Objective To clear, amplify and detect the activity of the recombinant adeno-assoeiated virus vector with adiponectin( rAAV2/1-Aerp30 ). Methods Recombinant plasmid pSNAV2.0-Acrp30 was obtained. The recombinant plasmid was then transfected into BHK21 cells using LipofectAMINETM 2000. The G418 resistant cells were obtained consequently. These cells were infected with HSVI-rc/△UL2 which has the function of packaging and copying recombinant AAV. After purification, the construction of recombinant rAAV2/1-Aerp30 was collected. Results The construction of recombinant pSNAV2.0-Acrp30 was confirmed by PCR electrophoresis and digestion with restriction enzyme. The gene sequencing showed that the recombinant pSNAV2.0-Acrp30 was correct. The virus titer was about 1.0×1012 μg/ml. The purity f the recombinant AAV2/1 was fairly high using the SDS-PAGE method. Conclusion With this method, rAAV2/-Aerp30 with high virus titers and purity can be acquired successfully and it can meet the demands of the experimental study of Acrp30 gene therapy of GK rats.

Objective To study the effects of exercise after myocardial infarction on the survival rate and the dynamic functioning of the left ventricle ( LV) . Methods Ninety-six male Sprague-Dawley rats were randomly assigned to a sedentary sham operation group ( Sed-Sh) , a previous exercise and sham operation group ( PreE-Sh) , a sedentary myocardial infarction ( MI) group ( Sed-MI) , a previous exercise MI group ( PreE-MI) , a post exercise MI group (PostE-MI) and a combined exercise MI group (ComE-MI), each of 16.All of the rats underwent either MI in-duction or a sham MI operation, the PreE groups after exercising for 5 weeks on a treadmill. The Sed groups did not exercise. The Post E-MI group did not exercise before the operation, but performed treadmill exercises for 4 weeks be-ginning 8 weeks after the operation. The ComE-MI group exercised for 5 weeks before and 4 weeks after the operation. All the exercising rats ran on the treadmill for 60 minutes daily, 5 d/wk. Life situation and spontaneous mortality were recorded, and echocardiographic measurements were performed on the4thday and 2, 4, 8 and 12weeks after the oper-ation. All of the rats were sacrificed at the end of the experiment. Results Compared with the Sed-Sh group, the Sed-MI group had significantly larger average LV dimensions at the end of both the diastol ( LVEDd) and the systol (LVEDs, but it had lower average fractional shortening (FS) and a smaller average ejection fraction(EF) at all of the different measuring times. The PreE-MI group had significantly lower average LVEDd and LVEDs, but a signifi-cantly higher FS and EF than the Sed-MI group on the 4th day. However, 2 weeks after the intervention the difference in LVEDd between the two groups was no longer significant. Significantly higher FS and EF were observed in the PostE-MI group at 12 weeks compared to the Sed-MI group. The ComE-MI group had significantly lower average LVEDd and LVEDs, but significantly higher FS and EF at all the time points than the Sed-MI group. The ComE-MI group had a significantly lower average LVEDs but a higher average FS and EF than the PreE-MI group at 12 weeks after the intervention. Conclusion Pre-MI exercise improves LV function significantly at the early stage, though the difference is temporary. Post-MI exercise improves LV function in the longer term. Combined exercise improves LV function the most effectively.

Objective To investigate maternal risk factors for fetal congenital heart diseases (CHD). Methods A case-control study was conducted on 16 645 pregnant women who underwent cardiovascular malformation screening for fetal cardiovascular system,whose pregnancy outcomes were recorded,and whose newborns were scanned by an echocardiography in Peking University People's Hospital,Haidian,Changping,Mentougou and Daxing Maternal and Child Health Hospital from Nov.2006 to Oct.2009.One hundred and twelve pregnant women whose babies were found to be CHD (40 severe CHD and 72 simple CHD) before or after delivery were taken as study group.Women in control group (n =304) were randomly selected from those pregnant women who had infants without CHD.Logistic regression analysis and x2 test were used to analyze the maternal risk factors for fetal CHD. Results (1) The average age of women whose infants had severe CHD was 28.3 years (21-40 years),and it was 29.9 years (22-39 years) for women whose infants had simple CHD.There were no significant differences between the control group (29.5 years,20-44 years) and the above two groups (t=1.511 and -0.826,P=0.138 and 0.410 respectively).(2) Single factor analysis:during first trimester,the rate of upper respiratory infection (18/39,46.2 ％) and exposure to certain chemicals (13/40,32.5％) of severe CHD group were higher than those of control group [(14.9％ (45/303) and 2.0％ (6/304)] (x2 =22.399 and 62.678,OR=4.895 and 23.753,95％CI:2.419-9.905 and 8.358-67.506,P =0.000 respectively).Compared with control group (0.0％,0/304),the rate of pregnant women with CHD family history in simple CHD group was significantly higher (4.2％,3/72)(Fisher exact test,P=0.007).(3) Logistic regression analysis:maternal upper respiratory infections (OR =5.120,95％CI:2.340-11.206,P =0.000) and exposure to certain chemicals (f)R=23.030,95％CI:7.506-70.665,P=0.000) during first trimester were risk factors for fetal severe CHD. Conclusions Upper respiratory infection and exposure to certain chemicals during first trimester might play important roles in the occurrence of fetal severe CHD.Maternal family history of CHD might associate with fetal simple CHD.

Abstract: Objective　To genotype and analyze whole genomic features of Coxsackievirus B3 (CVB3) isolated in Tianjin, to improve evolution information of CVB3 virus in Tianjin, and to provide basis for surveillance and early warning of related diseases. Methods　Viral RNA was extracted from five CVB3 strains isolated in Tianjin, whole genome sequence of the virus was amplified by RT-PCR and sequenced by next-generation sequencing method, and phylogenetic and recombinant analysis were carried out. Results　The open reading frame 1(ORF) of the five CVB3 strains contained 6 555 nucleotides and encoded 2 185 amino acids, and ORF2 was composed of sequences encoding 68 amino acids. The nucleotide sequence similarity ranged from 78.3%-100%, and the amino acid sequence similarity ranged from 95.7%-100%. Compared with the CVB3 prototype strain, the nucleotide sequence similarity of the five viruses was between 78.2%-79.1%, and the similarity of amino acid sequences was 94.9%-95.3%. All five viruses exhibited a T151A mutation on the VP2 protein. Additionally, the encephalitis isolate showed a K158E mutation on the VP2 protein, while one of the sewage isolates had a C234T mutation in 5' noncoding region. The five strains belonged to two different genotypes, among which the encephalitis isolate in 2016 belonged to the D genotype, while the sewage isolates in 2021 belonged to the E genotype. This is also the first report of E genotype CVB3 in northern China. The CVB3 strain may have recombinant events in non-structural protein regions, in which encephalitis isolate may recombine with a Coxsackievirus B5 (CVB5) strain, while sewage isolates may have recombinant events with a strain of ECHO virus 18 (E18). Conclusions　The CVB3 isolates in Tianjin belong to D and E genotypes, and recombination events may exist in non-structural protein region of the viral genome. The results of CVB3 virus genome analysis in sewage suggests presence of CVB3 infection in the population of Tianjin, and its epidemic dominant genotype may have changed.

Objective To introduce a new modified Bristow operation with mini-open incision and evaluate jts clinical outcomes in treatment of recurrent anteroinferior shoulder dislocation. Methods A total of 11 patients with recurrent anteroinferior shoulder dislocation were treated with modified Bristow op-eration with mini-open incision.The anterior skin incision with 3-5 mm was made to explore the coracoid process with"moving window"technique.Then.the coracoid process and its short head of biceps were os-teotornjed.transferred and fixed on the anteroinferior potion of scapular neck through the rotator cuff inter-space.All patients were followed up for an average period of 15.8 months(6-48 months)by using modified Rowe score. Results The average operation time was 45 minutes(40-65 minutes).The modified Rowe 8core ranged from 80 to 95 points.excellent in all patients, with SUCCESS rate of 100% and without any re-currence or complication. Conclusions Compared with traditional operation.the modified Bristow oper-ation has smaller incision,less trauma.shorter operation duration and better clinical results.

Objective To evaluate the clinical results of proximal humeral internal locking sys-tem(PHILOS)self-locking plates in treatment of proximal humeral fractures. Methods There were 35 patients including 19 males and 16 females(at mean age of 53.5 years,ranging from 29 to 92 years)with proximal humeral fractures treated with PHILOS plate.According to the Neer's classification sys-tem,there were two-part fractures in 19 patients,three-part fractures in 15 and four-part fractures in 1.Surgery was performed with PHILOS serf-locking plate via dehopectoral approach.All patients were fol-lowed up for mean 18.9 months(6-46 months)and evaluated by using Constant score and X-rays. Re-suits X-rays proved that all fractures got good reduction and complete healing during 8-12 weeks.The latest Constant scores was mean 89.5 points(82-94 points).There were no complications including plate loosing.humenral head necrosis or gleno-humeral ioint osteoarthritis. Conclusion PHILOS plate is a good choice for treatment of proximal humeral fractures because of its advantages including easy operation,stable fixation,good clinical results and few complications.

Objective:To investigate the effect of exercise both before and after myocardial infarction (MI) on microvessel density in the left ventricle, and on the mRNA and protein expression of vascular endothelial growth factor (VEGF) and its receptors.Methods:Ninety-six male Sprague-Dawley rats were randomly divided into a sedentary sham operation group (Sed-Sh), a pre-MI exercise sham operation group (PreE-Sh), a sedentary MI group (Sed-MI), a pre-MI exercise group (PreE-MI), a post-MI exercise group (PostE-MI) and a combined exercise group (ComE-MI). The rats of the exercise groups underwent 60min of treadmill training daily five times a week for five weeks, while those of the Sed-Sh and Sed-MI groups remained sedentary. Then all of the rats underwent either an operation to induce an acute MI (Sed-MI, PreE-MI, PostE-MI and ComE-MI groups) or a sham MI operation (Sed-Sh and PreE-Sh groups). Four weeks after the operation, the exercise groups underwent five days adaptive training, followed by 8 weeks of 60min of treadmill training daily, five times a week. All of the rats were then sacrificed. The microvessel densities in the infarcted and uninfarcted volumes were detected using Factor VIII-related antigen staining, while the mRNA expressions of VEGF and its receptors were examined using a real-time polymerase chain reaction. The protein expressions of VEGF and its receptors were measured using western blotting.Results:Compared with Sed-MI group, the average microvessel densities of the PostE-MI and ComE-MI groups had increased significantly, with the average density of the PostE-MI group significantly higher than that of the PreE-MI group, but significantly lower than the ComE-MI group′s average. Compared with the Sed-MI group, the average mRNA expression of VEGF and its receptors in the PostE-MI and ComE-MI groups was significantly higher, without significant differences between the two groups.Conclusions:Pre-MI exercise does not significantly improve left ventricular function or microvessel density, while post-MI exercise does, as it can upregulate the expression of VEGF and its receptors to activate VEGF signaling. The increase in microvessel density may benefit left ventricular remodeling and cardiac function.

Objective:To explor the effects of mentalization-based family therapy (MBFT) on the emotions of anxiety and depression, reflective functions, and related brain regions of adolescents with depression.Methods:Thirty-two adolescent patients with depression were selected in Beijing Huilongguan Hospital.Among them, fifteen patients treated with conventional drugs were defined as the control group, the other seventeen patients served as the observation group.On the basis of routine drug treatment, patients in the observation group were treated with 1.5 h MBFT once a week, for a total of 3 months.Among them, 10 patients underwent resting functional magnetic resonance imaging(rs-fMRI) scans before and after the intervention.SPSS 23.0 was used for statistical analysis of scale evaluation data by using chi-square test and repeated-measure analysis of variance, and pairwise Pearson correlation was used to construct a brain network matrix for the MRI results.The patient's brain network matrix was put into Gretna to calculate, and then use repeated measures analysis of variance and t test for judgment. Results:(1)Repeated measurement analysis of variance showed, over time and group, before and after intervention, the group×time interactions of patients' reflective function, anxiety, depression were statistically significant ( F=5.113, 9.350, 5.264, all P<0.05). Further simple effect analysis showed, in the observation group, there were statistically significant differences in patients' anxiety ((55.09±14.35) vs (30.58±3.62)), depression ((61.58±16.81) vs (31.00±3.48)) before and after intervention (all P<0.01). There was no significant difference in reflective function ((8.71±0.51) vs (8.87±0.50) ) in the observation group before and after intervention( P> 0.05). The reflective function((8.92±0.33) vs (8.73±0.35)) and anxiety((50.67±13.88) vs (45.78±12.89)) scores of the control group were not statistically significant different before and after intervention(all P>0.05), and the depression scores of the control group before and after inter-vention((69.33±9.11) vs (50.94±7.99)) were statistically significant different( P<0.01). (2)rs-fMRI showed that, using the posterior parietal lobe and the medial prefrontal lobe as seed points, the group×time interactions with right inferior frontal gyrus, right fusiform gyrus, right superior temporal gyrus, left middle temporal gyrus and right superior frontal gyrus were statistically significant ( F=29.56, 20.63, 8.69, 9.49, 8.62, all P<0.05). The further simple effect analysis showed in the observation group, the pre-test was less than the post-test, and the difference was statistically significant( P<0.05). In the control group, there were not statistically significant differences in the functional connection strength before and after the test(all P>0.05). (3)The brain network analysis of 264 regions of interest (ROIs) in the whole brain showed that the number of modules in the observation group was less than that in the control group, and the difference was statistically significant ( t=-2.64, P=0.046). Conclusion:MBFT can improve the anxiety and depression of adolescent patients with depression.It enhances the functional link between mentalization related brain regions and the whole brain, and enhances the activity of the whole brain link to become more integrated.