In this study,the VP2 gene of feline panleukopenia virus(FPV) isolated from tiger was successfully cloned and expressed in Pichia pastoris yeast.The VP2 gene was amplified by PCR with a pair of specific primer.Then PCR products was purified and cloned into pGEM-T for sequencing.The interesting gene fragment was recovered after the double enzyme digestion of EcoRI/NotI,then subcloned into pPICZ?A for secretory expression.The recombinant pPICZ?AVP2 was linearized with SacI and then transformed into competence yeast GS115 for expression under the induction of 1% methanol.The positive recombinants were screened by PCR method.The expression product was identified by SDS-PAGE and western-blotting.The results showed that there was a molecular weight of 32 kD,which could be specifically recognized by polyclonic antibody against FPV.It revealed that the recombinant VP2 protein had correct three-dimensional structure,which would be used as a antigen protein for the diagnosis and prevention of FPV infection in tigers.

Objective To explore the effects of Taiji exercise on the psychology and the functions of the autonomic nervous systems of middle aged and elderly people. Methods Two hundred and seven middle aged and elderly Taiji exercisers and 166 age matched non Taiji exercisers were sampled. The Zung Self rating Anxiety Scale(SAS), Zung Self rating Depression Scale (SDS) and autonomic nervous system equilibrium index (ANSEI)were adopted to assess the subjects. Results Compared with those of the control group, the scores of SAS and SDS in Taiji group were significantly higher, and the equilibrium of autonomic nervous systems in Taiji group was also superior. There was no significant difference between the male and the female with regard to SAS, SDS and the equilibrium of autonomic nervous systems within Taiji group. The longer the exercise, the better the equilibrium of autonomic nervous systems. Conclusion Taiji exercise has positive effects on the psychology and the autonomic nervous system of the middle aged and the elderly.

Obesity is becoming a chronic epidemic worldwide. Persistent obesity, in addition to triggering changes in skeletal muscle function and structure, may also lead to the occurrence of skeletal muscle atrophy, that maybe associate with autophagy and ubiquitin protea-some, interleukin-6, leptin, adiponectin, interleukin-10, tumor necrosis factor-alpha, growth hormone, angiotensin II, glucocorticoid, ad-vanced glycation end-product and myostatin, etc.

Objective To study characteristics of hospital infection and bacterial drug-resistance in our new Intensive Care Unit (ICU) to provide evidence to clinical rational use of drugs. Methods The data of 142 patients in our ICU last year were analyzed retrospectively, and compared to other inpatients treated in general ward during the same time period. Results Among the 142 ICU inpatients,31 times of hospital infection occurred in 22 patients,gave rise to a infection rate of 15.49% and case time infection rate of 21.83% in ICU; but among the 15 978patients treated simultaneously in general ward,413 times of hospital infection occurred in 324 inpatients, gave rise to a infection rate of 2. 03% and case time infection rate of 2. 58% outside of ICU. The hospital infection rate and case time infection rate were significantly different between the ICU and general ward (P ＜ 0. 01 ). Among the ICU infections, the lower respiratory tract infection accounted for 70. 96% ;the main pathogens were Gram-negative bacilli (65. 12% ) and Gram-positive bacilli (23.26%), follwed by fungus (11.63%). The most common bacteria were Escherichia ( 32. 65% ) and Staphylococcus ( 11.63% ). Gram-negative microorganisms were highly sensitive to cefoperazone/sulbactam,imipenem, meropenem and piperacillin/tazobactam; all Gram-positive microorganisms were sensitive to vancomycin,linezolid and teicoplanin. Conclusions The constitution,distribution and drug-resistance of pathogens in new ICU are similar to previous domestic reports. Gram-negative organisms are the main photogenic bacteria of hospital infection in ICU, and Escherichia is the most popular type in nosocomial infection.

Objective To treat the patients suffering from intractable epilepsy after brain injury with low-dosage linac accelerator stereotactic radiosurgery through positron emission tomography (PET) localization. Methods PET examination based on 18 F-2-fluorine-2-deglucose (FDG) was carried out in 32 patients in order to localize epileptogenic foci followed by radiosurgery with peripheral radiation dose of 9-13 Gy. The follow-up was conducted over one year. The seizure frequency of epilepsy was recorded to evaluate the therapeutic effect. Results The PET imaging showed that the cortical area around the cerebromalacia caused by primary injury presented low metabolic change. The metabolic changes in different injury areas mainly concentrated in the epileptic foci. In some cases,the low metabolic areas not only was localized at the part of impact lesion but also at the part of contrecoup lesion. The seizure frequency gradually decreased after radiosurgical treatment in most patients,with obvious statistical significance compared with pretreatment. According to Wieser's classification of operative effect,the cases at the grades Ⅰ-Ⅱ accounted for 44% (14/32),grades Ⅲ-Ⅳ for 41% (13/32) and grades Ⅴ-Ⅵ for 16%(5/32). No fresh complications were found in all the cases. Conclusions Low-dosage stereotactic radiosurgery with the PET localization is a safe,effective and minimally invasive surgical approach to intractable posttraumatic epilepsy.

Objective To assess the efficacy and safety of ACD-A solution as anticoagulant during continuous renal replacement therapy (CRRT) in high risk of bleeding patients.Methods Forty high risk bleeding patients on continuous veno-venous hemofiltration (CVVH) were randomly divided into two groups:ACD-A group (22 patients,61 cases) and heparin-free group (18 patients,47 cases).Serum creatinine,function of the coagulation system,electrolyte and acid-base were monitored pre-and post-CVVH.The vital signs of the patients during treatment,dialyser clotting and the incidence of bleeding episodes were recorded.Results (1) The serum level of creatinine decreased significantly after treatment in both groups,but the rate of decrease was obviously higher in ACD-A group than that in heparin-free group[(55.4± 10.2)％ vs (42.0±5.2)％,P=0.031].(2) The average duration of CVVH treatment was (17.3±3.8) h in ACD-A group and (9.7±4.5) h in heparin-free group.There was significant difference between them (P =0.019).The frequency of dialyzer clotting was much higher in heparin-free group than that in ACD-A group (88％ vs 4.9％,P ＜ 0.001).(3) There was no significant difference in the function of the coagulation system between pre-and post-CVVH in either group (P ＞ 0.05).(4) Electrolyte,acid-base and glucose tended to be stable during the treatment in ACD -A group.(5) The vital signs were kept stable and no bleeding episodes were found in all patients of two groups.Conclusions Anticoagulation with ACD-A is safe,effective and convenient for CRRT in critically ill patients at high risk of bleeding.The occurrence of complications can be reduced by configurating appropriate replacement fluid and close laboratory monitoring.

Objective To assess the value of 18 F-FDG PET imaging in the localization of epileptic foci.Methods 18 F-FDG PET were performed in 22 patients with medically intractable epilepsy (15 males,7 females,mean years:19.0 years,mean duration:8.2 years). The open skull operations were undergone in 21 patients according to the results of 18 F-FDG PET, combining with scalp EEG and CT or/and MRI. 1 patient was refered for stereotactic operation. During operation,EcoG was detected for identifying the site of epileptogenic focus and lobectomy was performed . All specimens were examninated with the pathological tests. All patients were followed up for the curative effects. Results PET showed abnormalities in 21 patients(95.5%). Among them,single or multiple focal hypometabolisms were found in 20 patients and focal hypermetabolism in 1 patient. The scalp EEGs showed abnormalities in 17 patients(77.3%),but 8 of them did not showed clear latreralizing manifestations.The abnormalities were found on MR or/and CT in only 5 cases (22.7%).The epileptogenic foci detected by PET were consistent well with EcoG in 18cases(85.7%), consistent approximately in 2 cases and not consistent in 1 cases .Among 19 patients with abnormal PET findings , the pathological abnormalities were found in 18 cases (94.7%).No pathological abnormality was found in the patients with nomal PET imaing. After the open skull operations, 9 patients (42.9%) becomed seizure free . 9 patients (42.9%) had more than 50% seizure reduction and 3 patients (14.3%)showed no improvement. The patient treated with stereotactic operation also showed no significant improvement.Conclusion 18 F-FDG PET imaging is a sensitive and accurate modality to localizate the epileptic foci and has important clinical value in surgical therapy.

Objective To evaluate the effect of 3D printing models applied to Medical Imaging Di-agnosis Undergraduate Teaching. Methods Students in the year 2012/2013 of Shanghai Jiao Tong Univer-sity School of Medicine stomatology specialty were chosen for this study, including 38 students in the exper-imental group (2013) and 40 students in the control group (2012). All of them were taught by the same group of teachers, including 28 core curriculum hours and 5 practice class hours. 3D printing technology with PPT was applied to the experimental group in the medical imaging diagnostic teaching, and conventional teaching including PPT and usual models in the control group. Medical students were given a test about cardiovascular system and tasked with completion of drawing hand and foot bones. Students in experimental group were also asked to complete a questionnaire about their experience. Results The questionnaire showed that 3D printing teaching molds with PPT was of great significance to improve the students interest in learning imaging anatomy, to meet the vast majority of students' learning requirements, and to achieve satis-factory results. The average scores of the circulatory system test in the experimental group were higher than that of the control group [(8.32 ±2.352) vs. (6.15 ±1.284), P<0.05)]. Students in experimental group per-formed better than students in the control group on the Locomotor system test [(6.81 ±1.347] vs. (3.84 ± 0.985), P<0.01]. Conclusions 3D printing mold combined with PPT teaching method can promote the med-ical imaging anatomy structure, learning effect and teaching quality, and has practical value in the under-graduate teaching of medical imaging diagnosis.

Objective To investigate the effects of antisense recombinant euraryotic expression vector of HCCR-2 on the proliferation and apoptosis of HepG2. Methods The antisense recombinant eukaryotic expression vector of HCCR-2 was constructed. The vector was stably transfected to the HepG2 cells, and positive clones were selected by G418 (antiseuse vector group), pIRES2-EGFP vector was transfected into the HepG2 cells in the same way (pIRES2-EGFP group). The conditions of the nontransfected HepG2 cells were used as control (HepG2 group). Changes in cell growth curve, cell cycle, cell apoptosis and morphology of HepG2 cells after the transfec-tion were detected by MTT method, flow cytometry and transmission electron microscopy, respectively. All the data were analyzed by one-way ANOVA and chi-square test. Results The expression level of HCCR-2 mRNA was down-regulated to 0.39±0.04 in antisense vector group, and the expression level of HCCR-2 mRNA in pIRES2-EGFP group and HepG2 group were 0.62±0.06 and 0.72±0.03, respectively, with significant difference among the 3 groups (F=43.701, P<0.05). The apoptotic rate of HepG2 cells in antisense vector group, pIRES2-EGFP grop and HepG2 group were 13.30%, 2.51% and 2.07%, respectively, with significant difference among the 3 group (χ2=6.793, 8.721, P<0.05). The growth of HepG2 cells in antisense vector group was retarded, and was blocked in G0/G1 stage. Conclusions The HCCR-2 antisense recombinant eukaryotic expression vector can inhibit the mRNA expression of HCCR-2 and promote the apoptosis of cells. HCCR-2 may be involved in cell regulation and the proliferation of hepatocellular carcinoma cells.

Feline calicivirus (FCV) is an important and highly prevalent pathogen of cats that causes feline respiratory disease. The reverse genetic systems for FCV have been established in national and international laboratories since 1995. This technique has been used widely in FCV basic research and good progress has consequently been made to determine the relationship between viral genome structures and the function of their proteins, the expression of foreign proteins, virus-host interactions, and viral pathogenic mechanisms. In this article,we review the state of progress with regards to the establishment and application of the FCV reverse genetic operating system,which will provide a useful reference tool for future related research.
Animals ; Caliciviridae Infections ; veterinary ; virology ; Calicivirus, Feline ; genetics ; metabolism ; Cat Diseases ; virology ; Cats ; Reverse Genetics ; methods ; trends ; Viral Proteins ; genetics ; metabolism