CYP2E1， CYP3A2 and CYP2C11 in CT of the model group were significantly down-regulated （P<0.01）， and the protein expressions of CYP3A2 and CYP2C11 were significantly down-regulated in PCT （P<0.01）. Compared with the model group， the protein expressions of CYP1A2， CYP2E1， CYP3A2 and CYP2C11 in CT and PCT were down-regulated in the AD group， but the differences were not statistically significant.ConclusionAD can down-regulate the mRNA and protein expressions of CYP1A2， CYP2E1， CYP3A2 and CYP2C11 in rat liver tissues. In clinical use of AD， attention should be paid to drug interactions that may be caused by CYP450 enzyme inhibition.
Abstract：ObjectiveTo investigate the effect of Ginseng Radix et Rhizoma-Notoginseng Radix et Rhizoma-Chuanxiong Rhizoma extract on endothelial microparticles （EMPs）-induced vascular endothelial cell senescence， and explore the possible mechanism.MethodHuman umbilical vein endothelial cells （HUVECs） were used as the research objects， and the aged model was established with 10-12 passages of replicative senescence cells. The experimental cells were divided into young group （2-4 passage cells）， aged group （10-12 passage cells）， only EMPs intervention group （extract EMPs produced by aged cells to intervene young cells） and low dose， middle dose and high dose drug intervention groups （200， 300， 400 mg·L-1）. Senescence related β-galactosidase （SA-β-gal） staining and cell cycle propidium iodide （PI） staining were used to determine cell senescence. Cell counting kit-8 （CCK-8） assay was used to screen the drug concentration. EMPs were extracted by two-step centrifugation， EMPs labeled with phycoerythrin （PE） anti-human CD31 antibody or fluorescein isothiocyanate （FITC） annexin V were detected by flow cytometry， intracellular reactive oxygen species （ROS） were detected by 2'，7'- dichlorofluorescein diacetate （DCFDA） staining.ResultAfter treatment with the drug， SA-β-gal activity of the aged cells significantly decreased （P<0.01）， the S phase arrest was restored （P<0.01）， and the number of CD31+ EMPs and annexin V+ EMPs secreted by aged cells decreased （P<0.05）. Compared with the young group， only EMPs intervention group could induce increased SA-β-gal activity and S phase arrest in young cells （P<0.05，P<0.01）. However， after intervention of EMPs and the drug， EMPs-mediated increase of SA-β-gal activity was significantly inhibited and S phase arrest was restored （P<0.05）. The increase of intracellular ROS induced by EMPs was also significantly inhibited by the drug （P<0.05，P<0.01）.ConclusionGinseng Radix et Rhizoma-Notoginseng Radix et Rhizoma-Chuanxiong Rhizoma extract can delay the senescence of vascular endothelial cells by influencing EMPs， and the mechanism may be related to the inhibition of increased intracellular ROS induced by EMPs.
Keywords：endothelial microparticles;senescence;Ginseng Radix et Rhizoma-Notoginseng Radix et Rhizoma-Chuanxiong Rhizoma extract;reactive oxygen species （ROS）;human umbilical vein endothelial cells （HUVECs）
Abstract：ObjectiveTo study the effect of Fuzheng Qufeng prescription （FZQP） on transforming growth factor-β1 （TGF-β1）/Smad signaling pathway and epithelial-mesenchymal transition of podocyte in membranous nephropathy （MN） rats and to explore its molecular mechanism for podocyte protection.MethodThe rats were randomly divided into normal control group （NC） and modeling group. Rats in modeling group induced by bovine serum albumin （C-BSA） were randomly divided into model group （MN）， losartan potassium group （LP， 0.05g·kg-1）， and FZQP high dose （FZQPH， 41 g·kg-1）， medium dose （FZQPM， 20.5 g·kg-1）， and low dose （FZQPL， 10.25 g·kg-1） groups. The administration lasted for 4 weeks. In week 0， 2， and 4 of administration， the levels of 24 hours urine protein （24 h-Upro） were tested. At the end of 4th week， the levels of blood urea nitrogen （BUN） and serum creatinine （SCr） were detected， and the rats in each group were sacrificed and the renal pathological morphology changes were observed by light microscope with hematoxylin-eosin （HE）， Masson and periodic acid-silver metheramine （PASM） staining. The deposition of immune complex， the thickening of glomerular basement membrane （GBM） and podocyte foot process were observed by transmission electron microscope （TEM）. The distribution and expression intensity of Desmin in renal tissues were detected by immunohistochemistry （IHC）. The mRNA and protein expression levels of TGF-β1， Smad2/3， phospho（p）-Smad2/3， Smad7 and Desmin in renal tissues were respectively detected by Western blot （WB） and Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）.ResultCompared with NC group， the levels of 24 h-Upro， BUN and SCr significantly increased in model group （P<0.01）， with increased deposition of immune complex， significantly thickened GBM and fusion of foot processes， significantly increased Desmin mRNA and protein expression （P<0.01） and increased TGF-β1， Smad2， and Smad3 mRNA and protein expression （P<0.05）， and decreased Smad7 mRNA and protein expression （P<0.05，P<0.01）. Compared with model group， 24 h-Upro and BUN decreased in FZQP groups and LP group （P<0.05）， levels of serum SCr in FZQPM group decreased （P<0.05）， deposition of immune complex， thickening of GBM and fusion of foot process were all alleviated in FZQP groups and LP group. Distribution of Desmin along GBM decreased in FZQPH group， FZQPM group and LP group （P<0.05）. Both mRNA and protein expression levels of TGF-β1 and p-Smad2/Smad2 in FZQPM group decreased， while mRNA and protein expression levels of Smad7 increased （P<0.05）. Both mRNA and protein expression levels of p-Smad3/Smad3 in FZQPH group decreased （P<0.05）. Both mRNA and protein expression levels of Desmin in podocyte in FZQPH group， FZQPM group and LP group decreased （P<0.05）.ConclusionFZQP might realize podocyte protection effect in MN via suppressing EMT mediated by overactivated TGF-β1/Smad signaling pathway.
Abstract：ObjectiveTo explore the molecular mechanism of spleen-strengthening traditional Chinese medicine （TCM） Weichang'an granule in inhibiting the invasion and metastasis of human gastric cancer MKN45 cells.MethodMKN45 cells were cultured in vitro and incubated with different concentrations（600， 900， 1 200， 1 500 mg·L-1）of Weichang'an granule for 24， 48， 72 h. Cell counting kit-8 （CCK-8） assay was used to detect its effect on the cell proliferation. Western blot was used to detect the expression of RUN and FYVE domain containing 3（RUFY3） in normal gastric mucosa cells and different gastric cancer cell lines. The expression of RUFY3 in the gastric cancer cells after Weichang'an granule intervention （600， 900， 1 200 mg·L-1） was detected by Western blot. Lentivirus transfection technique was used to achieve the stable and silenced expression of RUFY3 in gastric cancer MKN45 cells. Transwell assay was used to evaluate the influence of Weichang'an granule and silenced RUFY3 on the metastasis and invasion ability of MKN45. E-cadherin，N-cadherin，Vimentin，Zinc-finger transcription factor （SNAIL1 and SNAIL2） protein expression levels were detected by Western blot.ResultRUFY3 expression in human gastric cancer cells was significantly higher than that in normal gastric mucosa．The protein expression of RUFY3 in MKN45 cells of silenced RUFY3 group was significantly lower than that in Lentivirus negative group （P<0.01）． Weichang'an granule inhibited the expression of RUFY3 in human MKN45 gastric cancer cells （P<0.05， P<0.01） in a concentration-dependent manner. As compared with the blank group， both Weichang'an granule and silenced RUFY3 inhibited the metastasis and invasion ability of MKN45 （P<0.01）. After Weichang'an granule and silenced RUFY3 treatment， the protein expression of epithelial marker gene E-cadherin was up-regulated （P<0.01）， the protein expression of Vimentin and N-cadherin decreased， but with no statistical difference，while SNAIL1 and SNAIL2 were both significantly down-regulated （P<0.01）.ConclusionBy targeting RUFY3 to regulate epithelial mesenchymal transformation， the spleen-strengthening TCM compound Weichang'an granule can inhibit the invasion and metastasis of gastric cancer.
Keywords：Weichang'an granule;spleen-strengthening traditional Chinese medicine;gastric cancer;epithelial mesenchymal transformation（EMT）;invasion and metastasis;RUN and FYVE domain protein 3
Abstract：ObjectiveTo investigate the effect of Huayu Jiedu prescription medicated serum（HJRMS）on the proliferation， invasion and migration of human lung cancer cells （H1299 cells） and its mechanism.MethodCell counting kit-8 （CCK-8） method was used to detect the inhibitory effect of HJRMS on the proliferation of lung cancer cells， the effect of HJRMS on the invasion and migration of H1299 cells were determined by Transwell assay and wound healing assay. The protein expressions of Janus kinase 2 （JAK2）， signal transduction and activation transcription factor 3 （STAT3）， phosphorylated JAK2（p-JAK2） and phosphorylated STAT3 （p-STAT3） were detected by Western blot， the mRNA expression levels of JAK2 and STAT3 were detected by Real-time quantitative polymerase chain reaction（Real-time PCR）.Result① Compared with control group， the proliferation of H1299 cells was significantly inhibited after treatment with 1%~16%HJRMS serum for 24， 48 h， respectively（P<0.01）， and showed a certain concentration dependence. ② After treatment with HJRMS for 24 h， the scratch healing ability of cells in the 4%，8%HJRMS serum groups was inhibited（P<0.05，P<0.01）. ③ Compared with control group， the membrane permeability of H1299 cells in invasion and migration experiments in 2%，4%，8%HJRMS serum groups was decreased significantly（P<0.05，P<0.01）. ④ Western blot showed that compared with control group， 4%，8%HJRMS serum groups inhibited the expression of JAK2/STAT3 signaling pathway related proteins （JAK2， p-JAK2， STAT3， and p-STAT3） in lung cancer H1299 cells（P<0.05， P<0.01）. ⑤ Compared with control group， the mRNA expression levels of JAK2 and STAT3 in lung cancer H1299 cells treated with 8%HJRMS for 24 h decreased significantly （P<0.05）.ConclusionThe HJRMS can inhibit the proliferation， invasion and migration of lung cancer H1299 cells， and its mechanism may be related to the inhibition of JAK2/STAT3 signaling pathway.
Abstract：ObjectiveTo observe the clinical efficacy of Shenxie Zhitong capsule in the treatment of diabetic peripheral neuropathy（DPN） of stagnant blockade of collaterals， and evaluate its effectiveness and safety.MethodThe 104 patients were randomly divided into the Shenxie Zhitong capsule treatment group （the treatment group， 53 patients） and the alpha lipoic acid group （control group， 51 patients）， and two groups were compared by random and contrast test. The changes of the Toronto clinical scoring system （TCSS）， utah early neuropathy scores （UENS）， traditional Chinese medicine（TCM）syndrome scores， visual analysis scale （VAS）， ankle brachial index （ABI）， vibrating perception threshold （VPT） before and after treatment were compared between two groups， and the endpoint events， such as foot ulcers， percutaneous coronary intervention （PCI）， death and composite endpoint events， related indicators of glucose and lipid metabolism and safety indicators were recorded among patients.ResultCompared with the data before treatment， the scores of TCSS， UENS， and TCM syndromes in two groups were significantly reduced （P<0.01） after treatment， and VAS and glycosylated haemoglobin A1c （HbA1c） were significantly reduced （P<0.05）， during follow-up visit， the levels of right ABI，total cholesterol （TC）， and low-density lipoprotein （LDL） in two groups were significantly reduced （P<0.05）， and high-density lipoprotein （HDL） level was significantly increased （P<0.05）. control group in control group， the 2-hour postprandial plasma glucose （2 h PG） and HbA1c levels were significantly increased （P<0.05）. Compared with control group， the VAS of the treatment group after treatment was significantly reduced （P<0.05）. After treatment and during follow-up visit， compared with control group， the 2 h PG levels of the right toe in the treatment group were significantly reduced （P<0.05）. There was no statistically significant difference in endpoint events and safety indicators between two groups， but the incidence trend of composite endpoint events in the treatment group was lower than that in control group.ConclusionShenxie Zhitong capsule has definite clinical curative effect in treating diabetic peripheral neuropathy， which is more safe and effective than alpha lipoic acid in improving pain symptoms.
Keywords：Shenxie Zhitong capsule;alpha lipoic acid;diabetic peripheral neuropathy;stagnant blockade of collaterals
Abstract：ObjectiveTo observe the clinical efficacy modified Chaihu Shugansan combined with Xuanfu Daizhetang on anxiety and depression of patients with gastroesophageal reflux disease （GERD） and Qi stagnation and phlegm obstruction syndromes in clinic and the effect on neuropeptide factor and pro-inflammatory factor.MethodPatients 200 cases were divided into control group and observation group. Patients in control group got omeprazole enteric-coated tablets， 20 mg/time， 1 time/day， flupentixol and melitracen tablets， 1 tablet/time， 2 times/day， and Dalitong granules， 1 bag/time， 3 times/day. In addition to omeprazole enteric-coated tablets in control group， patients in observation group were also added with syndrome differentiation-based treatment of modified Chaihu Shugansan combined with Xuanfu Daizhetang， 1 dose/day. The treatment lasted for 8 weeks. Before and after treatment， scores of acid regurgitation， heartburn， poststernal pain， syndrome of Qi stagnation and phlegm obstruction， gastroesophageal reflux disease questionnaire （GerdQ）， Hamilton anxiety Scale-14 （HAMA-14） and Hamilton depression scale-17 （HAMD-17） were scored； And upper gastrointestinal endoscopy， levels of peripheral vasoactive intestinal peptide （VIP）， 5-hydroxytryptamine （5-HT）， tumor necrosis factor-α （TNF-α）， interleukin-1 （IL-1）， IL-6 were detected. All the patients （GerdQ<8） got follow-up for 16 weeks， and the recurrence and safety were recorded.ResultAfter treatment， scores of GerdQ， endoscope， main symptoms and syndrome of Qi stagnation and phlegm obstruction on observation group were lower than those in control group （P<0.01）， and scores of HAMA-14 and HAMD-17 decreased in both groups （P<0.01）， but with no statistically significant difference in both groups. The comprehensive efficacy of main symptoms in observation group was better than that in control group （Z=2.076， P<0.05）. The curative effect of traditional Chinese medicine （TCM） syndrome in observation group was superior to that in control group （Z=2.151， P<0.01）. The effect of endoscope was better than that in control group （Z=2.103， P<0.05）. And levels of VIP， 5-HT， TNF-α， IL-1 and IL-6 in observation group were lower than those in control group. During the 16-week follow-up， the recurrence rate in observation group was 19.23% （15/78）， which was lower than 35.38% （23/65） in control group （=4.741， P<0.05）. The average recurrence time of the observation group was lower than that of the control group（P<0.01）.ConclusionModified Chaihu Shugansan combined with Xuanfu Daizhetang can significantly improve the main symptoms and TCM syndromes， relieve depression， anxiety and other adverse emotions， promote the healing of gastroesophageal mucosa， reduce the recurrence rate and delay the recurrence time among patients with GERD and Qi stagnation and phlegm obstruction syndromes. The mechanism of action may be related to the expression of neuropeptide factor and the inhibition of pro-inflammatory factor.
Keywords：gastroesophageal reflux disease;syndrome of Qi stagnation and phlegm obstruction;anxiety;depression;Chaihu Shugansan;Xuanfu Daizhetang;vasoactive intestinal peptide;5-hydroxytryptamine;inflammatory factors
Abstract：ObjectiveTo observe the clinical efficacy of Didangtang combined with gemcitabine chemotherapy in postoperative patients with non-muscle invasive bladder ctraditional chinese medicineancer （NMIBC）.MethodA total of 100 patients with NMIBC in Jiangsu Provincial Second Chinese Medicine Hospital and Jiangsu Province Hospital of Chinese Medicine from January 2015 to September 2020 were randomly divided into control group and treatment group. 50 cases in the treatment group received Didangtang combined with intravesical infusion of gemcitabine， and 50 cases in the control group received placebo combined with intravesical infusion of gemcitabine. Before and after the treatment cycle， the blood biochemistry， immune cell subsets（CD3+，CD4+ and CD4+/CD8+） were detected to assess the safety and immune function changes in treatment group. Quality of life scale for patients with bladder cancer prepared by European Organization for the Research and Treatment of Cancer （EORTC QLQ-C30） and Specificity scale for patients with superficial bladder cancer （EORTC QLQ-BLS24） were scored to assess the changes in physical， role， emotional and social functioning of the patients， symptomatic conditions such as fatigue， pain， nausea and vomiting， shortness of breath， insomnia， loss of appetite， diarrhea and constipation， as well as bladder cancer-specific symptoms such as treatment-generated problems， future concerns， bowel symptoms and urinary tract symptoms.ResultThere were no differences in terms of age， sex， disease duration and tumor states. During the treatment period， both groups had no recurrence or adverse events. The traditional Chinese medicine（TCM） efficacy in the treatment group was significantly better than that in the control group（χ2= 7.23，P = 0.02）. After treatment， the number of CD3+，CD4+ ，CD8+ cells and the CD4+/CD8+ ratio in the treatment group were significantly higher than those in control group（P<0.05）. Both groups showed significant improvements in the domains of role functioning， social functioning， and general health after treatment （P<0.05）. In addition， fatigue， diarrhea， constipation， intestinal symptoms， and urinary tract symptoms in the treatment group were significantly better than those in the control group after treatment （P<0.05）.ConclusionDidangtang can reduce adverse clinical symptoms during postoperative gemcitabine infusion chemotherapy in patients with NMIBC， reduce urethral and intestinal discomfort reactions， improve immune function and enhance quality of life for patients.
Abstract：ObjectiveTo observe the regulatory effect of modified Erzhu Erchentang on metabolization of polycystic ovary syndrome （PCOS） with spleen deficiency and phlegm dampness syndrome.MethodPatients 140 cases were divided into control group and observation group. Both groups were given metformin hydrochloride tablets， 500 mg/time， 3 times/day. Control group was given Yuejun Erchen pills， 0.5 g/time， 3 times/day， while observation group was given modified Erzhu Erchentang， 1 dose/day. The course of treatment lasted for 24 weeks. Before and after treatment， levels of fasting blood glucose （FBG）， fasting insulin （FINS）， glycosylated hemoglobin Alc （HbA1c）， 2-hour postprandial blood glucose （2 h PG）， blood lipid， waist circumference （WC）， body mass index （BMI）， waist hip ratio （WHR）， luteinizing hormone （LH）， follicle stimulating hormone （FSH）， serum testosterone （T）， estradiol （E2）， dehydroepiandrosterone sulfate （DHEAS）， sex hormone binding globulin （SHBG）， leptin （LP）， adiponectin （APN）， resistin， visfatin and tumor necrosis factor-α （TNF-α） were detected. Homeostasis model assessment insulin resistance （HOMA-IR） was calculated， modified Erzhu Erchentang was scored， and recovery of menstruation and ovulation and ovarian volume were recorded.ResultLevels of FBG， 2 h PG， HbA1c， FINS， HOMA-IR， triglyceride （TG）， total cholesterol （TC）， low-density lipoprotein cholesterol （LDL-C）， LH， FSH， T， E2， DHEAS， LP， resistin， visfatin and TNF-α in observation group were lower than those in control group （P<0.01）， and levels of BMI， WC and WHR were lower than those of control group （P<0.05）. And levels of high-density lipoprotein cholesterol （HDL-C）， SHBG and APN were higher than those in control group （P<0.01）. Score of modified Erzhu Erchentang was lower than that in control group （P<0.01）， and ovarian volume was smaller than that in control group （P<0.01）. The normal rate of BMI was 49.23% （32/65）， which was higher than 30.30% （20/66） in control group （χ2=5.151， P<0.05）. The normal rate of blood lipid was 93.85% （61/65）， which was higher than 81.82 % （54/66） in control group （χ2=4.418， P<0.05）. The normal rate of blood glucose was 96.92% （63/65）， which was higher than 86.36% （57/66） in control group （χ2=4.474， P<0.05）.ConclusionIn addition to adipocytokines， modified Erzhu Erchentang could regulate adipokines of patients of PCOS with spleen deficiency and phlegm dampness， improve glucose， lipid metabolism and overweight， adjust endocrine hormone， reduce clinical symptoms and improve ovarian structure， so as to create conditions for conception.
Abstract：ObjectiveTo identify the chemical constituents of Platycladi Cacumen before and after being carbonized.MethodChemical constituents in 3 batches of Platycladi Cacumen and its carbonized products were identified and compared by ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry （UHPLC-QTOF-MS/MS）. Chromatographic separation was performed on an ACQUITY UPLC BEH C18 column （2.1 mm×100 mm， 1.7 μm） with 0.1% formic acid aqueous solution （A）- acetonitrile （B） as mobile phase for gradient elution （0-3.5 min， 5%-15%B； 3.5-6 min， 15%-30%B； 6-6.5 min， 30%B； 6.5-12 min， 30%-70%B； 12-12.5 min， 70%B； 12.5-18 min， 70%-100%B； 18-22 min， 100%B）. The flow rate was 0.4 mL·min-1 and the injection volume was 5 μL. Mass spectrometry was performed by an electrospray ionization， and the primary and secondary mass spectrometry data were collected with the full scan mode of positive and negative ions， the peaks containing MS/MS data were identified by self-established secondary mass spectrometry database and corresponding fragmentation law matching method.ResultA total of 77 and 76 substances with the same change trend were identified under positive and negative ion modes. After being carbonized， the disappeared components of Platycladi Cacumen were mainly amino acids， ketone aldehydes and other volatile components. Among newly produced components， there were 6 kinds of flavonoid aglycones （rhamnetin， 6，7，3'-trihydroxyflavone， 3，6，3'-trihydroxyflavone， 4'-hydroxy-2'-methyl-3，4，5-trimethoxychalcone， herbacetin and 3'，5'-dimethoxy-3，5，7，4'-tetrahydroxyflavone）， 3 kinds of coumarins （7-hydroxycoumarin， 7，8-dihydroxycoumarin and 8-acetyl-7-hydroxycoum-arin） and 3 kinds of benzoic acids （3-methylcatechol， pyrocatechol and chromone-3-carboxylic acid）. There were a total of 40 flavonoids （quercitrin， quercetin， kaempferol， etc.） among these identified chemical constituents.ConclusionThere are significant quantitative and qualitative changes in the chemical compositions of Platycladi Cacumen after being carbonized. The flavonoids， the identified main active ingredients， can provide data reference for further study on the material basis of efficacy changes of Platycladi Cacumen before and after being carbonized.
Abstract：ObjectiveTo investigate the antioxidant activity and chemical composition of 75% ethanol extract of Rosa cymosa roots and its different polar parts.MethodThe 75% ethanol extract of R. cymosa roots was divided into dichloromethane， ethyl acetate， n-butanol and water parts by organic solvent extraction. In vitro antioxidant activity of each fraction was determined by 1，1-diphenyl-2-picrylhydrazyl （DPPH） and 2，2'-azino-bis（3-ethylbenzothiazoline-6-sulfonic acid） （ABTS） free radical scavenging assays， as well as ferric reducing antioxidant power （FRAP） test. The contents of total triterpenes， total phenols， total tannins and condensed tannins in each fraction were determined by spectrophotometry. SPSS 24.0 software was used to conduct Pearson correlation analysis between the antioxidant activity of each fraction and the content of the main components， and then the main active fraction and the main active components were determined. The chemical constituents of the active fraction was analyzed by ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF-MS/MS）， and the structures of the main chromatographic peaks were predicted.ResultEach fraction of R. cymosa roots had certain antioxidant activity， and there was a significant dose-effect relationship within a certain concentration range， but the antioxidant activity of different polar parts was different. In DPPH and ABTS free radical scavenging tests， the antioxidant activity of each fraction and vitamin C （VC， the positive drug） was ranked as ethyl acetate fraction>VC>n-butanol fraction>ethanol extract>water fraction>dichloromethane fraction. In FRAP test， the activity of ethyl acetate fraction was weaker than that of VC， and the other order was unchanged. The contents of total triterpenes， total phenols， total tannins and condensed tannins in ethyl acetate fraction were 3.81%， 50.33%， 3.32%， and 39.79%， in n-butanol fraction were 0.88%， 41.42%， 2.25% and 23.55%， in ethanol extract were 2.90%， 41.95%， 3.43% and 20.14%， in water fraction were 0， 26.80%， 16.90% and 7.57%， and in dichloromethane fraction were 21.23%， 12.90%， 1.59%， and 6.17%， respectively. Correlation analysis results showed that the contents of total phenols and condensed tannins were positively correlated with the antioxidant activity， the contents of total triterpenes were negatively correlated with the antioxidant activity， and the correlation between total tannins and antioxidant activity was not obvious. A total of 26 compounds were identified from the ethyl acetate fraction by UPLC-Q-TOF-MS/MS， including 11 condensed tannins， 4 hydrolysable tannins， 6 triterpenes， 3 flavonoids， 1 benzoic acid derivative and 1 chlorogenic acid analogue.ConclusionEthyl acetate fraction is the main antioxidant active site of R. cymosa roots， and phenols mainly composed of condensed tannins are the main active components. The results can provide experimental basis for the development of natural antioxidants.
Keywords：roots of Rosa cymosa;antioxidant activity;total phenols;condensed tannins;total triterpenes;free radicals;vitamin C
Abstract：ObjectiveTo clone the full-length glycosyltransferase genes （PpUGT1，PpUGT7） related to saponins biosynthesis in Paris polyphylla var. yunnanensis，and perform bioinformatics analysis，relative expression analysis and prokaryotic expression analysis.MethodTotal RNA was isolated from P. polyphylla var. yunnanensis with use of the Eastep® Super Total RNA Extraction Kit and converted to cDNA. Specific primers were designed according to the transcriptome data to clone the full-length gene. Relevant software was then used for bioinformatic analysis of the protein sequences. The relative gene expression levels were detected by real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） and the prokaryotic expression vectors were built to heterologously express recombinant protein in Escherichia coli.ResultThe open reading frame （ORF） of PpUGT1 was 1 827 bp，encoding 608 amino acids，and was predicted as a steroid glycosyltransferase；the ORF of PpUGT7 was 1 380 bp，encoding 459 amino acids，and was predicted as a triterpenoid glycosyltransferase. The calculated relative molecular mass of two proteins were 67.6 kDa and 51.3 kDa respectively，and both of them were hydrophilic proteins，no transmembrane domain，no signal peptides，both showing high similarity and conservativeness with homologous sequences. The results of Real-time PCR showed that the expression level of PpUGT1 was root>leaf>flower>stem；the expression level of PpUGT7 was stem>leaf>flower>root. In addition，PpUGTs proteins were expressed in E. coli. in a soluble form.ConclusionThe genes of PpUGT1 and PpUGT7 were cloned successfully. Real-time PCR showed the genes were expressed differently in different plant organs， and their recombinant proteins were successfully expressed in Escherichia coli. This study lays a foundation for functional characterization of PpUGTs and analysis of the biosynthesis pathway of saponins in Paris polyphylla var. yunnanensis.
Abstract：ObjectiveTo investigate the effect of the production process of Zushima Guanjie Zhitong Gao from solvent method to hot-pressed method on in vitro kinetic behavior of this preparation.MethodSolvent and hot-pressed methods were used to prepare three batches of samples above pilot scale， and in vitro release and percutaneous penetration of the index components （7，8-dihydroxycoumarin and methyl salicylate） in Zushima Guanjie Zhitong Gao were investigated by modified Franz diffusing cells.ResultThe contents of 7，8-dihydroxycoumarin and methyl salicylate in Zushima Guanjie Zhitong Gao prepared by solvent method were 73.72， 494.67 μg/patch， and their contents in hot-pressed method samples were 159.21， 2 638.99 μg/patch， respectively. In the solvent method samples， the average cumulative release amounts of 7，8-dihydroxycoumarin and methyl salicylate in 24 h were 2.04， 12.21 μg， and their average cumulative release amounts in 24 h of hot-pressed method samples were 2.16， 36.24 μg， respectively. In the solvent method samples， the average cumulative permeation amounts of 7，8-dihydroxycoumarin and methyl salicylate in 24 h were 0.38， 2.79 μg， and they were 0.40， 7.49 μg in hot-pressed method samples. The cumulative release and permeation amounts in 24 h of 7，8-dihydroxycoumarin in the hot-pressed method samples were basically the same as those of the solvent method samples， but the cumulative release and permeation amounts in 24 h of methyl salicylate in the hot-pressed method samples were significantly higher than those of the solvent method samples （P<0.05）.ConclusionThe retention of 7，8-dihydroxycoumarin and methyl salicylate by hot-pressed method is better than that of the solvent method. The process change has no significant effect on the in vitro kinetics of 7，8-dihydroxycoumarin in Zushima Guanjie Zhitong Gao， however， after the change from the solvent method to the hot-pressed method， the methyl salicylate in this preparation has a higher cumulative release and permeation amounts.
Abstract：Textual research on Chinese herbs is the preliminary work for the preparation of famous classical prescriptions. Through literature review，it was found that the researches of Persicae Semen focused on chemical compositions，pharmacological mechanism and medical record analysis in the recent years，and few researches based on the ancient literature were about the origin，concocting methods，flavor，meridian tropism，effects and indications. Textual research shows that the most commonly used names are Taoren and Taoheren，with a wide range of producing areas. The plant origin of Persicae Semen has not changed much since ancient times. Before the Qing dynasty，the plant origin of Persicae Semen was from the seeds of a kind of fruit named Shantao or Maotao，and in modern times，the seeds of Amygdalus persica or A. davidiana have become the major source. While different books have different views on Latin names for the origin of the Persicae Semen. We suggest that the Latin names of A. persica and A. davidiana should be more reasonable for Tao and Shantao respectively .In the concocting methods of Persicae Semen for activating vital energy and blood circulation，raw Persicae Semen should be used with peel and tip，while for moisturizing dryness，it should be fried into yellow without peel. Therefore，in the concocting methods of Persicae Semen for Taohe Chengqitang and Taohong Siwutang，the raw materials should be fried into yellow without peel or tip，while for Shentong Zhuyutang，raw Persicae Semen materials should be used with peel and tip. The indications of Persicae Semen include amenorrhea，lump，parasite，obstruction of chest，cough and asthma，constipation，etc.，and the people with blood deficiency，blood dryness and the pregnant women should use it with caution or should not use it. The modern clinical application of Persicae Semen is only a partial inheritance of ancient literature，which means that the Persicae Semen still has many effects to be verified and studied，and it is worthwhile for further exploration in order to expand its clinical application. The records of ancient literature on flavor，meridian tropism，and quality evaluation of Persicae Semen were consistent with those in Chinese Pharmacopoeia. Because of the similar appearance，it is especially difficult to distinguish Persicae Semen and Armeniacae Semenis Amarum after crushing ，requiring much time and money for identification. It is recommended that medical institutions should purchase Persicae Semen without crushing as far as possible，then decide the best concocting methods according to the clinical requirement.
Keywords：Persicae Semen;famous classical prescriptions;origin;processing;flavor and efficacy;herbal textual research
Abstract：ObjectiveTo investigate the effective constituents and mechanism of Tingli Dazao Xiefeitang for the treatment of heart failure through network pharmacology and high resolution mass spectrometry technique as well as molecular docking technique.MethodChemical components and potential targets in Descurainiae Semen Lepidii Semen and Jujubae Fructus were searched by referring to literature and BATMAN-TCM database. The disease targets were searched in GeneCards database with ''heart failure'' as the key word. STRING platform was used to construct protein-protein interaction（PPI） network based on common targets of drugs and disease. The network topology was analyzed by using Cytoscape 3.7.2 software to obtain the core targets eventually. Kyoto encyclopedia of genes and genomes（KEGG） enrichment analysis of the core targets was conducted through DAVID database to draw a network of “herb-compound-target-pathway”. Based on the results of the network pharmacology research above，high resolution mass spectrometry was used to analyze the decoction and confirm the selected active components. AutoDock 4.2.6 software was used for molecular docking verification of key active components and related targets.ResultA total of 85 components were obtained in Descurainiae Semen Lepidii Semen，and 49 components were obtained in Fructus Ziziphi Jujubae. A total of 1 078 drug targets and 1 549 disease targets were identified. After PPI analysis and network topology analysis，23 core targets and 33 active components were obtained，involving 19 signaling pathways （P<0.05）. Mass spectrometry analysis results indicated that 18 components such as isovanillic acid，descuraininA and kaempferol-7-O-β-D-glucopyranoside were confirmed in decoction. Molecular docking analysis results indicated that 6 core components （degree value top 6），such as isovanillic acid，descurainin A and kaempferol-7-O-β-D-glucopyranoside，had good binding activity with silent information regulatory factor 1（SIRT1），interleukin（IL）1B，protein kinase Bα（AKT1） and tumor necrosis factor（TNF）. The compound recipe for heart failure mainly involved mitogen-activated protein kinase（MAPK），hypoxia inducible factor-1（HIF-1），Mammalian target of rapamycin（mTOR） and other signaling pathways.ConclusionThis study preliminarily investigated the effective constituents and mechanism of Tingli Dazao Xiefeitang in the treatment of heart failure. It can provide references for the precise clinical medication and screening of quality control markers，as well as the discovery of active components in the treatment of heart failure.
Abstract：ObjectiveTo analyze the research status of the external use of medicinal materials and decoction pieces in the 2020 edition of Chinese Pharmacopoeia （volume 1）， so as to provide reference for the clinical external use research of Chinese medicines.MethodThe external function， usage， dosage and other information of medicinal materials and decoction pieces in the 2020 edition of Chinese Pharmacopoeia （volume 1） were analyzed and compared.ResultIn the 2020 edition of Chinese Pharmacopoeia （volume 1）， there were 165 kinds of Chinese medicines that could be used externally， including 25 kinds of Chinese medicines that clearly recorded their functions for external use， such as Euphorbiae Semen， Euphorbiae Semen Pulveratum， Arisaematis Rhizoma， Arisaematis Rhizoma Preparatum， Crotonis Fructus， Crotonis Semen Pulveratum， Nardostachyos Radix et Rhizoma， Artemisiae Argyi Folium， Alumen， Stemonae Radix， Genkwa Flos， Cera Flava， Mel， Psoraleae Fructus， Gardeniae Fructus， Calomelas， Bruceae Fructus， Drynariae Rhizoma， Euphorbiae Ebracteolatae Radix， Cuscutae Semen， Phytolaccae Radix， Meretricis Concha， Talcum， Talci Pulvis and Propolis. The amount of Chinese medicine for external use recorded in the 2020 edition of Chinese Pharmacopoeia was mostly appropriate for external use， the amount for external use was not clear. The application method for external use focused on beating powder and applying the affected areas， decocting for fumigation and washing， simmering to paste， etc. Clinical external use was used for the treatment of eczema， tinea pedis， chapped skin and other skin diseases.ConclusionThe number of Chinese medicines recorded in the 2020 edition of Chinese Pharmacopoeia for external use is increasing， but the function， dosage and usage of these Chinese medicines for external use still need to be improved.
Keywords：Chinese medicines for external use;Chinese Pharmacopoeia;external functions;functions and indications;usage;dosage;clinical application
Abstract：ObjectiveTo explore the potential targets and related mechanism involved in the paclitaxel resistance to ovarian cancer.MethodOvarian cancer A2780 cells and A2780 paclitaxel-resistant cells （A2780/T） were treated by 2， 4， 8， 16， 32， 64， 128， 256 μmol·L-1 paclitaxel （PTX） for 24 h or 48 h respectively in vitro. The proliferation rate of A2780 cells and A2780/T cells treated with paclitaxel was determined by methyl thiazolyl tetrazolium （MTT） colorimetric method assay. A2780 and A2780/T cells were analyzed by LC-MS/MS Label-Free quantitative proteomics to identify and screen differentially expressed proteins in the two groups of cells. Gene ontology （GO） annotation and Kyoto encyclopedia of genes and genomes （KEGG） pathway analysis were used to determine the potential biomarkers of paclitaxel resistance in ovarian cancer. Conventionally cultured A2780 cells were used as a control group， and A2780/T cells were treated with 0， 1， 4 μmol·L-1 PTX. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot methods were used to detect and verify the mRNA and protein expression levels of potential target transforming growth factor-β-activated kinase 1 binding protein 1 （TAB1） and its downstream related molecules transforming growth factor-β-activated kinase （TAK1） and p38.ResultAfter PTX treatment for 24 h and 48 h， the cell viability of A2780 and A2780/T cells decreased. The inhibitory rate of PTX on A2780 cells was significantly higher than that of A2780/T cells. In A2780 cells， the IC50 of PTX treatment for 48 h was 0.002 μmol·L-1， while in A2780/T cells， the IC50 of PTX was greater than the maximum concentration of 128 μmol·L-1， indicating that A2780/T cells were resistant to PTX compared with A2780 cells. 441 differentially expressed proteins and 421 special differentially expressed proteins between A2780/T and A2780 cells were screened by label-free quantitative proteomic analysis. GO function enrichment analysis showed that the binding proteins accounted for the majority （80%） among the differentially expressed proteins. According to the results of KEGG pathway analysis and expression site analysis， TAB1 might be a potential biomarker in paclitaxel-resistant ovarian cancer. Compared with A2780 cells， mRNA and protein expression levels of TAB1 in A2780/T cells were significantly reduced （P<0.01）. mRNA expression of TAK1 and p38 that interacted with TAB1 were also significantly reduced （P<0.05， P<0.01）， while there was no significant change in protein expression.ConclusionTAB1 may be a potential biomarker of paclitaxel resistance to ovarian cancer ， and its mechanism may be related to the TAB1/TAK1/p38 MAPK pathway.
Abstract：ObjectiveBased on the holistic view of traditional Chinese medicine（TCM） and the characteristics of syndrome differentiation and treatment，methods of distance clustering and distance clustering combined with pseudo-bit clear（BIC） criteria are adopted in this study to simulate the doctor's clinical prescription process，and excavate a clinician's core prescription for the treatment of a specific disease.MethodIn this study， it was believed that the clinical prescriptions of TCM can be divided into the core prescription for the treatment of the main syndrome and the addition and subtraction part of the medication for the concurrent syndrome. The hospital information system （HIS）was used to export the medical records of specific diseases，and then the methods of distance clustering and distance clustering combined with pseudo-BIC criteria were adopted to analyze professor WANG Jun-hong's 103 prescriptions for the treatment of Qi and Yin deficiency type of attention deficit hyperactivity disorder（ADHD）. The core prescriptions were screened by finding the optimal balance between the number of herbs and the proportion of population using the herbs.ResultTest4 consisted of 15 herbs such as Acori Tatarinowii Rhizoma，Crataegi Fructus，Lycii Fructus，Gall1 Gigerii Endothelium Corneum，and Pseudostellariae Radix were selected as the core prescription. In analysis by TCM theory and confirmation by professor WANG Jun-hong， it was proved to meet the characteristics of TCM treatment for ADHD.ConclusionCombination of distance clustering with pseudo-BIC criteria method can effectively sort and screen the core clinical prescriptions of TCM，and compared with the current core prescription screening methods，it is characterized by conforming to the doctor's clinical prescription process. This research is helpful to provide certain support for inheriting the experience from famous TCM doctors and for the development of new Chinese medicines.
Keywords：clinical prescription of traditional Chinese medicine;core prescription;methodology;inheritance of experience
Abstract：ObjectiveTo evaluate the effect of Xuebijing injection on inflammatory indexes and immune function in patients with acute exacerbation of chronic obstructive pulmonary disease （AECOPD）.MethodPubMed，Cochrane Library，Embase，Wed of Science，CBM，CNKI，VIP and Wanfang Data Online Knowledge service platform were searched by computer，all of which were up to February 2020. After literature screening and quality evaluation by two researchers independently，relevant data were extracted and Meta-analysis was carried out by RevMan 5.3 software.ResultA total of 21 randomized controlled trials（RCT） were included，involving 1 618 patients. The results of Meta-analysis showed，Xuebijing injection adjuvant therapy was significantly better than the control group in total effective rate ［relative risk（RR）=1.20，95% confidence interval（CI）（1.14，1.27），P<0.000 01］，C-reactive protein（CRP）［standard mean difference（SMD）=-1.58，95% CI（-1.98， -1.19），P<0.000 01］，total white blood cell （WBC）［mean difference（MD）=-1.44，95% CI（-1.84，-1.04），P<0.000 01］，procalcitonin（PCT）［SMD=-0.57，95% CI（-0.74，-0.41），P<0.000 01］，interleukin-6（IL-6）［SMD=-1.51，95% CI（-2.07，-0.96），P<0.000 01］，percentage of neutrophils（N%）［MD=-5.35，95% CI（-7.13，-3.58），P<0.000 01］，and tumor necrosis factor-α（TNF-α）［SMD=-1.52，95% CI（-2.23，-0.81），P<0.000 1］， and had positive effects in regulating cellular immune disorders.ConclusionXuebijing injection combined with routine treatment can improve the immune function of AECOPD patients，reduce the number of inflammatory markers，neutrophils and CD8+ T cells， thus modulating the small airway microcirculation to promote inflammatory absorption and inhibit the progression of the disease， with high safety.
Abstract：Medicinal plants have a long history and play an important role in the prevention and treatment of human diseases. However， the interaction between bioactive components and specific biological targets in many medicinal plants is still unclear. Conventional separation and activity evaluation model of chemical constituents from natural products was time-consuming and laborious， and cannot truly reflect the interaction between the natural conformation of compounds and receptors， so these methods could not meet the urgent needs of modern drug development. Affinity ultrafiltration coupled with liquid chromatography-mass spectrometry （AUF-LC/MS） is a rapid， efficient and precise method focused on identification of bio-components from natural products， it can make up for many shortcomings found in conventional discovery of bio-constituents. However， AUF-LC/MS technology also has some drawbacks in experiments. In this paper， the principle， characteristics and application of AUF-LC/MS in the screening of active ingredients from medicinal plants are systematically reviewed， and its development prospect is prospected， so as to provide scientific basis for rapid and targeted screening of active components from medicinal plants.
Keywords：affinity ultrafiltration;liquid chromatography-mass spectrometry;medicinal plants;active components;targeted;traditional Chinese medicine （TCM）
Abstract：Fatigue refers to the manifestation of disorders in the process of carrying out or maintaining random activities， which can be regarded as an independent disease or as a symptom in a variety of chronic diseases. The high incidence of fatigue has seriously affected people's physical and mental health， and the prevention and treatment of fatigue has become an important problem to be solved urgently. The pathogenesis of fatigue mainly includes energy consumpation， accumulation of metabolites， abnormal secretion of neurotransmitters， decline of mitochondrial function， dysfunction of hypothalamus pituitary adrenal axis， etc. At present， there is no unified understanding about the pathogenesis of fatigue at home and abroad. The gene research of fatigue is the current research frontier. Gene expression profiling provides a new method for the study of the mechanism of fatigue. The combination of gene chip technology and traditional Chinese medicine（TCM） theory is expected to bring a breakthrough in the study of the pathogenesis of fatigue. In the study of fatigue gene chip， messenger RNA（mRNA） and microRNA（miRNA） are the common research objects， but few explorations are focused on the gene expression rule of fatigue by a specific signaling pathway and the effective regulation targets of TCM for treating fatigue. In recent years， the dysfunction of reward and inhibition mechanism in the central nervous system has become a research hotspot. In particular， gamma amino butyric acid （GABA） and dopamine （DA） have attracted much attention as the main substances of inhibition and reward mechanism， respectively. GABA and DA are used as inhibition and reward mechanisms to maintain the balance， and the body will not feel fatigue. Once the balance is broken， the fatigue will be formed. At the same time， DA and GABA receptors can also regulate cyclic adenosine monophosphate signaling pathway（cAMP） to affect fatigue. The research on key genes in GABA/DA balance mechanism and related cAMP signaling pathway by gene chip technology is expected to reveal the pathogenesis of fatigue in depth. The gene chip method is used to detect the changes of key genes in GABA/DA pathway and the related cAMP signaling pathway in the fatigue population and the normal population， so as to further explore the pathogenesis of fatigue. In this paper， the key genes in GABA/DA balance mechanism and cAMP signaling pathway related to fatigue were summarized by using the review method， so as to provide the basis for further study on the pathogenesis of fatigue and effective prevention and treatment from the perspective of genetics.
Abstract：Ovarian cancer is a kind of malignant tumor in female reproductive system with a high incidence. This disease is insidious at its early stage and the symptoms are not typical. Most of the patients have reached advanced stage by the time of diagnosis， seriously threating women's life and health. Many signaling pathways are involved in the formation and development of ovarian cancer， among which the phosphatidylinositol 3-kinase（PI3K）/protein kinase B（Akt） signaling pathway is one of the key regulatory pathways. In recent years， traditional Chinese medicine （TCM） has obtained wide attention in treatment of tumors due to its advantages of high safety and less adverse reactions， and more and more attention has been paid to the study of TCM monomers. Molecular biology studies have shown that TCM monomers can play a role against tumor by regulating multiple targets. By reviewing the literature and searching several databases， we found that TCM monomer can play an important role in the growth， proliferation， invasion and migration， apoptosis， autophagy and reversal of drug resistance of ovarian cancer cells by regulating PI3K/Akt signaling pathway. According to the existing studies， TCM monomers have a certain effect on ovarian cancer， but there are still many problems. Although the mechanisms of some TCM monomers have been clarified in the treatment of ovarian cancer， such TCM monomers are only limited to the tumor-bearing nude mice in vivo and experimental studies on in vitro cells， and further studies are needed in the future. In addition， in the future researches， ovarian cancer syndrome differentiation and targeted therapy can be linked to the TCM flavors， efficacy and indications to further develop the advantages of TCM. Based on the current research situation at home and abroad， this paper summarized the research progress of targeted intervention of TCM monomers in ovarian cancer by regulating PI3K/Akt signaling pathway， in order to provide reference for further research of TCM monomers， and provide important ideas for the development of targeted treatment of ovarian cancer with TCM.
Keywords：traditional Chinese medicine monomer;ovarian cancer;phosphatidylinositol 3-kinase（PI3K）/protein kinase B（Akt） signaling pathway;research progress
Abstract：Uterine leiomyoma （UL）， the most common benign tumor of the reproductive system in women of childbearing age， is characterized by clinical symptoms such as increased menstrual flow， prolonged menstrual period， breast tenderness，backache， lower abdominal pain and mass in the lower abdomen. With the continuous progress of modern society， the age of women's marriage and childbirth is gradually pushed back， which to a certain extent has led to an increase in the probability of modern women suffering from UL. Relevant literature shows that the incidence of UL is about 70%， and 25%-50% of the patients have clinical symptoms， seriously endangering women's physical health. The prevention and treatment of UL by modern medicine is currently limited to two aspects： drug control of estrogen and progesterone levels and surgical removal. Traditional Chinese medicine（TCM）has shown obvious advantages in improving the clinical symptoms of UL patients， with very broad application prospects as it can regulate body's Qi and blood on the basis of syndrome differentiation， treatment and overall concepts. Lichongtang， as a famous TCM prescription for replenishing Qi， activating blood and removing blood stasis， was created by ZHANG Xi-chun， a famous Chinese medicine doctor in the Qing dynasty， and recorded in the Records of Tradition Chinese and Western Medicine in Combination. It is widely used in the field of gynecological diseases in clinical practice. Studies have shown that Lichongtang is effective in treating UL. Clinical observations show that Lichongtang can significantly relieve the clinical symptoms of UL patients such as prolonged menstrual period， dysmenorrhea， waist and abdomen swelling and irregular vaginal bleeding， with the characteristics of stable curative effect， high safety， less side effect and low recurrence rate. The experimental results show that Lichongtang has a comprehensive regulatory effect on UL through inhibiting the proliferation of UL cells and inducing apoptosis， reducing serum estrogen and progesterone level， regulating the apoptosis pathway of tumor cells， and promoting the degradation of extracellular matrix（ECM）. After retrieval in PubMed， CNKI and other databases， the authors made a review by summarizing the theories， clinical efficacy and action mechanisms of Lichongtang in the treatment of UL， in order to provide reference for the follow-up in-depth study of pharmacological mechanism of Lichongtang and its further clinical application and promotion.
Abstract：Based on the clinical characteristics of thromboembolic vasculitis， the diagnostic criteria in western medicine and the dialectical standard in traditional Chinese medicine （TCM） were analyzed and established by consulting relevant literature， and the modeling method， modeling objects as well as modeling advantages and disadvantages for the animal models of thromboembolic vasculitis were summarized in this paper. By analyzing its coincidence with the characteristics of clinical symptoms in traditional Chinese and western medicine， it was found that the animal model of thromboembolic vasculitis had a higher degree of coincidence with the clinical symptoms in western medicine， as well as cold and dampness blocking collaterals syndrome and heat toxin injury yin syndrome in TCM， but lower degree of coincidence with damp-heat toxin syndrome and Qi-blood deficiency syndrome. There is no animal model consistent with blood stasis syndrome. The pathological and hemorheological indexes （blood viscosity， erythrocyte sedimentation rate） were the most common indexes. Compared with a large number of cases reported in clinical treatment of thromboangiitis obliterans， the experimental research was relatively weak. It is the focus of future research to establish a reasonable model to judge the quantitative standard， and to establish the animal model with higher coincidence degree with TCM syndrome.
Keywords：thromboangiitis obliterans （TAO）;gangrene;animal model;combination of disease and syndrome
Abstract：Osteonecrosis of the femoral head （ONFH） is a serious orthopedic disease caused by many factors. It is difficult to cure clinically and has a poor prognosis， which poses a serious threat to human health. The pathogenesis of this disease is still unclear. The ONFH caused by different causes involves the disorder of a variety of metabolic pathways in vivo. Abnormal proliferation and differentiation of bone mesenchymal stem cells （BMSCs）， imbalance of bone metabolism， and increased destruction of bone trabeculae caused by abnormal transduction of bone metabolism-related signaling pathways may be the important causes of ONFH. BMSCs are pluripotent stem cells with self-renewal and multidirectional differentiation ability， which have good regeneration rate. Improving the osteogenic and differentiation ability of BMSCs is the key to inhibit bone absorption and promote bone matrix reconstruction， which plays an important role in bone remodeling. In recent years， there are many studies on the prevention and treatment of ONFH in traditional Chinese medicine（TCM）， and it has been found that a variety of single herbs， monomers and compounds can regulate the differentiation direction and process of BMSCs by targeting signal molecules， with great potential for bone defect repair and anti-femoral head necrosis activity. Nowadays， prevention and treatment of ONFH by regulating bone metabolism signaling pathways has become a hot research topic. In this paper， the mechanism and related signal transduction pathways of TCM in preventing and treating ONFH were reviewed to explore some mechanisms of alleviating the rate of bone loss， promoting bone formation， and repairing bone defects， so as to provide reference for further research on the prevention and treatment of ONFH by TCM. The related clinical application studies also provided specific targets for gene-assisted therapy of ONFH.
Keywords：osteonecrosis of the femoral head;traditional Chinese medicine;signaling pathways;bone mesenchymal stem cells;review