Objective To investigate the impacts of psychosocial factors on the development of breast cancer.Methods Thirty-seven patients with confirmed breast cancer and 37 healthy women were enrolled in this study to complete psychological assessment,including ENRICH marital inventory,Life Event Scale(LES),Social Support Rate Scale(SSRS),and Trait Copying Styles Questionnaire(TCSQ).Results In comparison with the healthy controls,the patients experienced more stress events and negative response,and lowered quality of marriage over the last 5 years.The logistic regressive analyses indicated that the important contributors to breast cancer were stress events,objective social support and negative coping styles.Conclusion Stress events,insufficient access to available psychosocial support and negative responses are important risk factors of breast cancer in women.

Objecfive To investigate the change of V-ATPase B subunits on epithelial to mesenchymal transition (EMT)in rat renal tubular epithelial cells (NRK52E) stimulated by transforming growth factor β1 (TGF-β1). Methods NRK52E cells were stimulated by TGF-β1 (10 μg/L)for O h(control),12 h,24 h,48 h,72 h after sefrum-free culture for 24 h.The mRNA and protein expression of E-cadherin,α-SMA,B2 and B1 subunits of V-ATPase were detected by real-time PCR,Western blotting and immunofluorescence. Results After stimulated by TGF-β1 (10 μg/L)for 48 h,the expression of α-SMA was markedly increased(P<0.05),but the expression of E-cadherin was dramatically decreased(P<0.05).Meanwhile,the expressions of V-ATPase subunit B2 was significantly increased (P<0.05).However,the B1 subunit distributed rarely in NRK 52E cells,and did not increase after TGF-β1 stimulation.Double-label immunofluoerscence staining also showed that the V-ATPase B2 subunit was increased in the cytosol.tending to accumulate to the cell membrane after TGF-β1 stimulation. Conclusions The main isoform of V-ATPase distributed in NRK52E cells is B2 subunit.B2 subunit is increased alone with TGF-β1-induced EMT.It may suggest that V-ATPase B2 subunit may play a potential role in TGF-β1-induced tubular EMT and renal fibrosis.

We optimized the medium for recombinant dextransucrase expression in engineering strain Escherichia coli BL21 (DE3)/pET28-dexYG by an Orthogonal experiment. After the medium had been decided, we studied the effect of temperature, sucrose concentration and pH value on the yield. The results indicated that optimal conditions were adding IPTG of 0.25 mmol/L when OD600 reached 2.0 and cultivation lasted for 4 h at 25 degrees C. Under the selected medium and these conditions, the dextransucrase activity expressed by the engineering strain was high activity. Maximal activity reached 110.16 U/mL sucrose concentration effects the dextran yield grately. The results for dextransucrase expression would provide foundation for industrial application of dextransucrase.
Culture Media ; Culture Techniques ; methods ; Dextrans ; biosynthesis ; Escherichia coli ; genetics ; growth & development ; metabolism ; Glucosyltransferases ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Sucrose ; metabolism

To improve the efficiency of targeted gene replacement (TGR), a dual screen (DS) system with gusA gene as negative selective marker (GUS-DS) was developed in Magnaporthe oryzae. First, we tested the endogenous beta-glucuronidase (GUS) activities of 78 fungal strains. All tested strains were GUS-, only with 3 exceptions. Whereas, after the gusA being introduced in, M. oryzae, Fusarium oxysporum and Colletotrichum lagenarium acquired high GUS activities. The gusA is thus usable as a selective maker in fungal species. With gusA as the negative marker, HPH gene as the positive marker, and the peroxisomal targeting signal receptor genes MGPEX5 and MGPEX7 as 2 instances of target genes, we established the GUS-DS system. After transformation, we collected the transformants from hygromycin B screen media and then tested the GUS activities of them. The GUS- ones were selected as potential mutants and checked in succession by PCR and Southern blotting to identify the true mutants and calculate the efficiency of GUS-DS. As a result, GUS-DS improved the screen efficiency for delta mgpex5 from 65.8% to 90.6%, and for delta mgpex7 from 31.2% to 82.8%. In addition, we established a multiple PCR (M-PCR) method for mutant confirmation. By amplifying the different regions at the targeted locus, M-PCR differentiated the wild type, the ectopic transformants and the mutants effectively and rapidly, and had the same reliability as Southern blotting. In conclusion, GUS-DS and M-PCR are useful tools to improve the efficiency of TGR and would be helpful for fungal genomics.
Escherichia coli ; enzymology ; genetics ; Gene Expression Regulation, Enzymologic ; Genes, Fungal ; Glucuronidase ; genetics ; Magnaporthe ; genetics ; Mutagenesis, Insertional ; methods ; Mutation ; Recombination, Genetic ; Transformation, Genetic

OBJECTIVETo analyze deaf-related genes in patients with nonsyndromic hearing loss (NSHL) and set up a prenatal diagnosis system for such patients.

METHODSNine NSHL families were collected. Potential mutations of GJB2 (35delG, 176del16, 235delC, 299delAT), SLC26A4 (2168A> G, IVS7-2A> G), GJB3 (538C> T) and mtDNA (1494C> T, 12S rRNA 1555A> G) were detected by direct sequencing. Maternal blood contamination was excluded prior to the testing.

RESULTSSixteen patients from 4 families were detected with GJB2 mutations, 8 patients from 2 families were found with SLC26A4 mutations, and 4 patients from 2 families were found with mutations in mtDNA. For 2 patients from one remaining family, no mutations were found with above genes.

CONCLUSIONA diagnostic system for NSHL has been established, which may provide a basis for prenatal diagnosis and genetic counseling to NSHL families.

Connexin 26 ; Connexins ; genetics ; DNA Mutational Analysis ; DNA, Mitochondrial ; chemistry ; genetics ; Deafness ; diagnosis ; genetics ; Family Health ; Female ; Genetic Predisposition to Disease ; genetics ; Hearing Loss ; diagnosis ; genetics ; Humans ; Male ; Membrane Transport Proteins ; genetics ; Molecular Sequence Data ; Mutation ; Pedigree ; Pregnancy ; Prenatal Diagnosis ; methods ; RNA, Ribosomal ; genetics ; Reproducibility of Results ; Sensitivity and Specificity

OBJECTIVETo establish individualized prenatal diagnosis program for families affected with Duchenne/Becker muscular dystrophy (DMD/BMD) and different clinical background using a variety of methods.

METHODSMultiplex ligation-dependent probe amplification (MLPA) was performed on 50 patients suspected for DMD/BMD. For single exon deletions of the DMD gene, PCR was used for validating the results. For those without any deletion or duplication, Sanger sequencing was used to screen for DMD gene mutations in the children and their mothers. Prenatal genetic testing was provided to female carriers using chorionic villus, amniocentesis or cord blood samples. To ensure the accuracy of diagnosis, all prenatal specimens were also subjected to linkage analysis.

RESULTSAmong the 50 patients with DMD/BMD, 23 harbored large deletions, 11 only had single exon deletions, 10 harbored duplications, and 5 had small scare mutations. No mutation was detected in one family. For 37 women undergoing prenatal diagnosis, 10 fetuses were identified as affected males, 6 were female carriers, while 21 were not found to carry any mutation. Testing of creatine kinase was consistent with the results of prenatal diagnosis. For a patient harboring exon 51 deletion, the same mutation was found in a fetus but not in their mother. The proband and fetus had inherited the same haplotype, which suggested that the mother probably has germline mosaicism for the mutation.

CONCLUSIONApplication of individualized methods for analyzing pregnant women with different clinical background can minimize the risk for giving birth to further children affected with DMD/BMD.

Adolescent ; Child ; Child, Preschool ; DNA Mutational Analysis ; Exons ; Female ; Humans ; Male ; Multiplex Polymerase Chain Reaction ; Muscular Dystrophy, Duchenne ; diagnosis ; genetics ; Mutation ; Pedigree ; Pregnancy ; Prenatal Diagnosis

Objective:To evaluate the effect of uncoupling protein 1 (UCP1) expression of perirenal fat on the prognosis of clear cell renal cell carcinoma (ccRCC) .Methods:From Feb. 2013 to Oct. 2013 and Mar. 2015 to Oct. 2015, 98 patients with ccRCC who underwent retroperitoneal laparoscopic radical nephrectomy were analyzed. UCP1 mRNA of perirenal fat around tumor was detected by RT-qPCR. Preoperative Computed tomography (CT) images were used to evaluate the thickness and adhesiveness of perirenal fat. According to the UCP1 mRNA value, the patients were divided into high UCP1 group (42 cases) and low UCP1 group (56 cases) . The general clinical data, perirenal fat thickness and adhesiveness were compared, and Kaplan Meier curve was used to evaluate the difference of progression free survival (PFS) between the two groups. Univariate and multivariate Cox analysis were used to determine the potential independent prognostic factors of PFS.Results:In the high UCP1 group, the renal fat thickness, the ratio of fat adhesion, the ratio of Ⅲ to Ⅳ in Fuhrman grade and the ratio of >T2 in T stage were higher than those in the low UCP1 group[ (13.84±2.41) vs (10.75±1.99) , 42.86% vs 16.07%, 28.57% vs 8.93%, 21.43% vs. 5.36%; P=0.000, P=0.003, P=0.011, P= 0.037]. During the follow-up period (median, 62.0 months) , 15 cases (12 cases of high UCP1 group, 3 case of low UCP1 group) developed tumor progression. Kaplan Meier curve showed that PFS of high UCP1 group was worse than that of low UCP1 group (71.43% vs 94.64%, P=0.001) . Cox regression analysis showed that high UCP1 expression and high T stage were significantly correlated with low PFS ( β=1.334, RR=3.796, 95% CI=1.009-14.280, P= 0.048; β=2.886, RR=17.930, 95% CI=5.538-58.047, P=0.000) . Conclusions:The increased UCP1 expression of perirenal fat may be an independent risk factor of tumor progression in ccRCC. Combined with the assessment of browning of perirenal adipose tissue may be helpful for risk stratification of ccRCC patients after surgery.