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 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 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.

BACKGROUND: Previous results of our study show that oyster extract has some protective effects on apoptosis of the neuroepithelium in neural tube defects induced by hyperthermia in vivo.It remains unclear whether the extraet also protects in vitro cultured neural stem cells.OBJECTIVE: To investigate the protective effect of oyster extract on apoptosis of cerebral neural stem cells induced by hyperthermia.METHODS: The cerebral neural stem cells of embryonic mice of 13 days were cultured in vitro.Nestin expression was detected by immunofluorescence method to identify neural stem cells.The neural stem cells of passage 3 were divided randomly into 4groups: hyperthermia control group and oyster treated Ⅰ,Ⅱ,Ⅲ groups(mass concentration 2.5,5,10 g/L oyster extract solution).In addition,culture solution control group(no cells),and culture solution+oyster extract control group(no cells)were designed.All oyster extract groups and control groups were treated by hyperthermia over 39 ℃.The survival rate and the vitality of neural stem cells were detected by trypan blue staining and MTT assay.Western-blotting was employed to explore the expression of p53 in cerebral neural stem cells of each group.RESULTS AND CONCLUSION: The survival rate and the value of MTT assay in oyster treated groups Ⅱ and Ⅲ were significantly greater than hyperthermia control group(P < 0.05),but the expression of p53 in oyster treated groups Ⅱ and Ⅲ were weaker than hyperthermia control group.Oyster extract plays an important protective role in the apoptosis of neural stem cells induced by hyperthermia.

Objective To investigate the clinical features and the management of primary pigmented nodular adrenocortical disease PPNAD) and to evaluate its relationship with Carney complex. Methods One case of PPNAD reported. The patient was a 52 years oldmale. He was hospitalized because of hypertension for one year. The patient had a Cushing's face with elevated plasma and urine cortisone levels which could not be suppressed by both low dose and high dose dexamethasone tests. Ultrasonography howed normal bilateral adrenal glands. CT scan found a 1.6 cm × 2.0 cm mass in the left adrenal gland and normal on tralateral adrenal gland. Results The patient had accepted left laparoscopic adrenalectomy. The pathological examination onfirmed the diagnosis of PPNAD. Micro scopic study showed that there were black-gray spots in the center of the specimen. Hyperplasia was found in all the three adrenal zones. Lipofuscin was observed in the cytoplasm of reticular zone cells. The patient's blood pressure had returned to normal level after the surgery. Conclusions PPNAD is a rare type of ypercortisolism. As there is no specific feature in clinical manifestation and radiological examination of this disease, it is very easy to make a misdiagnosis in clinical practice. PPNAD itself can be the comorbidity of Carney complex, careful differentiation is needed.

Objective To investigate the correlation between chronic hepatic diseases and small intestinal inflammation.Methods Patients who received capsule endoscopy in The First Affiliated Hospital of Guangdong Pharmaceutical University were divided into groups of liver cirrhosis,non-alcoholic fatty liver disease(NAFLD),chronic hepatitis and non-hepatic disease according to clinic data from August 2011 to August 2015.The severity of small intestinal mucosal inflammation was graded according to Lewis Scoring system and incidence of small intestinal lesions in different groups and Lewis scores were compared.The liver function was also graded with liver noninvasive scoring systems.Then the correlation between liver function damage and small intestinal lesions was investigated.Results A total of 338 cases were enrolled in the study,including 25 cases of liver cirrhosis,47 cases of NAFLD,20 cases of chronic hepaitis and 246 cases of non-hepatic disease.There were 22 (88.0％),36 (76.6％),12 (60.0％) and 78 (31.7％) cases with lesions in small intestine in the four group respectively with significant differences(P＜0.001).Rate of small intestinal villi edema was significantly higher in liver cirrhosis group,NAFLD group,chronic hepatitis group than that in non-hepatic disease group(all P＜0.017).Small intestinal villi edema was found mainly in the upper and one third of middle parts in small intestine (P =0.033).Lewis scores of liver cirrhosis group (190.80±228.42)and NAFLD group(125.38± 191.31) were higher than those of non-hepatic disease group (42.91±97.69,P=0.021,P =0.034).Forns score,FIB-4 score,NAFLD-FS score and Child-Pugh score were positively correlated with Lewis score (correlation coefficient:0.247,0.244,0.223,0.284respectively,all P＜0.001).Conclusion Chronic hepatic diseases such as liver cirrhosis,NAFLD,chronic hepatitis might be risk factors for small intestinal mucosal inflammation,and the severity of chronic hepatic diseases may be positively correlated with that of small intestinal mucosal lesions.