Objective To observe the clinical outcome of maintenance hemodialysis patients with hepatitis C viruses(HCV)infection.Methods A retrospective investigation about hepatitis C viruses infection was performed in our hemodialysis centre on December 31,2007.The clinical data including demography,serum transaminase,bilirubin,albumin,anti-HCV antibody,HCV RNA,infection-control procedures associated hemodialysis and ultrasound of the liver was analyzed.Results The prevalent rate of HCV infection in our hemodialysis centre was 20.2%(20/99),The mean age of maintenance hemodialysis patients with HCV infection was(63.3?10.9)years old,the duration on dialysis was(79.9?38.7)months,and the duration of positive serum anti-HCV was(32.6?22.6)months(the longest duration was 103.5 months).16 patients(80%)had blood transfusion history in the patients with HCV infection.Only four cases had viremia of HCV,the serum HCV RNA became negative in one of them spontaneously and in another one after interferon therapy for half a year,the other two of them have viremia now.Most of patients found their serum transaminase elevated transiently and mildly.The age and duration on dialysis of the patients with positive serum anti-HCV were significantly longer than those of patients with negative serum anti-HCV.After strict infection-control procedures were carried out,the new onset cases of HCV infection was significantly decreased(1.2% vs 5.3%,P

Objective:To explore the feasibility of the optimized cytokinesis-block (CB) assay on radiation-induced nucleoplasmic bridge (NPB), and to provide a scientific basis for the application of NPB in biological dose estimation.Methods:Human peripheral blood in vitro was irradiated with 2 Gy 60Co γ-rays at a dose rate of 1 Gy/min (0 Gy control group). According to the culture time after irradiation, blood samples were divided into group 48, 56, 68 and 72 h. Cytochalasin-B (Cyt-B) with a concentration of 6 μg/ml was added into the samples at 28 h and harvested at 48, 56, 68 and 72 h after irradiation, respectively. On the other hand, the blood samples were treated with different concentration of Cyt-B i. e., 0.6, 1, 2, 6 and 10 μg/ml at the beginning of culture (0 h) and harvested at 68 h after irradiation. The proportion of mononucleated, binucleated and multinucleated cells, radiation-induced NPB and micronucleus (MN) frequencies were analyzed. Results:The nuclear division index (NDI) and proportion of binucleated cells at 2 Gy and 0 Gy had tendency of increasing with cell culture time. NPB frequencies (0.023 0-0.033 0/cell) and MN frequencies had no significantly difference ( P> 0.05). With the increase of Cyt-B concentration, NDI and the proportion of binucleated cells in group 2 Gy and 0 Gy also increased, but NPB frequencies (0.023 0-0.047 0/cell) had no significant difference ( P> 0.05). MN frequencies of group 10 μg/ml were significantly lower than that of group 6 μg/ml ( U=2.74, P< 0.01). Conclusions:Cell culture time and Cyt-B concentration had no significant influence on radiation-induced NPB frequencies, suggesting that NPB could be obtained by appropriately reducing cell culture time and Cyt-B could be added into blood samples at the beginning of culture. But this protocol reduced the number of cells for further analysis, and thus its feasibility for dose estimation still need to be studied.

OBJECTIVETo explore the diagnostic value of serum levels of BamHI-W fragment, latent membrane protein-1 (LMP-1), BZLF1 and ZEBRA protein in patients with natural killer (NK)/T-cell lymphomas (NKTCLs), and to evaluate their relationship with clinical features.

METHODSA total of 144 cases were analyzed in this study, including 48 NKTCLs patients, 48 other types of non-Hodgkin's lymphomas (NHL) patients and 48 healthy individuals as controls. Fluorescent quantitative real-time polymerase chain reaction (RQ-PCR) was used to measure the copy number of BamHI-W, LMP-1 and BZLF1 in serum. Enzyme linked immunosorbent assay (ELISA) was applied to measure the serum levels of ZEBRA protein. The relative operating characteristic (ROC) curve was applied in the evaluation of the tested markers in diagnosis of NKTCL patients, and the correlations among the tested markers and clinical feature were analyzed.

RESULTSCompared with the controls, NKTCL group showed significantly higher levels of all the tested markers (P < 0.01). The median values of serum BamHI-W, LMP-1 and BZLF1 DNAs level were 1870, 394 and 499 copies/ml, respectively. And the median value of ZEBRA protein level was 73.3 µg/L. Furthermore, the ROC curves analysis revealed that all the area under curve (AUC) of LMP-1, BZLF1 and ZEBRA were more than 0.70, which were probably helpful in the diagnosis of NKTCL. To predict the presence of NKTCL, BamHI-W showed a high sensitivity of 81.3%, while BZLF1 showed a high specificity of 81.2%. Untreated patients seemed to have a significantly higher level of serum LMP1 DNA than that of treated patients (median value 898 copies/ml vs 0 copies/ml, P = 0.050). Correlation analysis showed that serum BamHI-W DNA level was correlated with the presence of B symptoms. All the three genes expressed in 94.4% of the untreated cases. On the other hand, none of them expressed in treated cases.

CONCLUSIONSIt suggested that combined measurements of BamHI W, LMP1 and BZLF1 DNA levels might be helpful to the diagnosis and therapeutic monitor of NKTCL.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Deoxyribonuclease BamHI ; blood ; Female ; Herpesvirus 4, Human ; Humans ; Lymphoma, T-Cell ; blood ; diagnosis ; Male ; Middle Aged ; Real-Time Polymerase Chain Reaction ; Sensitivity and Specificity ; Trans-Activators ; blood ; Viral Matrix Proteins ; blood ; Young Adult

SUMMARY Objective:To investigate the influence of cryoprotectants and cooling rates in vitrificationmethod on the spindles of rabbit M Ⅱ oocytes.Methods:Rabbit oocytes were verified by using cryoloopwith ethylene glycol(EG)singly or EG combined with dimethyl sulphoxide(DMSO)as cryoprotectants,and cooled by taking oocytes directly into liquid nitrogen or by vitrification machine.After frozen rabbit o-ocytes thawed,the microtubulin and chromosome of the spindles were fixation and stained by immunofluo-rescent method.Confocal microscope was used to reveal spindle configuration.Results:In the two proto-cols of single EG used and EG combined with DMSO,the spindles were severely injured.But in protocolof EG combined with DMSO and at ultra-rapid cooling rate,the normal configuration of spindle rate ofthawed rabbit oobytes was similar to that of the control group.Conclusion:The protocol of EG combinedwith DMSO as cryoprotectants and with extremely high cooling rate by vitrification machine can producethe best effect on conservation of spindle configuration in vitrification of rabbit oocytes.

Objective: To investigate the inhibitory effect of adenovirus-mediated VEGF-siRNA on transplanted osteo- sarcoma in nude mice.Methods: VEGF-siRNA gene was cloned into the genome of replication-deficient adenovirus to construct Ad-VEGF-siRNA;the latter was then used to infect osteosarcoma MG63 cell line in vitro;and the expression of VEGF gene was detected by RT-PCR.Osteosarcoma transplantation model was established in nude mice;VEGF expression in tumor tissue was analyzed and the inhibitory effect on tumor growth and lung metastasis were also observed.Results: The recombinant adenovirus vector Ad-VEGF-siRNA was successfully constructed.In vivo and in vitro experiment both showed that Ad-VEGF-siRNA significantly downregulated VEGF expression in MG63 cells and transplanted tumor tissue. It was found that Ad-VEGF-siRNA significantly inhibited transplanted osteosarcoma growth(P

Objective:To screen radiosensitive lipid metabolites in rat small intestine and analyze their metabolic pathways, in order to provide scientific basis for radiation enteropathy biomarkers.Methods:The total body irradiation of 60Co γ rays was performed to rats with different doses of 0, 1, 2, 3, 5 and 8 Gy. The changes of lipids in small intestine were studied by targeted lipidomics method based on liquid chromatography coupled mass spectrometry (LC-MS). Results:Fifteen lipids in small intestine were screened as radiosensitive metabolites at 3 d after irradiation, including 4 up-regulated lipids and 11 down-regulated lipids( t=-6.395, 5.998, 5.836, -5.503, -5.449, -5.422, 4.841, 4.802, 4.621, 4.457, 4.426, 4.373, 4.110, 3.945, 3.902, P< 0.05 and FDR < 0.05). The metabolic pathways of sphingolipid, glycerophosphoplipid were significantly enriched. Four phosphatidyl serines (PS)increased while 1 phosphatidic acid(PA), 2 sphingomyelins(SM) and 4 fatty acids(FA)decreased in a good dose-response manner( R2> 0.80, P< 0.05), which were more potential radiation enteropathy biomarkers. Conclusions:Lipid metabolites in rat small intestine were significantly changed after the rat was total body irradiated with 60Co γ-rays.Eleven lipids with good dose-response relationship were more potential to be radiation enteropathy biomarkers.

Objective To compare the application and clinical significance of the liquid based cytology examination and the DNA quantitative analysis in female cervical lesions.Methods The cervical cell samples were collected from 879 women participating in the comparison by the cervical brush and performed the the liquid-based thin layer section preparation for conducting Papanicolaou staining and DNA staining respectively.The liquid based cytology examination was performed on the Papanicolaou staining section and the fully automatic scanning diagnosis was performed on the DNA staining section.Results The cases of above atypical squa-mous cells of undetermined significance(ASCUS)detected by the liquid based cytology examination and the partial cases of hetero-ploid cell detected by the fully automated DNA ploidy analysis system were recommended to further perform colposcopy and cervi-cal biopsy.28 women were performed the pathological biopsy.With the cytological examination result as the standard,the detection rate of above ASCUS cervical lesions detected by the cellular DNA quantitative analysis was calculated.Conclusion The combined application of the cellular DNA quantitative analysis method and the liquid based cytology examination can obviously increase the positive detection rate of cervical cancer and precancerous lesion,which has important significance for the prevention and treatment of female cervical cancer in our country.

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 study the genotypes of human papillomavirus (HPV)infection in female anus and anal canal condylo-ma acuminata(CA)tissues and their clinical significance.Methods 23 kinds of HPV-DNA were extracted from the paraffin-embed-ded anus and anal canal tissue samples in 140 cases of female CA and detected by using PCR combined with the gene-chips tech-nique.Furthermore the related clinical pathological data of the patients were analyzed.Results Among 140 female anus and anal ca-nal CA tissue samples,103 cases were HPV positive and the total HPV infection rate was 73.57%(103/140).Among them,68 ca-ses were single type HPV infection,the positive detection rate was 48.57%(68/140)and 35 cases were multiple types HPV infec-tion,the positive detection rate was 25.00% (35/140).In single type HPV infection,34 cases were HPV11 and the positive detec-tion rate was 24.29% (34/140),HPV11 was the main infection type,followed by HPV 6 in 27 cases,its positive detection rate was 19.29%(27/140).In the multiple types HPV infection,13 cases were HPV 6 + 11,accounting for 37.14% (13/35 )of multiple types infection,followed by HPV11 +18 in 3 cases and HPV 6+11+16 in 3 cases,each accounting for 8.57%(3/35)of the multi-ple types infection.Conclusion HPV 6,11 ,6+11,11 +18 and 6+11+16 are the main infection genotypes in female anus and anal canal CA.PCR combined with the gene-chips technique is a diagnostic method more suitable for clinical development of HPV geno-typing detection,which has high sensitivity and good specificity and is especially suitable for the molecular epidemiology study of 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.