Objective To evaluate the relationship of small heterodimer partner(SHP) gene and birth weight in China. Methods A cohort study of 191 normal pregnant women was conducted. Both maternal and cord blood samples were collected. PCR-RFLP was used to detect the polymorphism of SNP-rs7504 of SHP. Results (1) The frequency of both neonatal and maternal C allele and (TC+CC) genotype increased significantly with birth weight (P=0.004, OR=3.168; P=0.005, OR=3.315; P=0.013, OR=2.495; P=0.013, OR=2.495). (2) The babies were heavier if they were C allele carrier. The average increase of birth weight was 246.3 g comparing the neonates with TC+CC genotype with those with TT genotype [(3658.7?400.94)g vs (3412.4?444.4)g, P=0.005]. The average birth weight of those maternal C allele carriers was 210.3 g heavier than those non-C allele carriers[(3628.9?405.5) g vs (3418.6?449.0 g]. (3) The fetal C allele was associated with maternal weight in pregnancy, prepregnant BMI, paternal height and weight. Women with C allele were heavier and had higher BMI without statistical significance comparing with those non-C allele carriers. Neither neonatal nor maternal SHP gene was associated with blood glucose and insulin level. (4) Multiple factors analysis showed that birth weight was related to maternal height, weight gain during pregnancy, prepregnant BMI, maternal and cord blood insulin level. After adjustment, the neonatal birth weight remained significantly correlated with cord blood SHP (P=0.0354), but not with maternal SHP gene (P=0.0711). Conclusions SHP gene is associated with newborns birth weight and may affect fetal growth.

Flow injection on-line sorption preconcentration and separation in a knotted reactor (KR) was coupled to cold vapor atomic fluorescence spectrometry for the determination of trace mercury in mineral water. Mercury was preconcentrated by on-line formation of mercury diethyldithiocarbamate complex (Hg-DDTC) and absorption of the resulting neutral complex on the inner walls of a knotted reactor. A 20%(V/V) HNO3 solution heated by electromagnetic induction heating technique was used as eluent to remove the absorbed Hg-DDTC from the KR, and then the vapor mercury generated by mixing the resulting solution and KBH4 was determined on-line by cold vapor atomic fluorescence spectrometry. The 20% HNO3 was employed as both the efficient eluent and the required acidic medium for subsequent mercury vapor generation in our work. Using 20% HNO3 instead of conventional organic solvent as eluent, the proposed method is simple, easy operational and environmentally friendly. Under the optimal experimental conditions, the sample throughput was approximatively 30/h with an enhancement factor of 35. The detection limit of mercury was 2.0 ng/L. The precision(RSD, n=11) was 2.2% at the 0.1 μg/L Hg2+ level.

Objective To research the effect of the Kiss-1 gene promoter methylation on the Kiss-1 gene expression in colorectal carcinoma tissue,and to analyze the relationship between the Kiss-1 gene methylation and the clinical pathological features of colorectal carcinoma and its clinical significance.Methods The Kiss-1 gene promotor region methylation,Kiss-1 gene mRNA and protein expressions were detected respectively by methylation-specific PCR, Real-time PCR and Western blotting method in 126 cases of colorectal carcinoma tissue and para-cacinoma normal colorectal tissue.Results The positive rate of Kiss-1 gene methylation in colorectal carcinoma tissue (83.33%)was significantly higher than that in normal tissue (30.16%)(P<0.05).In the cancer tissue,the expression levels of Kiss-1 gene mRNA and protein in Kiss-1 gene promoter methylation positive group was lower than that in Kiss-1 gene promoter methylation negative group (P < 0.05 ). The positive rate of Kiss-1 gene promoter methylation in T3+T4 stage group was higher than that in T1+T2 stage group (P<0.05),but the Kiss-1 gene mRNA and protein expression levels were lower than those in T1+T2 stage group (P<0.05).In the poorly and moderately differentiated group,the positive rate of Kiss-1 gene promoter methylation was higher than that in well differentiated group (P<0.05 ), but the mRNA expression level of Kiss-1 gene was lower than that in well differentiated group (P<0.05 ). In lymph node metastasis group, the positive rate of Kiss-1 gene promoter methylation was higher than that in lymph node negative group (P<0.05),but the mRNA and protein expression levels were lower than those in lymph node negative group (P<0.05 ). And the positive rate of Kiss-1 gene promoter methylation in distant metastasis group was higher than that in non-distant metastasis group (P<0.05), but the mRNA and protein expressions levels were lower than those in non-distant metastasis group (P<0.05). There were no significant associations between the positive rate of Kiss-1 gene promoter methylation and the gender, age,tumor location, and tumor diameter of the patients (P>0.05 ). Conclusion The Kiss-1 gene promoter methylation in colorectal carcinoma tissue is associated with the Kiss-1 gene expression level and the malignant characteristics of colorectal carcinoma;Kiss-1 gene promoter methylation may be used as a reference indicator for predicting the risk of metastasis of colorectal carcinoma.

Objective To investigate the apoptosis of rat glioma C 6 cells induced by defective in-terfering( DI) particles of Sendai virus strain Tianjin .Methods Rat glioma C6 cells were treated with dif-ferent titers of DI particles of Sendai virus strain Tianjin in vitro with culture media as negative control and intact virus as positive control .At different time point , cells were collected and their apoptosis was detected by DNA gel electrophorsis , TUNEL assay and AnnexinⅤ/PI double-labeled flow cytometry .The C6 glioma-bearing rat model was established and then treated with three intratumoral injections of DI particles , intact virus or saline three times at interval of two days .The antitumor effects of ID particles were evaluated through daily measuring of the tumor size .Hematoxylin-eosin( HE) staining was used to observe the patho-logical changes in tumor tissues .TUNEL assay was performed to detect the apoptosis of tumor tissues .Re-sults Rat glioma C6 cells treated with DI particles or intact virus in vitro showed typical DNA ladder pattern in agarose gel electrophoresis in a time-and dose-dependent manner .With the intervention of DI particles , the apoptosis rate of C6 cells showed a time-and dose-dependent manner and was significantly higher than that of the control group (P<0.01) as indicated by flow cytometry assay and TUNEL assay .In vivo, DI par-ticles could markedly inhibit the growth of the tumors in comparison with saline control group .There were fe-wer tumor cells in tumor nodules in DI particles group or intact virus group as shown by histological examina -tion.The TUNEL assay showed that the apoptosis rate of tumor tissues injected with DI particles or intact vi -rus was much higher than that of the saline group (P<0.01), but there was no significant difference between the DI particles group and the intact virus group (P>0.05).Conclusion The DI particles of Sendai virus strain Tianjin could induce apoptosis of rat glioma C 6 cells in a time-and dose-dependent manner both in vitro and in vivo, suggesting that the DI particles might be applicable for the treatment of neurogliocytoma in the future.

Objective:To explore the effects of KISS1 gene transfected by lentivirus on the proliferation,invasion and migration abilities of the colorectal cancer HCT116 cells,and to clarify their mechanisms.Methods:The human colorectal cancer cells with the lowest expression level of KISS1 gene were selected.The lentiviral vectors were builted and transfected the KISS1 gene,and the cells were divided into control group (treated with PBS),empty vector group (treated with empty vector) and over-expression group(treated with KISS1 gene vector).The multiplicity of infection (MOI) of the cells was detected by fluorescence microscope.Real-time PCR and Western blotting methods were used to detect the expression levels of KISS1 mRNA and protein(metastin);CCK-8 method was used to detect the proliferation ability of the cells;Transwell chambers method was used to detect the invasion and migration abilities of the cells.Results:Among LoVo,SW620,SW480,HCT-116,and HT29 cells,the expression levels of KISS1 mRNA and protein were lowest in HCT116 cells,so they were chosen as the research carrier.After transfected with lentiviral vectors,the HCT116 cells could stably express the enhanced green fluorescent protein(EGFP) gene,and the MOI was over 80%.Compared with control group and empty vector group,the expression levels of KISS1 mRNA and protein in the cells in over-expression group were significantly increased (P<0.05);the proliferation abilities of the cells in over-expression group were decreased (P<0.05);the invasion ability and migration ability of the cells in over-expression group were decreased (P<0.05).But the differences of proliferation ability,invasion ability and migration ability of the cells between control group and empty vector group were not statistically significant (P>0.05).Conclusion:The KISS1 gene transfected by lentivirus vector can over-express KISS1 protein and inhibit the proliferation,invasion and migration abilities of the colorectal cancer cells,and the mechanism may be related to the expression of KISS1 protein.

Objective To compare the genotypes distribution of human papillomavirus (HPV ) infection in tissues of cervical cancers and cervical intraepithelial neoplasias (CIN ) and its clinical significance .Methods The polymerase chain reaction (PCR) and the gene-chips technique were utilized for the detection of 23 kinds of HPV genotypes in the tissue specimens from 192 cases of cervical intraepithelial neoplasia (CIN) and 85 cases of cervical cancers .And the related data of all subjects were analyzed .Results In 192 cases of CIN ,the total positive rate of HPV was 82 .29% (158/192) ,the positive rate of single genotype infection was 46 .88% (90/192) and the positive rate of multiple genotypes infection was 35 .42% (68/192);In 85 cases of cervical cancers ,the to-tal infection rate of HPV was 88 .24% (75/85) ,the positive rate of single genotype infection was 65 .88% (56/85) and the positive rate of multiple genotypes infection was 22 .35% (19/85) .Conclusion PCR combined with the gene-chips technique can be used in the detection of the tissue samples of cervical lesions ,once detection can detect 23 kinds of HPV genotypes with high sensitivity and strong specificity ,which has very important significance to the prevention and treatment of cervical cancer and precancerous lesions and the their vaccine research .

OBJECTIVETo investigate the effect of Kiss-1 gene suppression on the metastatic capacity of HCT116 human colorectal carcinoma cells in vitro and the involvement of nuclear factor-κB (NF-κB) signaling pathway.

METHODSA recombinant lentiviral vector of Kiss-1 gene pGC-LV-Kiss-1-EGFP or the empty vector was transfected in HCT116 cells. Cell Counting Kit-8 (CCK8) and Transwell chamber assay were used to detect the changes in cell proliferation, invasion and migration ability after the transfection. Western blotting was used to detect the expression of I-κB, the inhibitive protein of NF-κB signal pathway, and the expression of the downstream effector MMP-9 before and after transfection.

RESULTSIn cells over-expressing Kiss-1, I-κB expression increased and MMP-9 expression decreased significantly compared to those in the blank control and vector-transfected cells (P<0.05). Kiss-1 gene over-expression resulted in significant inhibition of HCT116 cell proliferation, invasion, and migration as compared to the control cells (P<0.05).

CONCLUSIONLentivirus-mediated Kiss-1 gene over-expression can inhibit the proliferation, invasion, and migration of HCT116 cells via the NF-B signaling pathway.

Cell Movement ; Cell Proliferation ; Colorectal Neoplasms ; pathology ; Genetic Vectors ; HCT116 Cells ; Humans ; I-kappa B Kinase ; metabolism ; Kisspeptins ; genetics ; Lentivirus ; Matrix Metalloproteinase 9 ; metabolism ; NF-kappa B ; metabolism ; Neoplasm Invasiveness ; Signal Transduction ; Transfection