Abstract：ObjectiveTo explore the effect of the serum containing Huanglian Wendantang on pyroptosis in vitro model of insulin resistance and its mechanism.MethodSD rats were randomly divided into two groups， namely Huanglian Wendantang-containing serum group and blank serum group， and given 7.8 g·kg-1·d-1 Huanglian Wendantang and equal volume of normal saline by intragastric administration according to body surface area. Blank serum and medicated serum with different concentration were extracted and prepared. HepG2 cells were treated with sodium palmitate to construct the model of insulin resistance （IR）， and they were randomly divided into control group， model group， metformin hydrochloride group， blank serum group， and Huanglian Wendantang-containing serum high-， medium-， and low-dose groups. After 24 h of cultivation， the cells of each group were treated with insulin for 15 min at concentration of 1×10-7 mol·L-1， and the cell supernatant was collected. The glucose oxidase （GOD-POD） kit was used to determine the glucose content of each group， and calculate the glucose consumption and inhibition rate. The methyl thiazolyl tetrazolium （MTT） assay was used to detect the cell proliferation， thus screening out the optimal dose of serum containing Huanglian Wendantang. HepG2 cells were randomly divided into control group， model group， and Huanglian Wendantang-containing serum group. The levels of interleukin-1β （IL-1β） and interleukin-18 （IL-18） in each group were determined by enzyme-linked immunosorbent assay （ELISA）， and the mRNA and protein expression levels of NOD like receptor protein 3 （NLRP3） in each group were determined by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot. In terms of the mechanism， HepG2 cells were randomly divided into control group， empty vector group， NLRP3 overexpression group， empty vector + IR group， empty vector + IR + Huanglian Wendantang-containing serum group， NLRP3 overexpression + IR group， and NLRP3 overexpression + IR + Huanglian Wendantang-contain serum group. GOD-POD method was used to measure the glucose content of each group cells， and calculate the glucose consumption. ELISA was used to determine the release of IL-1β and IL-18 in each group. Real-time PCR and Western blot assay were used to determine the mRNA and protein expressions of cysteinyl aspartate specific proteinase-1 （Caspase-1）， gasdermin D （GSDMD）， and NLRP3. Immunofluorescence assay was used to detect NLRP3， GSDMD， and Caspase -1 expressions.ResultAs compared with the control group， the glucose consumption in the model group was significantly decreased （P<0.01）. As compared with the model group， the increase of the glucose consumption of IR-HepG2 cells was the most significant in the Huanglian Wendantang-containing serum high-dose group （P< 0.01）. As compared with the control group， the IL-1β and IL-18 release levels and the mRNA and protein expressions of NLRP3 in IR-HepG2 cells were significantly increased （P<0.05， P<0.01）. Huanglian Wendantang effectively reduced IR-HepG2 cell supernatant IL-1β， IL-18， and NLRP3 mRNA and protein expressions as compared with the model group （P<0.05， P<0.01）. Overexpression of NLRP3 significantly reduced the cell glucose consumption as compared with the control group and the empty vector group （P<0.01）， and significantly up-regulated the IL-1β and IL-18 levels and the mRNA and protein levels of NLRP3， Caspase-1， and GSDMD as compared with the empty vector + IR group （P<0.05， P<0.01）. Huanglian Wendantang-containing serum effectively reversed the above indicators as compared with the NLRP3 + IR group （P<0.05， P<0.01）.ConclusionHigh fat-induced insulin sensitivity of IR-HepG2 cells is closely related to inflammation and NLRP3 expression. Huanglian Wendantang-containing serum improves IR-HepG2 cell pyroptosis through the targeted inhibition of NLRP3/Caspase-1 signaling pathway， which provides new therapeutic targets for the prevention and treatment of IR and type 2 diabetes mellitus （T2DM）.
Keywords：pyroptosis;NOD like receptor protein 3 （NLRP3）;Huanglian Wendantang;insulin resistance;HepG2 cell line
Abstract：ObjectiveTo investigate the protective effect of Zuoguiwan against 60Co-γ ray-induced premature aging of rats based on the phosphatidylinositol-3-kinases/protein kinase B/mammalian target of rapamycin （PI3K/Akt/mTOR） signaling pathway.MethodSixty sexually mature female SD rats were irradiated with 60Co-γ rays （6.0 Gy， LD40） for 24 h at one time. Then they were randomized into model group， Bujiale group （0.18 g·kg-1·d-1）， Bujiale （0.09 g·kg-1·d-1） + high-dose Zuoguiwan group （23.625 g·kg-1·d-1）， high-dose Zuoguiwan group （23.625 g·kg-1·d-1）， medium-dose Zuoguiwan group （9.45 g·kg-1·d-1）， and low-dose Zuoguiwan group （4.725 g·kg-1·d-1）. The administration （once a day） lasted 21 days. Serum indexes ［follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， and estradiol （E2）］ of rats were detected by enzyme-linked immunosorbent assay （ELISA）， and morphological changes of ovarian tissues were observed based on hematoxylin and eosin （HE） staining. The apoptosis rate of granulosa cells was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling （TUNEL） and the protein expression of phosphorylated （p）-PI3K， p-Akt， p-mTOR， B-cell lymphoma-2 （Bcl-2）， and Bcl-2-associated X protein （Bax） in ovarian tissues by Western blot.ResultCompared with normal group， model group demonstrated increase in serum FSH （P<0.01）， decrease in E2 （P<0.05）， and reduction of follicles and luteum in early ovary （P<0.01）. Moreover， the elevation of apoptosis rate of granulosa cells （P<0.01）， down-regulation of p-PI3K， p-Akt， p-mTOR， and Bcl-2 in ovarian tissue， and increase in expression of Bax were also observed in the model group as compared with the normal group （P<0.01）. In comparison with the model group， the administration groups showed rise of the number of early ovarian follicles， decrease in the apoptosis rate of granulosa cells， increase in the expression of p-PI3K， p-Akt， p-mTOR， and Bcl-2， and down-regulation of Bax， particularly the Bujiale + high-dose Zuoguiwan group（P<0.05，P<0.01）.ConclusionZuoguiwan protects radiation-damaged ovary by activating the expression of PI3K/Akt/mTOR protein in ovarian tissue， increasing Bcl-2， and inhibiting the expression of Bax.
Keywords：radiation damage;premature ovarian failure （POF）;Zuoguiwan;terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling （TUNEL）;phosphatidylinositol-3-kinases/protein kinase B/mammalian target of rapamycin （PI3K/Akt/mTOR） signaling pathway;B-cell lymphoma-2;Bcl-2-associated X protein
Abstract：ObjectiveTo explore the effect of Jiangtang Xiaozhi tablet （JTXZT） on metabolic dysfunction-associated fatty liver disease and to study the mechanism from the perspective of circadian clock-related genes such as circadian locomotor output cycles kaput （CLOCK）， brain and muscle aryl hydrocarbon receptor nuclear translocator-like protein 1 （BMAL1）， reverse-eritroblastosis receptor （REV-ERB）α and β.MethodA total of 50 male SPF C57BL/6J mice were randomized into normal group （n=10） and modeling group （n=40）. The normal group was fed with normal diet， and the modeling group with high-fat diet for 4 weeks. Then the model mice were randomly classified into model group， high-dose （12.5 g·kg-1） and low-dose （6.25 g·kg-1） Jiangtang Xiaozhi tablet groups， and orlistat group （70 mg·kg-1）， with 10 mice in each group. The normal group and model group received equivalent volume of distilled water （8 weeks）. Then， the body weight of mice was measured， and the content of serum triglyceride （TG）， total cholesterol （TC）， low-density lipoprotein cholesterol （LDL-C）， high-density lipoprotein cholesterol （HDL-C）， aspartate aminotransferase （AST）， and alanine aminotransferase （ALT） was determined with biochemical method. Serum content of free fatty acid （FFA） and leptin was detected by enzyme-linked immunosorbent assay （ELISA）. Pathological changes of liver tissue and epididymal adipose tissue were observed based on hematoxylin-eosin （HE） staining. Liver fibrosis was examined based on Masson's trichrome staining， and changes of lipids based on oil red O staining. The expression of CLOCK， BMAL1， REV-ERBα， and REV-ERBβ was detected by Western blot and immunohistochemistry assay.ResultCompared with the normal group， the model group had high content of TG， TC， LDL-C， HDL-C， AST， ALT， FFA， and leptin （P<0.05， P<0.01）， showed ballooning degeneration and focal microvesicular steatosis of liver cells， enlarged adipocytes， and inflammatory cell clusters and fibrous tissue hyperplasia， and displayed increased protein expression of sterol regulatory element binding protein （SREBP） 1 and peroxisome proliferators-activated receptor （PPAR）γ （P<0.01） and decreased protein expression of PPARα （P<0.05）， CLOCK， BMAL1， REV-ERBα and β （P<0.05， P<0.01）. Compared with the model group， JTXZT-H group down-regulated the content of TG， TC， LDL-C， HDL-C， AST， ALT， FFA， and leptin in mice （P<0.05， P<0.01）， and the JTXZT groups demonstrated reduction in the degree and range of ballooning degeneration of liver tissue， alleviation of the compression of hepatic sinusoidal tissue， unobvious inflammatory cell infiltration and fibrous tissue proliferation， reduction in the expression of SREBP1 and PPARγ （P<0.05， P<0.01）， and rise of the protein expression of PPARα （P<0.01）， CLOCK， BMAL1， REV-ERBα， and REV-ERBβ （P<0.05， P<0.01）.ConclusionJTXZT can significantly alleviate the metabolic dysfunction-associated fatty liver disease in mice caused by high-fat diet. The mechanism is the likelihood that it regulates downstream related lipid metabolism proteins （such as SREBP1， PPARγ， and PPARα）.
Abstract：ObjectiveTo investigate the protective effect of Zhizi prescription （ZZP） on carbon tetrachloride （CCl4）-induced acute and subacute liver injury and its mechanism.MethodAcute and subacute liver injury animal models were induced. C57 mice were randomly divided into a normal group， model group， obeccholic acid group， ZZP high-dose （0.5 g·kg-1） group， and ZZP low-dose （0.25 g·kg-1） group. According to the experiment design， the serum and liver tissue of mice were collected after the last administration. Hematoxylin-eosin （HE） and Sirius staining was used to observe the liver pathological changes. Serum alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， total bilirubin （TBIL）， liver homogenate hydroxyproline （Hyp）， malondialdehyde （MDA）， and superoxide dismutase （SOD） levels were determined by kit. The levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in the liver tissue were determined by enzyme-linked immunosorbent assay （ELISA）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expressions of collagen 1A1 （Col1a1）， collagen 3A1 （Col3a1）， fibronectin （FN）， transforming growth factor β receptor Ⅱ （Tgfbr2） and α-smooth muscle actin （α-SMA） in the liver tissue.ResultIn terms of the acute liver injury， as compared with the normal group， the levels of ALT， AST， TBIL and MDA in the model group were significantly increased （P<0.01）， while the activity of liver SOD was significantly decreased （P<0.01）. Compared with model group， the ZZP high-dose and low-dose groups both significantly reduced the degree of liver cell injury， and protected the acute liver injury induced by CCl4. The ZZP high-dose group had a better effect than the ZZP low-dose group. In terms of the subacute liver injury， the levels of ALT， AST， MDA，TNF-α and IL-6 in the model group were significantly increased （P<0.01）， while the activity of liver SOD was significantly decreased （P<0.01）. As compared with the model group， liver Hyp content in the ZZP high-dose and low-dose groups was significantly decreased （P<0.01）， and the collagen deposition in liver of both groups was significantly reduced. The ZZP high-dose group also significantly down-regulated the mRNA expressions of α-SMA， Col1a1， Col3a1， FN， and Tgfbr2 in the liver of mice （P<0.05， P<0.01）.ConclusionZZP effectively protects the acute and subacute liver injury induced by CCl4， and the protective effect is proportional to its concentration. The mechanism may be related to the increase of the activity of antioxidant enzymes in the liver tissue， the decrease of the level of lipid peroxidation， and the inhibition of inflammatory response， thus reducing collagen deposition and improving early liver fibrosis.
Keywords：homology of medicine and food;carbon tetrachloride （CCl4）;liver injury;antioxidant;collagen deposition;liver fibrosis
Abstract：ObjectiveTo explore the pharmacodynamic effects of total flavonoids of Paeonia suffruticosa flower （TFPFs） on rats with hyperuricemia and provide scientific data support for the research and development of therapeutic drugs for hyperuricemia.MethodThe hyperuricemia model was induced by adenine combined with ethambutol in rats. The rats were randomly divided into a blank control group， a model group， two positive control groups （allopurinol at 42 mg·kg-1 and Tongfengshu tablets at 600 mg·kg-1）， and high-， medium-， and low-dose TFPFs groups （260， 130， and 65 mg·kg-1）. The general conditions of rats were observed and recorded， and the body weight was recorded once every 5 days. The 24-hour urine volume， water intake， uric acid （UA）， and urinary protein of rats were determined after the last administration. The kidney index was calculated. The pathological changes in thymus and spleen tissues of rats were observed by hematoxylin-eosin （HE） staining. The serum activities of UA， creatinine （Cr）， blood urea nitrogen （BUN）， malondialdehyde （MDA）， superoxide dismutase （SOD）， and total antioxidant capacity （T-AOC） of rats were determined. The xanthine oxidase （XOD） and adenosine deaminase （ADA） activities in the liver were detected. The content of uric acid transporter 1 （URAT1）， organic anion transporter 1 （OAT1）， and glucose transporter 9 （GLUT9） in the kidney was detected by enzyme-linked immunosorbent assay （ELISA）.ResultCompared with the results in the model group， TFPFs could improve the mental state of rats， increase the body weight（P<0.01）， promote UA excretion（P<0.01）， reduce the content of urinary protein（P<0.05）， relieve renal glomerular atrophy， renal tubular epithelial cell status， and urate crystal deposition in renal tubules， dwindle 24-hour urine volume， water intake， kidney index（P<0.05）， serum levels of UA， Cr， BUN， and MDA（P<0.05，P<0.01）， inhibit the activities of XOD（P<0.05） and ADA（P<0.05，P<0.01）in the liver， diminish the expression of GLUT9 in the renal homogenate（P<0.05）， and increase serum SOD and T-AOC activities as well as OAT1 expression（P<0.01） in the kidney. The pathological changes of thymus and spleen were improved.ConclusionTFPFs possess a protective effect on the kidney of rats with hyperuricemia， which is achieved by promoting uric acid excretion， inhibiting oxidation and the activity of key enzymes in uric acid synthesis， and regulating the expression of uric acid transporters.
Abstract：ObjectiveTo study the inhibitory effect of Celastrus orbiculatus extract （COE） on gastric cancer cells， to clarify the specific mechanism of COE promoting the apoptosis of gastric cancer cells by affecting the mitochondrial structure and function， and to provide an experimental basis for the further development and clinical application of C. orbiculatus.MethodBrdu staining combined with flow cytometry and Annexin V-fluorescein isothiocyanate （AnnexinV-FITC） staining combined with flow cytometry were employed to detect the effects of COE （20， 40， 80 mg·L-1） on the proliferation and apoptosis of gastric cancer cells， respectively. The changes in mitochondrial membrane potential were detected with JC-1 mitochondrial membrane potential assay kit. The expression of apoptosis-associated proteins including B-cell lymphoma-2 （Bcl-2）， B-cell lymphoma-xL （Bcl-xL）， Bcl-2-associated X （Bax）， and cysteine aspartutespecific protease-3 （Caspase-3） in gastric cancer cells was determined by Western blot. Transmission electron microscopy was employed to detect changes in the mitochondrial microstructure of gastric cancer cells exposed to COE. Western blot was employed to measure the expression of mitochondrial marker proteins ［superoxide dismutase 1 （SOD1）， voltage-dependent anion channel （VDAC）， prohibitin 1 （PHB1）， and heat shock protein 60 （HSP60）］ in gastric cancer cells.ResultCompared with the control group， COE （40， 80 mg·L-1） inhibited the proliferation and promoted the apoptosis of gastric cancer cells （P<0.05）. Furthermore， COE reduced the mitochondrial membrane potential of gastric cancer cells. Compared with the control group， COE （20， 40， 80 mg·L-1） up-regulated the expression of Bax and Caspase-3 which promoted apoptosis of gastric cells （P<0.05， P<0.01）， and COE at 40 and 80 mg·L-1 down-regulated the expression of Bcl-2 and Bcl-xL which inhibited the apoptosis of gastric cancer cells （P<0.01）. The results of transmission electron microscopy showed that COE changed the microstructure of gastric cancer cells， which led to the appearance of vacuoles in the cell membrane and mitochondria and damaged the mitochondrial structure. Compared with the control group， COE （20， 40， 80 mg·L-1） changed the expression of mitochondrial marker proteins. Specifically， it up-regulated the expression of SOD1 involved in stress response （P<0.05， P<0.01） and down-regulated that of VDAC， PHB1， and HSP60 associated with mitochondrial stability and permeability （P<0.01）.ConclusionCOE can significantly inhibit the proliferation and promote the apoptosis of gastric cancer cells. It may activate the mitochondrial apoptosis pathway by destroying the mitochondrial structure and function of gastric cancer cells.
Abstract：ObjectiveTo investigate the effect of calycosin-mediated glucoprotein130/Janus kinase/signal transducer and activator of transcription factor （GP130/JAK/STAT） signaling pathway on oxidative injury of astrocytes in spinal cord.MethodAstrocytes in rat spinal cord were isolated and identified by immunofluorescence detection of glial fibrillary acidic protein （GFAP）. The cells were respectively pre-treated with 5， 10， 20 μmol·L-1 calycosin for 12 h， and then 100 μmol·L-1 H2O2 （24 h） was added to induce oxidative injury. Cell counting kit-8 （CCK-8） assay was employed to detect cell proliferation and select the optimal concentration of calycosin. The following experimental groups were designed： control group， model group （100 μmol·L-1 H2O2）， calycosin group （20 μmol·L-1 calycosin）， calycosin + LY294002 group （20 μmol·L-1 calycosin + 10 μmol·L-1 LY294002）， and calycosin + Stattic group （20 μmol·L-1 calycosin + 3 μmol·L-1 Stattic）. CCK-8 assay and immunofluorescence method were used to detect the proliferation of cells and flow cytometry was applied to detect cell apoptosis and cycle. The protein expression of phosphorylated （p）-JAK2， p-STAT3， p-protein kinase B （Akt）， GP130， and interleukin-6 （IL-6） was detected by Western blotting.ResultCompared with the control group， the model group showed low proliferation activity and high apoptosis rate of cells （P<0.05）. Compared with the model group， calycosin （20 μmol·L-1） group displayed high proliferation activity and low apoptosis rate of cells （P<0.05）. Compared with calycosin （20 μmol·L-1） group， both phosphatidylinosirtol-3-kinases （PI3K） inhibitor LY294002 and STAT3 inhibitor Stattic significantly reduced the proliferation activity and increased the apoptosis rate of cells （P<0.05）. The protein expression of p-JAK2， p-STAT3， p-Akt， GP130， and IL-6 in the model group was higher than that in the control group （P<0.05）， and the expression of the above indicators was lower in each treatment group than in the model group （P<0.05）.ConclusionCalycosin can promote the proliferation and inhibit the apoptosis of astrocytes with oxidative injury by inhibiting the phosphorylation of PI3K/Akt pathway and JAK2/STAT3 pathway.
Keywords：astrocytes in spinal cord;calycosin;oxidative injury;glucoprotein130/Janus kinase/signal transducer and activator of transcription factor （GP130/JAK/STAT） signaling pathway;spinal cord injury;Buyang Huanwutang
Abstract：ObjectiveTo explore the anti-abortional effect of Pangshi Antai Zhixue decoction and its mechanism in helper T lymphocyte 1 （Th1）/Th2 balance in the decidual tissues of spontaneous abortion rats with heat syndrome， based on the p38 mitogen-activated protein kinase （p38 MAPK） signaling pathway.MethodAconiti Lateralis Radix Praeparata， Zingiberis Rhizoma， and Cinnamomi Cortex decoction was used to replicate the rat model of spontaneous abortion with heat syndrome. The spontaneous abortion rats with heat syndrome were randomly divided into model group， aspirin group （5.25 mg·kg-1）， dydrogesterone group （3.02 mg·kg-1）， Pangshi Antai Zhixue decoction high-dose （44 g·kg-1）， medium-dose （22 g·kg-1）， and low-dose （11 g·kg-1） groups， with ten rats in each group. Ten normal rats were divided into a normal group. Rats in each group were given corresponding drugs， Once a day for 12 d. After 24 h of the last administration， blood was collected from the abdominal aorta. The enzyme-linked immunosorbent assay （ELISA） was used to determine the levels of β-human chorionic gonadotropin （β-HCG）， progesterone （P）， estradiol （E2）， γ interferon （IFN-γ）， and interleukin-4 （IL-4） in rat serum. The uterus and meconium tissues of rats were collected to determine the number and rate of miscarriages. Western blot was used to detect GATA3， T-bet， p38 MAPK， and its phosphorylation in the decidual tissue.ResultAs compared with the normal group， the number of live births， the β-HCG， P， E2， and IL-4 in the serum， and the GATA3 protein expression in the decidual tissue in the model group were reduced （P<0.01）， whereas the number and rate of miscarriages， IFN-γ in the serum， and the expression of p-p38 MAPK and T-bet protein levels in the demolded tissues increased （P<0.01）. As compared with the model group， the number of live births， the β-HCG， P， E2， and IL-4 in the serum， and the GATA3 protein expression in the decidual tissue in the Pangshi Antai Zhixue decoction medium-dose group increased （P<0.01）， whereas the number and rate of miscarriages， IFN-γ in the serum， and the expression of p-p38 and T-bet protein levels in the demolded tissues reduced （P<0.01）. As compared with the aspirin group， the P， E2， and IL-4 in the serum of rats in the dydrogesterone group and the Pangshi Antai Zhixue decoction high-dose and medium-dose groups increased （P<0.01）， the number of live births in the Pangshi Antai Zhixue decoction medium-dose group increased （P<0.01）， and the β-HCG and IFN-γ in the serum of rats in the dydrogesterone group decreased （P<0.01）. The number and rate of miscarriages， IFN-γ in the serum， and T-bet and GATA3 levels in the decidual tissues of rats in the Pangshi Antai Zhixue decoction medium-dose group decreased （P<0.05）. Compared with the Pangshi Antai Zhixue decoction medium-dose group， the low-dose group， high-dose group， and dydrogesterone group showed increased number and rate of miscarriages （P<0.05）， and the high-dose group and dydrogesterone group decreased the number of live birth （P<0.01）. The IFN-γ in the serum and p-p38 MAPK and T-bet protein in the decidual tissue in the low-dose group， and the p-p38 MAPK and T-bet protein in the decidual tissue in the high-dose group all increased （P<0.05）. The β-HCG， P， and E2 in the serum of rats in the Pangshi Antai Zhixue decoction low-dose group， dydrogesterone group， and aspirin group decreased （P<0.01）， and the IL-4 in the serum and GATA3 in the decidual tissue of rats in the Pangshi Antai Zhixue decoction low-dose and high-dose group and the dydrogesterone group decreased （P<0.01）.ConclusionPangshi Antai Zhixue decoction realizes the effect of fetal protection by regulating the activation of p38 MAPK signal pathways and Th1/Th2 balance.
Keywords：Pangshi Antai Zhixue decoction;p38 mitogen-activated protein kinase （p38 MAPK） pathway;helper T lymphocyte 1 （Th1）/Th2 balance;spontaneous abortion with heat syndrome
Abstract：ObjectiveTo investigate the therapeutic effect of Qingmei compound on acute gouty arthritis （AGA） in rats and preliminarily clarify its mechanism.MethodForty male SD rats were randomly divided into a blank group， a model group， a colchicine group （0.3 mg·kg-1）， and low- and high-dose Qingmei compound groups （200 and 400 mg·kg-1）， with eight rats in each group. The AGA model was induced by injecting 50 g·L-1 monosodium urate （MSU） into the ankle joint of the rats except those in the blank group. The ankle swelling index was measured before and 6， 24， and 48 h after modeling. The pathological changes in the joint tissues of AGA rats were observed by hematoxylin-eosin （HE） staining. The expression of tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β） in the joint tissues of rats was detected by immunohistochemistry. The protein expression of NOD-like receptor protein 3 （NLRP3） pathway and key proteins in the joint tissues of rats was detected by Western blot.ResultCompared with the blank group， the model group showed increased ankle swelling index， synovial hyperplasia， and inflammatory infiltration， and up-regulated expression of IL-1β， TNF-α， and NLRP3 proteins in the ankle joint and the ratio of Caspase-1 shear body to Caspase-1 precursor protein （Caspase-1 p20/Caspase-1） （P<0.01）. Compared with the model group， the Qingmei compound groups showed reduced ankle swelling index of AGA rats， especially the low-dose Qingmei compound group （P<0.01）. Meanwhile， Qingmei compound inhibited synovial hyperplasia and inflammatory infiltration （P<0.01） and reduced the levels of IL-1β， TNF-α， and NLRP3 proteins and Caspase-1 p20/Caspase-1 in joint tissues （P<0.01）.ConclusionQingmei Compound can significantly alleviate the joint swelling and inflammatory infiltration of AGA， and its mechanism may be related to the inhibition of the NLRP3 signaling pathway.
Keywords：Qingmei Compound;acute gouty arthritis;NOD-like receptor protein 3 （NLRP3） pathway;rat;inflammatory factor
Abstract：ObjectiveTo observe the therapeutic effect of Yiyuan Qiwei pills （YYQW） on diabetes mellitus-induced induced erectile dysfunction （DMED） in rats and explore its regulation on the nitric oxide （NO）-cyclic guanosine monophosphate （cGMP） signaling pathway.MethodFifty-five healthy SD male rats of clean grade aged 2-3 months underwent intraperitoneal injection of streptozotocin （STZ） to induce the DMED model， and another 10 healthy SD male rats of clean grade aged 2-3 months were assigned to the control group. The model rats were randomly divided into a model group， a sildenafil group （5 mg·kg-1， ig）， and low-， medium-， and high-dose YYQW groups （1.5， 3.0， 6.0 g·kg-1， ig）. The rats in the model group and the control group were given normal saline by gavage at 10 mL·kg-1， once a day for two months. After intervention， the penile erectile function of rats in each group was measured by a pressure detection system. The pathological changes and ultrastructure of penile corpus cavernosum were observed by hematoxylin-eosin （HE） staining and transmission electron microscopy， respectively. The level of NO in the corpus cavernosum was detected by nitrate reductase. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of cGMP and advanced glycation end products （AGEs）. Real-time quantitative polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of endothelial nitric oxide synthase （eNOS）， neurogenic nitric oxide synthase （nNOS）， total nitric oxide synthase （NOS）， and phosphodiesterase type5 （PDE5） in rat penile tissues. The expression of above proteins was detected by Western blot.ResultCompared with the control group， the model group showed decreased intracavernous pressure （ICP）， NO， and cGMP levels， reduced mRNA and protein expression of nNOS and NOS， and increased PDE5 mRNA and protein expression （P<0.05）. Compared with the model group， the sildenafil group and the YYQW groups displayed increased ICP， NO， and cGMP levels， elevated mRNA and protein expression levels of nNOS and NOS， and reduced PDE5 mRNA and protein expression levels （P<0.05）. There were no pathological changes in the tissues and cell ultrastructure of the corpus cavernosum in the control group， while serious pathological changes were observed in the model group. Additionally， the sildenafil group and the YYQW groups were superior to the model group， the optimal effect was observed in the high-dose YYQW group.ConclusionYYQW can improve the penile erectile function of DMED rats and reduce the pathological damage of corpus cavernosum. The mechanism may be related to the promotion of nNOS and NOS expression， the inhibition of PDE5 expression， and the activation of the NO/cGMP signaling pathway.
Abstract：ObjectiveTo collect common prescriptions from multiple channels based on the mainstream guidance materials combined with ancient books to provide references for the selection principles of common prescriptions in clinical practice guidelines of traditional Chinese medicine （TCM） and western medicine.MethodWith the research on the selection of commonly used prescriptions in Clinical Practice Guidelines of Chinese and Western Medicine for Cough in Children as an example，the nation-recommended Schedule Ⅲ drugs and prescriptions for cough in children in the clinical diagnosis and treatment guidelines of TCM （TCM and western medicine）/expert consensus/clinical pathway/diagnosis and treatment protocols，three TCM pediatric textbooks， the experience of famous experts， and ancient books were searched for the establishment of a prescription database and analysis of medication regularity.ResultThe results showed that there were 71 commonly used prescriptions for the treatment of cough in children. The top 3 clinically indicated syndromes were the syndrome of phlegm-heat blocking the lung，the syndrome of wind-heat invading the lung，and the syndrome of wind-cold attacking the lung，and the top 5 efficacies corresponding to high-frequency drugs were reliving cough and dyspnea， clearing heat/resolving phlegm， dispersing wind/dissipating cold， clearing heat/purging fire， and dispersing wind/dissipating heat. The most commonly used Chinese medicines included Armeniacae Semen Amarum， Platycodonis Radix， Bolbostemmatis Rhizoma， Ephedrae Herba， Scutellariae Radix， Pinelliae Rhizoma， and Citri Reticulatae Pericarpium. Some drugs showed high internal correlations with Armeniacae Semen Amarum and Platycodonis Radix. There were 228 prescriptions for the treatment of cough in children based on the ancient books in the Chinese Medical Code. The most commonly used Chinese medicines were Pinelliae Rhizoma， Armeniacae Semen Amarum， Ginseng Radix et Rhizoma， Poria， Citri Reticulatae Pericarpium， Platycodonis Radix， Zingiberis Rhizoma Recens， and Bolbostemmatis Rhizoma. The top 5 efficacies corresponding to high-frequency drugs were dispersing wind/dissipating cold， relieving cough and dyspnea， clearing heat/resolving phlegm， warming cold/resolving phlegm， and tonifying Qi. Some drugs showed high internal correlations with Pinelliae Rhizoma and Platycodonis Radix. As revealed by the comparison of ancient and modern medication， the medication principles could be roughly classified into two categories： ventilating the lung/descending Qi and resolving phlegm. In ancient medication， the drugs for cough in children were mainly used for warming， tonifying， dissipating cold， excreting water， alleviating edema， and astringing lung and intestines， while in modern medication， the prescriptions were mainly for dispersing wind， clearing heat， tonifying yin， and moistening dampness.ConclusionThis study systematically analyzed the commonly used prescriptions for the treatment of cough in children and provided references for the selection of TCM in clinical practice guidelines.
Keywords：cough in children;traditional Chinese medicine;clinical practice guidelines for traditional Chinese and western medicine
Abstract：ObjectiveTo observe the clinical effect of Mahuang Xixin Fuzitang combined with acupuncture and moxibustion in the treatment of localized scleroderma.MethodA total of 95 patients with localized scleroderma treated in Wuhan No. 1 Hospital from September 2019 to October 2021 were assigned into a control group （47 patients） and an observation group （48 patients） by random number table method. The control group was treated with Centella triterpenes tablets and heparin sodium cream， and the observation group was additionally treated with Mahuang Xixin Fuzitang combined with acupuncture and moxibustion. Both groups were treated for 8 weeks， and the clinical effect was compared between groups. The traditional Chinese medicine（TCM） syndrome score （local skin sclerosis， loss of skin texture， darkening of skin pigment， scaly dry skin， etc.）， serum levels of soluble interleukin-2 receptor （sIL-2R） and tumor necrosis factor-alpha （TNF-α）， erythrocyte sedimentation rate （ESR）， and eosinophil count （EO） were compared between before and after treatment as well as between groups. Additionally， the adverse reactions were recorded.ResultThe observation group had higher total effective rate than the control group ［95.83% （46/48） vs. 82.98% （39/47）， χ2=4.166 4， P<0.05］. Before treatment， the TCM syndrome score， sIL-2R， TNF-α， ESR， and EO showed no significant differences between the two groups. The 8 weeks of treatment improved the TCM syndrome score， sIL-2R， TNF-α， ESR， and EO. Moreover， the observation group was superior to the control group in these indicators （P<0.05）. During the treatment， the observation group showed 1 case of abnormal liver function and 1 case of nausea and vomiting， and the control group had 1 case of nausea and vomiting， 1 case of abnormal renal function， and 1 case of abnormal liver function. The total adverse reactions of the observation group （4.17%） and the control group （6.38%） had no significant difference （χ2=0.233 9， P=0.062 86）.ConclusionMahuang Xixin Fuzitang combined with acupuncture and moxibustion is safe and effective in the treatment of localized scleroderma.
Keywords：localized scleroderma;Mahuang Xixin Fuzitang;acupuncture and moxibustion;traditional Chinese medicine（TCM） syndrome score;therapeutic effect;adverse reactions
Abstract：ObjectiveTo study the inhibitory effect of aloe-emodin （AE） on aluminum ion （Al3+）-induced β-amyloid protein 42 （Aβ42）aggregation and its depolymerization on formed Aβ42-Al3+ aggregates in vitro， and to investigate the effect of AE on the cytotoxicity of Aβ42 aggregation in the presence of Al3+.MethodThe Aβ42 group， Aβ42+Al3+ group， Aβ42+AE group， Aβ42+Al3++AE group and the depolymerization test group were set up in the experiment. The aggregation fibrosis process， aggregation morphology， aggregation size and cytotoxicity of Aβ42 in each group were detected by thioflavin T （ThT） fluorescence assay， transmission electron microscopy （TEM）， dynamic light scattering （DLS） experiment and thiazolyl blue （MTT） cytotoxicity assay.ResultCompared with the Aβ42 group， Al3+ could promote Aβ42 aggregation， increase the fluorescence intensity of ThT by 124.48%， induce the aggregation of Aβ42 to form fiber bundles with larger particle size， and significantly reduce the cell viability of human neuroblastoma SH-SY5Y cells （P<0.01）， thus reducing the cell survival rate to 51.05%. AE not only inhibited Aβ42 aggregation， but also inhibited Al3+-induced Aβ42 aggregation in a concentration-dependent manner. Compared with the Aβ42+Al3+ group， high concentration of AE could reduce the ThT fluorescence intensity to 41.66%， and change the polypeptide aggregation pathway to form amorphous aggregates with small particle size. Besides， it significantly inhibited the cytotoxicity of Aβ42 induced by Al3+ （P<0.01）， and restored the cell survival rate to 84.87%. Further depolymerization was conducted， AE could depolymerize Aβ42-Al3+ aggregates to make the formed aggregates disappear and form some small-particle short fibers and amorphous structure aggregates with low toxicity.ConclusionAE can inhibit Aβ42 aggregation and cytotoxicity in the presence of Al3+， depolymerize the formed Aβ42-Al3+ aggregates and alleviate the cytotoxicity， thus laying the foundation for exploring the mechanism of AE in the treatment of Alzheimer's disease.
Keywords：Alzheimer's disease;β-amyloid protein;aloe-emodin;aggregation inhibitor;aluminum ion （Al3+）;chelation;cytotoxicity
Abstract：ObjectiveModern scientific methods and techniques were used to scientifically characterize the traditional softening process of Corydalis Rhizoma， so as to clarify the scientificity and rationality of the traditional process， and provide reference for inheriting the processing methods and experience of traditional Chinese medicine.MethodLow-field nuclear magnetic resonance imaging （LF-NMR/MRI） was used to characterize the water types and distribution in the softening process of Corydalis Rhizoma. Samples during the softening process was cut into thick slices and its section was observed by stereoscopic microscope. High performance liquid chromatography （HPLC） was employed to determine the content change of tetrahydropalmatine during the softening process with the mobile phase of methanol-0.1% phosphoric acid solution （60∶40， triethylamine regulated to pH 6.5） and detection wavelength at 280 nm. The determination method of softening endpoint of Corydalis Rhizoma was simulated by texture analyzer （hand pinch method）， and the softening degree of the finished products was determined after optimizing the relevant parameters.ResultLF-NMR/MRI showed that the water could penetrate through the core and distribute evenly in Corydalis Rhizoma softened by Zhangbang method. The water first entered into the medicinal material from the epidermis and stem marks in the soaking stage as the form of free water， and then penetrated into the inner core to achieve redistribution in the moistening stage. Under stereoscopic microscope， it was observed that Corydalis Rhizoma softened by the Zhangbang method could be sliced well， but the core bursting slices were easy to appear if the softening time was not enough， and the softening of samples was caused by the keratine-like powder after absorbing water. HPLC measurement showed that the loss of tetrahydropalmatine in the softening method was small， its content decreased about 5% in the soaking process， and its content was almost unchanged during the moistening process. The softening degree of Corydalis Rhizoma could be quantified by the texture analyzer， and the optimum parameters were 2 mm·s-1 of speed before test， test speed and speed after test， 20 g of the trigger force， 20% of compression degree. The compressive force of the qualified softened Corydalis Rhizoma was 12.75-15.69 N with the relative standard deviation （RSD） of 6.8%.ConclusionModern scientific methods and techniques can characterize the scientificity and rationality of the traditional processing methods， and confirm that the Zhangbang softening method has the advantages of high efficiency， convenience and small loss of index components. The texture analyzer can simulate the softening endpoint judgment method （hand pinch method）， and realize the goal from subjective experience judgment to objective technology quantification， which has a good demonstration role for the modern inheritance of traditional processing technology.
Keywords：Corydalis Rhizoma;low-field nuclear magnetic resonance imaging （LF-NMR/MRI）;Zhangbang;texture analyzer;tetrahydropalmatine;high performance liquid chromatography （HPLC）;traditional Chinese medicine processing
Abstract：ObjectiveTo analyze the sequence variation and genetic diversity of 47 Isatis indigotica germplasm materials， and carry out the study on the genetic differentiation and structure.MethodGenomic DNA of 47 I. indigotica germplasm materials were extracted by kit extraction method. Two chloroplast DNA （cp DNA） sequences and five inter-simple sequence repeat （ISSR） primers were used for amplification and sequencing. Chromas， Mega 7.0， DanSP5， and GenALEx were used to calibrate， splice， and analyze the sequence characteristics. PERMUT and PopGen 1.31 were used to analyze the genetic diversity parameters and genetic structure， and NTSYS was used to obtain the unweighted pair-group method with arithmetic means（UPGMA） clustering tree plot of 47 I. indigotica germplasm materials.ResultA total of 129 samples from 47 I. indigotica germplasm materials were successfully amplified and sequenced. The length of 2 cp DNA sequences after spliced was 1 412 bp， and there were 377 polymorphic variation loci， and 36 haplotypes. Fu and Li's D* test was significant （P<0.01）. The values of Pi， HS， and HT based on cp DNA were 0.119 89， 0.787， and 0.891， respectively. The genetic differentiation coefficients of gene differentiation coefficient（Gst）， nucleotide differentiation coefficient（Nst）， and fixation index（Fst） were 0.117， 0.468， and 0.488， respectively， and the gene flow （Nm） was 0.615. The mean values of PPB， Shannon information diversity index（I）， Nei's genetic diversity index（H）， and Gst based on ISSR were 78.85%， 0.334 8， 0.218 6， and 0.754 4， respectively， and the Nm value was 0.162 8.ConclusionI. indigotica has high genetic diversity and abundant haplotypes at the species level， with abundant haplotypes. Genetic differentiation among different germplasm materials is obvious， and gene exchange is not frequent. Genetic variation mainly exists among populations. The population has accumulated various low-frequency gene mutations recently， suggesting that it has experienced significant regional expansion in the history.
Keywords：Isatis indigotica;germplasm;chloroplast DNA （cp DNA）;inter-simple sequence repeat （ISSR）;genetic variation
Abstract：ObjectiveBy comparing the composition and content changes of the volatile components in Atractylodis Rhizoma before and after processing with rice-washed water， the effect of rice-washed water processing on volatile components in Atractylodis Rhizoma was investigated.MethodHeadspace-gas chromatography-mass spectrometry （HS-GC-MS） was used to detect the volatile components in rhizomes of Atractylodes chinensis and A. lancea， and their processed products of rice-washed water. Chromatographic conditions were programmed temperature （starting temperature of 50 ℃ for 2 min， rising to 120 ℃ with the speed of 10 ℃·min-1， then rising to 170 ℃ at 2.5 ℃·min-1， and rising to 240 ℃ at 10 ℃·min-1 for 3 min）， the inlet temperature was 280 ℃， the split ratio was 10∶1， and the solvent delay time was 3 min. The conditions of mass spectrometry were electron bombardment ionization （EI） with ionization temperature at 230 ℃ and detection range of m/z 20-650. Then the relative content of each component was determined by the peak area normalization method. SIMCA 14.1 software was used to perform unsupervised principal component analysis （PCA） and supervised orthogonal partial least squares-discriminant analysis （OPLS-DA） on each sample data， the differential components of Atractylodis Rhizoma and its processed products were screened by the principle of variable importance in the projection （VIP） value>1.ResultA total of 60 components were identified， among which 40 were rhizomes of A. chinensis and 38 were its processed products， 46 were rhizomes of A. lancea and 47 were its processed products. PCA and OPLS-DA showed that the 4 kinds of Atractylodis Rhizoma samples were clustered into one category respectively， indicating that the volatile components of the two kinds of Atractylodis Rhizoma were significantly changed after processing with rice-washed water， and there were also significant differences in the volatile components of rhizomes of A. lancea and A. chinensis. The compound composition of Atractylodis Rhizoma and its processed products was basically the same， but the content of the compounds was significantly different. The differential components were mainly concentrated in monoterpenoids and sesquiterpenoids， and the content of monoterpenoids mostly showed a decreasing trend.ConclusionAfter processing with rice-washed water， the contents of volatile components in rhizomes of A. lancea and A. chinensis are significantly changed， and pinene， 3-carene， p-cymene， ocimene， terpinolene， atractylon， acetic acid and furfural can be used as difference markers before and after processing.
Keywords：Atractylodis Rhizoma;rice-washed water;traditional Chinese medicine processing;volatile components;headspace-gas chromatography-mass spectrometry （HS-GC-MS）;terpenoids;principal component analysis （PCA）
Abstract：Taohe Chengqitang is a classical prescription published in The Catalogue of Ancient Classic Recipes (The First Batch). This study systematically summarized traditional Chinese medicine (TCM) ancient books and modern clinical reports on Taohe Chengqitang and investigated its origin， composition， basis, and historical evolution in processing requirements， dosage， preparation and administration， and functions. The findings indicated different opinions on drug basis， preparation of decoction pieces, and drug dosage conversion. Based on sufficient literature review, the actual development needs of classical prescriptions, and the usage habits of modern clinical practice， this study recommended dried mature seeds of Prunus persica or P. davidiana for Persicae Semen， dried twigs of Cinnamomum cassia for Cinnamomi Ramulus， honey-fried Glycyrrhiza uralensis for Glycyrrhizae Radix et Rhizoma, raw Rheum palmatum, R. tanguticum, or R. offìcinale for Rhei Radix et Rhizoma, and mirabilite for Natrii Sulfas. In terms of drug dosage， the doses directed converted from weights and measures in the Han dynasty were large. According to the common doses in modern clinical practice, conversion was carried out based on 6 g of Cinnamomi Ramulus, 6 g of Natrii Sulfas, 6 g of Glycyrrhizae Radix et Rhizoma, and 12 g of Rhei Radix et Rhizoma. The weight of Persicae Semen should be determined according to the actual measurement. In terms of preparation and administration, the drugs in the pot were decocted with 1 400 mL of water to obtain 500 mL of decoction, which was filtered and slightly boiled in the presence of Natrii Sulfas. About 100 mL of warm decoction was taken before meals， three times a day. It was recorded in the ancient books that Taohe Chengqitang was indicated for the syndromes of heat invading the bladder and accumulating inside in the case of released exterior syndrome in Taiyang. After symptom-based flexible application by doctors, it is currently commonly used in the treatment of diabetic nephropathy， constipation， chronic renal failure, and other diseases.
Keywords：Taohe Chengqitang;famous classical formulas;literature research;indications and syndromes;dose
Abstract：ObjectiveTo explore the rules of syndrome and treatment of cardiovascular event chain based on the results of data mining of medical records under the guidance of vessel-collateral theory.MethodMulti-source database research method was used to retrieve the medical records related to metabolic syndrome， angina pectoris of coronary heart disease， post-intervention of myocardial infarction， arrhythmia after myocardial infarction and heart failure after myocardial infarction between database inception and January 2022 from PubMed， China National Knowledge Infrastructure （CNKI）， Wanfang Data Knowledge Service Platform （Wanfang Data）， VIP and China Biomedical Literature Service System （SinoMed）. The fields of name， age， gender， symptoms， tongue image， pulse image， drug and disease location were extracted from these medical records， and drug efficacy classification and syndrome classification were determined based on vessel-collateral theory. The rules of syndrome and treatment of cardiovascular event chain were analyzed by data mining methods such as frequency deconstruction， systematic cluster analysis and FP-Growth association analysis.ResultThe above five key pathological links in the chain of cardiovascular event chain all showed symptoms related to the disorder of Yingwei. The proportion of dampness， phlegm， blood stasis and water secondary to the disorder of Yingwei in the above five key pathological links was different， and the frequency of corresponding drug combinations was also different.ConclusionThe disorder of Yingwei is the initiating factor of cardiovascular event chain and runs through the whole process. The abnormality of Yingwei at the terminal of vessel-collateral may lead to the disorder of Qi， blood， body fluid and essence transportation and metabolism. Body fluid accumulates into dampness， dampness accumulates into phlegm， blood stagnation accumulates into blood stasis， blood stasis accumulates into water， which becomes an internal driving factor to promote the development and aggravation of cardiovascular event chain. It leads to the transmission law of Ning-Yong-Sai-Butong， which constitutes the core pathogenesis of the development and evolution of the five key pathological links of cardiovascular event chain.
Keywords：vessel-collateral theory;Yingwei theory;cardiovascular event chain;rules of syndrome and treatment;FP-Growth correlation analysis;frequency deconstruction;cluster analysis
Abstract：ObjectiveTo investigate the effects of Asari Radix et Rhizoma-Zingiberis Rhizoma herb pair （XGHP） on lung and liver lipid metabolism in rats with chronic obstructive pulmonary disease （COPD）.MethodForty SD male rats were divided into a normal group （10 rats） and a model group （30 rats）. The method of cigarette smoke + tracheal injection of lipopolysaccharide（LPS） + cold stimulation was used to replicate COPD model with the syndrome of cold phlegm obstruction in lung. A COPD group， a XGHP group （5.4 g·kg-1·d-1）， and an aminophylline group （0.5 g·kg-1·d-1） were established after successfully inducing the model， with 10 rats in each group. After treatment， the serum triglyceride （TG）， total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， and low-density lipoprotein cholesterol （LDL-C） levels of rats in each group were measured. Gas chromatography-mass spectrometer （GC-MS） was used to detect the differential metabolites in the lung and liver tissues of rats in each group， and the relevant targets of the differential metabolites were predicted by network pharmacology. Molecular docking was used to verify the binding ability of key components in XGHP to the relevant targets in network pharmacology. The mRNA and protein expression levels of peroxisome proliferator-activated receptor α （PPARα） and fatty acid binding protein 4 （FABP4） in lung and liver tissues of rats in each group were detected by real-time polymerase chain reaction （PCR） and Western blot.ResultXGHP significantly increased the levels of TG， TC， and LDL-C in serum （P<0.05）， and decreased the level of HDL-C （P<0.05） in rats with COPD. GC-MS results showed that there were 8 lung differential metabolites and 17 liver differential metabolites in the COPD group and XGHP group. Network pharmacology predicted 59 common targets for the two differential metabolites， mainly enriched in the PPAR signaling pathway. Molecular docking results showed that the main components in XGHP were well combined with both PPARα and FABP4. Real-time PCR showed that XGHP effectively up-regulated the expression levels of PPARα and FABP4 mRNA （P<0.05）， and Western blot showed that XGHP effectively up-regulated the expression levels of PPARα and FABP4 proteins （P<0.05） in lung and liver tissues of rats with COPD.ConclusionXGHP effectively improves the blood lipid levels of rats with COPD， which may be related to the increase of the expression levels of PPARα and FABP4 mRNA and proteins in the PPAR signaling pathway， thus regulating lung and liver lipid metabolism.
Keywords：Asari Radix et Rhizoma-Zingiberis Rhizoma herb pair;chronic obstructive pulmonary disease （COPD）;lipid metabolism;peroxisome proliferator-activated receptor （PPAR） signaling pathway;metabolomics
Abstract：ObjectiveTo explore the mechanism of Xianglian Huazhuo prescription in the treatment of chronic atrophic gastritis （CAG） based on network pharmacology and animal experiments，so as to provide scientific basis for clinical application.MethodThe possible targets and pathways of Xianglian Huazhuo prescription in the treatment of CAG were obtained based on the prediction of network pharmacology. The CAG rat model was induced by sodium salicylate，N-methyl-N'-nitro-N-nitrosoguanidine （MNNG） and hunger and satiety disorder. Then the CAG rats were treated with Xianglian Huazhuo prescription and morodan for 60 days. After administration，the rats were sacrificed，and the content of interleukin-6 （IL-6），tumor necrosis factor-α （TNF-α），interleukin-1β （IL-1β） and vascular endothelial growth factor （VEGF） in serum was determined by enzyme linked immunosorbent assay（ELISA）. In addition， the protein expression of Bad and Bcl-2 in gastric mucosa was detected by immunohistochemistry （IHC）.ResultA total of 241 active components of Xianglian Huazhuo prescription and 53 core targets were obtained. Xianglian Huazhuo prescription affected multiple biological processes，such as cell proliferation and apoptosis，inflammatory reaction，regulation of DNA metabolism，and cell response to redox，as well as phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt），TNF，mitogen-activated protein kinase （MAPK），cancer and cancer-related signaling pathways. The animal model verification showed that Xianglian Huazhuo prescription lowered the levels of IL-6，TNF-α，IL-1β and VEGF in serum of CAG rats，and reduced the protein expression of Bad and Bcl-2 in gastric tissue.ConclusionXianglian Huazhuo prescription could regulate PI3K/Akt signal pathway and improve gastric mucosal injury in CAG by participating in biological processes such as cell proliferation，apoptosis and inflammation.
Abstract：ObjectiveTo investigate the targets and mechanism of Baofeikang granules in the treatment of pulmonary fibrosis based on network pharmacology and verify the predicted mechanism based on animal experiment.MethodThe active ingredients and targets of Baofeikang granules were screened via the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform， and the targets of pulmonary fibrosis were searched in various disease databases. The common targets shared by Baofeikang granules and the disease were extracted for the establishment of the protein-protein interaction （PPI） network in STRING. Cytoscape 3.8.0 was used to analyze the network topology of the key targets and to establish the ''active ingredient-target'' network. Gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analyses were performed on the core targets to explore their possible molecular mechanisms. The rats were assigned into four groups： normal group， model group， prednisone acetate group， and Baofeikang granules group. The rat model of interstitial lung fibrosis was established by tracheal instillation of bleomycin. After 21 days of gavage， the lung tissues of rats were stained with hemotoxylin and eosin （HE） for the observation of morphological changes， and phosphatidylinositol 3-kinase （PI3K） and protein kinase B （Akt） were detected via immunohistochemical （IHC） staining.ResultBased on network pharmacology， 18 key targets of Baofeikang granules were identified for the treatment of pulmonary interstitial fibrosis， including Akt1， mitogen-activated protein kinase （MAPK） 1， myelocytomatosis oncogene （MYC）， hypoxia-inducible factor-1α （HIF-1α）， cyclin-dependent kinase inhibitor 1A （CDKN1A）， epidermal growth factor receptor （EGFR）， and Runt-related transcription factor （RUNX2）. KEGG pathway enrichment predicted that Baofeikang granules exerted anti-fibrotic effect mainly through PI3K/Akt， tumor necrosis factor （TNF）， and interleukin-17 （IL-17） signaling pathways. The IHC results in animal experiment showed that the protein levels of PI3K and Akt were lower in the Baofeikang granules group than in the model group （P<0.05， P<0.01）.ConclusionBaofeikang granules has low toxicity， multiple targets， and multiple pathways in the treatment of pulmonary fibrosis. It may alleviate pulmonary fibrosis through regulating PI3K/Akt signaling pathway， so as to improve the lung function.
Abstract：Network pharmacology （NP） is a novel interdisciplinary subject based on the combination of systems biology， multi-omics theory， computer synthesize， network database， and so on. It is widely used in traditional Chinese medicine （TCM） for active component screening， compatibility rule， pharmacological mechanism， and toxicity-efficacy network. Domestic NP-related papers began to boost from 2017， but some research showed abnormal "homogenization" in the screening of key components. Due to the non-standard and unreasonable situation in early NP analysis， the screening results always contained the same and widely-existed 200-500 Chinese medicine substances as the key components for TCM compounds， regardless of the various “disease-treatment-medicine” approaches to the study. If the "homogenization" phenomenon cannot be promptly clarified and corrected， it will lead to the misunderstanding that the components such quercetin， kaempferol， and sitosterol are "guaranteed to cure all diseases"， which overestimates the pharmacological weights of the components in each disease. This phenomenon seriously interferes the selection and quality control of Q-Markers related to TCM compounds. It violates and negates the scientific connotation of "treating the same disease with different therapies" and "treating different diseases with the same therapy" guided by the holistic view and the theory of syndrome differentiation and treatment. In the long term， the "homogenization" phenomenon may even hinder the healthy development of NP in TCM. Based on TCM theory and modern medicine， this paper started with the "thomogenization" of component screening in NP， and analyzed from the three module of "tphenomenon consequences"， "tcause exploration"， and "tsolving strategies"， thus providing important references for NP research methods.
Keywords：network pharmacology;key components;homogenization;Q-Markers of traditional Chinese medicine;traditional Chinese medicine compounds
Abstract：With the acceleration of aging and the changes of people's lifestyle and environment， the incidence of chronic non-communicable diseases （chronic diseases） in China is increasing， which has become a major public health problem. The number of sub-health people is even more and rising. The thought of treating disease before its onset emphasizes taking precautions against disease and preventing change after disease onset， which is one of the core contents of traditional Chinese medicine （TCM） theory. According to the theory of TCM， health as a dynamic balance of yin and yang and diseases can be prevented and treated by maintaining this balance. The "state adjustment" strategy suggests to treat diseases by adjusting the state of exuberance or decline based on recognizing and distinguishing "state"， which has important guiding significance for developing and evaluating the healthcare food produced with Chinese medicinal materials. Healthcare food plays a vital role in health promotion， chronic disease prevention， and quality of life improvement. At present， the healthcare food produced with Chinese medicinal materials in China generally lacks TCM guidance and fails to fully reflect the advantages of TCM. In this study， the development strategy， functional claims， evaluation methods， and existing problems of healthcare food were discussed. Furthermore， with diabetes as an example， the "state adjustment" strategy was introduced into product development and evaluation to provide ideas for promoting the rapid growth of healthcare food with TCM characteristics.
Keywords：state adjustment;traditional Chinese medicine （TCM）;healthcare food;development and evaluation
Abstract：Chronic heart failure is a serious heart disease with dyspnea and limited activity tolerance as the main clinical manifestations. Activation of the inflammatory system can significantly stimulate cardiac fibrosis and remodeling and promote the progression of heart failure， playing a key role in the development of the disease. Studies have confirmed that inflammation is involved in the development of different types of heart failure. "Toxic pathogen theory" is an important basic theory of traditional Chinese medicine （TCM） to explain the occurrence of diseases. We concluded the similarities between TCM toxic pathogens and inflammation in concept， disease location， etiology， syndrome differentiation， and clinical characteristics. Chronic heart failure is manifested by the toxic pathogens of turbid phlegm， stagnated blood， and accumulated fluid. Heart vessel obstruction is the main pathological factor， and the inflammatory factors produced by necrotic cardiomyocytes are the microscopic manifestations of the obstruction. Therefore， based on the "toxic pathogen theory"， this study aimed to clarify the role of inflammation in the development of chronic heart failure from both macroscopic and microscopic perspectives. Moreover， this paper proposed that the stagnated blood has not been transformed into toxin in the early stage of the disease and thus the products of clearing heat and detoxification should not be used. At the development stage of the disease when the transformation tends to begin， treatment should be based on syndrome differentiation， and detoxifying Chinese medicine should be used in order to achieve the goal of "removing toxin without harming the healthy Qi". At the late stage of heart failure， toxins have been accumulated and detoxifying medicines and therapies should be applied to eliminate the toxic pathogens. This study is expected to lay a foundation for the modern research on the role of inflammation in the development of chronic heart failure with TCM theory and guide the diagnosis and treatment of this disease.
Abstract：Traditional Chinese medicine （TCM） theory holds that "kidney governing water" plays a leading role in maintaining the metabolism of water and fluid in the body. The opening and closing of kidney qi determines the distribution， retention， and excretion of water and fluid. The treatment of some diseases caused by the imbalance of water and fluid metabolism is often carried out based on the kidney. Aquaporins are channel proteins that specifically transport water. They act as a regulator of intracellular and intercellular water flow and maintain the dynamic balance of water and fluid. Because of the differences in their location and expression， they have different physiological functions. Numerous studies have shown that aquaporins are widely distributed in the kidney， and their altered expression is of important significance to reveal the imbalance of water and fluid metabolism caused by different diseases. Doctors of all dynasties have accumulated rich experience in the continuous exploration of TCM regulating water and fluid metabolism disorders. TCM has the unique advantages of holistic view involving multiple pathways， components， and targets， and has achieved satisfactory regulatory effect， but the specific mechanism of action is not fully revealed. Therefore， this study explored the internal correlation of "kidney governing water"， the expression of aquaporins， and water and fluid metabolism disorders， summarized the regulatory effect and mechanism of Chinese medicinal extract， single Chinese medicine， and Chinese medicinal compound on water and fluid metabolism disorder， and interpreted the scientific connotation of "kidney governing water"， in order to provide new ideas and new directions for the TCM treatment of diseases due to water and fluid metabolism disorders.
Keywords：kidney governing water;aquaporins;water and fluid metabolism;traditional Chinese medicine
Abstract：The research on the essence of triple energizer has not reached a consensus. The correspondence between the existing understanding and the classical theory of triple energizer is still limited in terms of structure and function. According to the traditional theory， nutrient-defense takes channels as the main circulatory system， while the operation of nutrient-defense in the triple energizer remains unclear. Since little is known about the physical structure of the triple energizer， the role of triple energizer as a collection of other Zang-fu organs has been ignored in most cases. The new progress in anatomy paves the way for the research on the essence of triple energizer. The function and structure of triple energizer are similar to those of interstitium and interfacial fluid flow， which enriches our understanding of the macro and micro structures of triple energizer. The triple energizer is distributed throughout the body and composed of membrane and interstitial space. The material structure of triple energizer includes fiber scaffold， collagen fiber， mesenchymal stem cells， histiocytes， pericytes， and interstitial fluid. The functions of triple energizer include passing body fluids， operating nutrient-defense， distributing original Qi， and transmitting and changing pathogenic Qi. According to the available theories and research achievements， we put forward the concept of vertical and horizontal triple energizer， pointed out that triple energizer had independent structure and the features of Zang-fu organs， and preliminarily defined the spatial distribution of triple energizer. The relationship between channels and triple energizer is essential for discussing the operation of nutrient-defense. Telocyte （Tc） and telopod （Tp） has the characteristics of channels in function and structure. The connective tissue with the distribution of Tc and Tp belongs to the same material as the basic structure of interstitial/interfacial fluid flow system and the fibrous skeleton of interstitium. It is clear that channels and triple energizer have material commonality. From the operation paths of nutrient-defense， we proposed that channels may be soaked and attached in triple energizer and put forward the model of channels soaked and attached in triple energizer. By combining the circulation of nutrient-defense with the vertical and horizontal triple energize， we developed the theory of triple energizer-nutrient-defense loop to comprehensively describe the generation， transport， and metabolism of nutrient-defense in channels and triple energizer， aiming to provide a theoretical model for future studies of disease transmission and change from exterior to interior.
Abstract：Ischemic stroke， also known as cerebral infarction， is the most common type of stroke. Ischemic stroke is extremely harmful with high rates of morbidity， incidence， disability， and mortality， bringing a huge burden on society and families. As a result， finding new and effective prevention and treatment methods is critical. The pathological mechanism of ischemic stroke is very complex and superimposed， with inflammatory response serving as a critical pathological link in the ischemic stroke cascade injury process. NOD-like receptor 3 （NLRP3） is an intracellular sensor， and the inflammatory cascade mediated by the activated NLRP3 inflammasome can exacerbate ischemic stroke injury through the release of inflammatory factors. Taking the NLRP3 inflammasome as the entry point， a large number of experimental studies on the intervention of traditional Chinese medicine （TCM） in the assembly and activation of NLRP3 inflammasome have been carried out， which proved that Chinese medicinal monomers or prescriptions with the main functions of tonifying deficiency， clearing heat and removing toxin， eliminating phlegm， promoting circulation and resolving stasis can interfere with the assembly and activation of NLRP3 inflammasome， reduce the inflammatory response， and relieve ischemic stroke. This study reviewed the assembly and activation of NLRP3 inflammasome， the mechanism of NLRP3 inflammasome in ischemic stroke， and the prevention and treatment of ischemic stroke by TCM through regulation of the NLRP3 inflammasome， which provides a new entry point for the pathological mechanism of ischemic stroke and a direction for the development of new treatments for ischemic stroke.
Keywords：NOD-like receptor 3 （NLRP3）;inflammasome;ischemic stroke;traditional Chinese medicine
Abstract：With the gradual aggravation of aging in China， the prevalence of osteoporosis is increasing year by year. Osteoporosis has become a major public health problem threatening the health of middle-aged and elderly people， especially middle-aged and elderly women. There are many predisposing factors and complex pathogenesis of osteoporosis. The interpretation of osteoporosis has been the focus of clinical research in recent years. How to prevent and treat osteoporosis more effectively has also become a major problem faced by researchers. In recent years， the balance and homeostasis of calcium and phosphorus regulated by intestinal absorption， renal excretion and bone have become one of the hot topics， and the balance and homeostasis of calcium and phosphorus in vivo are the key to normal bone homeostasis. At the same time， as a complex microbial community living in the gastrointestinal tract， intestinal flora can produce a variety of regulators affecting metabolism. It has been widely confirmed that it acts on the body indirectly or directly， in multiple ways and targets to prevent and treat osteoporosis. Therefore， further exploring the role and mechanism of intestine kidney bone axis in osteoporosis plays a far-reaching significance for the prevention and treatment of osteoporosis. In recent years， scholars have made a lot of exploration on the prevention and treatment of osteoporosis with traditional Chinese medicine （TCM）， and found that TCM can intervene the expression of intestinal flora and play the effect of prevention and treatment of osteoporosis. Based on the "intestine kidney bone axis"， this paper briefly discusses the integrated traditional Chinese and western medicine of kidney and osteoporosis， intestine and osteoporosis， intestine kidney axis， the treatment of kidney from intestine， intestine and osteoporosis， and the application of TCM in regulating intestinal flora in osteoporosis， in order to provide new ideas for the prevention and treatment of osteoporosis.
Abstract：Autophagy is a lysosome-mediated catabolic process that captures and degrades dysfunctional organelles and useless proteins during cellular stress process， which plays a dual role in cervical cancer. In the early stage of cervical cancer， autophagy inhibits the occurrence and development of cervical cancer by prohibiting the accumulation of oncogenic p62 protein. In the advanced stage of cervical cancer， inhibition of autophagy of cancer cells enhances the sensitivity of cancer cells to chemotherapeutic drugs， thus inhibiting their proliferation. In recent years， the research on Chinese medicine monomers regulating autophagy in the treatment of cervical cancer has attracted extensive attention from scholars at home and abroad. Chinese medicine monomers regulate the autophagy of cervical cancer cells through multiple pathways and multiple targets， so as to increase the apoptosis rate and reduce the resistance of cancer cells to chemotherapeutic drugs. Therefore， this paper reviewed the mechanism of Chinese medicine monomers in inhibiting cervical cancer through autophagy， expecting to find new breakthroughs in the discovery and development of preventive and therapeutic drugs for cervical cancer. By reviewing the literature， it was found that in the early stage of cervical cancer， Chinese medicine monomers activated autophagy to promote apoptosis of cancer cells， and the main mechanism was to increase lysosomal membrane permeability and chemotherapeutic sensitivity and activate intact autophagy flow. In the advanced stage of cervical cancer， inhibition of autophagy reduced the sensitivity of cancer cells to chemotherapy drugs by inhibiting the formation of autophagosomes and autolysosomes. The treatment of cervical cancer by Chinese medicine monomers regulating autophagy has achieved certain effect， but there are few clinical experimental studies and lack of reliable clinical theoretical basis. Therefore， it is essential to carry out more clinical experimental studies on Chinese medicine monomers regulating autophagy to treat cervical cancer， thus finding more reliable theoretical basis for the treatment of tumors.
Keywords：cervical cancer;Chinese medicine monomers;autophagy;anticancer effect;research progress
Abstract：Bladder cancer is the most prevalent malignant tumor in the urinary tract in China. Western medical treatments， including long-term regular endoscopy， intravesical chemotherapy， immunotherapy， and radical cystectomy， are effective， whereas the high recurrence rate still plagues both doctors and patients. Among the classical signaling pathways involved in the formation and progression of bladder cancer， the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway is a key one. Modern pharmacological studies have demonstrated that Chinese herbal medicines and their monomer components can alleviate the discomfort， prolong the survival， and improve the quality of life of the patients undergoing tumor treatment. The relevant literature in the past decade has revealed that flavonoids， terpenoids， polysaccharides， gambogic acids， bibenzyls， and alkaloids from Chinese herbal medicines regulate the PI3K/Akt signaling pathway to play a role in the proliferation， apoptosis， invasion， migration， drug resistance， and autophagy of bladder cancer cells， thereby exerting the activity against bladder cancer. Although some targets and the potential mechanisms of the monomer components in the treatment of bladder cancer have been clarified， the research on the monomer components is limited to in vitro cellular experiments and animal experiments. Researchers face the great challenge in the application of the monomer components from Chinese herbal medicines into clinical practice. We summarized the recent studies about the regulatory effects of monomer components from Chinese herbal medicines on the PI3K/Akt signaling pathway in bladder cancer， aiming to give insights into the research on the drug therapy of bladder cancer and the underlying mechanism.
Keywords：bladder cancer;extracts of Chinese herbal medicines;phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway;research progress
Abstract：Epilepsy is a recurrent neurological disease with synchronous abnormal high discharge of neurons in the brain. The pathogenesis of this disease is extremely complex， which is closely related to neurotransmitter regulation， oxidative stress response， inflammatory factors， neuroglial cell， and abnormal gene expression. Western medicine mainly uses phenobarbital， phenytoin sodium， carbamazepine， and other drugs， but long-term use also produces certain toxic and side effects. Traditional Chinese medicine （TCM） believes that the pathogenesis of epilepsy is related to wind， fire， phlegm， and blood stasis， which leads to dysfunction of viscera， disorder of Qi movement， and finally uncontrolled spirit. In recent years， TCM has achieved certain curative effects on the treatment of epilepsy. As a high-frequency antiepileptic drug， Acori Tatarinowii Rhizoma has the effects of opening orifices and eliminating phlegm， awakening spirit and benefiting intelligence， and removing dampness and opening stomach， which has been widely used in clinic. In this paper， the pathogenesis of epilepsy and the pharmacological mechanism of Acori Tatarinowii Rhizoma extract and chemical components in the treatment of epilepsy were expounded by referring to relevant pharmacological studies and animal experiments. It was found that Acori Tatarinowii Rhizoma played a role in regulating the neurotransmitter level， antioxidant stress response， scavenging oxygen free radicals， regulating the expression of c-fos gene， reducing the level of inflammatory mediators， resisting neuronal apoptosis， and regulating the neuroglial cells and the permeability of blood-brain barrier. This paper summarizes the positive effects of Acori Tatarinowii Rhizoma on the treatment of epilepsy， and provides a scientific basis for the popularization and application of Acori Tatarinowii Rhizoma in the prevention and treatment of epilepsy.
Abstract：The quality of Chinese materia medica is the premise to ensure its safety and effectiveness in clinical application， and the standardization of Chinese materia medica quality is the most important to realize the sustainable development of traditional Chinese medicine （TCM）. At present， the quality control system of Chinese materia medica has been transformed from a single chemical evaluation to the overall quality control guided by clinical efficacy. However， some quality control items of decoction pieces are still lacking or imperfect in the drug standard of prescription， which makes it difficult to guarantee the effectiveness and safety of Chinese materia medica in clinical application. Based on this， the quality control models and innovative ideas of Chinese materia medica were analyzed and discussed from the perspectives of chemical analysis， biological evaluation and clinical application in this paper. Aiming at the existing problems and actual needs in the control system of Chinese materia medica， this paper proposed the improvement strategies in accordance with the characteristics of TCM， in order to provide theoretical basis for the related research on quality control of Chinese materia medica.
Keywords：traditional Chinese medicine （TCM）;quality control;control extract;fingerprint;quality marker （Q-Marker）;grade evaluation;integrated quality view
Abstract：In this paper， by consulting the historical herbs and medical classics coupled with related literature in modern research， the historical edition of Chinese Pharmacopoeia and local chronicles of traditional Chinese medicine （TCM） along with the ancient historical evolution of the processing methods of Pinelliae Rhizoma origin as well as the related processing methods of Pinelliae Rhizoma origin from 1959 to 2020 were systematically collated and summarized. It was found that the main processing methods of Pinelliae Rhizoma origin were peeling， decoction washing， lime wrapping and sun-drying. However， stacking， peeling， sun-drying or oven-drying are the primary methods in modern local chronicles of TCM. Meanwhile， washing， peeling， removing fibrous roots and sun-drying are the main methods in Chinese Pharmacopoeia. In addition， there were some changes in the quality evaluation of Pinelliae Rhizoma in different historical periods. Round and white were the best in the quality evaluation of Pinelliae Rhizoma in ancient times， while the evaluation indexes were further refined to size， color， texture， powder property， purity and evenness in modern herbal works. In modern studies， the quality of Pinelliae Rhizoma was mostly evaluated by the chemical components such as alkaloids， total organic acids， polysaccharides， nucleosides， fingerprint and pharmacodynamics. At present， the purification and drying stages of Pinelliae Rhizoma are in the transitional stage between the traditional manual peeling and natural drying methods as well as the modern mechanized and large-scale production. Therefore， a reasonable and feasible modern processing methods and guiding standards of Pinelliae Rhizoma are developed urgently to normalize the processing of Pinelliae Rhizoma and ensure the quality of medicinal materials.
Keywords：Pinelliae Rhizoma;harvesting;producing area processing;historical evolution;quality evaluation;Chinese Pharmacopoeia;chemical composition