Objective To determine the correlation between Smad4 mRNA,protein expression,abnormal methylation of CpG islands of Smad4 and colorectal cancer.Methods The expression of Smad4 in 43 cases of colorectal cancer and its adjacent tissue.30 cases of colorectal adenoma and 12 normal colon mucosa were analyzed by RT-PCR,sequencing,semi-quantitative RT-PCR,methylation-specific PCR(MSP)and immunohistochemistry.Results The mRNA expression of Smad4 was detected in 25 cases of colorectal cancer(58.14%),with expression levels at 0.73±0.25,which was significantly lower than those in corresponding adjacent tissues(88.37%,0.95±0.29),eolorectal adenoma(90.63%,1.01±0.37)and normal mucosa(100.00%,1.18 ±0.33)(P＜0.05).The positive rates of Smad4 gone promoter methylation in colorectal caner(60.53%)was significantly higher than that in other tissues(27.03%,25.00%and 16.67%,respectively,P＜0.05).The expression of Smad4 protein was significantly lower in colorectal cancer(44.19%)than that in other tissues(81.40%,87.50%,91.67%,respectively,P＜0.05),which was correlated with tumor invasion and lymph node metastasis.Conclusion Down-regulation of Smad4 expression may be associated with the development,the biological behavior and prognosis of colorectal cancer,and it can be an important biological marker in evaluation of disease progression.

Objective To evaluate the feasibility of bladder extracellular matrix (BACM) seeding with autologous cultured vaginal smooth muscle cells (VSMC) repaired rabbit vaginal defect.Methods This study included 24 female rabbits.BACM and vaginal acellular matrix (VACM) were obtained from 8 rabbits by decellularization process.The other 16 rabbits were randomly divided into experimental and control groups.Vaginal tissue biopsies ( ～ 1 cm2) were harvested from female New Zealand rabbits.VSMC were cultured and stained with a-smooth muscle actin antibodies. Cultured VSMC cells were seeded on BACM ( experimental group) or VACM ( control group) at a cell density of 1 × 107 cells/cm2.The cell-seeded matrixes were cultured for 5days.In experimental group of 8 rabbits,a 2 cm segment of vagina was resected and replaced with BACM seeding with VSMC.Then the regenerative segment was studied with histological technique by hematoxylin-eosin staining after 3,6 and 12 weeks postoperative aud vaginography was performed at 12 weeks postoperative.The 8 rabbits in control group underwent the exact same procedure as above but the vaginal defect was repaired with VACM seeding with VSMC,Results The prepared BACM and VACM were transparent,HE staining and scanning electron microscopy showed the two acellular matrix was both consisted of abundant network of fibers with the regular arrangement,without cellular debris.Primary culture of VSMC was successfully established and passaged,and was uniformly spindle-shaped in the confluent state and showed a characteristic hill and valley'formation.Immunohistochemical staining showed VSMC in culture stained positively with a-smooth muscle actin antibodies.VSMC began to adhere to the BACM 5 hour after implanting and the number of the adhered cells increased with time.Cells gradually expanded and showed the typical morphology of smooth muscle cells.Three weeks after implantation in vivo,the luminal surface of matrix was completely covered by vaginal epithelial tissue,a layer of smooth muscle cells was formed in the outer surface of the matrix.Multilayered vaginal epithelial and improved development of organized muscle bundles was observed after 6 weeks.The regenerative tissue was equivalent to the normal vaginal tissue at 12 weeks postoperatively.Vaginography demonstrated the maintenance of full patency and a wide vaginal caliber without fibrosis and graft rejection.There was no significant difference in all evaluated items between experimental and control groups.Conclusions BACM has the same regenerative process as VACM in the replacement of vaginal defect.Moreover,BACM has wider source and appears to be an suitable material for vaginal replacement.

ObjectiveTo evaluate the impacts of pereutaneous nephrolithotomy (PCNL) on perioperative renal hemodynamics.MethodsThe hemodynamics of operated renal arteries of 30 patients who underwent unilateral PCNL with single pole access were observed 1 day before and 5-7 days after operation with CDFI.Parameters were analyzed statistically.ResultsAfter operation,resistance index (RI) of renal arteries decreased (P＜0.05).The diastolic flow statistically increased at main renal artery (MRA) of renal hilus,interlobar renal artery and interlobular renal artery (all P＜0.05).After PCNL,in serious hydronephrosis patients,RI decreased (P＜0.05) at segmental renal artery (SRA) and interlobar artery,end-diastolic flow velocity (Vmin) increased at interlobar renal artery (P＜0.05).In moderate hydronephrosis patients,RI decreased at all renal arteries (P＜0.05) after PCNL,Vmin increased at MRA and interlobular renal artery (P＜0.05).In minor hydronephrosis patients,RI decreased at MRA and SRA,Vmin increased at SRA.In patients without hydronephrosis,RI changeed like serious hydronephrosis patients.ConclusionAfter PCNL,ipsilateral renal perfusion improves,renal diastolic flow increases and RI decreases.CDFI can be used to observe the blood perfusion of kidney,and provide quantitative information of renal hemodynamics.

Objective To establish the normal reference ranges of T lymphocyte subsets in healthy Zhuang ethnic minority across different age/ gender groups in Guangxi for providing the scientific guidance for clinical diagnosis and treatment evaluation of immunodeficiency diseases and viral infections as well as relat-ed researches. Methods Peripheral blood samples were collected from 405(196 male and 209 female) healthy subjects of Zhuang ethnic group including 82 adolescents and 323 adults. Single platform FACSCali-bur flow cytometry analysis was performed to detect the absolute counts of T lymphocyte subsets(CD45+, CD3+,CD4+ and CD8+)in peripheral blood samples. Results Reference ranges of T lymphocyte absolute counts in healthy Zhuang Chinese aged 14 to 59 were 2 377±696,1 503±459,802±253,581±243 and 1. 52±0. 57 for CD45+,CD3+,CD4+,CD8+ and CD4+ / CD8+. Significant differences in CD45+,CD3+ and CD8+ T lymphocytes were found between adolescents and adults(t = 0. 000,P﹤0. 05),but no significant difference in CD4+ T lymphocytes was observed between the two groups of people(t = 0. 373,P﹥0. 05). Significant difference between male and female groups was observed only in the absolute counts of CD8+ T lymphocytes(t= 0. 036,P﹤0. 05). The absolute counts of T lymphocyte subsets were on the decline along with the growth of the age. Conclusion This study indicates that the counts of T lymphocyte subsets vary with ethnic group,age and gender. Therefore,it is important to establish normal reference ranges of T lym-phocyte subsets for local healthy people based on gender and age for accurate evaluation of local patients' im-mune system status and treatment efficacy.

Objective To investigate the genetic polymorphism of DXS8378 STR locus of chromosome X in Chinese Lisu,Pumi and De'ang populations in Yunnan and construct relative standard allelic ladders.Methods After being amplified by PCR,different STR allelic fragments were isolated from the PAG electrophoresis.The STR allelic fragments were extracted by kit and reamplified by PCR to obtain purified allelic fragments.Next,the purified allelic fragments were subcloned individually into the PUC plasmid vectors,and the size and structure of the inserts were confirmed by the analysis of their DNA sequences.Then we transfected it into competent E.coli DH5?TM cells,and finally,the recombinant plasmids DNA with the inserts were used as template for reamplification to generate the standard ladders.Results The standard allelic ladder for DXS8378 locus was obtained,with which the genetic polymorphisms of DXS8378 locus in three Chinese populations in Yunnan were studied.Conclusion The standard ladder made by this method is excellent,and DXS8378 is powerful for forensic practice in Chinese population.

Objective:To increase the critical relative humidity(CRH) of Qubi granule,thus to improve its humidity resistance. Methods:The best prescription of Qubi granule was optimized by comparative method,and to determine its critical relative humidity. Results:The old prescription of the Qubi granule's CRH was 50%.With new prescription,the Qubi granule’ s CRH was 61%. the latter has increased by 11% as compared with the former . Conclusion:Considering above result,there is a hope to improve the hygroscopicity of Qubi granule.

Objective · To observe the change of serum complement C1q tumor necrosis factor-related protein1 (CTRP1) level and explore its relationship with serum adiponectin (APN) level in elderly male metabolic syndrome (MS) patients. Methods · Clinical data of 279 male objects (60-90 years old) were analyzed retrospectively, serum CTRP1 and APN levels of all objects were tested by enzyme-linked immuno sorbent assay (ELISA). The patients were divided into three groups, i.e. 105 MS patients (MS group), 90 MS patients combined with hypertension (HMS group), and 84 non-MS patients (control group). All general information, including height, body weight, systolic blood pressure (SBP), diastolic blood pressure, fasting blood glucose, fasting insulin and serum three acyl glycerin (TAG) were recorded, in order to calculate body mass index (BMI) and homeostasis model assessment of insulin resistance (HOMA-IR). Results · Compared with the control group, serum CTRP1 levels of patients in MS group and HMS group both increased, and the latter was more obviously. Serum APN levels of patients decreased obviously in HMS group and MS group. The level of serum CTRP1 was negatively related with the level of serum APN. Conversely, serum CTRP1 level was positively related with blood glucose, BMI, SBP, TAG and HOMA-IR. Conclusion · The level of serum CTRP1 is elevated, while the level of serum APN declines in elderly male MS patients. The serum level of CTRP1 is even higher in HMS patients. Serum CTRP1 level is related to many risk factors of atherosclerosis.

Background and purpose:Pterostilbene is a natural antioxidant, whose role in retinoblastoma remains unclear. The aim of this study is to probe the effects of pterostilbene on the proliferation, apoptosis and autophagy in retinoblastoma WERI-Rb-1 cell lines.Methods:Cell counting kit-8 (CCK-8) assays were used to analyze the effects of pterostilbene on the proliferation of WERI-Rb-1 cells. Apoptosis rate was determined by Annexin V/PI. Autophagic vacuoles were observed by acridine orange staining. LC3 and P62 protein expressions were determined using Western blot.Results:Pterostilbene significantly inhibited the proliferation of WERI-Rb-1 cells (P<0.01). The cell viability were (93.02±0.47)%, (55.10±2.04)% and (30.33±1.45)% after WERI-Rb-1 cells were treated with 25, 50 and 100 μmol/L pterostilbene for 24 h, and the cell viability were (88.38±3.70)%, (53.37±1.17)%, (29.60±1.05)% after WERI-Rb-1 cells were treated with 50 μmol/L pterostilbene for 12, 24 and 48 h. Pterostilbene induced cell apoptosis (P<0.01), the apoptosis rates of control group, 24 h treated group and 48 h treated group were (4.08±0.79)%, (13.44±2.12)% and (23.49±2.01)%. Pterostilbene induced autophagy of WERI-Rb-1 cells, increased LC3 expression, downregulated P62 expression and increased the number of autophagic vacuoles in WERI-Rb-1 cells (P<0.01). 3-MA and Beclin1 were able to rescue pterostilbene-induced cell death (P<0.01). After 3-MA was used to blunt autophagosome formation, the apoptosis rate markedly decreased in 3-MA+pterostilbene-treated cells compared with cells treated with pterostilbene alone [(12.97±2.09)%vs (8.35±1.11)%], and after siRNA was used to knockdown Beclin1, the apoptosis rate had the same change [(13.80±2.19)%vs (9.62±0.52)%].Conclusion:Pterostilbene can inhibit the proliferation of WERI-Rb-1 cells and induce cell apoptosis via autophagy activation.

Objective To investigate the effect of cyclosporine A(CsA)on viral protein syn- thesis and hepatitis B virus(HBV)DNA replication in vitro.Methods The HBV DNA transfected cell line HepG2.2.15 was treated with different concentration of CsA(0.6-20.0?g/ml)for 4 days. Hepatitis B surface antigen(HBsAg)and hepatitis B e antigen(HBeAg)in supernatant were detected by enzyme-linked immunosorbentassay(ELISA);intracellular hepatitis B core antigen(HBcAg)mR- NA and HBV DNA were analyzed by RT-PCR and slot blot hybridization,respectively;the phospho- rylation at tyrosine acid position 402 of PyK2 kinase(PyK2 Y402)was detected by Western blot.Re- sults CsA could suppress the expression of HBsAg and HBeAg,and inhibit the HBV DNA replica- tion in a dose-dependent manner.The suppression rate of HBsAg and HBeAg under the action of CsA at the concentration of 10.0?g/ml for 4 days was 49.7% and 34.3%,respective;similar effect was observed on HBV DNA replication,HBV DNA was only 34.9% of the control at the concentration of 10.0?g/ml of CsA.The phosphorylation level of PyK2 Y402 was declined under the action of CsA at the concentration of 2.0?g/ml.Conclusions CsA can inhibit the expression of HBsAg,HBeAg and HBV DNA replication in the HepG2.2.15 cell line in a dose-dependent manner.Suppression of the phosphorylation level of PyK2 Y402 maybe involved in the mechanism of the inhibitory activity of CsA on HBV replication.

This study was aimed to investigate the distribution and implication of tap1 (transporter associated with antigen processing) and tap2 loci allelic and genotypic frequencies. The distribution of tap1 and tap2 loci allelic and genotypic frequencies in 339 random samples of healthy Chinese Hans was analyzed by TaqMan PCR. Several genetic information about power of discrimination, cumulative DP, polymorphism information content, expected heterozygosity and observed heterozygosity were calculated. The results indicated that 5 tap1 alleles (tap1*0101, 020101, 020102, 0301 and 0401) and 4 tap2 alleles (tap2*0101, 0102, 0103 and 0201) were detected in all samples. 8 tap1 genotypes were found which account for 53.3% of the theoretic genotype and 6 tap2 genotypes were found which account for 60% of the theoretic genotype. The genotyping results of tap1 and tap2 both conform to the Hardy-Weinberg expectations (p > 0.05). Tap1*0101 (79.79%) and tap2*0101 (82.74%) are the most common alleles in Chinese Hans. It is concluded that tap1*0101 and tap2*0101 are most common alleles in Chinese Hans, tap1 and tap2 loci carry some power of individual discrimination and polymorphism information content. These two locl can be used for the research in the fields of human genetics, linkage analysis of genetic disease genes, paternity test and individual identification and so on.
ATP-Binding Cassette Sub-Family B Member 2 ; ATP-Binding Cassette Transporters ; genetics ; ATP-Binding Cassette, Sub-Family B, Member 3 ; Alleles ; Asian Continental Ancestry Group ; genetics ; Gene Frequency ; Genotype ; Haplotypes ; Humans