Objective To construct a fusion minigene expression vector of seven HCV NS3 epitopes and express the fusion gene in the eukaryotic cell line, in order to form a foundation for the investigation of the DNA immunization for HCV prevention. Methods Three pairs of complementary oligonucleotides primers covering all seven HCV NS3 epitopes were synthesized. Overlapping extension PCR were performed to construct the fusion minigene and cloned in pEGFP-N3 and pBuDCE vector respectively. The fusion protein was detected by Western blotting and flow cytometric analysis. The transfection efficiency was assessed with by flow cytometric analysis. Results The HCV NS3 epitope fusion minigene was obtained and inserted into pEGFP-N3 and pBuDCE vector respectively. The recombinant plasmid pEGFP-DR4 and pBuDCE-DR4 were expressed in 293T and 046W cell lines respectively. The Western blot result indicated that the expected 36 kDa minigene/EGFP fusion product was expressed from pEGFP-DR4, while 13kDa product from pBuDCE-DR4 respectively. Conclusion The HCV NS3 epitope fusion minigene was expressed in the eukaryocyte expression system.

Physician-patient communication skill is the core clinical capability,and its im-portance is more and more recognized by educationists.Introducing the communication skill education into the standardized resident training can improve the residents'clinical and communicative ability and train them to be the comprehensive quality talents.

Objective:To investigate the correlation between human herpesvirus-6(HHV-6) infection and pathogenesis of oral squamous cell carcinoma.Methods:Samples of saliva, peripheral blood mononuclear cells (PBMCs),tumor tissue and tumor adjacent tissue were collected from 82 patients with oral squamous cell carcinoma, 25 patients with oral precancer lesion and 40 healthy young adults(controls). Nested PCR assay was used to examine HHV-6 infection.Results:1)In control, precancer lesion and carcinoma groups, HHV-6 positive rates(%) were 45.0,56.0, 72.0 in saliva, 22.5,48.0,57.3 in PBMCs, and general rates(%) were 47.5,56.0 and 80.5 respectively(control vs carcinoma,P0.05).Conclusions:Saliva gland is the position of HHV-6 latency/proliferation and saliva is the predominant route of HHV-6 transmission; HHV-6 infection correlates to the pathogenesis of oral squamous cell carcinoma.

Objective To investigate the effect of EMG-triggered stimulation on drop foot for poststroke hemiplegia. Methods 36 hemiplegic patients were randomized into routine group (n=18) and EMG-triggered stimulation group (triggered group, n=18). 2 groups were assessed by Fugl-Meyer Assessment (FMA), range of motion (ROM) of ankle dorsiflexion and iEMG values before and after treatment. Results The scores of FMA, degrees of ROM, iEMG values of the tibialis anterior, and co-contraction ratio improved in 2 groups after treatment (P<0.05), and the triggered group had better effect than the routine group (P<0.05). Conclusion EMG-triggered stimulation can improve muscle strength, and decrease co-contraction ratio.

Objective To measure the quality of life (QoL) of gestational trophoblastic neoplasia ( GTN ) survivors after chemotherapy by using a self-invented scale, and to explore the factors associated with QoL Methods The design of questionnaire was based on a series of internationally valid QoL scales,which was tested by epidemiology and showed good reliability and validity. A total of 100 survivors of GTN patients from Peking Union Medical College Hospital participated in this survey from December 2008 to May 2009. Results Patients with disease-free more than three months after chemotherapy enjoys a good QoL,while only 16% (16/100) of survivors feel general overall QoL, but no one feels bad Qol. As refer to sexual function, more than half of these patients (70%, 70/100) satisfied with their sexual life, while there were still 47% (47/100) and 45% (45/100) of the patients complaining of decreased sexual desire and dryness of vagina. 66% (66/100) of the GTN survivors expressed depression, and 50% (50/100) of patients complained anxiety, which were potential factors influencing QoL of GTN survivors. Relevant analysis explored the possible predictors of QoL for GTN patients, including physical function ( r = 0. 609,P ＜0. 01 ), sexual function ( r = 0. 473, P ＜ 0. 01 ), and social psychology ( r = 0. 294, P ＜ 0. 01 ).Conclusions GTN survivors have an overall good QoL after chemotherapy, the possible predictors of QoL for GTN patients include physical function, sexual function and social psychology. The sexual dysfunctions mostly present with short of sexual desire and dryness of vagina. Fear of recurrence may be a potential factor influencing QoL a long term after remission.

OBJECTIVE:To study the correlation between antibacterials amount and drug resistance of Echerichia coli,and to provide reference for clinical use of antibacterials. METHODS:Retrospective review was used to calculate DDDs of antibacterials and resistance rate of Escherichia coli to 11 kinds of antibacterials each quarter. The correlation analysis was carried out using the SPSS 13.0 statistical software. RESULTS:The resistance rates of E. coli to piperacillin/tazobactam,cefoperazone/sulbactam and le-vofloxacin were with upward trends,and the others showed downward trends. The resistance rates of E. coli to meropenem and imi-penem/cilastatin appeared in 2014,increasing from 0 to 8.8% and 9.4%,respectively. DDDs of them were significantly correlated to drug resistance of E. coli,showing positive correlation(r=0.915,0.793,P<0.01). DDDs of piperacillin/tazobactam was signif-icantly correlated to resistance rate of E. coli(r=0.807,P<0.01),while that of ceftazidime was negatively correlated to resistance rate of E. coli(r=-0.672,P<0.05). There was no statistical significance in resistance rate of E. coli to other 7 kinds of antibacte-rials. CONCLUSIONS:There are some correlations between the DDDs and resistance rates. We should strengthen the monitoring of bacterial resistance and the management of rational application of antibacterials.

Objective To investigate the associations between single nucleotide polymorphisms in the gene encoding human leukocyte antigen-G ( HLA-G) and the genetic susceptibility to high risk human papillomavirus ( HPV) type 18 infection in the subjects from Taizhou, Zhejiang province. Methods The genetic polymorphisms of HLA-G gene (14 bp In/Del and +3142C/G) in cervical samples collected from HPV 18-positive and healthy women were analyzed by PCR and gene sequencing technology. Statistical anal-ysis was performed by using SPSS version 16. 0. Chi-squared test was used to analyze the differences with HLA-G gene allele and genotype frequencies between healthy subjects and patients. Results Compared with healthy subjects, women with oncogenic HPV18 infection showed lower frequencies of -14 bp allele,-14 bp/-14 bp genotype and -14 bp/+3142C haplotype (P<0. 05). Moreover, lower frequencies of+3142C/C genotype were detected in HPV18-infected women with pathologically normal cervix (6. 3% vs 21. 1%, OR=0. 25, P<0. 05) and higher percentages of +3142G/G genotype were detected in women with CIN2/3 stage HPV18 infection as compared with those of the control group (68. 8% vs 35. 1%, OR=4. 06, P<0. 05). Conclusion The HLA-G gene 3′ UTR polymorphisms were closely associated with HPV18 in-fection and cervical intraepithelial neoplasia.

Objective To investigate the changes of soluble CD105,transforming growth factor-beta1(TGF-β1) and vascular endothelial growth factor(VEGF) in esophageal cancer(EC) patients,analyzing its clinical significance.Methods Collecting 56 cases of gastric cancer(observation group) and 49 cases of healthy people(control group)from Nov.2014 to Jan.2016 as the research objects.The levels of soluble CD105,TGF-β1 and VEGF were detected in EC patients pre-and post-operation and healthy subjects.The data of soluble CD105,TGF-β1 and VEGF were analyzed in the two groups.Results The pre-operation levels of soluble CD105,TGF-β1 and VEGF in EC patients were significantly higher than those in the control group,the difference was statistically significant(P<0.05).The pre-operation levels of soluble CD105,TGF-β1 and VEGF in EC patients were significantly higher than post-operation levels,the difference was statistically significant(P<0.05).The post-operation levels of soluble CD105,TGF-β1 and VEGF in EC patients were not statistically different with control group(P>0.05).Conclusion The levels of soluble CD105,TGF-β1 and VEGF could be abnormal in patients with EC,which might be the index for monitoring the clinical disease condition and judging the prognosis of the EC.

ObjectiveTo observe the effect of parathyroid hormone on expression of matrix GLA protein (MGP) in ovariectomized SD rats and primary osteoblast,and to study the role of MGP on the possible mechanism of postmenopausal osteoporosis.MethodsThirty-six Sprague-Dawley female rats were allocated into 3 groups,12 in each:sham operation group,ovariectomized group( OVX group),ovariectomized and parathyroid hormone treatment group.Animals in the parathyroid hormone group were injected parathyroid hormone (20 μg/kg,three times a week for 12 weeks) three weeks after ovariectomy.All rats were sacrificed after 18 weeks.Urine and serum were collected every three weeks.Lumbar vertebral bones were observed by immunohistochemistry.Bone mineral density (BMD) of lumbar vertebra of rats was determined.The content of MGP in serum and urine was determined by enzyme-linked immunosorbent assay (ELISA).Expression of undercarboxylated Matrix GLA Protein (ucMGP) was detected by immunochistochemistry.Relative quantification of MGP mRNA expression in lumbar vertebra bone was detected by Fluorescent real-time quantitative polymerause chain reaction.Results ( 1 ) 18 weeks after ovariectomy,BMD of lumbar vertebra in OVX group was lower than those in sham group and parathyroid hormone group significantly ( P＜0.05 ).(2) The content of MGP in serum and urine was dynamic variation after treatment hy parathyroid hormone,and it was significant compared with OVX group ( P＜0.05 ).( 3 ) Immunohistochemical localization of ucMGP was seen in lumbar vertebra in OVX group.(4) Relative quantification of MGP mRNA expression in lumbar vertebra in OVX group was increased significantly compared with other groups ( P＜0.01 ).( 5 ) parathyroid hormone ( 1-34 ) in 10-7mol/L,10-8mol/L,10-9 mol/L up-regulated MGP mRNA expression in primary osteoblasts about 6.78,5.31,and 2.23 times than control respectively.It was in a dose-dependent manner.ConclusionThe effect of parathyroid hormone on the expression of matrix gla protein may play an important role in mechanism of postmenopausal osteoporosis

Objective To observe the expression of matrix gla protein(MGP) mRNA in primary osteoblasts of Sprague-Dawley (SD) rat in vitro after treatment with anti-osteoporosis agents [vitamin K2,PTH,1,25 (OH)2D3,and alendronate],and to investigate the potential role of MGP in the pathogenesis of osteoporosis.Methods Primary osteoblasts(OBs) were derived from sequential trypsin/collagenase-digested calvaria isolated from newborn SD rat (postnastal day 1-3).OBs of the second generation were identified by Van Gieson collagen staining,alkaline phosphatase(ALP) staining and calcified nodules staining.OBs of the fourth generation were selected to interfere with vitamin K2,PTH,1,25 (OH)2D3,and alendronate,then cultured for 24 h in mediums which contained various concentrations of vitamin K2 (10-7,10-6,and 10-5 mol/L),PTH (10-9,10-8,and 10-7 mol/L),1,25 (OH) 2D3(10-10,10-9,and 10-8mol/L),alendronate(10-6,10-5,and 10-4mol/L).After being cultured for 24 h,total RNA was extracted and examined by real-time quantitative RT-PCR.Results The primary cultured cells had typical morphological characters of osteoblast.van Gieson collagen staining,ALP staining,and calcified nodules staining were all positive.Vitamin K2,PTH,1,25 (OH)2D3,and alendronate could modulate the expression of MGP mRNA in osteoblasts in a dose-dependent fashion.MGP mRNA expressions were 2.56-fold,2.12-fold,and 1.57-fold with 10-5,10-6,and 10-7 mol/L of vitamin K2 treatment,respectively.The expressions were 6.78-fold,5.31-fold,and 2.23-fold with 10-7,10-8,and 10-9mol/L of PTH(1-34) treatment,8.93-fold,6.95-fold,and 3.47-fold with 10-8 10-9,and 10-10mol/L of 1,25 (OH)2D3 treatment,and 3.47-fold,2.49-fold,and 1.98-fold with 10-4,10-5,and 10-6mol/L of alendronate treatment.Conclusion Vitamin K2,PTH,1,25 (OH)2D3,and alendronate all canregulate MGP mRNA expression in calvarial osteoblasts in a dose-dependent manner.MGP seems to be a potent target of anti-osteoporosis agents,and involved in the pathogenesis of osteoporosis.