Objective To study the immunoreaction of the recombinant proteins encoded by the fragments of ORF2 ( second open reading frames) gene of hepatitis E virus ( HEV). Methods The aimed sequence from the full-length ORF2 in clone PEH2 , which was derived from a Chinese strain of HEV,was amplified and cloned it into vector pcDNAS. 1 which was then transfected to 293 cell. Results The ORF2 protein was present in the soluble fractions of the cell lysate. The expressed protein of HEV ORF2 in 293 cells by using a plasmid pcDNA3. 1-based system showed positive on immunoblots probed against antibodies raised in BALB/c mice. Conclusion The experimental results laid a foundation for developing diagnostic reagent to detect HEV by using the expressed products of ORF2 protein.

Objective To investigate the effects of genistein used alone and in combination with cisplatin on the treatment of human ovarian carcinoma transplanted subcutaneously in nude mice. Methods Nude mouse model of human ovarian carcinoma transplanted subcutaneously was established. A total of 30 mice were divided into 5 groups: control group (0 04% saline), genistein group (subcutaneous injection of genistein at 0 2 mg/kg and 0 4 mg/kg), cisplatin group (intravenous injection of cisplatin at 4 mg/kg), and genistein plus cisplatin group. The growth of the subcutaneously transplanted tumor and changes of mouse body weight in each group after treatment using different medication regimens were observed at 7, 14, 21, and 28 d. Histopathological examination was also conducted. Results Genistein (0 4 mg/kg) had significantly inhibitory effect on the transplanted xenograft growth in vivo . Tumor volume, tumor weight, and T/C ratio (mean volume of treated group/mean volume of control group) decreased. Significantly enlarged necrotic areas were found in genistein treated group. The weight loss of nude mice after treatment with genistein was not significantly different from in the control group. Conclusion Genistein has the inhibitory effect on the growth of human ovarian carcinoma transplanted subcutaneously in nude mice. Genistein (0 4 mg/kg) in combination with cisplatin (4 mg/kg) has a synergistic effect on xenograft of human ovarian cancer cell line SKOV 3 in nude mice. The results provide the evidence for the potential usefulness of genistein for the prevention and treatment of human ovarian carcinoma.

Objective To isolate and cultivate Long-Evans rat bone mesenchymal stem cells (BMSCs) and identify their biological characteristics. Methods BMSCs were obtained from femurs of Long-Evan rats and dealt with 0.85 NH4Cl. The cell growth curve and cell cycle were measured and surface antigens were detected by flow cytometry. Osteoblast differentiation was studied by Alizarin red staining. Results BMSCs had active proliferative ability. The growth velocity of passage 5 cells was slower than that of passage 3 cells. The cell cycle analysis showed that 93.79 of BMSCs was at G0/G1 phase. BMSCs were positive for CD29 and CD44, but negative for CD45. Alizarin red staining of BMSCs after osteoblast induction was positive. Conclusion Highly purified BMSCs obtained from Long-Evan rats by 0.85 NH4Cl show stable biological properties.

Objective To select the optimal registration method for on-line kilovoltage cone-beam CT (KVCBCT) guided lung cancer radiation and evaluate the reproducibility of the selected method. MethodsSixteen patients with non-small cell lung cancer were enrolled into this study.A total of 96 pre treatment KVCBCT images from the 16 patients were available for the analysis.Image registration methods were bone-based automatic registration,gray-based automatic registration,manual registration and semi-auto matic registration.All registrations were accomplished by one physician.Another physician blindly evaluated the results of each registration,then selected the optimal registration method and evaluated its reproducibili ty.Results The average score of the bone-based automatic registration,gray-based automatic registration, manual registration and semi-automatic registration methods was 2.4,2.7,3.0 and 3.7,respectively.The score of the four different groups had statistics significant difference (F = 42.20,P < 0.001).Using the semi-automatic registration method,the probability of the difference between two registration results more than 3 ram in the left-right,superior-inferior,and anterior-posterior directions was 0,3% and 6% by the same physician,0,14% and 0 by different physicians,and 8%,14% and 8% by physician and radiation therapist.Conclusions Semi-automatic registration method,possessing the highest score and accepted re producibility,is appropriate for KVCBCT guided lung cancer radiation.

Objective To explore the feasibility and effectiveness of the revised OSCE stations.Methods 106 sstudents of grade 2008 and 2009 from Xinhua clinical medical school of Shanghai Jiaotong University School of Medicine were involved,32 of whom (experimental group) tool modified OSCE examination and 74 of whom (control group) tool traditional OSCE examination.The revised OSCE exam was the one which on the basis of original OSCE test station setup,improved the operating part of the test stand,adopted clinical cases as stem,asked the students to make possible diagnosis,and perform the corresponding operation.We evaluated the students based on the OSCE scores from the school exam and the overall performances in the national clinical skills competition.Comparison between the two groups was performed by using t test.Results All students who have received the new training methods agreed that it stood closer to the clinical setting.Revised-OSCE trained group of students test scores than traditional OSCE trained group of students,though it didn't have statistical significance.Students from the revised-OSCE group had the score of (80.50 ± 15.45),while the traditional group had the score of (78.30 ± 17.68).By taking the revised-OSCE trainings,all the students have improved their clinical skills and two years in a row to win the Shanghai Jiaotong University School of Medicine Clinical Skills Competition.Conclusions Case-based OSCE stations are well accepted.The new OSCE stations are much closer to clinical teaching and can make better objective assessment of students,so it plays a good role in improving medical students' overall performances.

Objective To assess the clinical factors impacting on the effective time of endocrine therapy for patients with prostate cancer.Methods The chnical data of 432 patients with prostate cancer who accepted endocrine therapy were analyzed retrospectively.The endpoint of the study was failure of endocrine therapy which was defined as continuous elevation of prostate specific antigen (PSA) from nadir for 2 times and more than 0.2 μg/L.The clinical data such as age,clinical stage,lymph node metastasis,bone metastasis,Gleason score,initial PSA,and PSA nadir were collected and their rehtionship with the effective time of endocrine therapy were further assessed via COX regression model.Results Age of onset was 57-88(73.70 ± 7.28) years.Initial PSA was 10.30-588.10(27.15 ± 75.90) μ g/L.The effective time of endocrine therapy was 3-62 (27.01 ± 13.10) months.Univariate regression analysis showed that initial PSA,clinical stage,Gleason score,PSA nadir,lymph node metastasis,bone metastasis were correlated with the effective time of endocrine therapy (P ＜ 0.01).Multivariate regression analysis showed that only Gleason score was correlated with the effective time of endocrine therapy(P=0.001).Compared with patients with Gleason score equal to or less than 3+4,patients with Gleason score equal to or more than 4+3 showed 2.49 fold increased risk of therapy failure (OR =2.49,95％ CI 1.44-4.30).Conclusion Gleason score has close relationship with the effective time of endocrine therapy for patients with prostate cancer,Gleason score equal to or more than 4+3 is an indicator for poor response to endocrine therapy.

Background Normal ultrastructure is the anatomical basis of retinal pigment epithelial(RPE) cells to perform normal physiological function.At present the precipitation method is often used to detect the ultrastructure of RPE cells with transmission electron microscopy(TEM).Objective The aim of this study was to explore a simple and feasible approach to examine the ultrastructure of human embryonic stem cell-derived retinal pigment epithelial (hESC-RPE) cells.Methods hESCs were induced and differentiated into RPE cells by the spontaneous differentiation method,and the expressions of microphthalmia associated transcription factor MITF and paired-box gene 6 (PAX6),specific protein of RPE cells,in the cells were detected by immunofluorescence assay.hESC-RPE cells were inoculated into Transwell filter,and the ultrastructure of the cell sheet was examined under the TEM.Then the ultrastructure of the cell sheet specimens was compared with those of hESC-RPE cells from cell precipitation and RPE cell specimens of 90-day-old Long Evans rats.Results MITF and PAX6 were positively expressed in hESC-RPE cells.The normal ultrastructure were visible in the RPE cells of rats under the TEM,including apical microvilli,polarized melanin granules,cellular nucleus,basement membrane and intercellular junctions,and the ultrastructure of hESC-RPE cell sheet on Transwell was similar to the RPE cells in rats.However,only scatter melanin granules,nonpolar nucleus and scanty microvilli were observed under the TEM in the hESC-RPE cells by cell precipitation method.Conclusions Without digestion process,hESC-RPE cell sheet on Transwell can retain the normal ultrastructure of hESC-RPE cells under the TEM,with a more simple and reliable advantage.

Objective:To understand the prevalence and epidemiological characteristics of glaucoma among people over 40 years old in rural areas of Chongqing.Methods:From August to October 2018, a stratified cluster random sampling method was used to investigate the cases of glaucoma among people over 40 years old in a rural population of the Rongchang and Qijiang Districts in Chongqing.The subjects had been living in the local area for over ten years.Basic information for each patient, including gender and age was documented, and their visual acuity and intraocular pressure were routinely measured.The Van Herick method was used to evaluate the depth of the central and peripheral anterior chamber, a preset lens was used to examine the fundus, and the cup to disc ratio(C/D) of the optic disc was emphasized.All suspected glaucoma patients underwent further standard glaucoma examinations.This study protocol was approved by the Ethics Committee of the First Affiliated Hospital of Army Medical University.Results:A total of 4 680 people were identified, and 4 073 respondents participated in the survey, and the response rate was 87%.The prevalence of glaucoma was 1.72% (70/4 073). There were no significant differences in the prevalence of primary angle-closure glaucoma (PACG) and primary open angle glaucoma (POAG) between different genders ( χ2=0.042, P=0.837; χ2=2.838, P=0.092). The prevalence of glaucoma in people over 70 years old was 2.5%, significantly higher than that in other age groups.In patients with glaucoma, the rate of visual impairment was 68.57%(48/70), and the rate of blindness was 47.14%(33/70). There was no significant difference in the incidence of low vision between different types of glaucoma ( χ2=2.785, P=0.248), but there was a significant difference in the rate of blindness ( χ2=10.668, P=0.005). The highest rate of blindness was found in secondary glaucoma.The detection rate of intraocular pressure, anterior chamber depth and C/D was statistically significant ( χ2=43.325, P<0.001), and the positive rate of C/D was the highest.When the factors of intraocular pressure, and C/D were considered jointly, the detection rate of glaucoma could be increased to 55.88%. Conclusions:The prevalence of glaucoma is 1.72% among people over 40 years old in rural areas of Chongqing, the prevalence of PACG is lower than previously recorded, but the proportion of glaucoma-induced visual impairment is higher.In field screening, the morphological examination of the fundus optic nerve is very important for the diagnosis of glaucoma.

Objective To compare the clinical efficacy and safety following the topical application of pazufloxacin mesylate eye drops with levofloxacin eye drops (LOFX) for bacterial conjunctivitis.Methods A multicenter,randomized,investigator-masked,parallel-controlled clinical trial was performed.Five hundred and twenty eyes of 520 patients with bacterial conjunctivitis were enrolled from March to October 2008 in seven ophthalmic centers in China.The patients were randomized into trial group and control group in 3 ∶ 1 ratio with the 390 eyes in the trial group and 130 eyes in the control group.Pazufloxacin mesylate eye drops was topically used 4 times per day for consecutively 7-14 days in the trial group,and levofloxacin eye drops was used in the same way in the control group.Microbiological cultures for conjunctiva sac secretions and drug sensitive test were carried out before and at the end of the administration of eye drops.The signs and symptoms were observed and scored before treatment and 0,3,7 and 14days after treatment.The adverse events following the administration of the eye drops were evaluated and compared.Results The intention to treat analysis (ITT) showed that the cure rate was 59.38％ in the trial group and 60.47％ in the control group,with the effective rate 88.80％ and 86.05％,respectively,with an insignificant difference between the two groups (x2 =0.12,P =0.72).The clinically evaluable analysis (CE) exhibited that the cure rates were 63.48％ in the trial group and 63.87％ in the control group,with the effective rate 92.46％ and 88.24％,whichwas not significantly different between them (x2 =0.54,P=0.46).The modified-ITT analysis (mITT) showed that the cure rates were 60.57％ in the trial group and 62.07％ in the control group,with the effective rate 90.32％ and 88.51％,without significant difference between the two groups (P＞0.05).Based on microbiologically evaluable analysis (ME),the clinical cure rates were 63.71％ and 63.41％ in the trial group and control group,and the effective rates were 93.44％ and 90.24％,respectively.There was no significantly difference between the both groups (P＞0.05).In the trial group and control group,the bacterial eradication rate was 89.42％ and 90.80％ based on ITT,90.11％ and 92.77％ based on CE,respectively.There was no significant difference in incidences of adverse events following the administration of the drug between the trial group and control group,including ocular tolerance,burning sensation,pricking and itching (P =0.34).Conclusions The effectiveness and adverse response were resemble between Pazufloxacin mesylate eye drops and LOFV following the topical application for bacterial conjunctivitis,which indicate that Pazufloxacin mesylate eye drops is effective and safe.