To understand the genetic origin and variation of porcine epidemic diarrhea virus (PEDV) prevailing in Henan province,a total of 22 PEDV positive samples from suckling piglets with severe diarrhea were collected from 16 pig farms and amplified by RT-PCR.ORF3 genes were then cloned into pMD18-T vector and sequenced.The result of sequencing showed that the length of ORF3 genes all were 675 bp which encoded 224 amino acids.The strains in this study had eight amino acid substitutions when compared with CV777 strain.The nucleotide homologies of the 22 strains were 95.9％-100％ and amino acid homologies were 96.4％-100％.By comparing with those of European strain CV777 and vaccine strain truncated CV777,the nucleotide homologies were 97.1％-97.9％ and 94.8％-95.4％,respectively.Based on the phylogenetic relationship of ORF3 genes,the PEDV field strains and PEDV reference strains could be divided into two groups,and all the field strains identified in this study belong to group 2.It suggested that prevalent PEDV strains in this study seem to be closely related to Henan isolates in previous years,domestic strains,Janpanese strains,American strains as well as Korean strains but differ genetically from European strains or vaccine strains used in China.This experiment analyzed the ORF3 gene sequence features in prevailing of Henan region in recent years,which provides a new support of epidemiology study and protein function research of PEDV ORF3 gene.

Objective To investigate the change of aquaporin 2 (AQP2) mRNA and protein levels in renal collecting duct of SD rats after hypoxin caused by rising of the altitude to 4600 m. Methods Forty male SD rats were randomly divided into 4 groups (24 h, 48 h, 72 h and 1 week group), and 10 rats in Xining city were used as control group. All the 40 SD rats were transported to Kekexili Natural Reservation areas (4600 m) in Qinghai province. Rats of four experimental groups were sacrificed and renal tissue samples were harvested at different time point respectively, the control group rats were treated in Xining city (2260 m) as well. The concentration of plasma antidiuretic hormone (ADH) was measured by radioimmunity method. The expression of AQP2 mRNA and proteins was evaluated by real-time fluorescent quantitative-PCR, Western blot and immunofluorescence assay. Results The concentration of plasma ADH was decreased at 24 h and was only 28.5% of that of control group, reaching the lowest concentration at 48 h [(86.94±6.49) μg/L vs (302.5±310.48) μg/L], then it increased gradually and was similar to the control group at 7 d [(306.46±11.14) μg/L vs (302.53±10.48)μg/L, P＞ 0.05]. There were significant differences of the control group with 24 h, 48 h and 72 h groups, respectively[(302.53± 10.48) μg/L vs (142.46±10.57)μg/L, (86.94±6.49)μg/L, (169.65±11.15) μg/L respectively, P＜0.01]. The change of AQP2 gene expression level was consistent with the change of ADH. It was decreased at the begining when exposure to altitude and it reached its lowest level at 48 h. It was then returned to high level similarly to that of the control group at 7 d (0.09±0.01 vs 0.09± 0.008, P＞0.05 ). There were significant differences of the control group with 24 h, 48 h and 72 h group, respectively (0.09±0.008 vs 0.04±0.005, 0.03±0.002, 0.04±0.003 respectively, P＜0.01 ). Conclusions AQP2 expression in the renal collecting duct of SD rats is altered over the period exposed to altitude. It is decreased in the early hypoxia period, and is increased in later period. This change may be related to the intensity of hypoxia, which is mediated by a potential adaptation mechanisms against hypoxia caused by high altitude.

Objective To study the expression of mitochondrial tumor suppressor gene 1 (MTUS1) mR-NA in papillary thyroid carcinoma(PTC).Methods Reverse transcription polymerase chain reaction(RT-PCR) was used to detect MTUS1 mRNA level in 68 cases of human PTC and the adjacent non-cancerous epithelium (NCE) specimens.Results The relative expression value of MTUS1 mRNA in PTC was 0.31 ± 0.09,while it was 0.65 ±0.12 in NCE.The difference had statistical significance between the 2 groups(t =2.39,P ＜0.05).In PTC,mRNA expression of MTUS1 gene was related to the pathological grade,and lymph node metastasis(P ＜ 0.05)while it was not related to patients' age,sex,differentiation degree,tumor size or TNM stage(P ＞ 0.05).Conclusion mRNA expression of MTUS1 gene is lower in PTC than in NCE,which may play an important role in carcinogenesis and progression of PTC.

Objective: To evaluate the relationship between serum mRNA level of heparin binding epidermal growth factor (HB-EGF) and acute coronary syndrome (ACS) occurrence. Methods: Our research included in 2 groups: ACS group,n=50 patients and Control group,n=100 normal subjects. Serum HB-EGF mRNA level was examined by RT-PCR and the relationship between HB-EGF mRNA and ACS occurrence was assessed by Logistic regression analysis. Results: Compared with Control group, the serum HB-EGF mRNA level of ACS group was higher (0.22±0.73) vs (0.46±0.14),P<0.05. With adjusted meaningful factors of hypertension, smoking, TG, TC, LDL-C, HDL-C and BMI by single factor analysis, multi Logistic regression analysis indicated that serum HB-EGF mRNA was related to ACS occurrence (OR=5.813, 95% CI 2.342-14.426,P<0.001) which meant that upon 0.1 grey value of HB-EGF mRNA elevation, the risk of ACS occurrence may increase 4.813 folds accordingly. Conclusion: Serum HB-EGF mRNA level was related to ACS occurrence.

Objective To investigate the prevalence and the related factors of female breast disease in Zhengzhou City, Henan Province, and to provide a targeted prevention guide for female breast disease. Methods A total of 6310 women were enrolled in this study. In addition to breast ultrasound, mammography and pathology assays were performed. Finally, the prevalence and influencing factors for female breast disease were analyzed. Results The prevalence of breast cancer and the total prevalence of breast diseases was 0.06% and 24.94%, respectively. The prevalence of female breast diseases was significantly correlated to age, educational level, occupa-tion, menstruation, reproductive age and a history of abortion. Logistic regression analysis showed that the occupa-tional type had a significant effect on the prevalence of female breast. Conclusion The prevalence of female breast disease is relatively high in Zhengzhou City, and it is affected by many factors. The targeted surveys and breast dis-ease screening should be conducted, and the secondary prevention of female breast disease should be strengthened.

The onset of hepatocelluar carcinoma, one of the serious complications of primary Budd-Chiari syndrome, is associated with poor prognosis.Although so, the diagnosis and treatment of such disease has still not been standardized at recent.In this paper, we overviewed the recent advances on Budd-Chiari syndrome associated with hepatocelluar carcinoma.

Objective To explore the efficacy and safety of percutaneous transhepatic portal vein or transjugular intrahepatic portosystemie shunt (TIPS)to implant the portal vein metallic stent in treatment of cavernous transformation of portal vein (CTPV).Methods Clinical and imaging data of 8 patients with CTPV were retrospectively analyzed who were treated in our hospital.All patients were treated with metallic stent implantation in portal vein including 3 patients by TIPS and 5 by percutaneous transhepatic portal vein.Results All patients were successful in the stent implantation without any occurrence of serious complications such as intra-abdominal hemorrhage and so on.Intraoperative angiography showed blood circulated freely in these stents.1 day-2 weeks later,the patients symptoms of abdominal pain and gastrointestinal bleeding were obviously relieved or disappeared.Follow up 1 month-3 years,1 patient with stent occlusion after one year of operation,the blood flow recovery after stent reimplantation,and the remaining patients,color doppler ultrasound reflected patency of blood flows in their stents.No one suffered from gastrointestinal bleeding or abdominal pain again.Conclusion Implantation of portal vein metallic stent via percutaneous transhepatic portal vein or via TIPS in treatment of cavernous transformation of portal vein is safe and effective.

Objective:To explore the safety and efficacy of Castor branched stent-graft exclusion in the treatment of aortic dissection and aneurysm involving left subclavian artery.Methods:The clinical and imaging data of 88 patients with aortic dissection or aneurysm involving left subclavian artery diagnosed by CTA or DSA in our hospital from December 2017 to December 2019 were collected retrospectively, including 67 aorta dissections, 7 thoracic aortic aneurysms and 14 aorta penetrating ulcer. All patients were treated with branched stent-graft under the guidance of DSA. The postoperative curative effect and complications were observed. The paired t test was used to compare the maximum aortic diameter of the lesion before and 6 months after the operation. Results:Eighty-eight patients were successfully treated with integrated stent, the success rate of operation was 100%, and the disease-related symptoms of all patients were basically or completely relieved. The mortality rate within 1 month after the operation was 2.7% (2/88). The two deaths were complicated with other serious diseases before the operation, and the cause of death was not related the operation. All patients were followed up except 4 patients who lost contact after discharge. During the follow-up, there were 1 case of retrograde type A dissection, 1 case of new aneurysm of aortic arch, 2 cases of in-stent stenosis of left subclavian artery branch, 3 cases of mild stroke, no persistent endoleak and no death or other serious complications. The mean maximum aortic diameter at 6 months after operation [(34±4)mm] was significantly lower than that before operation [(38±6 mm)] ( t=6.63, P<0.05). Conclusion:Castor branched stent-graft is simple, mini-invasive and effective in the treatment of aortic dissection and aneurysms involving the left subclavian artery.

Aim TostudytheeffectsofCuO,ZnOand TiO2 nanoparticles on the viability and metastatic po-tential of EC-9706 and EC-109 esophageal squamous carcinomacelllineinvitro.Methods Characteristics of CuO,ZnO and TiO2 nanoparticles were detected u-sing transmission electron microscope (TEM)and dy-namic light scattering (DLS ).EC-9706 and EC-109 cells were treated with different concentrations of CuO, ZnO and TiO2 (5 ~80 mg · L-1 ).The cell prolifera-tion was analyzed by MTT assay.The cell cycle and apoptotic rates were determined by flow cytometry (FCM).The cell invasion was assayed in Transwell chambers.The expression of Bcl-2 and caspase-3 pro-tein in cells was detected by Western blot method.Re-sults CuO,ZnOandTiO2nanoparticleswerespheri-cal with primary particle size 12,20. 6,12 nm.The particles were agglomerated in water and cell culture medium with negative charge.CuO and ZnO nanoparti-cles induced decreases in EC-9706 and EC-109 cell vi-ability dose-dependently.After exposed to increasing concentrations of CuO and ZnO nanoparticles,the cell cycle analysis revealed a decreasing proportion of cells in G2/Mand S phase,and up-regulation of the cells in G0/G1 phase.Apoptotic cells also increased along with decreased cell invasion upon CuO and ZnO treatment. Nanoparticles treatment after 48 h, the activated caspase-3 expression quantity increased significantly and the Bcl-2 expression quantity decreased obviously (P<0. 05 )compared with control group.TiO2 nanop-articles had no obvious effect on the EC-9706 and EC-109 cell proliferation,cell cycle,apoptosis and inva-sion.Conclusion ComparedwithTiO2,CuOand ZnO nanoparticles can inhibit EC-9706 and EC-109 cell viability and metastatic potential,the mechanism of action involves cell cycle arrest in G0/G1 phase and apoptosis.These findings can help the development of nanoparticles as anti-cancer therapeutics for esophageal cancer.

Objective To investigate the effect of targeted silencing FGF8b gene on the proliferation and apoptosis of ovarian cancer SKOV-3 cells and its possible molecular mechanism.Methods The design and synthesis of small interfering RNA targeting FGF8b (siRNA),was transfected into ovarian cancer cell SKOV-3,using four methyl thiazolyl tetrazolium (MTT) method to detect cell proliferation and apoptosis of TUNEL kit cells.The expression of Bcl-2 and Bax protein by Western blot assay in cells.Results FGF8b siRNA could inhibit the expression of FGF8b protein in ovarian cancer SKOV-3 cells.The experimental group showed a high proportion of TUNEL positive cells(22.33±2.517) (P＜0.01).The down-regulation of FGF8b protein significantly inhibited the proliferation of ovarian cancer SKOV-3 cells,reduced the expression of Bcl-2(0.586±0.025) (P＜0.01),and increased the expression of Bax(0.334±0.019) (P＜0.01).Conclusion Down regulation of FGF8b expression can inhibit the proliferation of ovarian cancer SKOV-3 cells and induce cell apoptosis,which may be related to the expression changes of Bcl-2 and Bax protein,suggesting that FGF8b plays an important role in the development of ovarian cancer.