OBJECTIVE To select the superior one from two skin-cleaning disinfection methods so as to reduce the possibility of hospital infection.METHODS The water plus soaps(control group) and Daniel disinfectants(test group) were used separately to clean and disinfect the skin of patients in intensive care unit(ICU).The degree of skin-cleaning of patients and hand pollution of nurses were observed and analyzed.RESULTS The number of bacteria on the skin of patients of the test group was remarkably lower than that of the control group with a statistically significant difference(t=7.94,P

Objective To investigate the relationship between non muscle-invasive bladder can-cer and polymorphisms of DNA repair genes among Han nationality in Shanghai. Methods From January 2006 to June 2008, 94 patients with non muscle-invasive bladder cancer and 304 controls were enrolled. Known single nucleotide polymorphism(SNP) in the XPC, XPG, XRCC1 genes were detec-ted by TaqMan real-time PCR. After adjusted for age, sex, and smoking, interaction effects of the genotypes and non muscle-invasive bladder cancer risk, genotypes and clinical and pathological features of bladder cancer were analyzed using unconditional Logistic regression. Results After adjusted for age, sex, and smoking, the XPC 939 Lys/Gln, XPC 939Gin/Gin genotype and XPG 1104 Asp/His, XPG 1104 His/His genotype were more frequent in patients with non muscle-invasive bladder cancer, adjusted OR=1.89, 95%CI 1.14-3. 23 and OR=1.07,95%CI 0.86-1.87, respectively. The XRCC1 Arg399Gln polymorphisms were not significantly associated with non muscle-invasive bladder cancer, adjusted OR= 1.15, 95% CI 0.55-2.40. There were no significant associations between tumor clinical and pathological features in patients who possessed either the XPC or XPG polymor-phisms (P>0.05). Conclusion XPC Lys939Gln and XPG Asp1104His may modulate non muscle-invasive bladder cancer risk among Han nationality in Shanghai.

Objective To investigate the diagnostic technique of Alport syndrome(AS)by immunohistochemical staining of type Ⅳ collagen ? chains on paraffin-embedded renal sections.Methods Renal biopsies were obtained from 2 patients with autosomal recessive form of AS,2 female patients and 2 male patients with X-linked dominant form of AS and 2 patients with hematuria(1male and 1 female).AS was diagnosed according to symptoms,family history,pathology,immunofluorescence staining of type Ⅳ collagen ? chains on renal and skin biopsies and gene analysis.Normal portions of nephrectomized kidneys from 2 patients with renal tumor were used as controls.Type Ⅳ collagen ? chains were stained by two-step immunohistochemistry staining method on paraffin-embedded renal sections.Three antigen retrieval methods including autoclave heating,pepsin digestion and proteinase were investigated to find the best antigen retrieval method for type Ⅳ collagen ? chains.The findings were compared with those examined by immunofluorescence staining on fresh frozen sections.Results By immunohistochemistry staining,type Ⅳ collagen ?3 and ?5 chains showed continuous linear pattern along glomerular basement membrane on sections from the controls and the hematuria patients,intermittent linear pattern for X-linked dominant female AS patients,negative for X-linked dominant male AS patient.For patients with autosomal recessive AS,the staining of type Ⅳ collagen ?3 and ?5 chains were negative on glomerular basement membrane,but ?5 chain was positive on glomerular capsules and partial tubular basement membrane.The results were the same as those examined by immunofluorescence staining.Conclusion AS can be diagnosed by immunohistochemistry staining of type Ⅳ collagen on paraffin-embedded renal sections,which is a new technique for diagnosis of AS in China.

Objective To detect anti-clinically amyopathic dermatomyositis (CADM)-140 antibody in patients with dermatomyositis (DM) or CADM,and to estimate its clinical correlation.Methods Serum samples were collected from 22 patients with DM,16 patients with CADM,46 patients with other connective tissue diseases complicated by interstitial lung disease(including 8 cases of polymyositis,15 cases of systemic lupus erythematosus,5 cases of systemic sclerosis,6 cases of Sj(o)gren syndrome,6 cases of mixed connective tissue disease,6 cases of idiopathic pulmonary fibrosis),and 5 normal human controls.Enzyme-linked immunosorbent assay (ELISA) was performed with the recombinant melanoma differentiation-associated gene 5(rMDA)as a substrate to measure the anti-CADM-140 antibody in these serum samples.Clinical manifestations were compared between patients with anti-CADM-140 antibody and those without.Results The anti-CADM-140antibody was found in 43.8% (7/16) of patients with CADM and 9.1%(2/22) of patients with DM(P<0.05),but absent in the patients with other connective tissue diseases and in the normal human controls.A significant incroase was observed in anti-CADM-140 antibody-positive patients with DM/CADM in the incidence of cutaneous ulceration and necrosis,interstitial lung disease and rapidly progressive interstitial lung disease (8/9 vs.6.9%,P<0.01;9/9 vs.48.3%,P<0.01;5/9 vs.0,P<0.05),serum lactate dehydrogenase level(328.3±104.2 vs 241.1±100.3 IU/L P<0.05),erythrocyte sedimentation rate(40.8±23.1 vs.22.5±16.8 mm/1 h,P<0.05),high resolution computed tomography score(122.9±54.8 vs.70.0±59.8,P<0.05)compared with anti-CADM-140 antibody-negative patients with DM/CADM.The ereatine kinase level was significantly lower(156.3±260.8 vs.1806.2±3737.1 IU/L P<0.05)in anti-CADM-140 antibody-positive patients with DM/CADM than in anti-CADM-140 antibody-negative patients with DM/CADM,while no significant difference was noted in the positivity rate of antinuclear antibodies or incidence of malignancies between the antibody-positive and-negative patients with DM/CADM.Conclusions Anti-CADM.140 antibody not only is useful for the diagnosis of interstitial lung disease in patients with DM/CADM,but also may serve as a serum marker for rapidly progressive interstitial lung disease.Monitoring of serum anti-CADM-140 antibody might help to predict the progression of interstitial lung disease in patients with DM/CADM.

Objective To explore the association of Crohn's disease (CD) with T cell immunoglobulin and mucin domain 3 (Tim-3) gene polymorphisms in patients of Zhejiang Han population in China.Methods A total of 308 CD patients and 573 age-and sex-matched healthy controls were enrolled in our study.Two single nucleotide polymorphisms (SNPs) of Tim-3 (rs1036199 and rs10515746) were examined by the improved multiple ligase detection reaction technique (iMLDR).Analyses of linkage disequilibrium and haplotype were also performed by Haploview 4.2 software in all study subjects.Results In general,the allele and genotype frequencies of Tim-3 (rs1036199 and rs10515746) were not statistically different between CD patients and the controls (all P ＞0.05).According to the Montreal Classification,CD patients were divided into different subgroups.The variant allele (C) and genotype (AC + CC) of rs1036199 were more frequent in CD patients with penetrating diseases than in the controls (10.4％ vs 1.7％,P =0.002;20.8％ vs 3.5％,P =0.023).Similar conclusions were also drawn for the variant allele (A) and genotype (CA + AA) of rs10515746 in patients with penetrating diseases when compared with the controls (10.4％ vs 2.2％,P =0.000;20.8％ vs 4.2％,P =0.033,respectively).The two SNPs of Tim-3 were in strong linkage disequilibrium (D'=1.0,r2 =0.928).The haplotype (AC) formed by their wild-type alleles (A) and (C) was decreased in patients with penetrating CD compared with the controls (89.6％ vs 98.3％,P =0.000).However,the haplotype (CA) formed by their variant alleles was more frequent in patients with penetrating CD than in the controls (10.4％ vs 1.6％,P =0.000).Conclusions Tim-3 (rs1036199 and rs10515746) variations might be correlated with the enhanced risk of penetrating diseases in CD patients.Furthermore,the haplotype (AC) and (CA) formed by the two SNPs might be a protective and a risky factor for penetrating CD respectively.

AIM: To evaluate the alterations in calcium metabolism of the vascular smooth muscle in the late phase of septic shock and test the hypothesis that nitric oxide might be involved in sepsis-induced vascular hyporeactivity. METHODS: Male Sprague-Dawley rats were subjected to sepsis by cecal ligation and puncture (CLP). 18 hours post CLP,rat aortic rings were employed for measurement of contractile responses by using organ bath technique. RESULTS: In endothelium-denuded aortic rings from CLP rats,concentration-contraction curves to phenylephrine (PE) and KCl were significantly decreased when compared to that from sham control rats. The transient contraction induced by PE in calcium-free Krebs solution and the concentration-dependent contraction to CaCl_2 in KCl-depolarized medium were also markedly reduced. The hyporeactivity was partially reversed by treatment with aminoguanidine,a selective inducible nitric oxide synthase inhibitor. CONCLUSION: An impairment in calcium handling in vascular smooth muscle is involved in the vascular hyporeactivity during the late phase of septic shock,in which an excessive nitric oxide production might be the major mechanism.

Objective:To study the effect of mindfulness-based training intervention on clinical efficacy in patients with nitrous oxide(laughing gas) addiction.Methods:From June 2019 to June 2020, sixty-six patients with nitrous oxide addiction in Beijing Gaoxin Hospital were selected and randomly divided into experimental group( n=33) and control group( n=33). The control group received Taijiquan training and physical training, while the experimental group added mindfulness-based training intervention on the basis of Taijiquan training and physical training.Symptom checklist 90 (SCL-90) scores and visual analog scales (VAS) craving scores were compared between the two groups at admission and 8 weeks after treatment.SPSS 22.0 software was used for statistical analysis.Independent sample t test and paired sample t test were used to compare the differences between groups and within groups. Results:(1)Before treatment, there were no significant differences in subscale scores of SCL-90 between the two groups except for depression factor((2.45±0.86), (2.03±0.46), t=2.474, P<0.05). After treatment, the subscale scores of somatization((1.38±0.35), (1.68±0.34), t=-3.656, P<0.05), phobic anxiety((1.49±0.37), (1.81±0.30), t=-3.993, P<0.05), paranoid ideation((1.50±0.47), (1.88±0.31), t=-3.898, P<0.05) and psychotism((1.34±0.54), (1.55±0.27), t=-3.094, P<0.05) of SCL-90 in the experimental group were significantly lower than those in the control group.(2)Before treatment, there was no significant difference in VAS craving score between the two groups( t=0.857, P=0.395). After treatment, the score of VAS in the experimental group was significantly lower than that in the control group( t=27.427, P<0.05). Conclusion:Mindfulness training intervention can effectively improve the clinical symptoms of patients with nitrous oxide addiction, which is worthy of clinical application.

Objective:To evaluate the ability of multiplex competitive fluorescence polymerase chain reaction in detection of large deletion and duplication genotypes of X-linked Alport syndrome.Methods:Clinical diagnosis of X-linked Alport syndrome was based on either abnormal staining of type Ⅳ collagen α5 chain in the epidermal basement membrane alone or with abnormal staining of type Ⅳ collagen α5 chain in the glomerular basement membrane and Bowman's capsule/ultrastructural changes in the glomerular basement membrane typical of Alport syndrome.A total of 20 unrelated Chinese patients (13 males and 7 females) clinically diagnosed as X-linked Alport syndrome were included in the study.Their genotypes were unknown.Control subjects included a male patient with other renal disease and two patients who had large deletions in COL4A5 gene detected by multiplex ligation-dependent probe amplification.Genomic DNA was isolated from peripheral blood leukocytes in all the participants.Multiplex competitive fluorescence polymerase chain reaction was used to coamplify 53 exons of COL4A5 gene and four reference genes in a single reaction.When a deletion removed exon 1 of COL4A5 gene was identified,the same method was used to coamplify the first 4 exons of COL4A5 and COL4A6 genes,a promoter shared by COL4A5 and COL4A6 genes,and three reference genes in a single reaction.Any copy number loss suggested by this method was verified by electrophoresis of corresponding polymerase chain reaction amplified products or DNA sequencing to exclude possible DNA variations in the primer regions.Results:Genotypes of two positive controls identified by multiplex competitive fluorescence polymerase chain reaction were consistent with those detected by multiplex ligation-dependent probe amplification.Deletions were identified in 6 of the 20 patients,including two large deletions removing the 5'part of both COL4A5 and COL4A6 genes with the breakpoint located in the second intron of COL4A6,two large deletions removing more than 30 exons of COL4A5 gene,one large deletion removing at least 1 exon of COL4A5 gene,and one small deletion involving 13 bps.No duplication was found.Conclusion:Our results show that multiplex competitive fluorescence polymerase chain reaction is a good alternative to classical techniques for large deletion genotyping in X-linked Alport syndrome.

Objective : To investigate the willingness to receive HIV testing in primary health service institutions (PHSIs) among young students in Wuhan City, so as to provide the evidence for improving the detection of HIV testing among young students. Methods: Fifteen PHSIs were sampled using a stratified random sampling method in 14 districts of Wuhan City, and school students at ages of 15 to 24 years were sampled from each district using a convenience sampling method. Participants' demographics, awareness of AIDS-related knowledge, HIV testing and willingness to receive HIV testing were collected using questionnaires, and factors affecting the willingness to receive HIV testing in PHSIs were identified among school students using a multivariable logistic regression model. Results : A total of 301 questionnaires were allocated, and 299 valid questionnaires were recovered, with an effective recovery rate of 99.34%. The respondents included 143 men (47.83%) and 156 women (52.17%), and had a mean age of (19.36±2.40) years; there were 223 respondents with an educational level of diploma and above (74.58%). The awareness of AIDS-related knowledge was 71.57% among the respondents, and 144 respondents had received AIDS-related health education in PHSIs (48.16%). There were 34 respondents that had received HIV testing (11.37%) and 203 respondents that were willing to receive HIV testing in PHSIs (67.89%). The respondents that were unwilling to receive HIV testing in PHSIs were mainly attributed to considering to be unlikely to get HIV infections (82.29%). Multivariable logistic regression analysis showed that school students who knew AIDS-related knowledge (OR=2.797, 95%CI: 1.583-4.941), knew free HIV counseling and testing services in PHSIs (OR=2.070, 95%CI: 1.123-3.814), and had received AIDS-related health education in PHSIs (OR=2.814, 95%CI: 1.573-5.032) were more willing to receive HIV testing in PHSIs. Conclusions There were 67.89% of school students that were willing to receive HIV testing in PHSIs in Wuhan City, and the willingness to receive HIV testing was correlated with the awareness of risk of HIV infections, and awareness and experience of AIDS control services in PHSIs.

OBJECTIVEAlport syndrome (AS) is a progressive renal disease characterized by hematuria and progressive renal failure. X-linked dominance is the major inheritance form of the syndrome, accounting for almost 80% of the cases, caused by mutations in COL4A5 genes. There is currently no effective treatment that has been shown to favorably affect the outcome of AS, so early diagnosis and even prenatal diagnosis is very important.

METHODSIn this study mutation of COL4A5 was detected by amplifying the entire coding sequence mRNA of peripheral blood lymphocytes using nested PCR in two Chinese X-linked dominant Alport syndrome (XLAS) families, then the first prenatal diagnosis of XLAS in China was performed. Mutation analysis of the fetus was performed on both cDNA-based level and DNA-based level of amniocytes. Fetus sex was determined by PCR amplification of SRY as well as karyotypes analysis. Maternal cells contamination was excluded by linkage analysis.

RESULTSThere was a deletion mutation in the proband of the first family, 2696 - 2705 del gtatgatggg in the 32 exon of COL4A5, but the mother did not carry the mutation (de novo). There was a G to A substitution at position 4271 in exon 46 of COL4A5 gene (c.G4271A) in the second family, the mother also carried this mutation. After genetic counselling, only the second family accepted prenatal diagnosis. Both amniocytes cDNA level and amniocytes genomic DNA level based prenatal diagnosis showed that the fetus did not carry the same mutation as the mother. PCR amplification of SRY and karyotypes analysis showed a male fetus. Linkage analysis of X chromosome polymorphic microsatellite markers showed that there was no MCC in amniocytes.

CONCLUSIONBoth cDNA level and DNA level analysis could enhance the accuracy and reliability of prenatal diagnosis. PCR amplification of SRY was faster than karyotypes analysis in the fetal sex determination. Linkage analysis was useful in the detection of maternal cells contamination in amniocytes.

China ; Chromosomes, Human, X ; Collagen Type IV ; genetics ; DNA ; analysis ; DNA Mutational Analysis ; trends ; DNA, Complementary ; analysis ; Exons ; physiology ; Female ; Genetic Counseling ; Genetic Linkage ; Genetic Testing ; Humans ; Mutation ; Nephritis, Hereditary ; diagnosis ; genetics ; Pedigree ; Pregnancy ; Prenatal Diagnosis ; methods ; RNA, Messenger