Objective To establish a viable bacteria assay for Helicobacter pylori(H.pylori)by assessing the cgt gene expression,and to develop accordingly a rapid and novel testing method for clinical precision treatment.Methods Viable bacteria count was determined in bacterial cultures.The transcriptional expression level of cgt(hp0421),the conserved gene that encodes cholesterol-α-glucosyltransferase(CGT)in H.pylori,was measured by RT-PCR.The correlation between the number of colonies and cgt gene transcription expression was analyzed and the regression model was constructed.The linear range,sensitivity,and specificity of the new method were examined accordingly.The bactericidal action of clarithromycin was assessed using this method to verify the performance of the method in determining clinical bacterial drug resistance.Results The Ct values of cgt for H.pylori colony counts of 102,104,106,and 108 CFU/mL were 29.67±0.14,23.37±0.36,17.65±0.37,and 11.38±0.39,respectively.In the range of 101-108 CFU/mL,the regression equation for cgt gene expression and viable bacterial counts determined by RT-qPCR was y=-0.350 1x+12.49,with the correlation coefficient being R2=0.9992 and the sensitivity being 101 CFU/mL,showing no cross-reaction with 13 other bacteria.The lg values of live H.pylori bacteria treated with clarithromycin at 0,5,10,20,and 40 μg/mL for 12 h were 2.57±0.02,2.45±0.01,2.19±0.02,1.91±0.07,and 1.33±0.05,respectively.The corresponding cgt gene Ct values were 27.76±0.09,28.37±0.24,29.51±0.14,30.11±0.12,and 31.66±0.11.By applying the cgt gene expression in the equation,the estimated counts of viable bacteria were found to be 2.73±0.03,2.52±0.08,2.11±0.05,1.89±0.02,and 1.33±0.04,showing no significant difference in statistical analysis(P＞0.05).Conclusion The method for assessing viable bacteria account by evaluating cgt gene expression in H.pylori was successfully established,significantly reducing the time required to determine viable bacteria count and providing a new method for clinical viable bacteria testing.

Acceptance of disability(AOD)is a key factor in psychosocial adjustment of disabled groups,and a good level of AOD plays an important role in promoting health outcomes of patients.At present,studies on AOD have been carried out in many disease fields.Therefore,this article focuses on the relevant concepts,evaluation tools,manifestations,influencing factors,intervention strategies and other aspects of AOD in diabetic foot amputation patients,aiming to improve patients'prognosis,quality of life and life satisfaction and other aspects for references and help.

Objective To evaluate the effect of Wnt5A gene overexpression on cytoskeletal proteins of melanocytes after the plasmid containing the Wnt5A gene is transfected into primary melanocytes.Methods In vitro cultured primary human melanocytes were divided into three groups:blank control group receiving no treatment,negative control group transfected with endotoxin-free pcDNA3.1 (+)empty vector by Lipo3000 in Opti-MEM medium,Wnt5A plasnid group transfected with endotoxin-free pcDNA3.1 (+) vector containing the Wnt5A gene by Lipo3000 in Opti-MEM medium.After the transfection,quantitative PCR (qPCR) was performed to measure the mRNA expression of Wnt5A,ras-related C3 botulinum toxin substrate 1 (Rac1),filamentous actin (F-actin) and β-tubulin,Western blot analysis to determine the protein expression of Wnt5A,receptor tyrosine kinase like orphan receptor 2 (ROR2),Rac1,F-actin and β-tubulin,and an immunofluorescence assay (IFA) to observe the expression of cytoskeletal proteins.Results qPCR showed significant differences in the mRNA expression of the Wnt5A gene and its downstream genes Rac1 and F-actin among the Wnt5A plasmid group,negative control group and blank control group (F =1374.179,112.576,66.458,respectively,all P ＜ 0.01),but there was no significant difference in the mRNA expression of β-tubulin among the three groups (P ＞ 0.05).Additionally,the Wnt5A plasmid group showed significantly higher mRNA expression of Wnt5A,Rac1 and F-actin compared with the blank control group and negative control group (all P ＜ 0.05).As Western blot analysis revealed,compared with the blank control group and negative control group,the Wnt5A plasmid group showed significantly higher Wnt5A protein expression (both P ＜ 0.05),but significantly lower protein expression of Rac 1,ROR2 and F-actin (all P ＜ 0.05).However,no significant difference in β-tubulin protein expression was observed among the three groups (P ＞ 0.05).IFA showed no obvious difference in the fluorescence intensity of β-tubulin or F-actin between the Wnt5A group and the two control groups,but melanocytes showed larger size and increased number of dendrites,and the cytoskeleton changed dramatically with varying fluorescence intensity of F-actin,fuzzy texture,fractured or locally clustered tonofilaments in the Wnt5A group.Conclusion The overexpression of the Wnt5A gene in melanocytes can regulate the mRNA and protein expression of cytoskeletal proteins,nake melanocytes larger and more dendritic,and cause changes in the cytoskeleton,which may facilitate the transportation of melanosomes,and participate in the occurrence of hyperpigmented diseases.

Objective To analyze the clinical laboratory characteristic of Candida albicans caused antibiotic-associated diarrhea (AAD) .Methods 100 cases of AAD were selected .Based on the results ,they were divided into Candida albicans group and non-Candida albicans group .The levels of sodium ,potassium ,fasting blood-glucose and albumin ,investigation of antimicrobial usage were compared in two groups .Results Candida albicans were detected in 42 cases .The levels of sodium ,potassium and fasting blood-glucose in the Candida albicans group were significantly higher than those in non-Candida albicans group(P<0 .01) ,and al-bumin were significantly lower than that in non-Candida albicans group(P< 0 .01) .The rate of two or more antibiotic usage in Candida albicans group was significantly higher than that in non-Candida albicans group(P<0 .05) .The drug resistance rates of fluconazole ,itraconazole ,voriconazole ,amphotericin B and flucytosine in Candida albicans group were significantly lower than those in non-Candida albicans group(P<0 .05) ,and the antibiotic sensitive rates of fluconazole ,itraconazole ,voriconazole ,amphotericin B and flucytosine in Candida albicans group were significantly higher than those in non-Candida albicans group(P<0 .05) .The drug resistance rates of vancomycin and macrodantin in Candida albicans group were significantly higher than those in non-Candida al-bicans group(P<0 .01) ,and the antibiotic sensitive rates of vancomycin and macrodantin in Candida albicans group were signifi-cantly lower than those in non-Candida albicans group(P<0 .01) .Conclusion For AAD cases ,the fecal culture can provide exper-imental basis for clinical treatment and avoid the abuse of antibiotics .