High-risk human papillomavirus (HPV) is the principal cause of various cancers including cervical cancer, anal cancer, vulvar cancer, and some head and neck cancers. In the viral life cycle, by interacting with both viral and host DNA and proteins, the HPV E2 protein plays a pivotal role in viral transcriptional regulation and DNA replication, and it is also associated with modification of various cellular processes, including host gene transcription, RNA processing, apoptosis, ubiquitination, and intracellular trafficking, to create a convenient environment for a replicative cycle of the virus and contribute to the HPV pathogenesis. Elucidating the roles of E2 protein throughout the viral life cycle will improve our understanding of the viral life cycle and pathogenesis and help us identify novel antiviral agents with therapeutic potential. This article reviews the research progress in the structure, roles, and activity of high-risk HPV E2 protein, particularly that of HPV-16.
Animals ; DNA-Binding Proteins ; genetics ; metabolism ; Female ; Gene Expression Regulation, Viral ; Human papillomavirus 16 ; genetics ; metabolism ; Humans ; Oncogene Proteins, Viral ; genetics ; metabolism ; Papillomavirus Infections ; genetics ; metabolism ; virology ; Uterine Cervical Neoplasms ; genetics ; metabolism ; virology

Objective To clone the gene of autolysin(LytA) which are from different clinical strains of Streptococcus pneumoniae and express them in Escherichia coli. Methods The LytA gene was amplified by PCR from the total DNA of S.pneumoniae. Primers were designed according to the LytA gene sequence of R6. Recombinant plasmids were constructed and the sequences of different clinical strains were analyzed through method of bioinformatics. The cloned genes were expressed in E.coli and detected by SDS-PAGE. Results Complete LytA gene were amplified from all of the different clinical strains of S.pneumoniae and recombinant plasmids pGEX-4T-1-LytA were constructed successfully. After comparing the sequence of DNA and supposed protein, we find some differences. Induced by IPTG, LytA gene was expressed effectively in E.coli Jm109. Result of SDS-PAGE showed that the molecular weight of expressed protein was 62 kD, the same as calculated. Conclusions The sequences encoding LytA from different clinical strains of S.pneumoniae were cloned, the recombinant plasmids pGEX-4T-1-LytA was constructed successfully. Sequence analysis showed that there have difference among the gene and amino acid sequences of LytA from different clinical strains. Further studies should be focused on whether the difference contributes to activity of autolysin and the drug-resistance of S.pneumoniae.

Background Recombinant hirudin variant Ⅲ(rHV3) can effectively prevent galactose-induced human lens epithelial cells LECs injury,but little is known about the molecular mechanism of its action.Objective The present study was to investigate the effects of rHV3 on the expression of apoptosis-related genes in damaged LECs induced by galactose.Methods The rHV3 was extracted by our research group,and the biological activity of rHV3 was identified by titration of thrombase according to Markwardt's method.Human LECs (SRA01/04) were cultured using 125×10-3 mol/L D-galactose+10% FBS+D/F12 medium to establish the damaged human LECs model.rHV3 was added into the medium of the damaged human LECs model.Human LECs were cultured in D/F12 medium containing 10% FBS as normal control.The expression of apoptosis-related genes,such as aldose reductase (AR),bax,bcl2 and p53,in LECs at the mRNA level was detected using RT-PCR.The abundance ratio of target genes was presented with the absorbance (A) of gene mRNA/GAPDH mRNA.Results Compared to the normal control group,the A values of AR mRNA/GAPDH mRNA,bax mRNA/GAPDH mRNA and p53 mRNA/GAPDH mRNA were significantly elevated in model group (t=3.90E-06,t=8.44E-04,t=5.15E-08,P＜0.01).However,in the rHV3-treated group,the A values of AR mRNA/GAPDH mRNA,bax mRNA/GAPDH mRNA and p53 mRNA/GAPDH mRNA were lower than those of model group (t=5.90E-06,t=1.51E-04,t=3.42E-06,P＜0.01).The bcl2 mRNA/GAPDH mRNA was markedly downregulated in the model group when compared with the normal control group (t=1.86E-05,P＜0.01);while after rHV3 addition,bcl2 mRNA/GAPDH mRNA increased in comparison with the model group (t=8.56E-05,P＜0.01).Conclusion 125×10-3mol/L D-galactose induces the damage and apoptosis of human LECs.rHV3 likely plays a protective function on D-galactose-induced damage of human LECs by inhibiting the polyol pathway and mitochondria-mediated pathway.

AIM:To investigate the cause of blindness, except those caused by cataract, in Haimen city.METHODS:According to the WHO`s criteria of blindness, the blindness level was decided through ophthalmic tests by associate chief or chief ophthalmologists who were trained especially for disability evaluation.The analysis of the the leading cause were taken too.RESULTS:Totally 3 266 persons were blindness, in which 2 118 were first level blindness, 1 148 persons were second lever blindness, and 1 308 persons were male, 1958 were female.The leading cause of blindness were retina and uveitis diseases (31.58%), genetic diseases(23.47%), cornea disease(14.49%).CONCLUSION:The leading cause of blindness are retina and uveitis diseases, genetic diseases, cornea diseases in Haimen city of Jiangsu province.Early prevention and treatment should be strengthened to reduce the occurrence of blindness.

Objective To observe the expression change of HRG-1 gene between urinary bladder carcinoma and normal tissues,and to investigate the effect of HRG-1-siRNA on the proliferation and apoptosis of human bladder carcinoma cells.Methods Immunohistochemisty was used to detect the expression of HRG-1 in 85 cases of bladder carcinomas and 20 normal bladder tissues.The siRNA of HRG-1 was designed,synthesized,and transfected into bladder cancer cell line T24.Results The HRG-1 gene expression had significant differences between bladder carcinoma and normal bladder tissues (P ＜ 0.05).The positive expression of HRG-1 gene had significant differences between the pathological grades and clinical stages of bladder carcinomas (P ＜0.05).After treated with siRNA,the expression levels of HRG-1 protein and mRNA in T24 cells decreased obviously (P ＜ 0.05).The apoptosis rate of T24 cells transfected with HRG-1-siRNA was significantly different from control-siRNA group and blank group (P ＜ 0.01).Conclusions The high expression of HRG-1 gene may play an important role in bladder carcinoma,and siRNA targeting HRG-1 can suppress HRG-1 protein expression markedly and enhance apoptosis of T24 cells.

Objective To explore the expression of a new candidate tumor suppressor N-myc downstream regulated gene 2 (NDRG2) in bladder cancer tissues and to investigate its clinical and pathological significance.Methods Formalin-fixed,paraffin-embedded tissue sections from 62 cases of bladder carcinomas and 10 cases of normal bladder tissues were analyzed retrospectively with immunohistochemistry (S-P method).Results The NDRG2 gene was highly expressed in normal bladder tissues,but low expressed in bladder carcinoma tissues.Positive expression of NDRG2 was detected in 8 of the 10 (80.0％) normal tissues and 40.3％ in bladder carcinoma ones (x2 =3.98,P ＜0.05).Furthermore,with the degree of malignancy increased,the positive expression of NDRG2 in bladder carcinoma samples was decreased.The expression of NDRG2 in bladder carcinoma was negatively correlated(r =-0.288,P ＜0.05) with C-myc(r =-0.436,P ＜0.01) and positively correlated with p53 in bladder carcinoma tissues(r =0.717,P ＜0.01).Conclusions The level of NDRG2 expression was lower in bladder carcinomas than in normal tissues.NDRG2 may play an important role in bladder carcinogenesis and in the progress of bladder cancers.

Objective To investigate the correlation between the mutation in serum lipoprotein-associated phospholipase A2(Lp-PLA2) Ile198Thr, Val279Phe and cerebral infarction in Chinese Han population of Jiangsu Province. Methods One hundred fifty patients with cerebral infarction and 100 healthy controls in Chinese Han population of Jiangsu Province were recruited. The correlation between Val279Phe and Ilel98Thr mutation in Lp-PLA2 gene and cerebral infarction was analyzed using polymerase chain reaction and denaturing high performance liquid chromatography. Results The Val279Phe genotype and the mutant allele frequency in the cerebral infarction group were significantly higher than those in the control group (χ2 were 6. 31and 5. 32, respectively, all P ＜0. 05), and there was no significant difference between the Ile198Thr genotype and the mutant allele frequency in the control group (χ2 were 0. 039 and 0. 037, respectively, all P ＞0. 05). Conclusions The Val279Phe mutation in Lp-PLA2 gene may be a genetic risk factor for cerebral infarction in Chinese Han population of Jiangsu Province.

Objective To investigate the efficacy of postoperative concurrent chemoradiotherapy for early-stage cervical adenosquamous carcinoma and adenocarcinoma.Methods A total of 62 patients with cervical adenosquamous carcinoma,149 patients with cervical adenocarcinoma,and 2687 patients with cervical squamous cell carcinoma,all of whom were in stage Ⅰ B-Ⅱ A and were treated from 2006 to 2012,were enrolled,and some of them received postoperative pelvic radiotherapy ± para-aortic extended field radiation ±afterloading radiotherapy.The chemotherapy regimen consisting of DDP,TP,and FP was given to these patients.The chi-square test was used for comparison of general clinical data,the Kaplan-Meier method was used for calculating survival rates,and the log-rank test was used for survival difference analysis.Results Cervical adenosquamous carcinoma and adenocarcinoma had no significant differences in clinicopathological features (P=0.107-0.639).The high-risk patients with adenocarcinoma had a higher recurrence rate than their low-risk counterparts even after adjuvant radiotherapy or chemoradiotherapy (P=0.000).In the patients treated with surgery and radiotherapy,those with adenosquamous carcinoma had the shortest median survival time,followed by those with adenocarcinoma and squamous cell carcinoma (P =0.134,0.787);in the patients treated with surgery and concurrent chemoradiotherapy,those with adenocarcinoma had the shortest median survival time,followed by those with adenosquamous carcinoma and squamous cell carcinoma (P=0.131,0.643),and the median survival time showed a significant difference between the patients with adenocarcinoma and those with squamous cell carcinoma (P =0.000).In the patients with adenosquamous carcinoma and adenocarcinoma,the patients treated with postoperative concurrent chemoradiotherapy had higher incidence rates of short-term adverse reactions than those treated with postoperative radiotherapy (P=0.037,0.003),but the incidence rates of long-term adverse reactions showed no difference between the two groups of patients (P=0.861,0.655).In the patients with adenosquamous carcinoma,the patients treated with postoperative concurrent chemoradiotherapy had a lower rate of distant metastasis (P =0.003) and prolonged median overall survival and disease-free survival (both increased by 17 months) (P=0.811,0.799),as compared with those treated with postoperative radiotherapy,while in the patients with adenocarcinoma,the median overall survival and disease-free survival were reduced by 11 and 9 months,respectively (P=0.330,0.115).Conclusions Compared with postoperative radiotherapy,postoperative concurrent chemoradiotherapy for early-stage high-risk cervical adenosquamous carcinoma can reduce the rate of distance metastasis.Compared with radiotherapy,postoperative concurrent chemoradiotherapy for adenosquamous carcinoma and adenocarcinoma cannot improve survival time.

Objective To evaluate the effectiveness of discharge planning and home care for patients with peritoneal dialysis (PD).Methods A total of 60 PD patients were randomly divided into the intervention group and control group,30 cases in each group.Discharge planning and home care were carried out in the intervention group, while routine nursing and telephone follow -up service after discharge were implemented in the control group.The patientsˊscores of Zung Self -Rating Anxiety Scale(SAS),Self -Rating Depression Scale(SDS),Self management ability of patients undergoing peritoneal dialysis,complication rate and readmission rate were compared between the two groups at the next day admission,discharge,lth month and 3th after discharge.Results The scores of SAS and SDS in the intervention group were significantly lower than that in the control group at 3th month after discharge,the difference was statistically significant(tSDS =5.263,tSAS =3.812,P <0.05).The Self management ability of patients undergoing peritoneal dialysis was significantly higher than that of the control group at lth and 3th month after discharge,the difference was statistically significant (t =6.845,t =7.231,P <0.05).No significant difference in the re -admission rate (16.67%,6.67%)was found between the two groups (χ2 =0.387,P >0.05).The complication rate in the intervention group was significantly lower than in the control group at 3th month after discharge(χ2 =5.124,P <0.05).Conclusion Discharge planning and home care can effectively relieve depression and anxiety in patients with peritoneal dialysis and improve the abilities of daily life.It is worth promoting and applying in clinic.

Objective:To analyze the polymorphism of urease B gene(ureB) from different Hp strains. Methods:The sequences of ureB gene from 14 different Hp strains were amplified by PCR to analyze PCR-RFLP with HaeⅢ digestion.At the same time,the sequence of ureB gene was analyzed with biochemical software to compare the homology of nucleotide and amino acid sequence and to profile the phylogenetic tree. Results: The results of 1.7 kb ureB gene digested with HaeⅢ showed there were 2-5 band types in 14 different Hp strains and formed five distinct RFLP types.The two reference strains had the same RFLP type as 3 clinical isolates while the other clinical isolates and 3 isolates from animal model belong to 4 different RFLP types respectively.The nucleotide sequences and putative amino acide sequences of Hp ureB were compared.The various ureB sequences had high homologies(more than 96.6% and 98% in nucleotide sequences and amino acide sequences respectively) among 14 Hp strains.Particularly,there was the highest homologies(100%) between CCS9801、CCS9806、M3 and M10.Phylogenetic tree analysis showed that two reference strains and other isolates from clinical patients and Hp-infected mice model were located in two different lineage respectively in phylogenetic tree of nucleotide sequences while there were some variance in phylogenetic tree of amino acide sequences. Conclusion: Hp ureB was high conserved and homologous in the gene level as well as in the protein level.