Objective To establish a method to detect viral integrity of human papillomavirus in women cervical HPV infection. Methods We amplified E6/E7 gene and E2 gene of HPV16,then inserted them into a plasmid containing single copy HBB gene. HPV16 infected cervical epithelium samples were screened out by genotyping with RDB of flow-through hybridization assay.Fluo-rescence quantitive PCR data of HBB,viral E2 gene and viral E6 gene of all samples were standardized by compared with respective parameters of the plasmid.The ratio IHPV and CHPV were calculated to find out E2 gene disruption and viral copies per cell in the cer-vical samples,respectively.Results The plasmid constructed for standardization was proved effective to make the FQ-PCR data of E2 gene,E6 gene and HBB gene comparable.Thirty-seven HPV16 positive cervical epithelium samples included 22 cases from women whose TCT were normal,and 15 cases from women who confirmed HIL/CIN 2-3 or above through colposcopic examina-tion plus biopsy.Fifteen samples were detected E2 gene disruption,including 10 HIL/CIN 2-3 or above samples and 5 TCT normal samples.E2 gene integrity in different groups were statistically significant different(P <0.05).The average viral copies per cell dis-played a significant decline along with E2 gene disruption(P <0.05).Conclusion The tandem single copy gene plasmid standard-ized methord for the detection of E2 gene disruption caused by viral integration in HPV16 infected cervical cells is feasible and effec-tive.

Objective To investigate the distribution of human papillomavirus (HPV) type 16 in women cervical infection in eastern Guangzhou, polymorphism of E6/E7 gene and association of gene dosage with disease progression. Methods Flow-through hybridization and gene chips were applied in HPV sub-type identification to screen out HPV-16 positive samples from cervical epithelium samples. HPV-16 E6/E7 gene was amplified through PCR with specific primers. The PCR products were cloned into pMD18-T vectors and fragments were determined through sequencing. Polymorphism analysis were performed through align-ment tools. Fluorescence quantitive PCR were used for the detection of viral E6 gene and L1 gene. Results Thirty-six (4.5%) HPV-16 positive samples were screened out through flow-through hybridization from 806 cervical epithelium samples. HSIL and above happened in 18 (50.0%) of the 36 HPV-16 positive patients. Within E6/E7 gene sequences from 7 selected samples, we found 15 sites with variances and 8 of them would cause coding amino acid change. HIL group (A, 11 cases) and LSIL group (B, 14 cases) possess significantly different gene dosage of both viral E6 gene and LI gene (P <0.05). The ratios of L1/E6 be-tween the 2 groups was not significantly different(P=0.19). Conclusion HPV-16 cervical infection oc-curs in 4.5% women (17-62 years old) in eastern Guangzhou. HIL or above accompany with half of the HPV 16 infected women. Viral load is probably associated with cervical HSIL, though L1/E6 ratios do not suggest viral integration.

Purpose To study the effect of China cobra venom active factor(CCVAF) from China cobra venom on endothelial cells and its mechanism.Methods MTT experiment was adopted to evaluate the effect of CCVAF on bovine arteria pulmonalis vascular endothelial cells(BAVEC).The Eosin-Coomassie brillient blue and rhodamine-phalloidin method was used for actin cytoskeleton.Flow cytometry for [Ca~(2+)]_i and spectrophotometry were used for lactate dehydrogenase(LDH) and nitrogen oxide(NO) levels in cell culture supernatant.Results CCVAF(0.625-20 μg/mL) inhibited the proliferation of BAVEC in dose-dependent manner,and IC50 of CCVAF on BAVEC was 2.45 μg/mL. After CCVAF and BAVEC coincubation, it was showed that regression of intercellular conjunctions and disorder of F-actin distribution occurred. The content of [Ca~(2+)]_i, [LDH] and [NO] increased respectively.Conclusion CCVAF can inhibit BAVEC proliferation and it maybe associated with the change of cytoskeleton and increasing of [Ca~(2+)]_i,[LDH] aod [NO].

Objective To evaluate the effect of comprehensive schistosomiasis control measures based on infection source control in Hanchuan City marshland and lake endemic regions . Methods The data of comprehensive schistosomiasis control in Hanchuan City from 2004 to 2013 were collected and the change trends of the Oncomelania hupensis snail status and the in?fection situation of human and livestock were analyzed to evaluate the control effect. Results After the implementation of the comprehensive measures the infection rates of residents and cattle decreased from 6.38%and 8.11%in 2004 to 0.16%and 0 in 2013 respectively. There were no acute schistosomiasis patients since 2007 no new infection cases since 2011 and no infected snails since 2012. Compared with 2004 the occurrence rate of frames with living snails and the average density of living snails decreased by 56.78%and 68.35% respectively but the snail area and susceptible area increased by 0.62%and 7.10% respec?tively. In 2013 all the 367 endemic villages in 26 townships reached the criteria of transmission controlled. Conclusions The comprehensive schistosomiasis control measures based on infection source control can control the schistosomiasis transmission effectively in marshland and lake endemic regions. When consolidating the achievement the snail area in inner embankment should be compressed to prevent the schistosome re?infection in human and livestock.

Objective:To observe the efficacy of gemcitabine and oxaliplatin combined with targeted drug apatinib in the treatment of gallbladder cancer.Methods:The data of 82 patients with gallbladder cancer who were admitted to Haiyang People's Hospital, Yantaishan Hospital and Rongjun Hospital of Yantai City in Shandong Province from May 2017 to December 2018 were retrospectively analyzed. The patients were divided into control group and observation group according to the medication, with 41 cases in each group. Patients in the control group were treated with gemcitabine combined with oxaliplatin regimen (gemcitabine 1 000 mg/m 2 was injected intravenously for 30 minutes on the 1st and 8th day, and oxaliplatin 100 mg/m 2 was injected intravenously for 3 hours on the 1st day), the observation group was combined with apatinib mesylate 500 mg/d based on the treatment method of the control group. The treatment effect, serum tumor marker levels, adverse reactions and survival were compared between the two groups. Results:The disease control rate of the control group was 70.73% (29/41), which was significantly lower than that of the observation group [92.68% (38/41)] ( χ2 = 6.609, P < 0.05). After treatment, the levels of vascular endothelial growth factor (VEGF), carcinoembryonic antigen (CEA), soluble interleukin-2 receptor (sIL-2R) and Dickkopf-1 in the observation group were significantly lower than those in the observation group, and the differences were statistically significant ( t values were 6.011, 3.048, 4.444, and 4.514, all P < 0.05). The incidence of proteinuria and hand-foot syndrome in the observation group were significantly lower than those in the control group, and the differences were statistically significant ( χ2 values were 6.116 and 4.986, both P < 0.05). During the 1-year follow-up, 2 patients in the observation group died, and the survival rate was 95.13% (39/41); 8 patients in the control group died, and the survival rate was 80.49% (33/41). There was a statistical difference in the survival rate between the two groups ( χ2 = 4.100, P < 0.05). Conclusion:Gemcitabine and oxaliplatin combined with apatinib mesylate are effective in treating gallbladder cancer, which can effectively reduce serum tumor marker levels and improve the prognosis of gallbladder cancer patients.

Objective:To investigate the value of real-time shear wave elastography (SWE) in the assessment of splenic congestion in patients with chronic right heart failure.Methods:Sixty patients with chronic right heart failure with cardiac function grade Ⅱ-Ⅳ of New York Heart Association (NYHA) treated in Lianyungang First People′s Hospital from March 2020 to February 2021 were collected as the study group, and 20 healthy subjects in the same period were selected as the control group. Routine echocardiography was performed on all subjects; spleen stiffness measurement (SSM) was detected by SWE, and blood biochemical indicators related to patients with right heart failure were detected and recorded. SSM and other related parameters between the two groups were analyzed; the SSM in patients of different cardiac function classifications and course of disease were compared; 60 patients were divided into low SSM group (SSM≤15.0 kPa), middle SSM group (15.0 kPa