Objective To analysis the DNA sequence of HEV ORF3 cDNA and express in HepG2 cell .Methods Recombinant vector carrying HEV ORF3 cDNA was constructed and identified by DNA sequencing .Expression of recombinant vector in HepG2 cell was determined by immunofluorescence technique .Results The recombinant vector was constructed and expressed in HepG2 cell successfully .Conclusion The research laid experimental foundation for exploring the biological function of ORF3 protein in the pathogenesis of viral hepatitis E .

Objective To investigate the relationship of carotic intimal-medial thickness (IMT) to high sensitive C-reative protein (hs-CRP) and homocysteine (Hcy) in IGT patients. Methods Seventy patients with IGT were divided into two groups by carotic IMT levels: IMT normal group( IMT≤0.9 mm) and IMT increased group(IMT>0.9 mm). Sixty healthy people were chosen as controls. Blood-lipid, hs-CRP, Hcy, BMI, SBP and DBP were measured in all subjects. Results The Hcy and hs-CRP levels in IGT group were higher than those in controls (P<0.01) ,and also the carotic SBP,DBP,TG and TC levels were higher than those in controls(P<0.05). The Hcy and hs-CRP levels in carotic IMT increased group were higher than those in IMT normal group(P<0.05). The levels of the carotic IMT is positively correlated with hs-CRP(r=0.616, P<0.01), Hcy(r=0.557 ,P<0.01), TC (r=0.351,P<0.05), SBP(r=0.252, P<0.05). Hcy is positively correlated with hs-CRP in IMT increased group (r=0.364, P<0.05). Conclusions Compare with controls, the IMT was higher in IGT patients, and it had the early stage of atherosclerosis (AS). Hcy and hs-CRP levels may predict the early stage of AS in IGT. The high level of Hcy may increase the level of hs-CRP,then lead to the early stage of AS.

Objective To investigate the effect of palato-pharyngo-plastry on patients with coronary heart disease (CHD) plus obstructive sleep apnea hypopnea syndrome (OSAHS). Methods The 113 OSAHS patients with CHD after palato-pharyngo-plastry from January 2001 to December 2006 were enrolled and followed up. The other 125 cases treated with continuous positive airway pressure (CPAP) were as control group. Results The fundmental factors of CHD including cholesterol, triglyeride, fasting blood glucose and body mass index (BM1) showed no statistical differences between two groups (x2 or t= 4.788, 0.479, 0.541, all P>0.05) . Only the oxyhemoglobin saturation during sleep was higher in study group than in control group [(91.2± 2.5)% vs. (87.6±4.2)%, P=0.046]. The angiography showed that 87 cases (77.0%) retained stable, 24 cases (21.2%) became severe in study group. As compared with control group, there were no statistical differences (both P>0.05). 37 cases (32.7%) in study group and 58 cases (46.4%) in control group who suffered from angina became severe with statistical differences between two groups (x2=4.615, P=0.032). The incidences of myocardial infarction and sudden death were lower in study group (3.5% and 0.8%) and in control group (5.6% and 2.4%), showed no statistical, difterences be tween twogroups. x2=0.571 and 0.495, P>0. 05). Conclusions Compared with CPAP, the palato-pharyngo-plastry can improve oxyhemoglobin saturation and reduce angina symptom, but has no better effect on progress of coronary atherosclerosis and acute coronary disease than CPAP.

Objective To evaluate the effectiveness of using low-frequency repetitive transcranial magnetic stimulation (rTMS) along with donepezil in the rehabilitation of aphasia after cerebral infarction. Methods Fortythree aphasic patients with acute cerebral infarction were assigned into a treatment group or a control group.Both groups received standard basic treatment for acute cerebral infarction.From day 7 on,both groups received oral donepezil hydrochloride (5 mg/d) for 12 consecutive weeks.The treatment group received in addition 2 sessions of low frequency rTMS on the language center of the right hemisphere.The first session started at day 7 after onset,and each session consisted of rTMS on 10 consecutive days (once per day).There was a resting interval of 3 d between the two sessions.The rehabilitation of the aphasia was assessed prior to and after each session,and at the end of the entire 12 weeks of treatment using the western aphasia battery ( WAB ) scoring system and the aphasia quotient (AQ). Results There was no significant difference between the two groups after the first session in terms of the WAB or AQ results.However,the difference in improvement became statistically significant after the second session,as well as at the end of the entire 12 weeks.The average change in AQ was significantly greater in the treatment group. Conclusion Donepezil combined with rTMS early in the acute phase of cerebral infarction is helpful for the rehabilitation of aphasia.

Objective To discusses the methods of the case selection and monitoring and diagnosis and treatment to selective non-operative management of closed splenic injuries .Methods The clinical data and results of 98 cases from October 1999 to Octo-ber 2011 with closed splenic injuries managed non-operative treatment were analyzed retrospectively .Results There were 90 cases (91 .8% ) conservative successfully ,5 cases(5 .1% ) transit operation because hematoma increase and bleeding continuously during therapeutic observation and 2 cases(3 .1% ) died from multiple injuries merge infection and multiple organ failure .Conclusion Most of the closed splenic injuries below level Ⅱcan be treated by non-operative management ,the effect and prognosis determined by the reasonable case selection and monitoring conditions and diagnosis and treatment .

Objective To measure the normal sagittal and coronal diameters of thumb and great toe distal phalanx bottom by ultrasonography. Methods One hundred and twenty volunteers' sagittal and coronal diameters of thumb and great toe distal phalanx bottom were measured by ultrasonography. The measurements was analysed by the statistical method. Results The thumb and great toe distal phalanx bottom showed hyperechoic zone,clear boundary with the adjacent fascia and tendon tissue showed low echo area at cross section by high frequency ultrasound. The sagittal diameters of thumb distal phalanx bottom was (8. 07 ± 0. 67)mm in men, while that of great toe distal phalanx bottom was (8. 34 ± 1. 02) mm( t = 1.73, P =0.86).The coronal diameters of thumb distal phalanx bottom was (11.61 ±0.89)mm in men, while that of great toe distal phalanx bottom was (14. 25 ± 0. 84)mm( t = 16. 77, P = 0. 00). The sagittal diameters of thumb distal phalanx bottom was (7. 52 ± 0. 62) mm in women, while that of great toe distal phalanx bottom was (7. 72 ± 0. 67) mm( t = 1. 72, P =0. 14). The coronal diameters of thumb distal phalanx bottom was (10.94 ± 0.97) mm in women, while that of great toe distal phalanx bottom was (13. 51 ±0. 75) mm( t =16.21, P = 0.00). Conclusions The normal sagittal and coronal diameters of thumb and great toe distal phalanx bottom measured by the ultrasonography can guide the operation of wrap-around flap of a great toe with phalanx ungual for thumb II degree defect reconstruction.

Objective:To study the clinicopathologic characteristics and the prognostic factors of triple negative breast cancer,and to study the correlation between clinicopathologic features and EGFR expression.Methods:Clinical characteristics and prognosis of 200 breast cancer patients diagnosed between 1997 and 2004 were reviewed.The paraffin embedded tissues were selected and tissue microarray blocks were made.Immunohistochemical staining was used to evaluate the expression of ER,PR and HER2 for breast cancer molecular type.EGFR expression was evaluated to analyze its correlation with the clinical characteristics and prognosis.Results:Fourty-two cases(21%)of 200 were triple negative breast cancer(TNBC).There was no significant difference between TNBC and non-TNBC in clinicopathologic parameters such as age,tumor size,clinical stage,and lymph node status.However,there was significant difference in histological category and menostasis status between the two groups(P<0.05).Among TNBC,17 0f 42 died during the follow-up(the overall suivival rate was 59.52%),while 26 0f 158 non-TNBC died(the overall survival rate was 83.54%).EGFR was significantly overexpressed in TNBC(69.05%)and it was associated with lymph node status and histological category.Conclusion:Although the incidence of TNBC is low,it has a poorer prognosis than other subtypes.EGFR is highly expressed in TNBC and may serve as a prognostic index and an important therapatic target for breast cancer.

Objective To evaluate the impact of the re-modified Sugiura procedure on portal hemodynamics and liver function in cirrhotic patients with portal hypertension.Methods Forty patients with cirrhosis and portal hypertension who underwent the re-modified Sugiura procedure in the Yichang Second People's Hospital from June 2006 to October 2014 were studied.Changes in the free portal pressure (FPP),portal venous flow (PVF) and liver functions before and after operation were analyzed.Results (1) The FPP at different phases of the operation (after opening the abdomen,after splenectomy,and after devascularization) were (43.2 ± 1.8) cmH2O,(34.8 ± 1.6) cmH2O and (35.2 ± 1.7) cmH2O,respectively.There were significant differences in FPP between the phases of after splenectomy and after opening the abdomen,as well as after devascularization and after opening the abdomen (P ＜ 0.05).There was no significant difference in FPP between after devascularization and after splenectomy (P ＞ O.05).(2) The PVF,which were measured with Doppler sonography at 4 time points (preoperative 1 day,postoperative 10 days,postoperative 6 months,postoperative 18 months),were (1 420.4 ± 137.7) ml/min,(1 205.2 ± 126.7) ml/min,(875.8 ± 118.0) ml/min and (893.8 ± 114.7) n1/min,respectively.There were significant differences in PVF between postoperative 10 days and preoperative 1 day,between postoperative 6 months and postoperative 10 days,as well as between postoperative 18 months and preoperative 1 day (P ＜0.05).There was no significant difference in PVF between postoperative 18 months and postoperative 6 months (P ＞0.05).(3)The liver functions were evaluated using the Child-Pugh score at 4 time points (preoperative 1 day,postoperative 10 days,postoperative 6 months,postoperative 18 months).There were no significant differences among the time points,(P ＞ 0.05).Conclusion The re-modified Sugiura's procedure durably,appropriately and effectively reduced the PVF and FPP,but it did not have any negative effects on the liver functions of patients with cirrhosis.

Objective: To develop a real-time quantitative PCR (qPCR) assay for detecting Sendai virus gene copy number. Methods: The recombinant plasmid was constructed and a reference used to establish the real-time qPCR method with the TaqMan probe and verified for reproducibility and sensitivity. Results: The linear range of the developed real-time qPCR assay was 1.024 × 10³ ~5 × 10¹⁰ copies/μl, with an R² value of more than 0. 99. The standard recovery of the tested samples was 84.56%~119.48%. Conclusions Compared with traditional hernagglutination assay for particle number detection, this method has higher sensitivity, better repeatability and high quantitative accuracy. It can be used for the detection of particle number of vaccine with sendai virus as the carrier, so as to detect the specific activity of vaccine products.

Objective:To construct a recombinant herpes simplex virus 2 (HSV-2) expressing enhanced green fluorescent protein (EGFP) using clustered, regularly interspaced, short palindromic repeat/CRISPR-associated nuclease 9 (CRISPR/Cas9) technology.Methods:Four strategies for inserting exogenous EGFP gene into HSV-2 genome using CRISPR/Cas9 technology were designed: (1) conventional homology-directed repair: circular two homology arm donor-mediated gene knock-in; (2) linearized single homology arm donor-mediated gene knock-in; (3) homology-independent targeted integration; (4) conventional homology-directed repair-mediated by cell lines stably expressing Cas9 and sgRNA.Results:The recombinant virus HSV-2-EGFP was successfully constructed based on the second, the third and the fourth strategies. The second strategy was the most efficient, followed by the third and the fourth strategies. The purified recombinant virus could stably express green fluorescent protein in seven passages and shared similar growth characteristics in Vero cells to the parental virus.Conclusions:Linearized single homology arm donor could increase the efficiency of gene knock-in, and cell lines stably expressing Cas9 and sgRNA could increase the efficiency of gene knock-in mediated by homology-directed repair.