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 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 study the technical conditions and parameters of DiaionHP-20 macroporous adsorption resin for the purification of the anti-PMOP(primary Type I osteoporosis) active fraction from Camellia semiserrata seed.METHODS: With the elution ratios and purity of total flavonoids and polyphenol from Camellia semiserrata as indexes,the technical parameters of DiaionHP-20 macroporous adsorption resin including the adsorption quantity,the loading sample,the elution concentration and volume of alcohol were investigated.RESULTS: The adsorption quantity of DiaionHP-20 macroporous resin on total flavonoids and polyphenol were 59.5 mg?g-1 and 23.9 mg?g-1 respectively.The optimal technical conditions were as follows:the quantity of drug was 10.5 times that of resin;water-soluble foreign substance was washed off with distilled water weighing 3 times the weight of resin,and total flavonoids and polyphenol were eluted using 25% ethanol weighing 13.5 times the weight of resin.CONCLUSION:DiaionHP-20 macroporous resin had good purification effects on total flavonoids and polyphenol from Camellia semiserrata seeds.The process is simple,low cost,and suitable for lavge-scale production.

Objective To study the distribution status and clinical significance of human papillomavirus(HPV) infection geno‐types in condyloma acuminate(CA) tissues of vulva ,vagina and cervix .Methods The gene‐chips combined with polymerase chain reaction (PCR) technology were utilized for detecting 23 kinds of HPV genotypes in tissue specimens from 63 cases of vulval CA , 61 cases of vaginal CA and 65 cases of cervical CA .Their clinical pathological data were analyzed .Results In 63 cases of vulval CA ,56 cases were HPV positive with the HPV infection rate of 88 .89% (56/63) ,in 61 cases of vaginal CA ,55 cases were HPV positive with the HPV infection rate of 90 .16% (55/61) ,and in 65 cases of cervical CA ,62 cases were HPV positive with the HPV infection rate of 95 .39% (62/65) .Conclusion HPV infection is closely related to the CA pathgenesis in vulva ,vagina and cervix . HPV6 and HPV 11 are main stream genotypes ,in which vulval CA is most common .The gene‐chips combined with PCR technology is a method suitable for HPV typing diagnosis ,and has the characteristics of good sensitivity and high specificity ,which has an im‐portant significance for clinical diagnosis ,treatment and vaccine study of CA in femal vulva ,vagina and cervix .

Objective To compare the sensitivity of fluorescent quantitation polymerase chain reaction (fluorescent quantitation method) and gene‐chips typing method(gene‐chips method) in the detection of human papillomavirus(HPV) ,and to analyse differ‐ences and clinical significance .Methods A total of 246 women were selected as subjects ,among them ,111 cases of cervical exfolia‐ted cells and 135 cases of cervical tissues were collected and detected .15 kinds of high‐risk HPV genetypes were detected in all sub‐jects by using fluorescent quantitation method and gene‐chips method respectively ,and the detection results were compared . Results The sensitivity of the fluorescent quantitation method in detecting HPV was 55 .28% and that of the gene‐chips method was 55 .69% ,there was no statistically significant difference in sensitivity between the two methods (P>0 .05) .The two methods had relative high conformance(κ=0 .745) .The positive rate of HPV infection was increased with the progression of cervical dis‐ease .Conclusion The fluorescent quantitation method and the gene‐chips method have a relative high conformance ,and both with high sensitivity in detecting HPV .The severity degree of cervical cytological and histological changes may be positively correlated with HPV infection .

Purpose To compare the distribution of 23 kinds of human papillomavirus ( HPV) genotypes in tissues of condyloma acu-minata ( CA) of cervix in 120 women and its clinical significance. Methods Polymerase chain reaction ( PCR) and gene-chips tech-nology were utilized for the detection of 23 kinds of HPV genotypes in tissue specimens from 120 cases of CA in cervix and related ma-terials of all subjects were conducted and analyzed. Results There were 115 positive cases in 120 women with CA in cervix and the rate of total HPV infection was 95. 83% (115/120). The rate of single type was 70. 83% (85/120) and multiple types was 25. 00%(30/120). The predominant type of single infection was HPV11 and the infective rate was 45. 00% (54/120), followed by HPV6 (22. 50%, 27/120). Otherwise, the predominant type of multiple infections was HPV6+11 with the infective rate of 20. 00% (6/30), and HPV11+16 infection accounted for 10. 00% (3/30). Conclusions HPV11, 6, 6+11 and 11+16 are the main genotypes in the pathogenesis of CA in cervix in 120 women. PCR and gene-chip technology can detect single and multiple HPV genotyping in tis-sues of CA in cervix with high sensitivity and specificity. Detection of HPV genotypes could be used to understand the prevalence situa-tion of HPV infection in tissues of CA and tumors of cervix and further to provide references for the research and development of HPV vaccine in women.

Objective To investigate the distribution situation of human papillomavirus (HPV) genotypes profile in cervical cells among women in Xuzhou area and its clinical significance .Methods 23 kinds of HPV DNA were extracted in cervical cell samples from 8 010 women in Xiuzhou area .The gene‐chips technique of PCR combined with reverse dot blot was adopted to detect the HPV genotypes .Results Among 8010 cervical cell samples ,there were 1 852 HPV infected cases ,the total HPV infection rate was 23 .12% ,the HPV infection rates of single type accounted for 17 .17% and its predominant types were 16 type (4 .35% ) ,followed by 58 type (2 .12% ) and 52 type (1 .82% ) ,The detection rate of multiple HPV infection was 5 .96% ,in which the predominant types were HPV16+58(4 .40% ) ,16+52(2 .94% ) ,11+16(2 .52% ) .Conclusion The single HPV infection of HPV16 ,58 ,52 and the multiple HPV infection of HPV16+58 ,16+52 ,11+16 are the main genotypes of cervical cells among women in Xuzhou area , this gene chip technique is suitable for the cervical cell sample ,its once detection can detect 23 kinds of HPV genotypes with high specificity and high sensitivity ,which has an important significance for the molecular epidemiologic survey study of HPV genotypes distribution among women in our country .

Objective To explore the clinical distribution states of human papillomavirus genotypes in tissues of 691 women with vulva condyloma acuminates in Nanjing city and Zhenjiang city in Jiangsu Province and genotyping clinical significance.Methods Polymerase chain reaction(PCR)and gene-chips technology were utilized for the detection of 23 kinds of HPV genotypes in tissue specimens from 619 women of vulva condyloma acuminates in Nanjing city and Zhenjiang city in Jiangsu Province.And related materials of all subjects were analyzed.Results In 691 women of vulva condyloma acuminates,597 women of HPV infecton,total infection rate of HPV was 86.40％(597/691),including single genotype infection rate of HPV was 51.38％(355/691),11、6 and 16 genotypes are the most common in single genotypes,they are successively 51.55％(183/355)、41.97％(149/355)and 3.38％(12/355).multiple genotypes infection rate of HPV was 35.02％(242/691),6+11、11+18、6+16 and 11+16 genotypes are the most common in multiple genotypes,they are successively 9.92％(24/242)、9.09％(22/242)、4.96％(12/242)and 4.13％(10/242).Conclusion The low-risk HPV types are the main factors to cause the female vulva CA,a few high-risk HPV types may cause warts as well in tissues of women with vulva condyloma acuminates in Nanjing city and Zhenjiang city in Jiangsu Province.The vulva examine of HPV types should be held to the vulva CA patients.This precaution will has extremely important meaning to the prevention and treatment of the female vulva CA and cervical lesion in our nation.

Objective To investigate the distribution of 39 kinds of human papillomavirus(HPV)infection genotypes in female cervical cells and its clinical significance.Methods 39 types of HPV DNA were extracted from 434 samples of female cervical cells. The gene amplification combined with the gene chip technique was adopted to detect 39 kinds of HPV genotype.And the clinical da-ta of the patients were analyzed.Results Among 434 samples of female cervical cell,175 cases were HPV positive,the total HPV infection rate was 40.32%(175/434).Among them,105 cases were the single type HPV infection with the positive detection rate of 24.19%(105/434)and 70 cases were the multiple types HPV infection with the positive detection rate of 16.13%(70/434).Among single type HPV infection,31 cases were the HPV18 infection with the positive detection rate of 17.71%(31/175),which was the main HPV infection type;followed by HPV16 in 12 cases with the positive detection rate of 6.86%(12/175)and HPV52 in 11 cases with the positive detection rate of 6.29%(11/175).Among the multi-type HPV infection,each 2 cases were HPV 6+54,HPV 18+52,HPV 51+68 infection respectively,each accounted for 2.86% of the multi-type HPV infection,which were the main infection types.Conclusion HPV 16,18,52 and HPV 6+54,HPV 18 +52 and HPV 51 +68 are the main HPV infection genotypes of fe-male cervical cells.The gene amplification combined with the gene chips technique is a method suitable for clinically conducting the HPV genotyping diagnosis and the molecular epidemiologic research of HPV infection.Along with the increase of detected HPV genotypes,the HPV infection rate is also increased,its genotypes combinations trend towards diversification.

Objective To compare the distribution situation of human papillomavirus(HPV)infective genotypes in normal cells and atypical squamous cells of undetermined significance(ASC-US)in uterine cervix and its clinical significance.Methods The pol-ymerase chain reaction(PCR)combined with the gene-chips technology were adopted to detect 23 kinds of HPV genotype from 1 000 cases of normal cells specimens and 229 cases of ASC-US specimens.Results 106 cases of HPV-positive infection were de-tected from 1 000 cases of normal cells with the total HPV infection rate of 10.60%(106/1 000),in which the single genotype in-fection rate was 9.30%(93/1 000)and the multiple genotypes infection rate was 1.30%(13/1 000);116 cases of HPV-positive in-fection were detected from 229 cases of cervial ASC-US specimens with the total HPV positive rate was 50.66% (116/229 ),in which the single genotype infection rate was 34.06%(78/229)and the multiple genotypes infection rate was 16.59%(38/229).The total HPV positive rates,single and multiple genotype infection had statistically significantly differences between the two groups(P<0.05).Conclusion The HPV types 16,18,33,42,43,52,58 are the predominant genotypes in normal cervical cells and ASC-US. PCR combined with the gene-chip technology can be used in the HPV genotype detect in cervical cells,conduces to perform the fur-ther distribution management on ASC-US and has the important significance to prevention and control of cervical cancer.