AIM To study the chemical constituents from the water fraction of rhizoma of Smilax trinervula Miq.and their biological activities.METHODS Polyamide,silica gel,Sephadex LH-20,ODS and semi-preparative HPLC were used for isolation and purification,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The antitumor activities were determined by MTT mothod,and the inhibitory activities on α-glucosidase were determined by PNPG method.RESULTS Eleven compounds were isolated and identified as tyrosine(1),uridine(2),2-(2',3',4'-trihydroxybutyl)-6-(2",3",4"-trihydroxybutyl)-pyrazine(3),2-(1',2',3',4'-tetrahydroxybutyl)-6-(2",3",4"-trihydroxybutyl)-pyrazine(4),2-(1',2',3',4'-tetrahydroxybutyl)-5-(2",3",4"-trihydroxybutyl)-pyrazine(5),uracil(6),2-(1',2',3',4'-tetrahydroxybutyl)-5-(1",2",3",4"-tetrahydroxybutyl)-pyrazine(7),dioscin(8),shikimic acid(9),pyrazine(10),3,4-dihydroxyphenyethyl alcohol 8-O-β-D-glycopyranoside(11).The IC50 values of compounds 8 to human breast cancer cell MCF-7 was(2.36±0.26)μg/mL,and the IC50 values of compounds 3-5 and 7 to α-glucosidase were(1.54±0.15)-(10.53±0.38)μg/mL.CONCLUSION Compounds 1-7,10 are isolated from Smilax genus for the first time,and compound 9,11 are first isolated from this plant.Compound 8 has anti-tumor activity,and compounds 3-5,7 have α-glucosidase inhibitory activities.

As the world's second largest vector of pathogens,ticks can spread a variety of pathogens by sucking the host's blood.Ticks not only threaten human life and health,but also cause great economic losses in animal husbandry.Artificial breeding of ticks can provide a stable environment for the growth and reproduction of ticks,thereby generating sufficient exper-imental materials for understanding ticks'biological characteristics,studying tick-borne pathogens,and developing anti-tick drugs and vaccines.Current methods of breeding ticks in the laboratory can be roughly divided into two categories:breeding methods using host animals or artificial membranes.The selection of breeding method must be comprehensively considered,ac-cording to tick types,blood-sucking habits,living environments,and other aspects.The development processes of the two methods,and their respective advantages and disadvantages,are described and discussed,to assist laboratories in artificial breeding of ticks.

Objective To investigate the effects of chronic intermittent hypoxia(CIH)on hippocampal neuronal injury in rats,and to clarify the role of endoplasmic reticulum(ER)stress-related apoptotic pathways.Methods Male Sprague-Dawley rats were randomly assigned to a control group(n=8)or a CIH group(n=8).The CIH group was exposed to intermittent hypoxia for 8 h/day over 8 weeks.Hippocampal neuronal morphology was examined by hematoxylin-eosin staining and transmission electron microscopy.Neuronal apoptosis was assessed using the TUNEL assay.Intracellular Ca2? levels were measured by flow cytometry.The mRNA and protein expression of ER stress-related factors(GRP78,CHOP)and the apoptotic effector Caspase-3 were quantified by qPCR and Western blot.Results Compared with controls,rats in the CIH group exhibited marked hippocampal neuronal damage,including disrupted cytoarchitecture,cytoplasmic dissolution,and swollen rough ER.Ultrastructural analysis revealed nuclear deformation and organelle disruption.TUNEL assay demonstrated a significant increase in apoptotic cells(P<0.05).Flow cytometry showed elevated intracellular Ca2? levels(P<0.05).GRP78,CHOP,and Caspase-3 were significantly upregulated at both mRNA and protein levels in the CIH group(all P<0.05).Conclusion CIH induces pronounced hippocampal neuronal injury and apoptosis in rats,associated with Ca2? dysregulation and activation of ER stress-mediated apoptotic pathways.These findings provide experimental evidence for elucidating the mechanisms of OSAHS-related neuronal injury and identifying potential therapeutic targets.

Objective To investigate the clinical features of chronic hepatitis C(CHC)patients with hepatitis C virus genotype 3(HCV GT3)infection and the risk factors for disease progression.Methods A multicenter retrospective cohort study was conducted among 1 002 CHC patients from 11 clinical centers in Northwest China from December 2017 to November 2023,and according to their genotype,they were divided into GT1,GT2,GT3,and GT6 groups.Clinical features were compared between the patients with different genotypes.The one-way analysis of variance was used for comparison of normally distributed continuous data between groups,and the Scheffe test was used for further comparison between two groups.The Kruskal-Wallis H test was used for comparison of data with skewed distribution between groups;the chi-square test or Fisher test was used for comparison of categorical data between groups.The multivariate logistic regression analysis was used to explore the influencing factors for the progression of CHC to liver cirrhosis.Results In terms of the genotype,there were 427 patients with GT1 infection,242 with GT2 infection,299 with GT3 infection(210 patients with GT3a infection,87 with GT3b infection,and 2 with unclassified genotype),and 34 with GT6 infection.The patients with GT3 infection had a significantly younger age than those with GT1 infection(51.3±0.5 years vs 53.2±0.6 years,P<0.05)or GT2 infection(51.3±0.5 years vs 53.7±0.8 years,P<0.05),and for the patients with liver cirrhosis,the patients with GT3 infection had a significantly younger age than those with GT1 infection(52.1±0.5 years vs 59.4±0.9 years,P<0.001)or GT2 infection(52.1±0.5 years vs 58.1±1.1 years,P<0.001).Among the patients with GT3 infection,male patients accounted for 77.9%and the patients with liver cirrhosis accounted for 46.2%,which were significantly higher than those among the patients with GT1,GT2 or GT6 infection(all P<0.001).At baseline,the patients with GT3 infection had significantly higher levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)than those with GT1 or GT2 infection,significantly higher aspartate aminotransferase-to-platelet ratio index(APRI)and fibrosis-4(FIB4)than those with GT1,GT2 or GT6 infection,a significantly lower platelet count(PLT)than those with GT2 or GT6 infection,a significantly higher level of alpha-fetoprotein than those with GT2 or GT6 infection,and a significantly lower level of albumin(Alb)than those with GT6 infection(all P<0.05).There were no significant differences between the patients with GT3a infection and those with GT3b infection in age,sex,the proportion of patients with liver cirrhosis,comorbidities,HCV RNA quantification,PLT,ALT,AST,alkaline phosphatase,Alb,APRI,and FIB-4(all P>0.05).The multivariate logistic regression analysis showed that PLT≤150×109/L(odds ratio[OR]=10.72,95%confidence interval[CI]:5.76-35.86,P<0.001)and Alb≤35 g/L(OR=3.74,95%CI:1.22-11.45,P=0.021)were risk factors for liver cirrhosis.Conclusion Most CHC patients with GT3 infection are male in Northwest China,and compared with the patients with other genotypes,such patients tend to have a younger age of onset and higher degrees of liver inflammation activity and fibrosis.Low PLT and a low level of Alb are risk factors for progression to liver cirrhosis in CHC patients with GT3 infection.

Objective:Elucidating causal links between gut microbiota and breast malignancies using mendelian randomization(MR)to Identify specific causal microbial taxa.Methods:Genome-wide association studies(GWAS)summary statistics on gut microbiota and breast malignancies,MiBioGen studied GWAS(N=18473)as exposure samples,and GWAS(N=123579)of breast malignancies in the IEU Open GWAS database as outcome samples.The in-verse variance weighted(IVW),weighted median and MR-Egger methods were used to analyze the MR statistical re-suits.Results:Increased abundance of Pasteurellales increased the risk of breast malignancy(IVW PFDR＜0.05,OR=1.21,95％Cl:1.07～1.36),heterogeneity test and pleiotropy test showed no effect on causality,and the"leave-one-out"method showed that the identified causal association was not driven by any single instrumental variable.The causal relationship is positive.Conclusion:Elevated abundance of pasteurellales in the gut microbiota is associated with an increased risk of breast malignancies

Standardizing the management of drugs used in clinical trials is one of the important means to protect the safety of trial participants and ensure that the trial results are scientifically reliable.The management of trial drugs runs throughout the entire clinical trial process,with some hospitals centralizing the management and others managing it within departments.Participants in the process include doctors,nurses,and pharmacists,among others.Investigate the management of investigational drugs in 15 hospitals and combine domestic relevant regulations and guidelines to standardize the management process of investigational drugs.A central pharmacy staffed by dedicated pharmacy personnel should be preferred,and if a departmental management model is used,a research physician/nurse or a pharmacy personnel with the necessary qualifications and competencies should be appointed as the drug manager.The reception and inventory of clinical trial drugs,as well as the return of drugs to the sponsor,should be completed by the institutional drug manager.The distribution of clinical trial drugs can be authorized to individuals with a professional background in pharmacy who have obtained professional qualification certificates or licensed pharmacist certificates.If non-pharmacist personnel distribute clinical trial drugs,other equivalent documents should be used instead of dispensing drugs based on prescriptions.

Objective:To observe the clinical effect of Cangfu Daotan Decoction combined with ethinylestradiol and cyproterone acetate,metformin in patients with spleen deficiency phlegm-dampness type polycystic ovary syndrome(PCOS)accompanied by insulin resistance.Methods:80 patients with PCOS accompanied by insulin resistance who were admitted to our hospital from June 2023 to June 2024 were included.The patients were divided into control group(treated with ethinylestradiol and cyproterone acetate,metformin)and study group(treated with modified Cangfu Daotan Decoction combine with ethinylestradiol and cyproterone acetate,metformin)by the random number table method,with 40 cases in each group.The total clinical effective rate,TCM syndrome score,sex hormone levels[prolactin(PRL),estradiol(E2),follicle-stimulating hormone(FSH),testosterone(T),and luteinizing hormone(LH)],insulin resistance indicators[fasting insulin(FINS),fasting blood glucose(FBG),homeostatic model assessment of insulin resistance(HOMA-IR)],and endometrial receptivity[pulsatile index(PI),resistance index(RI),endometrial thickness(ET),ovulation rate]and the occurrence of adverse reactions were compared between the two groups.Results:Compared with the control group after treatment,the total clinical effective rate,PRL,E2,FSH level,ET and the ovulation ratein the study group were higher,while the main symptoms,secondary symptoms,total score,T,LH levels,FBG,FINS levels,HOMA-IR,PI and RI were lower(P＜0.05).There was no difference in the incidence of adverse reactions between the two groups(P＞0.05).Conclusion:Cangfu Daotan Decoction combined with ethinylestradiol and cyproterone acetate,metformin in patients with spleen deficiency phlegm-dampness type PCOS accompanied by insulin resistance,can improve the clinical symptoms,increase the total clinical effective rate,it may be related to the regulation of insulin resistance,sex hormone levels,and endometrial receptivity,it is safe and reliable.

AIM To study the chemical constituents from ethyl acetate fraction of Balanophora harlandii Hook.f.and their tyrosinase inhibitory activity.METHODS Separation and purification were performed using silica gel,MCI,ODS,Sephadex LH-20 and semi-preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The monophenolase inhibitory activity was determined by the tyrosinase-catalyzed oxidation of L-tyrosine.RESULTS Twenty-four compounds were isolated and identified as sesamin(1),methyl caffeate(2),quercetin(3),5,7-dihydroxychromanone(4),methyl 3,4-dihydroxybenzoate(5),esculetin(6),kaempferol(7),naringenin(8),pyrogallic acid(9),pinosylvin(10),methyl propionate(11),caffeic acid(12),saccharinol(13),ferulic acid(14),trans-p-hydroxycinnamic acid(15),cinnamic acid(16),vanillic acid(17),vanillin(18),4-hydroxyacetophenone(19),4-hydroxybenzaldehyde(20),apigenin(21),(-)-isolariciresinol(22),(-)-secoisolariciresinol(23)and meso-2,3-di(3′,4′-methylenedioxybenzyl)butane-1,4-diol(24).The IC50 values of compounds 3,5,7,8,19,and 20 ranged from(0.246 5±0.028 3)to(1.278 2±0.021 3)mmol/L.CONCLUSION Compounds 1-9、11、15、17-21、24 are isolated from this plant for the first time,and 1,6,9,17-19,24 are first isolated from genus Balanophora.Compounds 3、5、7、8、19 and 20 have tyrosinase inhibitory activity.

Objective:To observe the clinical effect of Cangfu Daotan Decoction combined with ethinylestradiol and cyproterone acetate,metformin in patients with spleen deficiency phlegm-dampness type polycystic ovary syndrome(PCOS)accompanied by insulin resistance.Methods:80 patients with PCOS accompanied by insulin resistance who were admitted to our hospital from June 2023 to June 2024 were included.The patients were divided into control group(treated with ethinylestradiol and cyproterone acetate,metformin)and study group(treated with modified Cangfu Daotan Decoction combine with ethinylestradiol and cyproterone acetate,metformin)by the random number table method,with 40 cases in each group.The total clinical effective rate,TCM syndrome score,sex hormone levels[prolactin(PRL),estradiol(E2),follicle-stimulating hormone(FSH),testosterone(T),and luteinizing hormone(LH)],insulin resistance indicators[fasting insulin(FINS),fasting blood glucose(FBG),homeostatic model assessment of insulin resistance(HOMA-IR)],and endometrial receptivity[pulsatile index(PI),resistance index(RI),endometrial thickness(ET),ovulation rate]and the occurrence of adverse reactions were compared between the two groups.Results:Compared with the control group after treatment,the total clinical effective rate,PRL,E2,FSH level,ET and the ovulation ratein the study group were higher,while the main symptoms,secondary symptoms,total score,T,LH levels,FBG,FINS levels,HOMA-IR,PI and RI were lower(P＜0.05).There was no difference in the incidence of adverse reactions between the two groups(P＞0.05).Conclusion:Cangfu Daotan Decoction combined with ethinylestradiol and cyproterone acetate,metformin in patients with spleen deficiency phlegm-dampness type PCOS accompanied by insulin resistance,can improve the clinical symptoms,increase the total clinical effective rate,it may be related to the regulation of insulin resistance,sex hormone levels,and endometrial receptivity,it is safe and reliable.

Immobilized enzyme-based enzyme electrode biosensors, characterized by high sensitivity and efficiency, strong specificity, and compact size, demonstrate broad application prospects in life science research, disease diagnosis and monitoring, etc. Immobilization of enzyme is a critical step in determining the performance (stability, sensitivity, and reproducibility) of the biosensors. Random immobilization (physical adsorption, covalent cross-linking, etc.) can easily bring about problems, such as decreased enzyme activity and relatively unstable immobilization. Whereas, directional immobilization utilizing amino acid residue mutation, affinity peptide fusion, or nucleotide-specific binding to restrict the orientation of the enzymes provides new possibilities to solve the problems caused by random immobilization. In this paper, the principles, advantages and disadvantages and the application progress of enzyme electrode biosensors of different directional immobilization strategies for enzyme molecular sensing elements by specific amino acids (lysine, histidine, cysteine, unnatural amino acid) with functional groups introduced based on site-specific mutation, affinity peptides (gold binding peptides, carbon binding peptides, carbohydrate binding domains) fused through genetic engineering, and specific binding between nucleotides and target enzymes (proteins) were reviewed, and the application fields, advantages and limitations of various immobilized enzyme interface characterization techniques were discussed, hoping to provide theoretical and technical guidance for the creation of high-performance enzyme sensing elements and the manufacture of enzyme electrode sensors.