Aim To understand the hemostatic effect of a non-polysaccharide fraction of Bletilla striata ( BS-80EE) and to clarify its mechanism of action .Methods The non-polysaccharide fraction ( BS-80 EE ) was prepared by passing the 95%ethanol extract of Bletilla striata through a D101 macroporous resin column elu-ted first with water and then with 80%ethanol.Bleed-ing time ( BT ) and clotting time ( CT ) of heparinized mice were employed as indicators for evaluating the he-mostatic effect of BS-80EE.The mechanism of action was investigated through observing the effect of BS-80 EE on platelet aggregation induced by adenosine diphosphate ( ADP) in rats with nephelometry and tes-ting the effect of BS-80EE on the thrombin time(TT), prothrombin time(PT), activated partial thromboplas-tin time(APTT), fibrinogen(FIB), P-selectin(P-S), thrombin-antithrombin complex ( TAT ) , D-dimer ( D-D) and plasminogen activator inhibitor-1 ( PAI-1 ) . Results BS-80 EE significantly shortened the CT and BT( P<0.01 or 0.05 ) of heparin mice in a dose-de-pendent manner; groups of all doses significantly re-duced the rat TT ( P <0.01 or 0.05 ) , and the high-dose group significantly increased the FIB content ( P<0.05); the mid-dose group and high-dose groups of BS-80EE significantly increased the contents of P-S, TAT and PAI-1 , while reduced the D-D production in rats ( P <0.01 ); although dose-dependent reductions of APTT and PT were observed for each treatment-group, no significance was observed .Conclusion BS-80EE possess pronounced hemostatic effect by promo-ting platelet aggregation and coagulation .

OBJECTIVETo develop an ultra performance liquid chromatography (UPLC) method for simultaneous determination of five ellagic acids in Euscaphis japonica.

METHODAnalysis was carried out on an Acquity BEH C18 (2.1 mm x 100 mm, 1.7 microm) column at 40 degrees C eluted with acetonitrile-0.1% phosphoric acid aqueous solution as mobile phases in gradient elution. The flow rate was 0.2 mL x min(-1), and the detection wavelength set was monitored at 245 nm.

RESULTAll calibration curves showed good linear relationship within test ranges (r >0.9997), and the overall recoveries were in the range of 97.2%-102.1%, with RSD less than 3.2% (n = 6). The overall RSD of precision test were less than 2.9%.

CONCLUSIONThe developed method was simple, rapid, accurate and reproducible, and can be used for the quality control of E. japonica.

A simple and selective ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS) assay was developed for the determination of the human plasma protein binding of four bioactive flavonoids (such as orientin and vitexin) in Polygonum orientale. Protein precipitation was used for sample preparation. Equilibrium dialysis technique was applied to determine the plasma protein binding under physiological conditions. The separation was achieved through a Waters C18 column with a mobile phase composed of 0.1%formic acid in acetonitrile and 0.1%aqueous formic acid using step gradient elution at a flow rate of 0.35 mL/min. A Waters ACQUITY?TQD system was operated under the multiple reaction monitoring (MRM) mode of positive electrospray ionization. All of the recovery, precision, accuracy and stability of the method met the requirements. Good correlations (r40.99) of the four compounds were found, which suggested that these compounds can be simultaneously determined with acceptable accuracy. Results showed that the plasma protein bindings of the four bioactive flavonoids were in the range of 74-89% over the six concentrations studied. The binding parameters containing protein binding affinity, protein binding dissociation constant, and protein binding site were studied. The maximum ability to bind with protein was also determined in the assay in order to understand the drug-protein binding of each compound better.

OBJECTIVETo study the chemical constituents of the essential substance from the Polygonum oriental.

METHODChromatographic techniques were employed for isolation and purification of the constituents and their structures were determined by spectral analysis and chemical evidence.

RESULTSeven compounds were obtained and identified as ombuine-3-O-beta-D-galactopyranoside (1), ombuine-3-O-rutinoside (2), tryptophan (3), quercetin-3-O-methyl ether (4), kaempferol-3-O-(2"-O-alpha-L-rhamnopyranosyl) -beta-D-glucuronopyranoside (5), quercetin-3-O-(2"-O-alpha-L-rhamnopyranosyl)-beta-D-glucuronopyranoside (6), quercetin-3-O-beta-D-glucuronide (7).

CONCLUSIONCompounds 4-7 were isolated from P. oriental for the first time and compounds 1-3 were firstly obtained from the genus Polygonum. The total 1H and 13C-NMR date of compound 1 were assigned for first time.

OBJECTIVE:To identify lipid metabolites of endophytic fungus Aspergillus terreus from Polygonum capitatum, and to investigate their anti-multidrug resistant bacteria and anti-inflammatory effects. METHODS:GC-MS was used to analyze and identify lipid metabolites of A. terreus. MIC of lipid metabolites and main composition to 10 strains of multidrug resistant bacteria (Klebsiella pneumoniae,Proteus common,Staphylococcus epidermidis,Escherichia coli,Staphylococcus aureus,Enterobacter cloacae,Pseudomonas aeruginosa,Proteus mirabilis,Enterococcus faecium and Acinetobacter baumanii) were determined by 96-well plate microdilution method. The spread plate method was used to determine MBC of samples to bacteria. LPS-induced RAW264.7 inflammatory model was used to investigate the effects of different mass concentrations(50,100,200 μg/mL)of lipid metabolites on the release of NO and TNF-α after treated for 24 h. RESULTS:A total of 13 compounds were identified in lipid metabolites of A. terreus,among which palmitic acid,stearic acid,linoleic acid and oleic acid were main components,and relative percentages of them were 29.35％,10.87％,21.94％,34.85％. The lipid metabolites displayed anti-bacterial activity against E. coli and K. pneumonia with MICs of 12.5 mg/mL and 50 mg/mL,MBC of 25 mg/mL and 100 mg/mL,respectively. The main 4 compositions could inhibit the growth of E. coli,with MIC of 0.5-1 mg/mL,among which palmitic acid showed significant antibacterial activity,especially to E. faecium(MIC of 0.25 mg/mL). 50,100 μg/mL lipid metabolites could signifiantly inhibit the release of inflammatory factor of NO and TNF-α in RAW264.7 in vitro. CONCLUSIONS:The lipid metabolites of endophytic fungus A. terreus from P. capitatum show anti- multi-drug resistant bacteria and anti-inflammatory effects.

Objective:To observe the application effect of mind mapping combined with interactive communication mode in clinical teaching of neurosurgery.Methods:A total of 40 students who practiced in the Department of Neurosurgery in the Affiliated Hospital of Guizhou Medical University from September 2019 to September 2020 were included in the control group, and traditional teaching was adopted; another 40 students who practiced from October 2020 to October 2021 were included in the observation group, and mind mapping combined with interactive communication mode was adopted for teaching. The two groups of students were taught for 2 weeks, and after the teaching, the teaching effect was compared between the two groups. SPSS 25.0 software was used to conduct t-test and Chi-square test. Results:After 2 weeks of teaching, the scores of theoretical knowledge (90.38±4.03) and practical operation skills (93.37±3.48) in the two groups were higher than those before teaching [(85.52±5.26) and (87.25±4.48)], with statistically significant differences ( t=4.63, 6.83, P<0.001). The case analysis score of the two groups was higher than that before teaching, and that of the observation group (86.03±6.07) was higher than that of the control group (79.13±5.57), with statistically significant differences ( t=5.30, P<0.001). The scores of interpersonal communication ability and cooperation ability of the two groups were higher than those before teaching. The scores of interpersonal communication ability (82.53±4.74), cooperation ability (169.73±7.55) of the observation group were higher than those of the control group [(77.93±4.45) and (158.42±8.01)], with statistically significant differences ( t=4.48, 6.49, P<0.001). Conclusion:Mind mapping combined with interactive communication mode can effectively improve the clinical basic knowledge and clinical practice ability of interns in the Department of Neurosurgery, and improve their communication and cooperation ability.

OBJECTIV E To investigate the effect s and mechanism of the ethanol extract of Tiarella polyphylla （“TPE”）on learning and memory impairment in mice. METHODS Male Kunming mice were randomly divided into normal group ，model group，positive group （donepezil hydrochloride 4 mg/kg）and TPE low-dose ，medium-dose and high-dose groups （150，300，600 mg/kg），with 10 mice in each group. Drug administration groups were given relevant medicine intragastrically once a day ，and normal group and model group were given water intragastrically once a day ，for consecutive 22 d. On the 17th day ，administration groups and model group were intraperitoneally injected with scopolamine hydrobromide （3 mg/kg）to establish a model of learning and memory impairment. The learning and memory ability of the mice were evaluated by the Morris water maze. Hematoxylin-eosin （HE） staining was used for morphological observation of hippocampus cells of the mice. The levels of acetylcholinesterase （AChE），choline acetyltransferase （ChAT），superoxide dismutase （SOD），malondialdehyde（MDA），interleukin-6（IL-6）and tumor necrosis factor-α（TNF-α）in cerebral tissue as well as the relative expression of phosphorylated Tau protein （p-Tau），β-site amyloid precursor protein cleaving enzyme 1（BACE1）and amyloid precursor protein （APP）in hippocampus tissue were all detected. RESULTS The escape latency of mice in positive group ，TPE medium-dose and high-dose groups were all significantly shortened than the model group on the 4th to 5th day of training ，while the times of crossing platform and the percentage of movement distance in target quadrant were significantly increased （P＜0.05）. Compared with model group ，the neurons in the hippocampal CA 1 region of mice were increased to var ying degrees in administration groups ，the ne urons in solidified and atrophic state decreased ，and the arrangement of neurons tended to be close；the levels of ChAT and SOD in cerebral tissue were significantly increased in positive group and TPE medium-dose and high-dose groups ；the levels of AChE ，MDA，IL-6，the levels of TNF-α and relative expression of p-Tau ，BACE1 and APP in hippocampus tissue were decreased significantly （P＜ 0.05）. CONCLUSIONS TPE can improve the learning and memory impairment induced by scopolamine in mice ，and the mechanism may be related to balancing the brain cholinergic system ，alleviating oxidative stress injury ，improving inflammatory response，and inhibiting the overexpression of p-Tau ，BACE1 and APP .

OBJECTIVE To investigat e the effects of water extract （WCS）and ethanol extract （ECS）from the root of Caragana sinica on hyperuricemia （HUA）in mice. METHODS Kunming mice were randomly divided into normal control group ， model group ，allopurinol group （positive control ，5 mg/kg），benzbromarone group （positive control ，7.8 mg/kg），WCS low-dose ， medium-dose and high-dose groups （38，75，150 mg/kg），ECS low-dose ，medium-dose and high-dose groups （50，100，200 mg/kg）， with 10 mice in each group. Except for the normal control group ，the other mice were given potassium oxazinate intraperitoneally and hypoxanthine intragastrically for consecutive 7 d to establish HUA model. On the third day of modeling ，mice in each administration group were given corresponding drugs intragastrically ，and normal control group and model group were given equal volume of normal saline once a day for 5 consecutive days.The body weight of mice were weighted during administration ；one hour after the last administration ，the organ indexes of liver ，kidney and spleen were calculated ；the contents of serum uric acid （SUA）， blood urea nitrogen （BUN）and serum creatinine （SCR）；the activity of xanthine oxidase （XOD）in serum and liver tissue were determined. Relative mRNA and protein expressions of XOD in liver tissue ，relative expre ssions of GLUT9，URAT1 and OAT 1 in renal tissue were all detected ；and the pathological changes of renal tissue were observed. RESULTS There were no significant differences in liver index and spleen index in each group （P＞0.05）. Compared with normal control group ， except for allopurinol group ， there were no significant differences in the body weight and the contents of BUN and SCR in mice of other administration groups （P＞0.05）；the renal index and SUA content of mice in the m odel group and allopurinol group were significantly increased （P＜0.05）；in the model group ，the XOD activity in serum and liver tissue ，the relative mRNA and protein expression of XOD in liver tissue ，the relative expressions of GLUT 9 and URAT 1 protein in renal tissue were significantly increased （P＜0.05），and the relative expression of OAT 1 protein in renal tissue was significantly decreased （P＜ 0.05）. Compared with model group ，renal indexes of mice were decreased significantly in WCS and ECS groups （P＜0.05），and the pathological damage of renal tissue was significantly improved ；SUA content ，XOD activity in serum and liver tissue ，the relative mRNA and protein expression of XOD in liver tissue ，and the relative expression of URAT 1 protein in renal tissue were decreased significantly in administration groups （P＜0.05）. The relative expression of GLUT 9 protein in renal tissue of mice in benzbromarone group and ECS high-dose group decreased significantly （P＜0.05）；relative expression of OAT 1 protein in renal tissue of mice in benzbromarone group ，WCS low-dose and high-dose groups ，ECS low-dose group were increased significantly （P＜0.05）. CONCLUSIONS WCS and ECS can significantly decrease the contents of SUA in HUA model mice ，and improve pathological state of renal tissue ，the mechanism of which may be associated with inhibiting XOD activity and uric acid reabsorption，and down-regulating protein and mRNA expression of XOD.

OBJECTIVE：To investigate the improvement effect of Tiarella polyphylla ethanol extract （TPME）on CCl 4-induced hepatic fibrosis in mice ，and to explore its possible mechanism preliminarily. METHODS ：Totally 60 male Kunming mice were randomly divided into normal group ，model group ，positive control group （colchicine 0.1 mg/kg），TPME low-dose ，medium-dose and high-dose groups （250，500，1 000 mg/kg）according to body weight ，with 10 mice in each group. Except for normal group ， other groups were given 20％ CCl4 olive oil solution intraperitoneally to induce hepatic fibrosis ，twice a week ，for consecutive 8 weeks. From the fifth week after modeling ，administration groups were given relevant medicine intragastrically ，normal group and model group were given constant volume of normal saline intragastrically ，once a day ，for consecutive 4 weeks. Twelve hours after last administration ，the liver weight of mice in each group was measured and the liver index was calculated. The serum contents of ALT，AST，SOD，MDA，PC-Ⅲ，C-Ⅳ，LN，TNF-α and IL- 6 were determined. Western blot assay was used to detect the protein expression of α-SMA，TGF-β1 and Smad 3 in liver tissue. HE and Masson staining were used to observe the pathological changes of hepatic tissue. RESULTS ：Compared with normal group ，the liver index ，the activities of ALT and AST and the contents of MDA ， LN，PC- Ⅲ ，C- Ⅳ ，LN，TNF-α and IL- 6 in serum were increased significantly ， while the activity of SOD was 6011） decreased significantly in model group （P＜0.01）；the protein expression of α-SMA，TGF-β1 and Smad 3 in liver tissues were hfjsznd8@126.com increased significantly （P＜0.01）. Obvious fibrosis lesions was observed in liver tissue. Compared with model group ，the live indexes ，the activities of ALT and AST ，the contents of MDA，PC-Ⅲ，C-Ⅳ，LN，TNF-α and IL-6 in serum were decreased significantly in positive control group and TPME groups ， while the activities of SOD were increased significantly （P＜0.05 or P＜0.01）. The protein expression of α-SMA，TGF-β1 and Smad3 in liver tissue were decreased significantly （P＜0.05 or P＜0.01），and liver fibrosis was improved to different extent. Compared with TPME low-dose group ，the contents of PC- Ⅲ，LN and IL- 6 in serum ，protein expression of TGF-β1 and Smad 3 in liver tissue were decreased significantly in TPME high-dose group （P＜0.05）. CONCLUSIONS ：TPME can improve hepatic fibrosis induced by CCl 4 in mice ，the mechanism of which may be associated with the inhibition of collagen synthesis and oxidative stress，the reduction of inflammatory factors ，and the down-regulation of the expression α-SMA and relative proteins of TGF-β1/ Smad signal pathway.

ObjectiveTo investigate the effect of phytoestrogen biochanin A （BCA） on liver fibrosis induced by CCl₄ in female mice with bilateral oophorectomy （ovariectomized） and its mechanism. MethodsA total of 50 ovariectomized Kunming mice were selected and given intraperitoneal injection of CCl₄ to establish a model of liver fibrosis， and then according to body weight， they were randomly divided into model group， positive control group， and low-， middle-， and high-dose BCA groups， with 10 mice in each group. In addition， 10 female mice in the same litter were given resection of a small amount of adipose tissue near both ovaries to establish the sham-operation group. The mice in the positive control group were given estradiol 2 mg/kg by gavage， and those in the low-， middle-， and high-dose BCA groups were given BCA by gavage at a dose of 25， 50， and 100 mg/kg， respectively， once a day for 7 consecutive weeks； the mice in the sham-operation group and the model group were given an equal volume of 0.5% sodium carboxymethyl cellulose solution by gavage. The mice were anesthetized and sacrificed after administration to collect samples. Liver index and uterus index were measured； HE staining and Masson staining were used to observe liver histopathological changes； the biochemical analysis was used to measure the activity of aspartate aminotransferase （AST） and alanine aminotransferase （ALT）； ELISA was used to measure the levels of interleukin-6 （IL-6） and tumor necrosis factor-α （TNF-α） in liver tissue， and Western blot was used to measure the relative protein expression levels of collagen Ⅰ， transforming growth factor-beta 1 （TGF-β₁）， alpha-smooth muscle actin （α-SMA）， estrogen receptor beta （ERβ）， and p-NF-κBp65/NF-κBp65 in liver tissue. The t-test was used for comparison of continuous data between two groups； a one-way analysis of various was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. The Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between multiple groups and further comparison between two groups. ResultsCompared with the sham-operated group， the model group had a significant increase in liver index and a significant reduction in uterus index， as well as significant increases in the activities of serum AST and ALT， the levels of IL-6 and TNF-α in liver tissue， and the protein expression levels of collagen Ⅰ， TGF-β₁， α-SMA， and p-NF-κBp65/NF-κBp65 in liver tissue （all P<0.05）， with no significant change in the expression of ERβ in liver tissue （P>0.05）， and the model group showed significant fibrosis lesions in the liver， such as hepatocyte edema， steatosis， and necrosis with inflammatory cell infiltration and hyperplasia， deposition， and staggered distribution of collagen fibers. Compared with the model group， the low-， middle-， and high-dose BCA groups had significant reductions in liver index， the activities of serum AST and ALT， the levels of IL-6 and TNF-α， and the protein expression levels of collagen Ⅰ， TGF-β₁， α-SMA， and p-NF-κBp65/NF-κBp65 in liver tissue （all P<0.05）， with no significant change in uterine index （P>0.05）， as well as a significant increase in the protein expression level of ERβ in liver tissue （P<0.05） and varying degrees of improvement in liver fibrosis lesions. ConclusionBCA can effectively improve CCL₄-induced liver fibrosis in ovariectomized female mice， possibly by upregulating ERβ to inhibit the NF-κB signaling pathway and then alleviating inflammatory response.