Objective To investigate the Influence of Survivin and hTERT gene on cell proliferation and apoptosis in human colorectal carcinoma cell line SW 480 and to find experiment evidence for gene therapy of colorectal carci-noma .Methods Plasmids carrying shRNAs targeting survivin and hTERT were designed , constructed and trans-fected into SW480 cells.SW480 cells were then divided into blank group , blank Plasmid control group , survivin RNAi group , hTERT RNAi group and Survivin-hTERT RNAi group .The telomerase activity was examined by TRAP-PCR-ELISA analysis 48h after hTERT-shRNA transfection.Survivin and hTERT mRNA and protein expres-sion was analyzed by RT-PCR and Western blot .Cell apoptosis , proliferation were measured by flow cytometry , CCK-8 assay.Results Telomerase activity of SW480 cells in Survivin-hTERT RNAi groups were significantly decreased compared with the blank group ( P<0.01 ) .The expression of survivin and hTERT mRNA, proteins in the Survivin-hTERT RNAi group was reduced by 82.8%and 73.6%( P<0.01 ) ,79.2%and 66.7%( P<0.01 ) respectively .The inhibitory rate of cell proliferation of Survivin-hTERT RNAi group was 43.6% ±0.1%( P <0.01 ) .The apoptosis rate was 39.2%±2.3%( P<0.01 ) in the Survivin-hTERT RNAi group .Conclusions The Survivin-hTERT RNAi group could significantly reduces the protein expression of survivin and hTERT mRNA, in-hibit cell proliferation and induces cell apoptosis in human colorectal carcinoma cell line SW 480 .

ObjectiveTo investigate the methods and clinical significance of detecting PLAC4 and COL6A1 gene on fetal chromosome 21 from maternal peripheral blood. Methods From Oct. 2008 to Nov. 2009 30 normal pregnancies in Weifang People's Hospital were selected as pregnant group, and 9 nonpregnant women were selected as control group. Quantitative real-time PCR was used to determine transcript levels of the target genes ( PLAC4 and COL6A1 ) in blood samples. Correlation between the expression level and gestational age was analyzed. Results ( 1 ) PLAG4 mRNA was detected in peripheral blood of all pregnant women. Its maximum level was 12. 760 × 103 copies/ml, whereas the minimum was 2. 105 × 103 copies/ml, and the average value is 6. 612 × 103 copies/ml. In control group the PLAC4 mRNA could not be detected. There was statistically significant difference ( P ＜ 0. 01 ) between the two groups. ( 2 ) COL6A1 mRNA is detected in pregnant group and control group, and the concentration was 6. 847 × 103 copies/ml and 7. 322 × 103 copies/ml respectively, with no statistically significant difference ( P ＞ 0. 05 ). ( 3 ) Correlation analysis: there was no relationship between the level of PLAC4, COL6A1 mRNA and the gestational age, the correlation coefficients (r) were 0. 29 and 0. 31, and the P values were 0. 121 and 0. 168 respectively. Conclusions COL6A1 mRNA can be detected in both pregnant group and control group, so it is not specific for pregnancy. PLAC4 mRNA can be detected only in pregnant women, so it has specificity in pregnancy and can be a discriminative marker gene for prenatal dignosis of trisomy 21 fetuses.

Objective To explore an optimal method of producing STZ-induced type 1 diabetic KM mice model by investigating the molded rate of single high dose and multiple low dose of STZ injection.Methods Sixty KM mice were randomly divided into 4 groups(n=15), two control groups and two model groups.In the two control groups, one was blank control and the other was negative control.Mice in the blank control group treated with no injection, but mice in the negative control group were treated with injection of citric acid salt buffer.For the two model groups, one was single high-dose group, 150 mg/kg STZ was injected only once.The other was multiple low-dose group, 50 mg/kg STZ was injected for 5 consecutive days.After the last injection, daily food and water intake were tested, blood glucose level and body weight were examined once a week for 4 consecutive weeks. Results Mice in the two model groups showed typical features of diabetes.The blood glucose levels in the two model groups were significantly higher than in the two control groups ( P <0.05 ) .For the two model groups, the molded rate of 150 mg/kg and 50 mg/kg group were 60% and 53.33%respectively at 1st week, but at the 4th week, they were 60% and 80% respectively.The mortality of these two groups at the 4th week was 33.33% and 20% respectively.Moreover, the blood glucose level in multiple low-dose group increased stably from the 2nd week to the 4th week.Conclusion The multiple low-dose STZ injection (50 mg/kg for 5 consecutive days) is an optimal method for producing KM mice model of type 1 diabetes mellitus.

OBJECTIVETo observe sperm motion parameters in pre-freeze and post-thaw semen samples using computer-assisted sperm analysis (CASA) system.

METHODSSemen analyses of 238 samples before freezing and after thawing were separately performed by Hamilton-Thorne Sperm Analyzer.

RESULTSSperm motility in post-thaw samples was significantly decreased. There was significant correlation and difference between pre-freeze and post-thaw samples in sperm motion parameters, including average path velocity (VAP), straight line velocity (VSL), curvilinear velocity (VCL), amplitude of lateral head displacement (ALH), straightness (STR) and linearity (LIN), except heat cross frequency (BCF). The percentage of sperm movement velocity parameters (VAP, VSL and VCL) and moving pattern parameters (ALH) significantly decreased, while that of LIN and STR significantly increased in post-thaw samples.

CONCLUSIONCASA system is of clinically applied value and is a useful tool for evaluating sperm motion parameters in pre-freeze and post-thaw semen samples.

Cryopreservation ; Diagnosis, Computer-Assisted ; Freezing ; Humans ; Male ; Sperm Motility

OBJECTIVETo evaluate the semen quality of the donors.

METHODSThe semen parameters of the primary samples of 512 donors were examined following the World Health Organization (WHO) guide and computer-aided sperm analysis (CASA, product by Harmilton Thorne) system.

RESULTSOf the 246 (246/512, 48%) donors with potential fertility, only 146 (146/512, 28.5%) came up to the semen standard set by the Chinese Ministry of Health, while 266 (266/512, 52%) were below the WHO reference values of semen parameters.

CONCLUSIONRepeat semen analyses may increase the success rate in screening semen donors. The semen quality of the donors suggests that it is necessary to pay more attention to male reproductive health.

Adult ; Humans ; Image Processing, Computer-Assisted ; Male ; Mass Screening ; Middle Aged ; Reference Values ; Semen ; physiology ; Sperm Banks ; Sperm Count ; Sperm Motility ; Tissue Donors

OBJECTIVE： To establish HPLC fingerprint of Miao medicine Hedyotis uncinella from Guizhou. METHODS： HPLC method was adopted. The determination was performed on Agilent Eclipse XDB C18 column with mobile phase consisted of acetonitrile-0.2％ phosphoric acid （gradient elution） at the flow rate of 1.0 mL/min. The detection wavelength was set at 235 nm， and column temperature was 25 ℃. The sample size was 15 μL. Using rutin as reference， HPLC chromatograms of 10 batches of H. uncinella from different areas of Guizhou province were determined. Similarity Evaluation System for TCM Chromatographic Fingerprints （2004A edition） was used to identify common peaks and evaluate similarity. RESULTS： There were 12 common peaks in HPLC fingerprints of 10 batches of H. uncinella， and the similarity was higher than 0.90. After validation， HPLC chromatograms of 10 batches of sample were in agreement with control fingerprints. CONCLUSIONS： Established HPLC fingerprints can provide reference for the identification and quality evaluation of H. uncinella.

Objective:To investigate the correlations of laminin subunit gamma 3 (LAMC3) expression with prognosis of ovarian cancer (OC).Methods:LAMC3 protein expression was measured using immunohistochemical streptavidin-peroxidase-biotin connection method (IHC). Gene expression and related clinical data in the cancer genome atlas (TCGA) cohort and clinical proteomic tumor analysis consortium (CPTAC) were applied to analyse the correlation between gene and protein expressions and clinical outcomes. Correlations between LAMC3 and clinicopathological factors were evaluated using the Pearson χ2 test (2-sided). The probability of survival and significance was calculated using the Kaplan-Meier plot. The functional clustering of biological pathways enriched from co-expressed genes of LAMC3 was used to explore the possible mechanisms that LAMC3 might contribute to poor prognosis. Results:Based on the IHC results of 216 OC tissues or ovaries (including 208 tumors and 8 normal tissues) and 51 OC tissues (including 24 chemotherapy-resistant and 27 sensitive tissues), and the protein expression data from CPTAC (including 100 primary tumors and 25 normal tissues), the results showed that the protein expression of LAMC3 was significantly decreased in OC tissues compared with normal, decreased in advanced-stage tissues compared with early-stage tissues, and decreased in drug-resistant tissues compared with sensitive tissues (all P<0.05). Furthermore, low expression of LAMC3 protein was significantly associated with poor disease-free survival (DFS) and overall survival (OS) in 51 OC tissues ( P<0.01), consistent with the results that the low levels of LAMC3 mRNA predicted short DFS and OS in 489 OC tissues of the TCGA cohort ( P<0.05). The results suggested that low expression of LAMC3 might be the adverse factors for OC development, such as drug resistance and advanced tumors, and might be a risk indicator for prognosis. Moreover, functional clustering of biological pathways enriched from the co-expressed genes of LAMC3 in TCGA ovarian cohort indicated that LAMC3 potentially involved in regulation of OC via oncogene-pathways such as Ras associated protein 1 (Rap1), mitogen-activated protein kinase (MAPK), Ras and cell adhesion-related pathways such as extra cellular matrix (ECM)-receptor interaction and focal adhesion. It indicated that LAMC3 might contribute to short survival and tumor progression by regulation of the above pathways. Conclusion:Low expression of LAMC3 is related to poor prognosis and malignant progression in OC, and thus it is expected to be a new prognostic marker and therapeutic target for clinical treatment.