To achieve efficient and rapid detection of duck astrovirus type 2(DAstV-2),RPA prim-ers and crRNA were designed and synthesized based on the conserved sequence of the ORF2 gene of DAstV-2.A detection method for DAstV-2 was constructed,integrating RPA nucleic acid ampli-fication,LwCas13a cleavage,and colloidal gold lateral flow dipstick visualization.The specificity,sensitivity,and concordance of this detection method were evaluated.The experimental results showed that the detection limit for the DAstV-2 recombinant plasmid standard was 1.2×101 cop-ies/μL,which is superior to the conventional RT-PCR method.The method can specifically detect DAstV-2 pathogenic nucleic acids without cross-reactivity with DAstV-1,DAstV-3,DAstV-4,duck plague virus(DEV),and duck tembusu virus(DTMUV).When testing liver tissue samples from ducks suspected of being infected with DAstV-2,the results obtained using this method were com-pletely consistent with those from real-time quantitative PCR,with a 100％concordance rate.How-ever,this method is simpler and faster to perform.The research indicates that the established RPA-CRISPR/Cas13a-LFD detection system has high sensitivity,strong specificity,and high accuracy,capable of completing rapid visual detection of DAstV-2 nucleic acids within 1 h at a constant tem-perature of 37 ℃,providing a new technical platform for the rapid diagnosis of DAstV-2.

BACKGROUND:Polycystic ovary syndrome is a common reproductive endocrine disease leading to infertility in women of reproductive age.Currently,there is no effective treatment.Human umbilical cord mesenchymal stem cells may help to repair ovarian function,but few studies have reported their role in the treatment of polycystic ovary syndrome.OBJECTIVE:To explore the effect and mechanism of human umbilical cord mesenchymal stem cells in the treatment of polycystic ovary syndrome.METHODS:3-week-old female ICR mice were divided into three groups(n=15).Normal control group was not treated.Model group was given letrozole for 21 days to induce polycystic ovary syndrome.Treatment group was given letrozole via intragastric administration for 21 consecutive days,and human umbilical cord mesenchymal stem cell suspension was injected through the tail vein.The body weight of mice was monitored before treatment,7 and 14 days after treatment.The estrous cycle of mice was detected by continuous vaginal smears for 10 consecutive days after treatment.Fourteen days after treatment,peripheral blood sex hormone levels of mice were detected by ELISA.Hematoxylin-eosin staining was used to observe ovarian morphological changes.Immunofluorescence and immunohistochemistry were used to detect the expression of Rab27A protein in ovarian tissues.RESULTS AND CONCLUSION:(1)Compared with the normal control group,the estrous cycle of mice in the model group was disturbed;body weight was significantly increased;follicle stimulating hormone was decreased;luteinizing hormone and testesterone were both increased;more follicles were found in the ovaries,and the relative expression level of Rab27A protein was decreased.(2)Compared with the model group,the treatment group had diminished body weight,increased follicle stimulating hormone,decreased luteinizing hormone and testesterone,decreased follicles with polycystic dilatation,and increased Rab27A protein relative expression level.(3)These findings suggest that human umbilical cord mesenchymal stem cells improve serum sex hormone levels and ovarian function in polycystic ovary syndrome mice by upregulating the expression of Rab27A.

BACKGROUND:Polycystic ovary syndrome is a common reproductive endocrine disease leading to infertility in women of reproductive age.Currently,there is no effective treatment.Human umbilical cord mesenchymal stem cells may help to repair ovarian function,but few studies have reported their role in the treatment of polycystic ovary syndrome.OBJECTIVE:To explore the effect and mechanism of human umbilical cord mesenchymal stem cells in the treatment of polycystic ovary syndrome.METHODS:3-week-old female ICR mice were divided into three groups(n=15).Normal control group was not treated.Model group was given letrozole for 21 days to induce polycystic ovary syndrome.Treatment group was given letrozole via intragastric administration for 21 consecutive days,and human umbilical cord mesenchymal stem cell suspension was injected through the tail vein.The body weight of mice was monitored before treatment,7 and 14 days after treatment.The estrous cycle of mice was detected by continuous vaginal smears for 10 consecutive days after treatment.Fourteen days after treatment,peripheral blood sex hormone levels of mice were detected by ELISA.Hematoxylin-eosin staining was used to observe ovarian morphological changes.Immunofluorescence and immunohistochemistry were used to detect the expression of Rab27A protein in ovarian tissues.RESULTS AND CONCLUSION:(1)Compared with the normal control group,the estrous cycle of mice in the model group was disturbed;body weight was significantly increased;follicle stimulating hormone was decreased;luteinizing hormone and testesterone were both increased;more follicles were found in the ovaries,and the relative expression level of Rab27A protein was decreased.(2)Compared with the model group,the treatment group had diminished body weight,increased follicle stimulating hormone,decreased luteinizing hormone and testesterone,decreased follicles with polycystic dilatation,and increased Rab27A protein relative expression level.(3)These findings suggest that human umbilical cord mesenchymal stem cells improve serum sex hormone levels and ovarian function in polycystic ovary syndrome mice by upregulating the expression of Rab27A.

To achieve efficient and rapid detection of duck astrovirus type 2(DAstV-2),RPA prim-ers and crRNA were designed and synthesized based on the conserved sequence of the ORF2 gene of DAstV-2.A detection method for DAstV-2 was constructed,integrating RPA nucleic acid ampli-fication,LwCas13a cleavage,and colloidal gold lateral flow dipstick visualization.The specificity,sensitivity,and concordance of this detection method were evaluated.The experimental results showed that the detection limit for the DAstV-2 recombinant plasmid standard was 1.2×101 cop-ies/μL,which is superior to the conventional RT-PCR method.The method can specifically detect DAstV-2 pathogenic nucleic acids without cross-reactivity with DAstV-1,DAstV-3,DAstV-4,duck plague virus(DEV),and duck tembusu virus(DTMUV).When testing liver tissue samples from ducks suspected of being infected with DAstV-2,the results obtained using this method were com-pletely consistent with those from real-time quantitative PCR,with a 100％concordance rate.How-ever,this method is simpler and faster to perform.The research indicates that the established RPA-CRISPR/Cas13a-LFD detection system has high sensitivity,strong specificity,and high accuracy,capable of completing rapid visual detection of DAstV-2 nucleic acids within 1 h at a constant tem-perature of 37 ℃,providing a new technical platform for the rapid diagnosis of DAstV-2.

Objective:To describe demographic,clinical and physiological characteristics,treatment between first-episode major depressive disorder(MDD)and relapse MDD,and to explore characteristics of relapse MDD.Methods:Totally 858 patients who met the diagnostic criteria for depression of the Diagnostic and Statistical Manual of Mental Disorders,Fifth Edition(DSM-5),were included by using the Mini International Neuropsychiatric Interview(MINI),Clinician-Rated Dimensions of Psychosis Symptom Severity,and Hamilton Depression Scale etc.Among them,529(58.6％)were first-episode depression and 329(36.0％)were relapsed.The differences of demographic characteristics,clinical and physiological characteristics,treatment were compared byx2test and Kruskal-Wallis rank sum test.Multivariate logistic regression was used to explore the characteristics of MDD recur-rence.Results:Compared to first-episode MDD,relapse MDD had more comorbidity(OR=2.11,95％CI:1.00-4.44),more days out of role(OR=1.26,95％CI:1.01-1.56),more history of using psychiatric drug more than one month(OR=1.41,95％CI:1.02-1.97)and electroconvulsive therapy(OR=3.23,95％CI:1.42-7.36),and higher waist-hip ratio(OR=33.88,95％CI:2.88-399.32).Conclusion:Relapse MDD has positive as-sociation with comorbidity of mental disorders,out of role,and higher waist-hip ratio.

OBJECTIVE To evaluate the effects of different functional dyspepsia(FD)modeling methods and explore the thera-peutic effect and potential mechanism of Banxia Xiexin Decoction on FD.METHODS BALB/c mice were randomly divided into the blank group,iodoacetamide group,loperamide group,tail clamp group and vinegar group.After 1 week of intervention,the status of mice in each group was observed and their gastrointestinal motility,hormone levels and pathological changes were detected.A more i-deal FD modeling method was evaluated and determined.After modeling,different doses of Banxia Xiexin Decoction were given to in-tervene,and the changes in the gastrointestinal function of mice were observed.The expression of related proteins was studied by im-munohistochemistry,ELISA,Western Blot and other experimental methods.RESULTS Comparing the four modeling methods,it was found that the mice in the iodoacetamide group,loperamide group,and vinegar group showed weight loss compared to the blank group;the gastric emptying rate and small intestinal propulsion rate of mice in the iodoacetamide group and vinegar group decreased;changes in gastrointestinal hormones were found in the serum of mice in the tail clip group and vinegar group.Finally,the iodoacetamide meth-od was evaluated as the optimal FD modeling method.The administration results showed that Banxia Xiexin Decoction had no signifi-cant effect on the food intake and body weight of FD mice,while medium and high doses could improve the physical condition of FD mice,increase their gastric emptying rate and small intestine propulsion rate.The experimental results of immunohistochemistry,West-ern blotting,and ELISA confirmed that medium and high doses of Banxia Xiexin Decoction can significantly reduce the expression lev-els of TNF-α and IL-6 in the duodenum and serum of FD mice.CONCLUSION The iodoacetamide method is a better FD modeling method.Banxia Xiexin Decoction can improve the condition of FD mice,increase gastrointestinal motility,reduce the secretion of in-flammatory factor,thereby achieving the therapeutic effect of treating FD.

Intestinal fibrosis is a complication of inflammatory bowel disease,and its refractory and recurrent nature impose a serious disease burden on patients.The disease's pathogenes is not clear,and there is no effective treatment.Moreover,there is still a lack of recognized intestinal fibrosis models.In this paper,we review the method used to establish animal models of intestinal fibrosis both at home and abroad,and consider the clinical relevance,key characteristics,and advantages and disadvantages of the procedures.Intestinal fibrosis models were summarized according to the modeling period and method.

Objective To systematically evaluate the risk prediction model for central venous catheter-related bloodstream infections and provide references for clinical practice.Methods Databases such as CNKI,Wanfang,CBM,VIP,PubMed,Web of Science,Cochrane Library,etc.were retrieved.The search period is from database establishment to June 2,2023.There are 2 researchers who independently screened and extracted the literature,and evaluated the quality of the literature using bias assessment tools of predictive model risk.Results A total of 9 articles were included,including 9 risk prediction models for catheter-related bloodstream infections.The total sample size was 80～11 901 cases;the number of outcome events was 19～403 cases;the C index of the included model was 0.81～0.93.The area under the curve of the subjects ranged from 0.73～0.90.The predictors that appear more frequently in the model mainly included the history of diabetes,albumin value,the number of days of catheterization,the location of catheterization.The evaluation results of the bias assessment tool of predictive model risk for research show that the overall applicability of the included risk prediction models is good,but the bias risk is high.The reasons are related to the improper source of research sample data,inappropriate processing of continuous variable methods,failure to process missing data,insufficient model performance evaluation,and non-standard evaluation indicators.Conclusion There are still some shortcomings in the risk prediction model for central venous catheter-related bloodstream infections.In the future,the quality of related model research should be further improved,especially in terms of predictive factor analysis,model evaluation indicators,etc.,which should be further standardized.

The aim of this study was to investigate the effect of norcantharidin (NCTD) on the proliferation and apoptosis of triple-negative breast cancer cell line MDA-MB-231.Western blot was used to detect the effect of NCTD on the expression levels of apoptosis-related proteins Bax/Bcl-2, cleaved-PARP/PARP/PARP, cleved-caspase-9, cleaved-caspase-3 and MCL-1 in MDA-MB-231 cells.Also, the expression levels of autophagy-related proteins LC3-II/LC3-I, Parkin and PINK1 in MDA-MB-231 cells were measured by Western blot.Flow cytometry was used to measure the effect of NCTD on the changes of mitochondrial membrane potential and mitochondrial reactive oxygen species (ROS).The effect of NCTD on autophagy flow in cells expressing mCherry-EGFP-LC3 was detected by a confocal microscope.Moreover, the effects of NCTD combined with chloroquine (CQ) or 3-methyladenine (3-MA) on the apoptosis of MDA-MB-231 cells were detected by flow cytometry.The results showed that NCTD significantly increased the expression levels of Bax/Bcl-2, cleaved-PARP/PARP, cleaved-caspase-9, cleasved-caspase-3 and LC3-II/LC3-I proteins, and promoted the mitochondrial translocation of Parkin, and blocked the autophagic flow in MDA-MB-231 cells. Moreover, NCTD combined with CQ accelerated apoptosis, while NCTD combined with 3-MA decreased apoptosis.These results suggest that NCTD can induce autophagy accumulation and lead to apoptosis of MDA-MB-231 cells.

ObjectiveTo observe the clinical effect of Danshenyin on chronic atrophic gastritis and explore the mechanism. MethodA total of 152 patients with chronic atrophic gastritis who were treated in Xining No. 1 People's Hospital were selected and randomized into control group （76 cases） and observation group （76 cases）. The control group was given conventional western medicine treatment， and the observation group was given Danshenyin （oral）. The treatment lasted 8 weeks for both groups. The clinical effect， traditional Chinese medicine （TCM） syndrome score， and pathological score of gastric mucosa were compared between two groups. The content of pepsinogenⅠ （PGⅠ）， pepsinogenⅡ （PGⅡ）， proinflammatory cytokines ［soluble interleukin-2 receptor （sIL-2R）， tumor necrosis factor-α （TNF-α）］， and gastric mucosa-proteting factors ［prostaglandin E₂ （PGE₂）， calcitonin gene-related peptide （CGRP）］ was determined， and the adverse reactions and recurrence rates were recorded. ResultThe effective rate （91.18%， 62/68） in the observation group was higher than that （7.61%， 52/67） in the control group （χ²=4.727， P<0.05）. The TCM syndrome score in the observation group was lower than that in the control group （P<0.05）. Compared with the control group， the observation group showed low pathological scores of glandular atrophy and intestinal metaplasia and dysplasia （P<0.05）. The observation group had higher serum PGⅠ content and PGⅠ/PGⅡ value and lower PGⅡ content than the control group （P<0.05）. The content of sIL-2R and TNF-α decreased （P<0.05） and that of PGE₂ and CGRP increased （P<0.05） in the observation group as compared with those in the control group. The incidence of adverse reactions in the observation group was 7.35% （5/68）， as compared with the 14.93% （10/67） in the control group. The recurrence rates in the 6-month and 12-month follow-up in the observation group were lower than those in the control group， but the difference was not statistically significant （Fisher's exact test）. ConclusionDanshenyin shows satisfactory effect in relieving TCM syndrome， regulating PG secretion， and improving gastric mucosal lesions in the treatment of chronic atrophic gastritis. The mechanism is the likelihood that it modulates sIL-2R， TNF-α， PGE₂， and CGRP to alleviate inflammatory damage of gastric mucosa and protect gastric mucosa. The medicine is safe and reduces the recurrence of chronic atrophic gastritis.