M.tuberculosis strains isolated strains from sputum specimens of 134 patients were analyzed by PCR SSCP and traditional drug susceptibility tests. The results sbowed that the gene mutation rate of PZA, SM, RFP,INH and EMB resistance in those clinically isolated strains was 42 7%,71 7%,78 9%,68 6% and 43 9%, respectively. The gene mutation was in relation with drug resistance level of M. tuberculosis. The gene mutation rate was higher in high concentration resistance strains than in low concentration resistance strains.

Objective To investigate the mutation of embB and pncA genes in mycobacterium tuberculosis isolates, and assess its clinical value. Methods The drug-sensitivity of 106 clinical isolates of mycobacterial species was identified by the tranditional method, and then analyzed the mutation of their embB and pncA genes with PCR-SSCP. Mycobacterium tuberculosis strain H 37 RV was used as a control. Results The mutational frequency of pncA gene in 94 PZA-resistant mycobacterium tuberculosis was 44 6%(42/94). The mutational frequency of embB gene in 83 EMB-resistant isolates was 54 2%. The frequency of both genes mutation was 11 8%(11/94). Conclusion EMB and PZA resistances in some mycobacterium tuberculosis isolates were due to mutations of embB and pncA genes. PCR- SSCP method might become a simple and rapid diagnostic test for genotypes of EMB and PZA resistant mycobacterium tuberculosis.

Objective To evaluate the clinic application value of detection of rPOB,katG and rPSL resistance genes to M tuberculosis by polymerase china reaction-single stranded conformation polymorphism (PCR-SSCP).Method The drug-resistance tests of M tuberculosis clinical isolates from 141 patients with the tuberculosis was analyzed by Bactec-960 rapid culture,drug-sensitive and PCR-SSCP techniques.Results As nearly As one of three patients with priliminary treatment had resistance to one kind of the drugs tested at least.The mutation rates of RFP,INH and SM drug-sensitive isolates were 6 5%,11 4% and 27 2% respectively.That of the drug-resistant isolates were 60 2%,45 7% and 62 7%.As drug-resistant concentration heightened,so mutation rate of genes increased.Conclusions Detecting the resistance genes was a new explore in guiding treatment,and multiple drug-sensitive test combinative use have cooperativity each other.It might become good test for clinical management.

Objective To investigate the pathogenesis role of aquaporin 3 and aquaporin 9 in idiopathic polyhydramnios by detecting their expression and distribution in fetal membranes and placenta.Methods Twenty-one of term pregnancy women with idiopathic polyhydramnios were enrolled as patient group matched with 30 women with normal term pregnancy as control group.The expression and localization of aquaporin 3 and aquaporin 9 in fetal membranes and placenta were detected by real-time polymerase chain reaction and streptavidin peroxidase immunohistochemiscal staining.Results (1)The mRNA expressions of aquaporin 3 and aquaporin 9 were detected in amnion,chorion and placental tissue in both patient group and control group.Both aquaporin 3 and aquaporin 9 were demonstrated positive staining in the amnion epithelia,chorion cytotrophoblasts and placental trophoblast.(2)The ratio of aquaporin 3 and aquaporin 9 mRNA expressions in amnion in patient group comparing to those in control group were 5.00 and 3.25,while in chorion they were 2.03 and 2.08.When compared with those in amnion and chorion of control group,there was a significant difference(P<0.01).However,the relative change fold of aquaporin 3 and aquaporin 9 in placental trophoblast in patient group were decreased in comparison of those in control group,which also showed statistical difference(P<0.01).(3)The expression of aquaporin 3 and aquaporin 9 protein in anmion were 7.5 ±2. 0 and 11.1 ± 1.8 in patient group, while they were 5.3 ± 1. 6 and 5.6 ± 2. 3 in control group. In chorion, the expression of aquaporin 3 and aquaporin 9 protein was 7.5±2. 0 and 10. 0 ±1.6 in patient group, respectively, while in control group, they were 5.4 ±2.2 and 5.6±2. 1. When compared with those proteins in control group, it exhibited statistical difference (P<0.05). However, in placental trophoblast of patient greup,the expression of aquaporin 3 and aquaporin 9 protein were 3.5±1.4and 4. 0±2. 5, respectively, which were significantly decreased than 5.6±1. 3 and 7. 1±2. 9 in control group(P< 0. 05). Conclusions The alterations of aquaporin 3 and aquaporin 9 expressions in fetal membrane and placenta might be an adaptive response to idiopathic polyhydramnios. Further investigation should be needed to clarify the regulatory mechanism of aquaporin 3 and aquaporin 9 expressions.

Objective To investigate the role of mitogen-activated protein kinases (MAPK)/extracellular signal regulated kinase1/2 (ERK1/2) signal transduction pathway in regulating the expression of aquaporin 3 (AQP3) in human amnion epithelial cells.Methods Primary cell culture of human amnion epithelial cells deriving from amnion of term pregnancy with normal amniotic fluid volume (AFV) or isolated oligohydramnios was conducted in the Second Affiliated Hospital of Wenzhou Medical College from January to November 2011.Either group included 10 elective cesarean cases.The primarily cultured cells were treated with different concentrations (0,5,10,20 and 40 mol/L) of ERK inhibitors (U0126) for 12 h,and then the optimal concentration of U0126 which resulted in the lowest expression of phospho~ERK1/2 (p-ERK1/2) was added for different durations(0,2,6,12 and 24 h).Immunocytochemistry was used to detect the localization of AQP3 and Western blot analysis was used to examine the expression of total ERK1/2,p-ERK1/2 and AQP3 in human amnion epithelial cells.Statistical analysis was performed by t-test and one-way ANOVA.Results (1) Compared with those in normal AFV group,p-ERK1/2 and AQP3 expression in human amnion epithelium cells decreased in oligohydramnios group,respectively (p-ERK1/2:3.46 ± 0.33 and 2.46±0.25;AQP3:2.34 ± 0.18 and 1.56±0.10,t=9.243 and 13.292,P＜0.01).(2) In oligohydramnios groups,after treated with different concentrations of U0126,the expressions of total ERK1/2 did not change (F=0.365,P＞0.05).The expression of p-ERK1/2 and AQP3 in 5,10,20,40 μmol/L U0126 (p-ERK1/2:0.96±0.16,1.12±0.13,0.98±0.17 and 1.02±0.26; AQP3:1.10±0.09,1.12±0.08,1.13±0.06 and 1.11±0.06) were all significantly lower than those in 0 μmol/LU0126 group (p-ERK1/2:2.46±0.25; AQP3:1.56±0.10,P＜0.05).However,the expression of p-ERK1/2 and AQP3 showed no significant difference among 5,10,20,and 40μmol/L U0126 groups (P＞0.05).The optimal concentration of U0126 was 5 μmol/L.After treated with 5 μmol/L U0126 for 2 h,the expressions of p-ERK1/2 and AQP3 (1.27±0.29 and 1.44±0.12)were lower than those after treated for 0 h (2.55±0.12 and 2.15±0.09,P＜0.05).However,there was no significant difference among groups treated for 2,6,12 and 24 h.Therefore,the optimal treatment time was 2 h.(3) The expression of AQP3 was distributed in both cell membrane and cytoplasm in amnion epithelial cells with normal amniotic fluid volume or isolated oligohydramnios,but mainly in cytoplasm.U0126 did not change the localization of AQP3 expression.Conclusions U0126 inhibits the phosphorylation of ERK1/2 and expression of AQP3 of women with oligohydramnios,indicating that the MAPK/ERK1/2 signal transduction pathway might regulate the expression of AQP3 in human amnion epithelial cells,and therefore affect the balance of amniotic fluid volume.

Objective To construct recombinant plasmid containing CFP-10 gene of Mycobacterium tuberculosis(MTB).Methods The gene fragment of CFP-10 was amplified by PCR from Mycobacterium tuberculosis H37Rv genomic DNA and cloned to pET-32a(+) vector.The recombinant plasmid pET-32a-CFP-10 was transformed into E.coli BL21 (DE3) and induced by IPTG.Results CFP-10 gene fragment was amplified from genomic DNA of Mycobacterium tuberculosis H37Rv strain,and thepET-32a(+) prokaryotic recombinant plasmid was constructed successfully.The recombinant protein was expressed with the induction of IPTG.Conclusions The prokaryotic expression vector for CFP-10 was successfully constructed and the recombinant protein was highly expressed in E.coli BL21 (DE3),which lays a foundation for its subsequent immunological function study.

Objective To explore the effect of continuing care on the intermittent catheterization compliance of patients with neurogenic bladder. Methods From January to December, 2015, 60 patients with neurogenic bladder after spinal cord injury receiving intermittent cathe-terization were randomly assigned to control group (n=30) and intervention group (n=30). The control group received routine discharge in-struction, while the intervention group received continuing care in addition. The intermittent catheterization compliance, residual urine vol-ume, urinary tract infection and quality of life were assessed at discharge and three months after intervention. Results After intervention, the intermittent catheterization compliance was better in the intervention group than in the control group (χ2=7.500, P=0.006). The residual urine volume significantly decreased in both groups (t>12.040, P<0.001), and was less in the intervention group than in the control group (t=-2.190, P=0.032), as well as the urinary tract infection rate (χ2=10.800, P=0.001). The score of quality of life increased significantly after intervention in both groups (t>4.572, P<0.001), and was higher in the intervention group than in the control group (t>5.505, P<0.001). Con-clusion Continuing care could improve the intermittent catheterization compliance, reduce the residual urine volume and the urinary tract in-fection rate, and improve the quality of life in patients with neurogenic bladder after discharge.

Objective To investigate the Mycobacterium tuberculosis infections in 2014 among Beijing army recruits, and evaluate a new method for screening latent tuberculosis infections.Methods A total of 194 army recruits were subjected to chest X-ray examination purified protein derivative(PPD) skin test, antibody detection, and interferon gamma release assay(IGRA) by ELISA combined with recombinant protein CFP10-ESAT6 and latent infection protein Rv2628.Results The positive rates of PPD skin test and antibody test were 49.7% and 15.5%, respectively.The latent infection rate of IGRA test was 22.2% in 194 cases after CFP10-ESAT6 stimulation.After stimulation of latent tuberculosis infection(LTBI) with Rv2628, IFN-γ level was significantly higher than that in healthy control group (P<0.05).The weak positive group of TST (5 mm≤diameter<15 mm) had a significantly higher level of IFN-γ than the strong positive group(diameter≥15 mm)(P<0.05),but after stimulation with CFP10-ESAT6,IFN-γ levels were not significantly different between the two groups(P>0.05).There was no significant difference between antibody negative and positive groups after stimulation by CFP10-ESAT6 and Rv2628 (P>0.05).The area under the ROC curve of Rv2628 diagnosis of tuberculosis infection was 0.84.When Youden index was 0.621,the specificity was 94.7% and sensitivity was 67.4%.ConclusionCombined detection of antigens Rv2628 and CFP10-ESAT6 specific IFN-γ values can be potentially used for differential diagnosis of active or latent tuberculosis infections.

This study aims to learn drug resistance situation of four first-line anti TB drugs among 251 Mycobacterium tuberculosis isolates from Qinghai and to explore their relationships with genotypes by Spoligotyping,so as to provide basis for effective prevention of tuberculosis.Isolates of 251 M.tuberculosis were tested susceptibilities of four first-line drugs including isoniazid (INH),rifampicin (RFP),streptomycin(SM) and ethambutol (EMB) by using conventional proportion method and genotyped by Spoligotyping.Relationship between drug resistance and genotypes was analyzed statistically.Results showed the total drug resistance rate was 56.2％ (141/251) among 251 M.tuberculosis isolates.Resistance rates of four first-line drugs were 43.0％ (108/251) for INH,37.1％ (93/251) for RFP,39.0％ (98/251) for SM,27.9％ (70/251) for EMB respectively.Rate of multidrug-resistant TB (MDR TB) was 31.5％ (79/251).All 251 isolates of M.tuberculosis were typed by spoligotyping.The 185 (73.7 ％) were Beijing genotypes and 66 (26.3.％) were non-Beijing genotypes,and no statistical association was found with drug resistance.This paper concludes that isolates of M.tuberculosis prevail in Qinghai have both high rates of drug resistance and MDR and dominant isolates are Beijing genotypes by spoligotyping.

Bone age is a quantitative representation of the skeletal development pattern.X-ray imaging of the wrist with the Greulich-Pyle method is commonly used to assess bone age in clinic.In adolescent children, the sensitivity and specificity of the the Greulich-Pyle method are not sufficient because the bones of the wrist are already mature.In contrast, epiphyseal morphological changes in the knee joint throughout adolescence can provide information for the assessment of bone age in adolescent children, and the feasibility of knee joint bone age assessment has been verified.With the application of artificial intelligence (AI) in the medical field, the accuracy of AI interpretation of bone age is also recognized.One of the important uses of bone age assessment in adolescent children is to predict the remaining growth potential.Based on knee images, exploring the use of AI to build a model for predicting residual growth potential is a more meaningful research direction for clinical purposes.This paper reviews the anatomical characteristics of the knee joint, the application of knee joint imaging and the research progress of AI in bone age assessment.