Abstract：ObjectiveTo observe the effect of Danggui Niantongtang on the protein and mRNA expression of key regulatory factors of the extrinsic and intrinsic apoptotic pathway in synovial tissue of adjuvant arthritis （AA） rats， and to further explore the mechanism of Danggui Niantongtang in the prevention and treatment of rheumatoid arthritis.MethodThe general condition of AA rats， including its body weight， were observed. The changes of toe volume were detected by toe volume meter. Histopathological changes of synovium of knee joint were observed by hematoxylin-eosin （HE） staining. The mRNA and protein expression levels of tumor necrosis factor receptor super family 6 （Fas）， Fas-associating protein with a novel death domain（FADD）， B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax） and cysteinyl aspartate specific proteinase Caspase-3 （Caspase-3） were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot.ResultCompared with the normal group， the toe volume of the model group increased significantly （P<0.01）， with significantly proliferated synovial cells， significantly reduced mRNA and protein expression levels of Fas， FADD， Bax and Caspase-3 in synovial tissues（P<0.05， P<0.01）， and significantly increased Bcl-2 level （P<0.01）. Compared with the model group， the swelling degree of toes in Danggui Niantongtang group and Tripterygium group was significantly alleviated （P<0.01）， with significantly improved synovial hyperplasia， significantly increased mRNA and protein expression levels of Fas， FADD， Bax and Caspase-3 （P<0.05， P<0.01）， and significantly decreased expression levels of bcl-2 mRNA and protein （P<0.05， P<0.01）.ConclusionDanggui Niantongtang can effectively reduce joint swelling and abnormal proliferation of synovial tissue in AA rats. Its mechanism may be related to regulating the expression of Fas， FADD， Bax， Bcl-2 and Caspase-3， and promoting the apoptosis of synovial cells.
Keywords：Danggui Niantongtang;rheumatoid arthritis;adjuvant arthritis rats;tumor necrosis factor receptor super family 6 （Fas）;B-cell lymphoma-2 （Bcl-2）;apoptosis
Abstract：ObjectiveTo explore the protective effect of Gegen Qinliantang on the intestinal mucosal epithelial barrier function of ulcerative colitis （UC） mice， and to explore its mechanism of action in the treatment of ulcerative colitis via matrix metallopeptidase-9 （MMP-9）/p38 mitogen-activated protein kinase （p38 MAPK） signaling pathway.MethodThe 48 female C57BL/6 mice were randomly divided into normal group， model group， sulfasalazine group （0.3 g·kg-1） and Gegen Qinliantang high， medium and low dose groups （2.84，1.42，0.71 g·kg-1）. The UC murine model was established by 3% dextran sulfate sodium （DSS）. Gegen Qinliantang and sulfasalazine were intragastrically administered on the 8th day after the model was established for 7 days， and the normal group was treated with the same amount of normal saline. Colon tissues were collected after the last administration， and the pathological changes of colon tissues were detected by hematoxylin-eosin （HE） staining. The expression of tight junction （TJ） proteins such as Occludin and zonula occludens-1（ZO-1） in colon tissues was detected by immunohistochemistry （IHC）， and the expression levels of tumor necrosis factor-alpha （TNF-α）， interleukin-1β （IL-1β）， and MMP-9 mRNA in colon tissues were detected by Real-time polymerase chain reaction （Real-time PCR）. The expression of phosphorylated p38 MAPK （p-p38 MAPK）， p38 MAPK and MMP-9 protein in colon tissues was detected by Western blot.ResultCompared with normal group， the body weight of mice decreased （P<0.01） and disease activity index （DAI） score increased significantly （P<0.01） in model group， the colon tissues of the model group were damaged more obviously， the expression of occludin and ZO-1 proteins in model group was significantly reduced （P<0.01）， and the relative expression levels of TNF-α， IL-1β， and MMP-9 mRNA in model group were significantly increased （P<0.01）， the expression of p-p38 MAPK and MMP-9 in model group was significantly increased （P<0.01）. Compared with model group， the body mass and DAI score of the sulfasalazine group and Gegen Qinliantang group were significantly improved （P<0.05，P<0.01）， the colonic tissues damage were significantly improved， and the expression of Occludin and ZO-1 protein was significantly increased （P<0.05，P<0.01）， the relative expression levels of TNF-α， IL-1β， and MMP-9 mRNA were significantly decreased （P<0.01）， and the expression of p-p38 MAPK and MMP-9 was significantly decreased （P<0.01）. The changes in the middle dose group were the most obvious among the various dose groups of Gegen Qinliantang.ConclusionGegen Qinliantang repairs the intestinal mucosal barrier function by inhibiting the expressions of MMP-9 and inflammatory cytokines such as TNF-α and IL-1β， blocking the activation of the p38 MAPK signaling pathway， and increasing the expressions of tight junction protein.
Keywords：Gegen Qinliantang;ulcerative colitis;Occludin;zonula occludens-1;matrix metallopeptidase-9;p38 mitogen-activated protein kinase
Abstract：ObjectiveTo discuss the effect of Sishenwan on phosphatidylinositol-3 kinase/protein kinase B/mammalian target of rapamycin（PI3K/Akt/mTOR） signaling pathway related genes and proteins in colon tissue and interleukin-1β（IL-1β） and interleukin-10（IL-10） expression levels in serum of ulcerative colitis （UC） model rats with spleen kidney Yang deficiency.MethodThe 120 SPF Wistar rats were randomly divided into normal group and model group after 7 days of adaptive feeding in SPF laboratory. The model group were given dinitrobenzene sulfonate （DNBS）/ethanol solution enema+hydrocortisone subcutaneously injection+senna leaf gavage to establish UC model of spleen kidney yang deficiency. The rats who successfully established the model were randomly divided into five groups：model group， mesalazine （0.36 g·kg-1） group， and Sishenwan high， medium and low dose （3.2，1.6，0.8 g·kg-1） groups， the volume of which was 10 mL·kg-1. The model group and the blank group were given distilled water of the same volume. Once a day for 21 days. Observe the general conditions of the rats daily， and record the weight， fecal traits and occult blood of the mice for disease activity index （DAI） scoring.Take the rat colon tissue to observe the gross morphology and colon injury， and score the colon mucosal injury index （CMDI）. Hematoxylin-eosin （HE） staining was used to observe pathological changes. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）was used to detect the expression level of PI3K， Akt， mTOR mRNA in colon tissue. The levels of IL-1β and IL-10 in serum were detected by enzyme-linked immunosorbent assay （ELISA）. Western blot was used to detect the expression level of PI3K， phosphorylation （p）-PI3K， Akt， p-Akt， mTOR， p-mTOR protein in colon tissue.ResultCompared with blank group， the general survival status of the rats in model group was relatively poor， the DAI score and the CMDI index were significantly increased （P<0.01）. The intestinal mucosa partially disappears， the glands disappear， and a large number of inflammatory cells infiltrate and gather in the mucosal layer and the base layer in the pathological sections of the model group. The expression levels of PI3K， Akt， and mTOR mRNA were significantly increased （P<0.01）. The IL-1β content was significantly increased and the IL-10 content was significantly decreased （P<0.01）. The expression levels of p-PI3K，p-Akt and p-mTOR protein were significantly increased （P<0.01）. Compared with the model group， the DAI scores of Sishenwan high and medium dose groups and mesalazine group decreased （P<0.05）. The CMDI index of mesalazine group and the high， middle and low dose groups of Sishenwan was significantly reduced （P<0.01）. Pathological sections of rats showed that the inflammatory cells in the drug group decreased， and the mucosal layer structure returned to normal to varying degrees. The mesalazine group and the Sishenwan medium-dose group had the best effects， and the mucosal structure was close to the blank control group. The expression levels of PI3K， Akt， mTOR mRNA in the high and medium dose groups of Sishenwan， mesalazine group and Akt mRNA in low dose group of Sishenwan were significantly reduced （P<0.05，P<0.01）. The expression levels of PI3K and mTOR mRNA in low-dose group of Sishenwan decreased， but the difference was not statistically significant. The IL-1β content was significantly reduced and the IL-10 content was significantly increased in high， medium dose groups of Sishenwan and mesalazine groups （P<0.05，P<0.01）. The level of IL-1β decreased and the level of IL-10 increased in the low-dose group of Sishenwan， but the difference was not statistically significant. The expression levels of p-PI3K， p-Akt and p-mTOR protein in the high， medium， and low dose groups of Sishenwan and mesalazine group decreased to varying degrees， and the differences were statistically significant（P<0.05，P<0.01）.ConclusionSishenwan has the effect of improving the general condition and intestinal mucosal damage of ulcerative colitis model rats with spleen and kidney Yang deficiency. The mechanism may be related to the inhibition of PI3K/Akt/mTOR signaling pathway.
Keywords：ulcerative colitis;warming the kidney and strengthening the spleen method;Sishenwan;phosphatidylinositol-3 kinase/protein kinase B/mammalian target of rapamycin signaling pathway
Abstract：ObjectiveTo observe the effect of Guizhitang （GZT） on peripheral blood monocytes， intestinal flora and AS plaque formation of ApoE-/- mice induced by Western diet （WD）.MethodIn this study， 40 12-week-old homozygous female ApoE-/- mice were randomly divided into chow diet （CD） group （ApoE-/-+CD）， WD group （ApoE-/-+WD）， GZT group （ApoE-/-+WD+GZT， 7.83 g·kg-1） and atorvastatin （Atr） group （ApoE-/-+WD+Atr， 3.33 mg·kg-1）. And 10 matched C57BL/6 mice were set as wild CD control group （C57BL/6+CD）. Except the CD group， the rest groups were given WD to induce models. Treatment groups were given Guizhitang or atorvastatin orally in addition to WD， while ApoE-/-+CD and ApoE-/-+WD model groups were treated with the same volume of double steam water at the same time. After 4 weeks of intervention， 5 mice in each group were selected to collect the eyeball blood samples. The levels of plasma lipids were detected by automatic biochemical analyzer， and the proportion of peripheral blood mononuclear cells and its subtypes， and the expression levels of surface receptors toll like receptor 4 （TLR4） and CD36 were detected by flow cytometry， the intestinal flora of mice was detected by 16S rDNA sequencing. The remaining 5 mice in each group were intervened for 12 weeks， and the aorta was taken to detect the formation of aortic plaque by oil red O staining.ResultAfter intervention for 4 week， compared with C57BL/6+CD group， the levels of plasma total cholesterol （TC） and low-density lipoprotein （LDL） levels in ApoE-/-+CD and ApoE-/-+WD groups were increased （P<0.01）. ApoE-/-+WD group showed increase in the proportion of monocytes， their inflammatory subtypes Ly6C， and TLR4 expression on monocyte surface in blood （P<0.05）. ApoE-/-+WD group induced the imbalance of intestinal flora， with increase of Firmicutes and decrease of Verrucomicrobia in ileum of ApoE-/- mice. Compared with ApoE-/-+WD group， there was no significant change in blood lipid level and monocyte proportion in ApoE-/-+WD+GZT group， but with decrease in the proportion of Ly6C， increase in the proportion of anti-inflammatory subtype Ly6C-， and decrease in the expression of TLR4 and CD36 on monocyte surface （P<0.05）. ApoE-/-+WD+GZT group showed decrease of Firmicutes and increase of Bacteroidetes and Verrucomicrobia in ileum of ApoE-/- mice. After 12 weeks of intervention， ApoE-/-+WD group showed increase in the number and area of aortic plaques in ApoE-/- mice. ApoE-/-+WD+GZT group showed decrease of the area of aortic AS plaques.ConclusionGZT can reduce the immune damage and imbalance of intestinal flora caused by WD， then inhibit the formation of AS plaque.
Abstract：ObjectiveThe purpose of this article was to observe the effect of modified Shengjiangsan on podocyte apoptosis in membranous nephropathy （MN） rats， to explore the molecular mechanism of its treatment of MN and to provide experimental basis for its clinical application.MethodThe MN rat model was established by injection of cationic bovine serum albumin into the tail vein of rats. The successfully modeled rats were then randomly divided into model group （equal volume of normal saline）， modified Shengjiangsan group （27.3 g·kg-1） and benazepril group （10 mg·kg-1）， with corresponding drug dosage once a day for 4 weeks of continuous intervention. After drug administration， the 24-hour urine protein （UTP） was detected. Real time fluorescent quantitative polymerase chain reaction （Real-time PCR） and immunohistochemical （IHC） methods were used to detect Podocalyxin， Nephrin， Podocin， Synaptopodin mRNA and protein expression levels in rat kidney tissue. terninal-deoxynucleoitidyl transferase medsated nick and labeling （TUNEL） method was used to detect cell apoptosis rate in rat kidney tissue， and Western blot was used to detect Notch1， Hes1， B lymphoblastoma-2 （Bcl-2） associated X protein （Bax）， and Bcl-2 protein expression levels in rat kidney tissue.ResultCompared with the normal group， UTP in the model group increased significantly， renal tissue cell apoptosis increased significantly， podocyte marker proteins podocalyxin， Nephrin， Podocin， Synaptopodin mRNA and protein expression levels decreased significantly， and Notch1， Hes1， Bax protein expression increased significantly， and Bcl-2 protein expression was significantly reduced（P<0.05）. Compared with the model group， UTP levels in MN rats were significantly reduced in modified Shengjiangsan and benazepril groups， with reduced rate of renal cell apoptosis， increased mRNA and protein expression levels of podocalyxin， Nephrin， Podocin， and Synaptopodin in renal tissue， decreased Notch1， Hes1， Bax protein expression， and increased Bcl-2 protein expression（P<0.05）.ConclusionModified Shengjiangsan can inhibit the Notch signaling pathway， reduce the apoptosis of rat kidney tissue podocytes， and reduce the kidney injury of MN rats.
Keywords：modified Shengjiangsan;membranous nephropathy;podocyte;podocyte marker;apoptosis;notch signal pathway
Abstract：ObjectiveThis studu aims to investigate the effect of aqueous extract of modified Xiao Xianxiongtang on the epithelial mesenchymal transition（EMT） and the change of its invasion and migration ability of human gastric cancer MGC-803 cells mediated by transforming growth factor-β1（TGF-β1），and to explore the mechanism of regulating Wnt5a/Ca2+/ activated T-cell nuclear factor（NFAT） signaling pathway to inhibit EMT and invasion and metastasis of MGC-803 cells.MethodTGF-β1（10 μg·L-1）was used to induce EMT and the invasion and metastasis model of human gastric cancer MGC-803 cells. Transwell chamber experiment， scratchhealing experiment， Western blot and immunofluorescence assay were used to detect cell invasion and migration ability， expression of EMT marker protein and key protein expression of Wnt5a/Ca2+/NFAT signaling pathway， and intracellular Ca2+ concentration.ResultCompared with the blank group， TGF-β1 could significantly enhance the invasion and migration ability of MGC-803 cells（P<0.01）， down-regulate the level of E-cadherin（P<0.01）， up-regulate protein expressions of N-cadherin， Snail and Vimentin（P<0.01）， and induce cell Wnt5a， calcineurin （CaN）， total protein of activated T-cell nuclear factor 1（NFAT1）， up-regulation of phosphorylated proteins p-NFAT1 and NFAT1 nucleoprotein and intracellular accumulation of Ca2+（P<0.01）. Compared with the TGF-β1 group， modified Xiao Xianxiongtang （10， 20， 40 mg·L-1） could significantly inhibit this phenomenon，and 40 mg·L-1 had the best effect（P<0.05，P<0.01）.The specific inhibitors of Wnt5a/Ca2+/NFAT signaling pathway （R）-（+）-Bay-K-8644 and modified Xiao Xianxiongtang （40 mg·L-1） could significantly inhibit theinvasion and migration of MGC-803 cells mediated by TGF-β1， up-regulate the level of E-cadherin， and down-regulate expressions of N-cadherin， Snail， Vimentin， Wnt5a， CaN and NFAT1 proteins and reduce the intracellular accumulation of Ca2+（P<0.05，P<0.01）.Moreover， （R）-（+）-Bay-K-8644 combined with modified Xiao Xianxiongtang （40 mg·L-1） had stronger inhibitory effect（P<0.05，P<0.01）.ConclusionThese results suggest that modified Xiao Xianxiongtang can inhibit the EMT mediated by TGF-β1 via Wnt5a/Ca2+/NFAT signaling pathway，thereby reducing the invasion and migration ability of MGC-803 cells.
Keywords：aqueous extract from modified Xiao Xianxiongtang;human gastric cancer MGC-803 cells;epithelial-mesenchymal transition;invasion and migration;Wnt5a/Ca2+/NFAT signaling pathway
Abstract：ObjectiveTo investigate the mechanism of Jianzhong Bushen Xiaozheng decoction in regulating the effect of miRNA139 on Wnt/β-catenin signaling pathway for renal interstitial fibrosis.MethodThe 120 mice were randomly divided into sham operation group， unilateral ureteral obstruction （UUO） group， Jianzhong Bushen Xiaozheng decoction low， middle， high dose group， and Niaoduqing group. The UUO animal model was established to observe the morphological changes in mice. Intragastic administration was started from day 3 after modeling. The sham operation group and UUO group received the same amount of distilled water every day. The low， medium and high-dose groups received Jianzhong Bushen Xiaozheng decoction solution at 6，12，24 g·kg-1·d-1 respectively. The Niaoduqing group received 6.2 g·kg-1·d-1 Niaoduqing granule solution. After 14 d and 21 d， 28 d ， the morphological changes， general signs and renal interstitial fibrosis index of the obstructed side were observed， hematoxylin-eosin （HE） staining and Masson staining were used to observe the pathological changes of renal tissue. Real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） method was used to detect the miRNA-139 expression in renal tissue volume， Western blot was used to detect expression of beta serial proteins （β-catenin） and fibrinolytic enzyme activators inhibitor-1 （PAI-1） in renal tissues， and immunohistochemical assay was used for detection of matrix metalloproteinases-7 （MMP-7） protein expression at the obstruction side.ResultAfter 14， 21 and 28 days， the expression levels of β-catenin and PAI-1 in UUO group were higher than those in sham operation group（P<0.05），while the expression levels of miRNA139 and MMP-7 protein were lower than those of sham operation group （P<0.05）. The expression levels of β-catenin and PAI-1 proteins in mice after treatment in Niaoduqing group and the traditional Chinese medicine groups were lower than those in the UUO group（P<0.05）， the expression of miRNA139 and MMP-7 proteins increased（P<0.05）， and the efficacy of high-dose Jianzhong Bushen Xiaozheng decoction group was better than that of other dosage groups or Niaoduqing group（P<0.05）.ConclusionJianzhong Bushen Xiaozheng decoction may regulate miRNA139 to mediate the process of renal interstitial fibrosis through the Wnt/ β-catenin pathway and delay the development of renal interstitial fibrosis to improve renal function.
Keywords：miRNA-139;Wnt/β-catenin signal pathway;renal interstitial fibrosis;Jianzhong Bushen Xiaozheng decoction;experimental study
Abstract：ObjectiveTo confirm the protective effect of Xiangsha Yuyang decoction on acetic acid-induced gastric ulcer model rats and explore its mechanism， so as to provide experimental basis for clinical drug use.MethodThe 60 SPF Wistar rats were randomly divided into 6 groups： group， model group， high， middle and low dose groups of Xiangsha Yuyang decoction and omeprazole control group. The rat model of gastric ulcer was induced by acetic acid. The rats in the high， middle and low dose groups of Xiangsha Yuyang decoction were intragastrically administered at the dose of 28，14，7 g·kg-1， and with omeprazole at the dose of 4.17 g·kg-1in normal saline， respectively. The rats in the blank group and model group were intragastrically infused with the same volume of normal saline once a day. After 14 days of continuous treatment， the rats were killed， the blood was collected， the area and inhibition rate of gastric ulcer were measured and calculated， the histopathological sections of gastric mucosa were made and the state of gastric mucosal injury was observed， and the changes of gastric mucosal repair factor， gastric tissue related protein， oxidative stress factor and inflammatory factor in serum were detected by enzyme-linked immunosorbent assay（ELISA）. Detected the expression of p62 Kelch-like epichlorohydrin-related protein 1 （Keap1）， nuclear transcription factor E2 related factor 2 （Nrf2） and heme oxygenase-1 （HO-1） signal pathway-related proteins in gastric mucosa by Western blot.ResultCompared with control group， the gastric mucosa of the model group showed obvious pathological changes and a large number of leukocytes infiltrated. In model group， the ulcer area was significantly increased（P<0.01）， the contents of mucin mucoprotein 5AC （MUC5AC）， epidermal growth factor （EGF）， superoxide dismutase （SOD） and increased prostaglandin E2 （PGE2） were significantly decreased（P<0.01）， the gastrin （GAS）， 8-hydroxydeoxyguanosine （8-OHdG）， malondialdehyde （MDA）， tumor necrosis factor-α （TNF-α）， cyclooxygenase-2 （COX-2） were significantly increased. The expression of HO-1 and Nrf2 protein decreased significantly（P<0.01）， the content of Keap1 increased significantly （P<0.05）， and the expression of p62 protein decreased. Compared with model group， the hierarchical structure of cells in Xiangsha Yuyang decoction high dose group and omeprazole group were clearer and regular， middle and low dose groups could also repair gastric mucosa to a certain extent. The high and middle dose groups of Xiangsha Yuyang decoction could significantly reduce the gastric ulcer area of acetic acid-induced gastric ulcer rat model （P<0.01） and increase the ulcer inhibition rate. It can effectively promote the expression of MUC5AC and EGF in gastric mucosa， decrease the level of GAS（P<0.05，P<0.01）， decrease the level of 8-OHdG and MDA， increase the activity of SOD（P<0.01）， decrease the expression level of TNF-α and COX-2， increase the content of PGE2， and significantly increase the amount of Nrf2 and HO-1 protein in gastric mucosa（P<0.01）. The high dose group of Xiangsha Yuyang decoction could decrease the protein expression of Keap1（P<0.05） and increase the expression of p62 protein.ConclusionXiangsha Yuyang decoction is effective in the treatment of acetic acid-induced gastric ulcer model rats， which can effectively reduce the ulcer area， increase the ulcer inhibition rate and protect the ulcer tissue. Its mechanism may be related to activating p62/Keap1/Nrf2 signal pathway and regulating the expression of related genes so as to improve inflammatory response and regulate oxidative stress response.
Keywords：gastric ulcer;Xiangsha Yuyang decoction;inflammatory factors;oxidative stress;nuclear factor E2 related factor 2 （Nrf2）;Kelch-like epichlorohydrin-related protein 1 （Keap1）;malondialdehyde（MDA）;superoxide dismutase （SOD）
Abstract：ObjectiveTo observe the effect of tetrahydroxy stilbene glycoside （TSG） on the expression of glycogen synthase kinase 3β （GSK3β）， cyclic adenosine monophosphate-dependent protein kinase （PKA） and Serine/threonine phosphatase 2A（PP2A） in the brain of amyloid precursor protein/presenilin-1/Tau （APP/PS1/Tau） triple-transgenic mice dementia model.MethodA total of forty-five 8-month-old APP/PS1/Tau transgenic mice were randomly divided into model group， positive control group （Huperzine-A， 0.15 mg·kg-1）， low， medium and high dose TSG groups （TSG， 0.033，0.1，0.3 g·kg-1）， with 9 mice in each group， and another nine C5B7L/6J mice of the same age were selected as normal control group. After 60 days of intragastric administration， the general structure of hippocampal neurons was observed by hematoxylin-eosin （HE） staining， immunohistochemical （IHC） was used to detect the expression of PKA protein in the brain of mice in each group， the mRNA expression levels of GSK3β， PKA and PP2A were detected by real time quantitative reverse transcription polymerase chain reaction （Real-time PCR）， and protein expression levels of GSK3β and PP2A were detected by Western blot.ResultCompared with the normal control group， the apoptosis level of neurons in the model group was significantly increased， the protein and mRNA expression levels of GSK3β and PKA were significantly increased （P<0.05， P<0.01）， and the protein and mRNA expression levels of PP2A were significantly decreased （P<0.05， P<0.01）. Compared with the model group， the apoptosis level of neurons in each treatment group was significantly down-regulated， the protein and mRNA expression levels of GSK3β and PKA were significantly down-regulated （P<0.05， P<0.01）， and the protein and mRNA expression levels of PP2A were significantly increased （P<0.05， P<0.01）.ConclusionThe mechanism of TSG in the treatment of Alzheimer's disease （AD） may be related to lowering the transcription and expression of GSK3β and PKA， increasing the transcription and expression of PP2A.
Abstract：ObjectiveThis study aims to observe the effect of baicalein on the clonal formation of triple negative breast cancer MDA-MB-231 and MDA-MB-468 cells， and to explore the mediation role of Yes- related protein （YAP） in it.MethodMDA-MB-231 and MDA-MB-468 cells were treated with baicalein. Thiazole blue （MTT） colorimetric method was used to detect cell proliferation ability. Plate cloning experiments was used to detect the colony forming ability. Immunofluorescence method was used to detect the nuclear distribution of YAP， and Western blot test was used to detect the protein expression levels of YAP large tumor suppressor factor 1 （LATS1）， YAP， phosphorylated Yes- related protein（p-YAP） and phosphorylated YAP large tumor suppressor factor 1 （p-LATS1）.ResultCompared with the blank group， baicalein （40， 80， 160 μmol·L-1） significantly inhibited the proliferation ability of MDA-MB-468 and MDA-MB-231 cells （P<0.05， P<0.01）， and the inhibitory effect was dose-dependent. The half inhibit concentration（IC50） of baicalein against MDA-MB-468 and MDA-MB-231 cells were （80.3±7.2），（70.4±6.5） μmol·L-1， respectively. Compared with blank group， baicalein （5， 10， 20 μmol·L-1） had no significant effect on the proliferation of MDA-MB-468 and MDA-MB-231 cells， and the difference was not statistically significant. Compared with the blank group， baicalein （5， 10， 20 μmol·L-1） significantly dose-dependently reduced the cell colony formation rates of MDA-MB-468 and MDA-MB-231 cells （P<0.05， P<0.01）， and baicalein （10， 20 μmol·L-1） significantly inhibited the nuclear expression of YAP in MDA-MB-468 and MDA-MB-231 cells in a dose-dependent manner（P<0.01）. Also， baicalin （5， 10， 20 μmol·L-1） significantly up-regulated p-YAP and p-LATS1 protein expressions in MDA-MB-468 cells in a dose-dependent manner （P<0.05， P<0.01）. Baicalein （10， 20 μmol·L-1） significantly up-regulated p-YAP and p-LATS1 protein expressions in MDA-MB-231 cells in a dose-dependent manner （P<0.01）.ConclusionBaicalein can inhibit colony formation of triple negative breast cancer MDA-MB-468 and MDA-MB-231 cells by mediating the reduction of YAP entry into the nucleus.
Keywords：baicalein;Yes-related protein;proliferation;triple-negative breast cancer;mechanism
Abstract：ObjectiveTo explore the clinical efficacy and mechanism of Quyu Qingjintang in the treatment of stroke complicated with lung infection （SCLI） with phlegm-heat accumulation lung syndrome and blood stasis syndrome.MethodThe 60 patients with SCLI with phlegm-heat accumulation lung syndrome and blood stasis syndrome were selected and divided into control group （30 cases） and observation group （30 cases）. The patients in both groups received basic treatments such as thrombolysis and anticoagulation， and were injected with imipenem cilastatin sodium and ambroxol hydrochloride. The patients in control group additionally received Tanreqing capsule on the basis of the conventional treatment while those in observation group were additionally treated with Quyu Qingjintang on the basis of the conventional treatment. The clinical efficacy， lung function， inflammatory factor levels， time to symptoms disappearance， haptoglobin （HPT） and amyloid A （SAA） levels， T lymphocyte subset level and safety index were compared between two groups.ResultThe total effective rate was 93.33% （28/30） in observation group， significantly higher than 70.00% （21/30） in control group （χ2=5.450， P<0.05）. After treatment， the symptoms of both groups were improved （P<0.05）. The scores of fever， cough， wheezing， and expectoration in observation group were lower than those in control group （P<0.05）. The levels of interleukin-6（IL-6）， white blood cell（WBC）， procalcitonin（PCT）， C-reactive protein（CRP）， HPT， and SAA in observation group were significantly lower than those in control group （P<0.05）. The forced expiratory volume in one second （FEV1）， forced vital capacity （FVC） and peak expiratory flow rate （PEF） in observation group were better than those in control group （P<0.05）. The levels of CD3+， CD4+， CD4+/CD8+ in observation group were significantly higher than those in control group （P<0.05）. The recovery time of WBC count， hospitalization time， lung rales， the time to fever and cough disappearance in observation group were lower than those in control group （P<0.05）， no serious adverse reactions occurred in two groups.ConclusionQuyu Qingjintang in the treatment of stroke complicated with lung infection with phlegm-heat accumulation lung syndrome and blood stasis syndrome can significantly improve the treatment efficiency， improve the symptoms of pulmonary infection， reduce the level of inflammatory factors， and improve lung function， with less adverse reactions and high safety， so it is worthy of clinical application.
Abstract：ObjectiveTo evaluate the efficacy of addition and subtraction therapy of Huaganjian combined with Jinlingzisan for bile reflux gastritis （BRG） with stagnancy heat of liver and stomach syndrome， and to investigate its effect on inflammatory factors and gastrointestinal hormones.MethodOne hundred and fifty patients were divided into control group and observation group evenly according to random number table. The 68 patients in control group finished the treatment （5 cases of dropout， loss of follow-up and 2 cases of withdrawal）， and 69 patients in observation group completed the treatment （3 cases of dropout， loss of follow-up and 3 cases of withdrawal）. Patients in control group got oral Dalitong granules before the meal， 1 bag/time， 3 times/day. Patients in observation group got addition and subtraction therapy of Huaganjian combined with Jinlingzisan， 1 dose/day. The treatment continued for 4 weeks in both groups. Before the treatment， gastroscope was used to evaluate the degree of bile reflux and the condition of mucosa under gastroscope. Before and after treatment， scores of stagnancy heat of liver and stomach syndrome， patient reported outcome （PRO） scale of chronic gastrointestinal diseases， self rating anxiety scale （SAS）， and self rating depression scale （SDS） were recorded. A follow-up of 6 months was conducted to record the clinical recurrence. Levels of gastrin （GAS）， motilin （MTL）， cholecystokinin （CCK）， prostaglandin E2 （PGE2）， tumor necrosis factor-α （TNF-α）， and interleukin-8 （IL-8） were detected both before and after treatment. In addition， the safety was discussed.ResultScores of degree of bile reflux， mucosa under gastroscope and stagnancy heat of liver and stomach syndrome in the observation group were lower than those in control group （P<0.01）. Scores of 6 dimensions in PRO scale （dyspepsia， reflux， defecation， social， psychological， and general state） and scores of SAS and SDS in the observation group were lower than those in control group （P<0.01）. Levels of CCK， TNF-α and IL-8 in the observation group were all lower than those detected in control group （P<0.01）， while levels of GAS and MTL were higher than those in control group （P<0.01）. Observation group was superior to the control group in terms of efficacy for traditional Chinese medicine（TCM） syndrome and efficacy under gastroscope （Z=2.083， P<0.05； Z=2.104， P<0.05）. Clinical recurrence rate in observation group was 20.37% （11/54）， lower than 40.82% （20/49） in control group （=5.105， P<0.05）. No adverse reaction was found after oral administrationh of TCM.ConclusionAddition and subtraction therapy of Huaganjian combined with Jinlingzisan can ameliorate the clinical symptoms， relieve anxiety and depression， improve the quality of life in patients with BRG and stagnancy heat of liver and stomach syndrome， improve the degree of bile reflux， promote the healing of gastric mucosa， reduce inflammatory reaction and regulate gastrointestinal hormones. It has good efficacy for TCM syndrome and under gastroscopy in a short term， and can reduce the recurrence rate with high safety in a long term.
Keywords：bile reflux gastritis;stagnancy heat of liver and stomach syndrome;Huaganjian;Jinlingzisan;recurrence;inflammatory factors;gastrointestinal hormones
Abstract：ObjectiveTo observe the effect of different doses of Fufang Huangbaiye Tuji asin the treatment onof the inflammatory response in healing process for of skin with deep Ⅱ degree burn. Methods in healing process.MethodsThe 120 cses patients with deep Ⅱ degree burn of fire-toxin injuring fluid syndrome diagnosed in the affiliated hospital of Chengde Medical University between June 2019 and March 2020 were randomly divided into control group，low -dose treatment group and high -dose treatment group，with 40 cases in each group and once. They got a dressing change perevery day. Control group was locally administered with lodophor solution 35 mL per 1% on the body surface area. Low-dose treatment group was locally administered with compound cortex phellodendri fluid 17.5 mL per 1% on the body surface area，while high-dose treatment group was locally administered with compound cortex phellodendri fluid 35 mL per 1% on the body surface area. Observe theThe inflammatory reaction of wound surface in each group onwas observed at admission and after treatment. The pathological changes of each groupsgroup were observed， and determination of nuclear factor kappa-B（NF-κB） p65 expression inon the wound surface was determined by immunohistochemistry on the 4th day after the treatment. The levels of interleukin（IL）-2，IL-8 and tumor necrosis factor（TNF）-α in wound tissue were measured with ELISA and Bacterial culture and count were performed in each group on the 4th，10th and 21st daydays after treatment. The levels of IL-2，IL-8 and TNF-α in wound tissue were measured with ELISA.ResultsThere was no significant difference in the degree of wound inflammation in each group at admission，and the degree of relief after treatment was positively correlated with the treatment time. At the simultaneous phase point，the inflammatory reaction was severest in control group，which was followed by low-dose treatment group and high-dose treatment group. Bacterial growth were observed on the 4th day in control group，which was found in low-dose and high-dose treatment groups on the 10th day，the detection rates of Staphylococcus aureus and Pseudomonas aeruginosa were the highest. Compared with control group，the mean integrated optical density of NF-κB p65 in wound tissue decreased markedly in low-dose and high-dose treatment groups on the 4th day after treatment（P<0.05），the bacterial count decreased significantly in low-dose and high-dose treatment groups on the 10th and 21st days after treatment（P<0.05），and the levels of IL-2，IL-8 and TNF-α in wound tissue decreased markedly in low-dose and high-dose treatment groups on the 4th，10th and 21st days after treatment（P<0.05），with statistically significant differences between low-dose and high-dose treatment groups（P<0.05）. Histopathological examination showed that inflammatory granulocytes and edema were improved in low-dose and high-dose treatment groups compared with control group，with a more significant performance in high-dose treatment group.ConclusionThe external application of compound cortex phellodendri fluid can reduce thebacterial growth of bacteria in on the wound surface，which may reduce the inflammatory reaction by inhibiting the production and release of inflammatory mediators，with a certain dose-effcteffect relationship，and is worth clinical promotion.
Abstract：ObjectiveTo observe the clinical efficacy of Qingzhenfang for plasmoby （chronic urticaria）， and to investigate its effect on cellular immune function.MethodOne hundred and thirty-two cases patients were divided into control group and observation group evenly according to random number table. The 60 patients in control group finished the study because of 6 cases of dropout， loss of follow-up and withdrawal， and 62 patients in observation group finished the study. Patients in both groups got Yiebastine tablets， 10-20 mg/time， 1 time/day. Patients in control group additionally got Piminxiao capsule， 4 grains/time， 3 times/day， while patients in observation additionally got Qingzhenfang， 1 dose/day. The treatment continued for 8 weeks in both groups. Before the treatment， and at the second， fourth， and eighth week after treatment， scores of urticaria activity for 7 days （USA7） and total symptom score （TSS） were graded. Before and after treatment， scores of chronic urticaria quality of life scale （CU-Q2oL） and syndrome of rheumatic fever were graded. A follow-up of 3 months was conducted for the patients whose score of USA7 was less than 7 to record the recurrence. Complement 3 and 4 （C3， C4）， CD4+， CD8+ cells were detected， and Th17/ CD4+ and Treg/ CD4+， CD4+/CD8+ and Th17/Treg were calculated. Levels of peripheral blood interleukin-10 （IL-10）， IL-17 and IL-23 were detected， and safety was evaluated after the treatment.ResultAt the second， fourth and eighth week after the treatment， scores of USA7， TSS， CU-Q2oL and syndrome of rheumatic fever in observation group were lower than those in the control group （P<0.01）. Levels of C3， C4， CD4+， Treg， CD4+/CD8+and IL-35 in observation group were higher than those levels detected in control group （P<0.01）， while levels of CD8+， Th17， Th17/Treg， IL-10， IL-17 and IL-23 were lower than those in the control group （P<0.01）. Recurrence rate was 25.58% （11/43） in observation group， lower than 48.48% （16/33） in control group （=4.276， P<0.05）， and the clinical efficacy in observation group was superior to that in control group （Z=2.021， P<0.05）.ConclusionYaoyi Qingzhenfang can control the degree of disease and improve the quality of life for patients with chronic urticaria， with superior clinical efficacy. In addition， it can reduce recurrence rate， increase the levels of C3， C4， regulate cellular immune function， and reduce immune inflammatory response， so it is worthy of further clinical research and use.
Abstract：ObjectiveTo screen qualitative preparation quality markers of Yuliantang， in order to provide data support for the selection of indicator components， and establish the direct connection between indicator components and efficacy （Xiehuo Zhitong） for achieving the quantity-effect combination.MethodThe stability of preparation process of Yuliantang lyophilized powder was investigated by HPLC fingerprint technology， then， the components in Yuliantang lyophilized powder were identified by UHPLC-LTQ-Orbitrap-MS. By referring to the relevant literature， the pharmacological activities of these identified compounds were compared with the pharmacological effects corresponding to the efficacy of Yuliantang， and the composition of the qualitative preparation quality markers of Yuliantang lyophilized powder was determined.ResultThe similarities between HPLC fingerprint of 10 batches of Yuliantang lyophilized powder and the control fingerprint were >0.9， indicating that the preparation process was stable and feasible. A total of 29 components were identified from Yuliantang， of which 23 alkaloids， 3 phenylpropanoids， 2 sesquiterpenoids and 1 limonoid， and there were 15 ingredients of Coptidis Rhizoma， 12 ingredients of Euodiae Fructus， and 2 ingredients of Aucklandiae Radix. The composition of the qualitative preparation quality markers of Yuliantang was initially determined as magnoflorine or 10-hydroxy-2，3，9-trimethoxyberberine， phellodendrine， menisperine， thalifendine， groenlandicine， dehydroevodiamine， coptisine， jatrorrhizine， columbamine， methylcoptisine， berberine， epiberberine， palmatine， evodiamine， rutaecarpine， neochlorogenic acid， cryptochlorogenic acid， chlorogenic acid， limonin， costunolide， dehydrocostus lactone.ConclusionThe method for researching and screening the preparation quality markers in Yuliantang lyophilized powder is scientific， reasonable and feasible， it can provide reference for the determination of component indicators in the process research of Yuliantang and qualitative and quantitative indexes in its quality standard.
Abstract：ObjectiveTo quickly analyze and identify the components in raw and wine-processed products of Polygonatum cyrtonema （PC） dried rhizomes by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF-MS）， and then find out the differential components before and after processing.MethodThe ACQUITY UPLC BEH C18 column （2.1 mm×100 mm， 1.8 μm） was used with 0.1% formic acid aqueous solution-acetonitrile as the mobile phase for gradient elution at a flow rate of 0.25 mL·min-1. Electrospray ionization was selected for collection and detection in positive and negative ion modes， and the data were analyzed by PeakView 22.214.171.124. According to the retention time， accurate relative molecular weight and fragmentation ion information provided by MS， and combined with the reference substance and literature， the components were identified. After normalized treatment， the MS data of each sample were analyzed with principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA）， and then the differential components before and after processing were screened according to the principle that variable importance in the projection （VIP） value was >1.ResultA total of 38 components were identified from raw and wine-processed products of PC dried rhizomes， including 15 steroidal saponins， 6 alkaloids， 3 flavonoids， 2 amino acids， 2 organic acids and 10 others. The results of PCA and OPLS-DA showed that there were significant differences in the contents of components in PC dried rhizomes before and after processing， and 16 differential components such as kingianoside Z， disporopsin and linoleic acid were screened.ConclusionUPLC-Q-TOF-MS technique can accurately and comprehensively identify the components in PC dried rhizomes， these components are mainly steroidal saponins， flavonoids and alkaloids. It takes a great difference in the contents of components before and after processing， and transformation of the same category components is the main reason for the differences of raw and wine-processed products， which will provide reference for the researches on material basis and processing chemistry of PC dried rhizomes.
Keywords：Polygonatum cyrtonema;traditional Chinese medicine;processing;chemical compositions;raw products;wine-processed products;ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF-MS）
Abstract：ObjectivePolygalae Radix is mainly produced in Northeast，North，Northwest and Central China and Sichuan province. There are obvious differences in quality of Polygala tenuifolia from different areas. P. tenuifolia cultivation areas are relatively concentrated. Currently，there are only a few studies about the suitability zoning of P. tenuifolia nationwide，in order to determine the suitable zoning of P. tenuifolia in China. This paper made a habitat suitability evaluation of P. tenuifolia nationwide based on analysis of ecological factors in distribution areas of sampling points.MethodMaxEnt model was used to select ecological factors that affected the distribution of P. tenuifolia and construct an evaluation model. ArcGIS software was used to evaluate the suitability of different habitats of P. tenuifolia. The suitable areas were classified into high，medium，low suitability areas and unsuitable area of P. tenuifolia nationwide.ResultThe evaluation model was highly accurate，and concluded that P. tenuifolia's suitability distribution area in China was 3.21×106 km2，including 0.52×106 km2 of high suitability area，0.96×106 km2 of medium suitability area and 1.73×106 km2 of low suitability area. High suitability areas were mainly distributed in Shanxi province，Hebei province，Shandong province，Shaanxi province，Liaoning province and Henan province. The main ecological factors affecting the distribution of P. tenuifolia were annual average temperature change range，mean temperature of driest quarter，precipitation of wettest quarter，altitude，slope and topsoil （0-30 cm） calcium carbonate.ConclusionThe results of the study are consistent with the records in relevant literatures，and can provide a theoretical basis for protection and cultivation of Chinese medicine resource P. tenuifolia.
Abstract：ObjectiveTo establish a high performance liquid chromatography （HPLC） fingerprint of the substance benchmark of Xiao Chengqitang and evaluate its quality with chemical pattern recognition method.MethodDiamonsil C18 column （4.6 mm×150 mm， 5 μm） was used， mobile phase was consisted of methanol （A）-0.1% phosphoric acid solution （B） for gradient elution （0-60 min， 20%-90%A； 60-70 min， 90%-100%A）， the flow rate was 1 mL·min-1， the column temperature was 25 ℃， and the detection wavelength was 254 nm. The similarity evaluation system of chromatographic fingerprint of traditional Chinese medicine （2012 edition） was used to evaluate the similarity of HPLC fingerprint of 15 batches of substance benchmark of Xiao Chengqitang， and the chromatographic data were analyzed by cluster analysis， principal component analysis and orthogonal partial least squares-discriminant analysis， in order to evaluate the quality difference between different batches of substance benchmarks of Xiao Chengqitang and find out the main chemical components that caused the quality difference.ResultThe HPLC fingerprint of Xiao Chengqitang substance benchmarks was established， 31 common peaks were identified， and 18 components were identified by comparing with the reference substances. The similarities of 15 batches of HPLC fingerprint of Xiao Chengqitang substance benchmarks were >0.92. The samples could be divided into two categories by three chemical pattern recognition methods. Nine main components leading to the quality discrepancy of samples between batches were screened out， including rhein， chrysophanol-8-O-β-D-glucoside， aloe-emodin-8-O-β-D-glucoside， sennoside A， chrysophanol-1-O-β-D-glucoside， rhein-8-O-glucoside and others.ConclusionThe established fingerprint analysis method is accurate， stable and reproducible， which basically reflects the overall chemical composition characteristics of Xiao Chengqitang， and can be used for the quality control of Xiao Chengqitang preparations.
Abstract：ObjectiveTo screen out stable internal reference genes suitable for real-time quantitative polymerase chain reaction（Real-time PCR） analysis of different parts of Cinnamomum cassia and C. cassia var. macrophyllum，in order to provide stable internal reference genes for gene expression analysis of three different parts of and C. cassia var. macrophyllum branches and leaves.MethodWith 6 different tissues and organs， such as bark，branches and leaves of two plants of C. cassia and C. cassia var. macrophyllum as experimental materials，Real-time PCR technology was used to detect the five internal reference genes， namely glyceraldehyde-3-phosphate dehydrogenase（GAPDH），actin，ubiquitin-ligase enzymes（UBE），histone and tubin（TUB）. The analysis of the expression of the data. Furthermore， three commonly used internal reference gene analysis software，namely geNorm，NormFinder and BestKeeper，was used to analyze and evaluate the stability of the candidate internal reference gene.ResultThe internal five reference genes were expressed in the bark，branches and leaves of the two plants，but with differences in stability. Comprehensive analysis showed that the expression stability of candidate internal reference genes was in the order of GAPDH>actin>UBE>histone>TUB. The internal reference genes of the two plants were analyzed separately，and the optimal internal reference gene was still GAPDH，indicating that GAPDH was the most suitable internal reference gene. TUB and histone ranked low in the three software，and should be eliminated in the screening of reference genes. They were not suitable for gene expression analysis of C. cassia and C. cassia var. macrophyllum.ConclusionThe most suitable internal reference gene for different parts of cinnamon，branches，and leaves of C. cassia and C. cassia var. macrophyllum was GAPDH. In this study，a screening system for internal reference genes of Real-time PCR of C. cassia and C. cassia var. macrophyllum was established to provide theoretical basis for studying functional regulation and expression of genes during the accumulation of effective components in different parts.
Keywords：quantitative real-time polymerase chain reaction;Cinnamomum cassia;C. cassia var. macrophyllum;reference gene
Abstract：ObjectiveTo establish the grading standard of Curcuma phaeocaulis seed rhizoma，and explore the effect of seed rhizoma classification on the growth，yield and quality of C. phaeocaulis.MethodThe purity，diameter，weight，length，germination rate and water content of C. phaeocaulis seed rhizoma samples were determined. The grading index was determined by partial correlation analysis，variance analysis and cluster analysis. According to actual conditions of production，the classification standard of C. phaeocaulis seed rhizoma was established. Then the field experiments were carried out with different grades of seed rhizoma for treatment， so as to analyze the effect of seed rhizoma grade on plant growth，yield and quality of medicinal materials.ResultWith seed rhizoma diameter and seed rhizoma weight as classification indexes，C. phaeocaulis seed rhizoma were divided into three grades. The field experiments showed that with the growth and development of C. phaeocaulis，there were significant differences in plant height and leaf length among different grades of seed rhizoma during the root tuber expansion period （P<0.05）. The yield of Curcumae Radix and Curcumae Rhizoma produced by different grades of seed rhizoma was in the order of first-class ginger>second-class ginger>third-class ginger>substandard seed rhizoma. According to the analysis of the proportion of the commodity grade of medicinal materials，the higher the grade of seed rhizoma was，the higher the proportion of the first-grade Curcumae Radix occupied. According to the analysis of the quality of medicinal materials，there was no significant difference in the content of extract，volatile oil and germanone in Curcumae Radix and Curcumae Rhizoma produced by different grades of seed rhizoma.ConclusionThe grading standard of seed rhizoma was established based on the systematic study on the classification method of C. phaeocaulis seed rhizoma. According to the field experiment， grading planting of seed rhizoma can promote plant growth and development，increase the yield of medicinal materials，and improve the proportion of first-grade medicinal materials， with no impact on the internal quality of medicinal materials. Therefore， classification of C. phaeocaulis seed rhizoma is scientific and necessary，and can lay the foundation for the standardized cultivation of C. phaeocaulis.
Keywords：Curcuma phaeocaulis;Curcumae Radix;Curcumae Rhizoma;grading of seed rhizoma;growth;yield;quality
Abstract：ObjectiveThis paper aims to study the genetic diversity of Pogostemon cablin by amplified fragment length polymorphism （AFLP） markers.MethodThe 12 pairs of primers were used for AFLP analysis of 212 samples from 14 varieties，and biological analysis software such as POPGENE 32，Arlequinver 3.5，MEGA 7 and NTSYSpc 2.10e were used for polymorphism parameter calculation，principal coordinate analysis and cluster analysis.ResultA total of 2 238 loci were amplified by 12 pairs of primers. 2 226 of them were polymorphic loci， accounting for 99.38%. At the inter-population level，the values of effective alleles（Ne），Nei's gene diversity index（H），Shannon polymorphic information index（I） were 1.365 6±0.066 3， 0.220 7±0.036 4， and 0.343 7±0.050 2，respectively；and 1.118 5±0.038 7，0.071 3±0.023 0，0.109 4±0.035 0，respectively at the intra-population level. Analysis of molecular variance（AMOVA）showed that 71.57% of the total variation of P. cablin was of inter-population nature， and 28.43% was of intra-population nature. The 14 populations could be divided into four groups by cluster analysis.ConclusionThe results of AFLP molecular markers showed that abundant genetic diversity was present at inter-population level of P. cablin，however，relatively low at intra-population level； the genetic differentiation at the inter-population level was significant，which could provide a reference for the subsequent study of good germplasm selection of P. cablin.
Keywords：Pogostemon cablin;amplified fragment length polymorphism;germplasm resources;genetic diversity
Abstract：ObjectiveTo study the distribution of ischemic stroke treatment with data mining technology and evaluate its clinical efficacy.MethodChina National Knowledge Infrastructure Database（CNKI），China Science and Technology Reader's Digest Database（VIP），Wanfang Data，Chinese Biomedical Literature Database（Sino Med）were retrieved from January 1978 to December 2018. The clinical observation and study literatures on the treatment of ischemic stroke with the combination of traditional Chinese medicine and Western medicine were retrieved in the four databases. After standardized and hierarchical collection and processing of all syndromes，treatment methods，prescriptions and other information in the literatures，a database of syndrome elements and treatment of ischemic stroke was established. Syndrome factors and treatment methods were analyzed by scale evaluation and hierarchical classification methods. Kendall's tau-b correlation analysis，principal component analysis and other statistical methods were used to describe the correlation and distribution of syndrome factors and treatment methods of ischemic stroke.ResultThe results of heterogeneity analysis showed that the included literatures were homogeneous and could be combined with subsequent statistics. A total of 450 syndromes and treatment methods were included in this study，and 1 287 single syndrome elements and 1 562 single treatment methods were obtained after unified and standardized splitting. Besides the corresponding syndrome elements and treatment methods，phlegm-dampness-invigorating Qi（-0.52） and Qi deficiency-invigorating Qi（-0.56） were also highly correlated. The study team represented the importance of syndrome and treatment elements with class Ⅰ，Ⅱ，Ⅲ from high to low. Qi deficiency，blood stasis and fire heat，phlegm，viscera excess were class Ⅰ syndrome elements；Yin deficiency，endogenous wind were class Ⅱ syndrome elements；Yin deficiency and Yang deficiency were class Ⅲ syndrome elements；Removing phlegm dampness，clearing heat，clearing the hollow viscera and extinguishing wind，promoting blood circulation to remove blood stasis，tonifying Qi were class Ⅰ treatment of ischemic stroke，and removing phlegm dampness，clearing heat，clearing the hollow viscera were more likely to appear simultaneously； and extinguishing wind，activating blood circulation and removing blood stasis，and benefiting Qi were more likely to appear simultaneously. Nourishing Yin and regulating Qi were class Ⅱ therapies of ischemic stroke，which were highly correlated and often appear simultaneously. Inducing resuscitation，tonifying Yang and dredging collaterals were class Ⅲ，Ⅳ，Ⅴ therapies.ConclusionQi deficiency，blood stasis，phlegm dampness，fire heat and viscera excess were the main syndromes of ischemic stroke，while Qi deficiency and blood stasis，phlegm heat and viscera excess were the main syndromes. Eliminating phlegm and dampness，clearing heat，clearing the hollow viscera，promoting blood circulation and removing blood stasis，extinguishing wind and benefiting Qi were the main therapies for the treatment of ischemic stroke. In clinical treatment for ischemic stroke，the therapies for relieving phlegm and dampness，clearing heat and relieving organs are often used in combination，and the therapies for promoting blood circulation and removing blood stasis were often used in combination with the therapies for invigorating Qi and extinguishing wind for the synergistic effect.
Abstract：Xiebaisan is one of the 100 classical prescriptions published by the state， and the research and development of its compound preparation has gradually become an upsurge. The research and development of classical prescriptions should start from the theory of traditional Chinese medicine and clinical practice， laying a solid foundation for research and development. Based on the above thinking， this study adopts the methods of traceability and bibliometrics to analyze the source， pathogenesis， efficacy， Fangyi and clinical application of Xiebaisan in order to further promote its literature research work. The results show that the origin of Xiebaisan can be traced back to the prescription of Xiefeitang in Yinhai Jingwei（Volume 1）. Although there are more than 20 prescriptions with the same name of Xiebaisan developed in later generations， they either reflect the inheritance and development of Qian Yi's thought of composing prescriptions， or only the meaning of "Xiebai" in the name of the prescription. The main pathogenesis of Xiebaisan is adverse lung Qi of heat. Mori Cortex and Lycii Cortex are sweet and cold， which can clear away lung heat and reduce adverse Qi， and then cough and asthma can stop. Supplemented with Glycyrrhizae Radix et Rhizoma and japonica rice， which are endowed with the meaning of nourishing earth and generating gold， to invigorate spleen and replenish Qi. The compatibility of Xiebaisan can clear away lung heat， relieve cough and asthma， and norish erath and generate gold to treat lung heat， asthma and cough. Although the main treatment of Xiebaisan recorded in ancient books involves various diseases and syndromes of internal， external， gynecology and pediatrics， the pathogenesis of Xiebaisan is "adverse lung Qi of heat". Modern clinical application of Xiebaisan focuses on the respiratory system， skin and subcutaneous tissue system， ear， nose and throat system， digestive system， ophthalmic system， etc.， and it has more advantages in the treatment of pneumonia， cough， bronchitis， epistaxis， acne， bronchiectasis， postinfectious cough， constipation and other diseases. The syndrome differentiation of the above diseases is in accordance with the relevant syndrome types with lung heat as the main pathological factor， so we can modify and apply the prescription appropriately.
Abstract：Tumors are new organisms formed by uncontrollable cell proliferation of local tissues driven by various oncogenic factors. The cause of tumors is unknown with life-threating outcome. Tumors can be classified into benign tumors， borderline tumors， and malignant tumors according to their pathological properties. Among them， malignant tumor is commonly known as cancer， with no specific medicines or reliable cure means， so this is a hot spot and difficult point in current medical research. In ancient literatures， there are many records about the efficacy of Chinese herbal medicine in treating tumor， and modern pharmacological researches have shown that more and more active ingredients of traditional Chinese medicine（TCM） have gradually highlighted their inhibitory effect on various types of tumor. Caulis sinomenii has been used for treatment of rheumatic diseases in TCM for a long history. Sinomenine is a major bioactive alkaloid presented in C. sinomenii， which has demonstrated a wide range of pharmacological activities such as anti-inflammation， immunosuppression， analgesia and sedation， and due to its slightly soluble in water， it is commonly used in clinic in the form of hydrochloride， with its commercial name of Zhengqing Fengtongning. Recent studies show that sinomenine alone or combined with chemoradiotherapy can inhibit growth of several tumors significantly or in a synergistic way， so it is termed as an inhibitor of tumors. Anti-tumor effect of sinomenine involve inhibition of tumor cell proliferation， induction of tumor cell apoptosis， blockade of tumor cell cycle， suppression of tumor invasion and metastasis， induction of autophagy of tumor cells， and reversal of multidrug resistance of tumor cells. Upon combination with nanomaterials， it can enhance efficiency and reduce toxicity. Here we summarized and reviewed recent advances on basic anti-tumor research of sinomenine， and then made a classification and description according to its in vivo and in vitro pharmacological action and mechanism of action， so as to elucidate the great potential of sinomenine as a promising anti-tumor drug， and provide reference for further research on its anti-tumor mechanism.
Keywords：sinomenine;anti-tumor;mechanisms of action;signaling pathways;apoptosis
Abstract：Asari Radix et Rhizoma （ARR） is a traditional Chinese medicine for relieving exterior syndrome， and its roots and stems contain rich chemical components， including volatile oils （terpenoids， aromatics and aliphatics）， lignans， flavonoids， etc. Clinically， it has been traditionally used for the treatment of diseases such as phlegm and cough， anemofrigid cold， rheumatic arthralgia due to its ability to spread cold. Modern pharmacological studies have shown that ARR played beneficial roles in analgesic， anti-inflammatory， antitussive， antiasthmatic， antiviral， antibacterial， sedative， antioxidative， and antidepressant responses， antihypertension， as well as tumor suppression. The current studies on the chemical composition of ARR mainly focused on volatile components， and little information is available for the occurrence and pharmacological effects of non-volatile components. In addition， there is a lack of clear classification of chemical components and the distribution of chemical components in medicinal parts and the origin of species. Therefore， in this study， the authors reviewed a large number of literature on the chemical compositions and pharmacological effects of ARR， and hoping to provide a reference for further pharmacological research and the new drug development of ARR.
Keywords：Asari Radix et Rhizoma;volatile oil;non-volatile components;pharmacological effects;pharmacodynamic material basis;anti-inflammatory;antitussive
Abstract：The global incidence of breast cancer has increased year by year. Breast cancer has the highest mortality rate in female patients with malignant tumors. Traditional Chinese medicine（TCM） has made great contribution to health of human being， improving the overall curative effect， reducing the patients' pain， improving the quality of life and alleviating adverse reactions in patients. TCM and its active compounds can inhibit the proliferation of breast cancer cells by inducing cell cycle arrest， invasion， metastasis and reversing multidrug resistance. The effect of the compounds in TCM is obvious on inducing the arrest of the breast cancer cells cycle. It′s a novel method to fight against breast cancer by influencing the progress of the breast cancer cell cycle and inducing the cell cycle arrest in breast cancer cells. Lots of studies have shown that the G2/M phase checkpoint which transition from gap-phase （G2 phase） to mitotic phase （M phase） in the cell cycle is the key point for cell survival or death. Many antitumor drugs can inhibit the proliferation of tumor cells through the cell cycle arrest. We summarized the domestic and foreign literatures in recent years， and comprehensively explained the research progress on the related regulatory molecules in G2/M arrest. In addition， we summarized and sorted out the researches on the methods and ways of alkaloids， polysaccharides， terpenes， flavonoids， saponins and other active compounds of TCM in inducing the G2/M arrest of human breast cancer cells. By summarizing the active compounds of various Chinese medicines in inducing G2/M arrest of breast cancer cells， and reviewing the research progress on mechanism of active TCM compounds for inhibiting the proliferation of breast cancer cells， we will， in this paper， investigate the mechanism of active TCM compounds for inhibiting the proliferation of breast cancer cells through inducing G2/M arrest of human breast cancer cells， so as to provide a scientific basis for in-depth research on the anti-breast cancer mechanism of the active compounds in TCM.
Abstract：Intestinal Peyer's patches （PPs） are local immune tissues of the intestine， which are considered to be the main induction site of the intestinal mucosal immune response， and closely related to immune-related refractory enteropathies， such as ulcerative colitis and Crohn's disease. In recent years， more and more scholars have tried to find a new breakthrough for treating refractory enteropathies with a limited efficacy of clinical interventions through in-depth study of the relationship between PPs and enteropathy. Traditional Chinese medicine （TCM） polysaccharides are considered to be a key component for immune regulation with TCM. Modern studies show that TCM polysaccharides have a significant positive intervention effect on the structure and function of PPs， with good development prospects. Based on this， this paper focuses on PPs and intestinal-related diseases， and systematically introduces the physiological structure of PPs and their drug delivery mechanism， and summarizes the interactions of PPs with effect on immune-related enteropathies， analyze of current studies and prospects of effect of TCM polysaccharides in intervening intestinal disease and its dysfunction by regulating PPs， with the aim to provide new strategies for basic studies and clinical treatment of immune-related refractory enteropathies from the perspective of PPs， and new ideas for basic studies and clinical studies on effect of TCM polysaccharides in intervening enteropathies.
Keywords：traditional Chinese medicine polysaccharides;intestinal Peyer's patches;ulcerative colitis;mechanism
Abstract：Adequate sleep is an important factor to ensure the healthy functioning of the body. A type of chronic sleep diseases characterized by insufficient sleep can be collectively referred to as sleep deprivation （SD）， which is divided into primary and secondary sources in terms of sources. As one of the most frequent types of diseases in recent years， SD has received more and more attention and attention from the whole society. SD can have a wide-ranging and far-reaching impact on cognitive behavior， such as decreased wakefulness， decreased alertness， and inattention， decreased sensory perception， decreased learning and memory capabilities， et al， involving the impact on multiple system functions of the human body， and It is closely related to the occurrence of many diseases， and may cause serious troubles to the normal life of patients and even their relatives and friends. The cognitive impairment caused by SD has been fully verified in clinical tests and various animal behavior experiments， mainly involving pathological damage such as changes in synaptic plasticity， enhanced endoplasmic reticulum stress， circadian rhythm disorders， and energy metabolism imbalance. Western medicine treatments for SD mostly have negative factors such as high side effect and strong addiction. However， Chinese medicine intervention focuses on the overall concept， has long-lasting effectiveness， significant effects， and mild side effects. It has also been widely recognized clinically for improving the complications of sleep disorders. This article reviews the current status and classification of SD research， its pathological mechanisms that lead to cognitive impairment and its molecular-level exploration directions and results. In recent 5 years， the therapeutic effect and experience of traditional Chinese medicine intervention therapy such as compound Chinese medicine， acupuncture and moxibustion as well as auxiliary therapy such as exercise and five sounds， in order to further summarize and clarify the interaction mechanism between SD and cognitive behavior， and provide a theoretical basis for the study of the pathological mechanism of SD disease and future clinical treatment.
Keywords：sleep deprivation;cognitive behavior;synaptic plasticity;endoplasmic reticulum stress;circadian rhythm;energy metabolism;Chinese medicine treatment
Abstract：Gastric cancer is one of the most common malignant tumors in the digestive system， and precancerous lesion of gastric cancer （PLGC） represents a long-term stage in the process of malignant development of normal gastric mucosa into gastric cancer. Gastric cancer and precancerous lesions are difficult to cure clinically， leaving poor prognosis and a serious negative impact on the quality of daily life of patients. In recent years， studies on cell autophagy have been at the forefront of the natural life science. Regulating autophagy to treat precancerous lesions and prevent gastric cancer has become nowadays a hot topic. Autophagy is a process in which cells enclose some redundant or damaged cytoplasm， proteins and organelles to form autophagosomes， and bind to lysosomes to degrade the contents. Autophagy has bidirectional effect on different cells and different stages of the same cell. Autophagy at a lower level can kill cancer cells， while autophagy can promote the growth and proliferation of cancer cells under stress conditions such as hypoxia， hunger and infection， or when autophagy clears damaged proteins in cells and organelle function is abnormal. Traditional Chinese medicine（TCM）， which has low toxicity and easy acceptance by patients， has a positive effect on the treatment of gastric cancer and PLGC. At present， studies on the prevention and treatment of gastric cancer and PLGC by TCM have been carried out in depth with cell autophagy as the breakthrough point. More and more research results have confirmed that TCM can regulate the autophagy process of gastric cancer cells and play an anti-tumor role by interfering with various autophagy related genes， signal pathways and organelles. This paper summarizes the studies on the regulation of cell autophagy by TCM in the treatment of gastric cancer and precancerous lesions， so as to provide references for future studies on the regulation of autophagy by TCM.
Keywords：autophagy;traditional Chinese medicine;gastric cancer;precancerous lesion of gastric cancer;research progress
Abstract：Periplaneta americana is one of the important basic medicinal materials of traditional Chinese medicine "fei lian". The traditional functions mainly include promoting blood circulation， sore muscles， diuresis， spleen and phlegm. Because of its exact curative effect， proprietary Chinese medicines， which are mainly used as raw materials， are widely used in clinical practice， especially in the repair of various wounds. The drug has not been included in the Chinese Pharmacopoeia. The local standard is only based on the alanine content of total amino acids. The physiologically active small peptides， nucleosides， proteins and other substances have not been obtained. Qualitative or quantitative control. In recent years， peptide monomers isolated from the P.americana， such as antimicrobial peptides， neuropeptides， and diuretic peptides， have strong pharmacological activities such as antibacterial， antitumor， and muscular neurotrophic， and dihydroisocoumarins are also irritating. Dermal Dermal fibroblasts produce collagen. Based on this， this paper uses CNKI， Wanfang Database and Pubmed Database to search the relevant research literatures of P.americana from 1984 to 2019， and systematically analyzes the current research of P.americana from three aspects： chemical composition-pharmacological action-clinical application. Interpretation provides reference for the further development of the drug and the development of more specific and stable quality control standards for its proprietary Chinese medicines.
Keywords：Periplaneta americana;chemical composition;pharmacological action;clinical application;quality control
Abstract：Succus Bambusae， regarded as the holy medicine for the treatment of phlegm by traditional Chinese medicine experts， has the functions of clearing heat and resolving phlegm， resuscitating and relieving convulsions. In clinical practice， it is mainly used to treat phlegm-heat cough， phlegm yellow and shortness of breath， stroke with exuberant phlegm and other syndromes. The research of its processing procedure and quality standard is the key issue to realize the process modernization and quality standardization of this traditional medicine resource. On the basis of literature research， this paper conducted the herbal textual research on Succus Bambusae， sorted out its medicinal history， and compared the advantages and disadvantages of its traditional and modern preparation procedure. Based on the historical records， it is pointed out that there are still many problems in the traditional and modern processing procedure of Succus Bambusae， such as low-yield processing procedure and extensive production technology that cannot meet the requirements of large-scale green manufacturing. At present， there are a lot of literature on the comparative study of the origin， variety and processing procedure of Succus Bambusae， but most of them are limited to the chemical components， and lack of in-depth study on its pharmacodynamic substance basis and mechanism. Meanwhile， the pharmacological effects of Succus Bambusae are mostly limited to antitussive and expectorant， and few other pharmacological effects have been reported. Based on the above analysis， it is suggested that the follow-up research work can be carried out from these five aspects：①Clarifying the original source， enriching and perfecting the medicinal resources. ②Standardizing the processing procedure， establishing relevant quality standards. ③Clarifying the pharmacological mechanism， strengthening the basic research on the pharmacological effect and clinical application. ④Inheriting ancient processing， researching and developing special preparation equipment and standardizing production operation rules. ⑤Comprehensive utilization of the by-products in preparation process.
Keywords：Succus Bambusae;herbal textual research;processing;active ingredients;pharmacological effects;pharmacodynamic substance basis;quality standard