Objective To provide histology base for the microsurgical repair of the free gracilis muscle transplantation after brachial plexus.Methods Totally 6 fresh male adult cadaveric inferior extremities were obtained.The gracilis muscle nerve were exposed and divided with the microdissection.Specimens were got from different segment after marked direction.All specimens was faced in 4％ formaldehyde solution and then crossing sections were cut by cryoultramicrotome.All slides were stained use the technique of Kamovsky-Roots AchE histochemical.The result of never tissue staining and the distribution of individual functional fascicular group were observed on each cross-section.According to the result of staining combined with the microdissection and the order of different branches branching off the nerve trunk,the distribution of individual functional fascicular group were observed on each cross-section.The 5 μm-thick routine waxed crossing sections were made and stained according to the myelin technique of Loyez.These histological sections were analyzed by using image analysis system.For each histological section,the number of the medullated nerve fibers and the section areas of the each nerve tracts and trunks were measured and calculated.Then the proportion of nerve tracts and connective tissue were calculated.The proportion of each connective tissue was adopting paired-samples t test.Results Under low power lens most of the gracilis muscle nerve were positive reaction,only a few sparse block-shape enzyme staining regions were shown.Under high power lens,the gracilis muscle nerve have clear outline,enzyme staining limited at neuraxis,no staining at myelin sheath and connective tissue.Quantitative analysis shows that the total myelinated fiberscilis nerve was about (1958 ± 375) radix.The branches arising from the posterior subdivision were more than that from the anterior (P =0.000).There were statistical difference between the number of the medullated nerve fibers and the section areas of the anterior and posterior subdivisions,the posterior subdivision were more than that of the anterior (P ＜ 0.05).There also had statistical difference between different section areas of the connective tissue in the gracilis nerve trunk (P ＜ 0.05),the section areas of the connective tissue of the distal were more than that of proximate.Conclusions The motor fascicles characteristic of the gracilis muscle nerve can be distinguished clearly by using AchE histochemical staining,combin with the microanatomy results,we can gain the distribution of the fascicular groups on each crossing sections.Using Loyez staining and quantitative analysis,we can ensure the number of the medullated nerve fibers.It is helpful to select the suitable donor nerve and ensure the dialyneury matching each other.

Objective To investigate the clinical characteristics and the causative mechanism of acetabular fracture and dislocation of hip complicated with sciatic nerve injuries. Methods A retrospective analysis was done for 155 cases (159 sides) of acetabular fracture, dislocation of hip, or acetabular fracture combined with hip dislocation, 35 of which were complicated with sciatic nerve injuries. The epidemiological features of acetabular fracture and hip dislocation, the incidence, and relationship between sciatic nerve injury and classifications of acetabular fracture and dislocation of hip were analyzed. Results 81.3%of the patients were male. The ages of 86.5%of the patients ranged from 20 to 50 years old. 83.9%of the cases were injured in traffic accidents. The incidence of sciatic nerve injury was 22.01%in all the patients, 17.19%in patients with acetabular fracture, 12.90%in patients with posterior wall, and 36.36%in patients with acetabular fracture combined with posterior dislocation of hip. Conclusions A road traffic accident is the major causative factor for acetabular fracture and dislocation of hip. Most victims are male. Sciatic nerve injuries largely happen in cases of acetabular posterior wall fracture and posterior column fractures combined with posterior dislocation of hip. Peroneal nerve injuries are the most common type of sciatic nerve injury.

0.05).CD34 showed widespread expression in cholangio-carcinoma tissues,but limited in normal bile duct,which showed significant difference(P

Objective To analyze the 23S rRNA gene partial sequences of common bacteria, and establish molecular biologic techniques to identify bacteria by the difference of gene sequences. Methods Analyzing the sequences of variable region of bacterial 23S rRNA genes, primers and oligonucleotide probes were designed accordingly. Thereafter, bacteria were identified by PCR gel electrophoresis and PCR reverse hybridization. Results There exists significant sequence difference between Gram negative bacteria and Gram positive bacteria and it could be used to differentiate these 2 kinds of bacteria quickly with PCR gel electrophoresis. Meanwhile, sequence variety in different species of bacteria was also observed and PCR reverse hybridization could be used to identify different bacterial species further.Conclusions There exist significant sequence differences among 23S rRNA genes in different common bacteria. By the sequence differences, a specific, sensitive and rapid molecular biologic techniques could be established to quickly identify the pathogens of bacterial infections.

Objective To investigate the clinical and pathological characteristics of bacterial infectious diarrhea.Methods The clinical and pathological characteristics of 2380 cases of bacterial infectious diarrhea in Jinshan Hospital,Fudan University from 1998 to 2007 were analyzed retrospectively.Enumeration data were analyzed by X~2 test.Results Among the 20 169 patients who went to hospital because of diarrhea in 10 years,2380 cases fecal bacterial culture were positive,including Vibrio parahaemolyticus(2247 cases,94.4%),Shigella(99 cases,4.2%),Salmonella (29 cases,1.2%),Vibrio alginolyticus(3 cases),pathogenic Escherichia coli(2 cases).Patients with diarrhea were common from June to 0ctober in each year.The main manifestations of Vibrio parahaemolyticus infection were abdominal pain,diarrhea,nausea,vomit or dehydration.The main manifestations of Shigella infection were fever,abdominal pain and diarrhea.Conclusions The bacterial culture positive rate of stool samples from patients with bacterial infectious diarrhea is not high in Jinshan district.Shanghai.The major pathogens are Wbrio parahaemolyticus and Shigella.

Objective To develop a multiple polymerase chain reaction (PCR) technique based assay for rapid detection of vanA, vanB, vanD and vanM in high-level vancomycin-resistant enterococci.Methods After analyzing the uncleotide sequence divergence among D-Ala∶D-Lac ligase genes, an multiplex PCR assay for vanA, vanB, vanD and vanM genes in high-level vancomycin-resistant enterococci were designed.By using recombination plasmids containing vanA, vanB, vanD and vanM genes as positive control, and non-vancomycin resistant enterococci (non-VRE) common pathogenic bacterial DNA as negative control, the sensitivity and specificity of the assay were evaluated.Fifty vancomycin-resistant enterococci (VRE) isolates were detected by the assay.Fifty clinical strains of VRE were isolated from 9 hospitals in Shanghai from January 2006 to December 2014.The results were compared with the conventional PCR and sequencing methods.Results The identity of the D-Ala∶D-Lac ligase genes were 60.8%-71.3% of vanA, vanB, vanD and vanM genes.The multiplex PCR assay could identify the genotypes of the positive control samples accurately.No false positive results were found in negative control samples.Among fifty VRE strains detected by the assay, 18 were vanA genotype and 32 were vanM genotype.Comparison of the multiplex PCR assay and sequencing methods revealed sensitivity and specificity of 100%.The detection limit of the assay was 2×10 copies/PCR reaction.The experiment could be done within 3.5 h.Conclusions A multiplex PCR assay is developed to rapid identify the genotype of the high-level vancomycin-resistant enterococci, which can be used for the molecular epidemiology research and detection of VRE.

Objective To examine the pathological changes in the liver of an AIDS patient with complicated infection of Pneumocystis carinii(PC). \ Methods\ A liver biopsy was made. The tissue was stained with HE, PAS, Giemsa, GMS, and acid\|fast staining, and examined under light microscope and transmission electron microscope. \ Results\ Granulomas (acid\|fast negative) in the tissue and numerous pathogens (PAS positive) in hepatic sinusoids were detected. Giemsa and GMS staining and electron microscopy all confirmed that the pathogen was Pneumocystis carinii. \ Conclusion\ The pathological findings revealed a diffuse extrapulmonary infection of Pneumocystis carinii in the patient of AIDS.

Objective To understand the clinical features of candidemia.Methods A retrospective analysis was performed based on the data of 88 candidemia cases treated in Huashan Hospital during the period from 2007 to 2012.The clinical data were re-viewed in terms of species distribution,underlying diseases,clinical manifestations,treatment and outcomes.The prognostic factors were analyzed by chi-square test or Fisher exact probability test.Multivariate analysis was conducted by multiple Logis-tic regression.Results Candida albicans (40/88,45.5%)was the most common pathogen isolated from these candidemia ca-ses,followed by Candida tropicalis (20/88,22.7%),Candida parapsilosis (17/88,19.3%),Candida glabrata (10/88, 11 .4%),and Candida krusei (1/88,1 .1 %).Solid malignancy,diabetes,and surgical procedure were the most frequently identified underlying diseases.Fatal or deteriorative outcome was reported in 28 cases.The attributable mortality was 18.2%. Multivariate prognostic analysis indicated that presence of central venous catheter (OR:6.322,95% CI :1 .055-37.891 ,P =0.044)was independently correlated to increased mortality.Appropriate antifungal therapy was an independent predictor of de-creased overall mortality (OR:0.137,95% CI :0.039-0.480,P =0.002).Conclusions The pathogen distribution of candi-demia has changed slightly.Appropriate antifungal therapy plays a key role in the treatment of candidemia.

Objective To detect the clinical pathogenic bacteria and antimicrobial-resistant genes quickly and sensitively using DNA chip.Methods Based on the analysis of 23S rRNA gene se- quences and other genes sequences associated with antimicrobial resistance(SHV＜CTX_M),oligo nucleotide microarray was designed according to different bacteria and antimicrobial-resistant genes. The DNA fragments were amplified by labeling Cy5 fluorescence and detect clinical pathogenic bacte- rias and antimicrobial-resistant genes by hybridization.Results The result of detection(10~3-10~6 bac- teria/ml)was consistent with that of some documents in domestic and overseas under ideal circum- stances of detecting bacteria genomic DNA by the Reagent Box.And it was specific and reproducible when the detection system were evaluated with some clinical isolates and drug-resistant standard strain.DNA chip could identify 16 species and 7 generics including common diverse clinical pathogenic bacteria,and could detect the drug-resistant of extended spectrum?lactamase gene simultaneously. Conclusions The methods that we have established DNA chip is a sensitive,specific and reproducable tool for supplying routine methods.

AIM: To construct prokaryotic expression vector of human angiogenesis inhibitor arresten gene and express recombinant arresten in Escherichia coli. METHODS: Human arresten gene was amplified from recombinant plasmid pGEM-Arr with polymerase chain reaction (PCR), and then cloned into prokaryotic expression vector pRSET by means of recombinant gene technology. The recombinant plasmid pRSET-Arr was transformed into E.coli BL21(DE3), and recombinant arresten was expressed in the bacteria under induction of IPTG. The expressed products were detected by SDS-PAGE analysis. RESULTS: Restriction analysis indicated that the arresten gene was successfully inserted into the expression vector, and DNA sequencing verified that the reading frame of the recombinant vector was correct. Recombinant arresten was successfully expressed in Escherichia coli; its molecular weight was about 26 kD and its amount was approximately 30% of total bacterial proteins.CONCLUSION: The successful construction of prokaryotic expression vector containing human arresten gene and the effective expression of recombinant arresten in Escherichia coli laid the foundation for further study on its biological functions.