Objective:To construct of transfectant cells expressing ??TCR with specific CDR3 sequence.Methods:Specific CDR3 region sequence of DBS4.3,a known ??T cell clone,was inserted into ?9 and ?2 chain to substitute original CDR3 sequence using overlapping PCR method.After the full-length ?9 and ?2 chains were ligated with expression vector pREP7and pREP9 respectively,they were co-trasfected into a cell line of human Jurkat cells with TCR ? chain gene-defective mutant J.RT3-T3.5.When the transfectant cells expressing specific ??TCR were stimulated by antigen,production of IL-2 was detected by ELISA and Realtime PCR.Results:By ELISA and Realtime PCR,it was exhibited that the transfectant cells expressing DBS4.3 specific ??TCR secreted IL-2 under stimulation of iso-butylamine and anti ??TCR antibody.Conclusion:The transfectant Jurkat cells expressing ??TCR with specific CDR3 sequence are successfully constructed.It provides a platform for the research of recognition mechanism of ??TCR.

Objective To establish a canine model with acute thrombo-embolic pulmonary hypertension monitored by echocardiography. Methods Simulated the pathophysiologic process of acute thrombo-embolic pulmonary hypertension in human, the canine model was developed. The femoral vein puncture and the right heart catheterization to monitor pulmonary artery pressure were operated guided by echocardiography, while the autologous blood clot was injected to the contralateral femoral vein. The criteria of model was the pulmonary artery systolic pressure was higher than 30 mmHg. Results Echocardiography can accurately guide the placement of right heart catheterization and ancillarily observe the pressure testing of pulmonary artery. Of twenty seven healthy experiment dogs, twenty four models with acute thromboembolic pulmonary hypertension were successfully developed. The successful rate was 88.9 %. Conclusions This canine model with acute thrombo-embolic pulmonary hypertension guided by echocardiography is easy to operate and its repeatability is good.

Objective To study the prevalence and trend of obesity among primary and middle school students in Beijing from 1985 to 2005.Methods Morphological data of students aged 7-18 from the National Physical Fitness and Health Surveillances of Chinese School Students in Beijing in 1985,1995,2000 and 2005 was analyzed with methods of cross-sectional study.The prevalence of obesity was calculated,and its secular trends was characterized.Results The prevalence of obesity among students aged 7-18 in Beijing were 0.65%,3.40%,5.23% and 8.53% respectively in the four surveillance programs with obvious rising trend which had an increase.In 2005,it was indicated that the obesity prevalence of subgroups were significantly different from each other.It was higher for males(11.11%)than females(5.98%),also for students aged 10-12(11.63%).In rural schools(6.60%)it was much 10wer than in urban(10.38%),but the areas with high SES was deemed to have a 10wer prevalence.Conclusion In the last two decades,among primary and middle school students in Beijing,obesity had changed its characteristics from sporadic to epidemic with the features of both developed and developing countries.

A surgical technique of lateral crus strut graft to correct the deformity of nostril contour and improve asymmetric nostril shape by autologous rib cartilage was reported. Thirty-five patients (20 males and 15 females), with a mean age of 19 years (range of 16-26 years) were included in this study. All of the patients suffered moderate to severe unilateral cleft lip nostril deformity and underwent surgical technique of lateral crus strut graft by autologous rib cartilage to correct the asymmetric nostril contour from 2010 to 2012. The views of the nostril contour on both sides were reviewed and some parameters were measured at different time points (preoperatively, and 3 months and one year postoperatively) including the long axis and short axis of the nostrils. The differences in nostrils including long axis, short axis, elliptical area and eccentricity on the both sides were compared at different time points. The differences in the long axis and eccentricity of the nostrils on the two sides were decreased significantly between the preoperative view and postoperative view (P<0.05). The surgical technique of lateral crus graft with autologous rib cartilage is effective to correct the abnormal nostril contour and improve the asymmetry for unilateral cleft clip patients.

BACKGROUND: Inhibiting the degradation of extracellular matrix in the intervertebral disc can delay the degenerative process of intervertebral disc. Matrix metalloproteinases-3 (MMP3) is considered as a key enzyme for degradation of extracelular matrix components such as type II collagen and aggrecan.
OBJECTIVE:To construct the short hairpin RNA lentiviral vector targeting human MMP3 gene and to detect its efficiency of gene silence by infecting human degenerative nucleus pulposus cells.
METHODS:According to the human MMP3 mRNA (NM_002422.4) sequence, four groups of the short hairpin RNA gene sequences targeting MMP3 were designed, synthesized and annealed to form double stranded DNA fragments, which were connected with the LV3 vectors digested by BamHI andEcoRI enzymes, and then transfected into the competent cels. The positive clones were identified by PCR, and analyzed by sequencing. The packaging and titer of lentivirus were determined after transfecting 293T cells. Human degenerative nucleus pulposus cels were infected with lentivirus vector, and the transfection efficiency of each group was observed under inverted fluorescence microscope. The interfering efficiency was detected by real time-PCR and western blot at 72 and 96 hours.
RESULTS AND CONCLUSION:The ds-oligo DNA was successfully inserted into the lentiviral vector as confirmed by electrophoresis and sequence analysis. The recombinant lentivirus was harvested from 293T cels with a viral titer of 1-5 ×108 TU/mL. RNA interference targeting the GCC AGG CTT TCC CAA GCA AAT sequences with the highest interfering efficiency in MMP3 gene at 72 and 96 hours resulted in suppression of MMP3 mRNA expression by 98% and 72%, respectively; and at 96 hours, the interfering efficiency of protein expression was 57.2%. The recombinant lentivirus vector containing RNA interference targeting MMP3 gene is successfuly constructed, which lays a foundation for further studies on the MMP3 function and gene therapy.

Objective To evaluate the value of two-dimension and color Doppler flow imaging(CDFI) in diagnosis of deep venous thrombosis(DVT) for different segments of lower extremity compared with digital subtract angiography(DSA). Methods Fifty-eight patients suspected of DVT enrolled in the study. Every segment venous of affected extremities was observed by CDFI. All patients underwent DSA examination. CDFI findings were compared with the results of DSA. Statistical analysis was according to quadruple tabular form. The indexes include sensitivity, specificity, accuracy, false positive rate and so on. Every segment of lower extremity deep venous was evaluated and analysis. Results Total of 150 vessels of fifty-eight patients were detected with CDFI and DSA. Evaluation findings of statistical analysis with quadruple tabular form as follows= sensitivity of common lilac vein was 82.6%, specificity was 66.7%; sensitivity of external lilac vein was 83.3%, specificity was 80.0%; sensitivity of hypogastria vein was 80.0 %, specificity was 66.7%;sensitivity of popliteal vein was 92.6 %, specificity was 92.2 % ;sensitivity of femoral vein was 96.2%, specificity was 93.8% ; sensitivity of anterior tibial vein was 71.4%, specificity was 75.0%; sensitivity of posterior tibial veins was 72.7%, specificity was 75.0%. Conclusions As the first-choice examination for DVT, ultrasonography has different diagnostic value for different segment of venous. It is more sensitive and accurate for femoral, popliteal and external iliac veins thrombus, but less sensitive for anterior,posterior tibial veins and bypass circuit vessels.

Objective To investigate the effect of tea polyphenols on global cerebral ischemia reperfusion injury in rats.Method Forty-five pathogen-free male SD rats weighing 180-220 g were randomly divided into 3 groups( n =15 each):sham operation group (group S),cerebral ischemia reperfusion group (group IR) and tea polyphenols group (group TP).Global cerebral ischemia reperfusion injury was establish by four-vessel occlusion method.At 24 h of reperfusion,five rats were chosen and Evan's blue(EB) was injected iv,and then sacrificed and brain was removed for determination of EB content; another five rats were sacrificed and brain was removed for determination of water content; five rats were chosen for Morris water maze test.Result Compared with group S,EB content and water content in brain tissue were increased in groups IR and 'rP,and escape latency was prolonged,frequency of crossing the original platform was reduced in group IR ( P ＜ 0.05 ).Compared with group IR,EB content and water content in brain tissue were decreased,and escape latency was shortened,frequency of crossing the original platform was increased in group Tp ( P ＜ 0.05).Conclusion Tea polyphenols can attenuate global cerebral ischemia reperfusion injury in rats.

Acquired Immunodeficiency Syndrome ; epidemiology ; Epidemiologic Methods ; Humans ; Risk Assessment

Objective To explore the agreement of anterior chamber angle examination by slit lamp optical coherence tomography (SL-OCT) and gonioscope.Design Case series.Participants Thirteen patients with primary glaucoma (26 eyes) and 8 normal persons(16 eyes).Methods Anterior chamber angle was measured with SL-OCT and gonioscope in turns for temporal,nasal,superior and inferior quadrant.Results of two methods were analyzed and the data were analyzed with Pearson correlation coefficient and Kappa value.Main Outcome Measures Anterior chamber angle.Results The Pearson correlation coefficient of the two methods was 0.86(P=0.00)and the Kappa value is 0.75 (P=0.00).The specificity and sensitivity of detecting occludable angle were 94.7% and 89.4%.Conclusions Anterior chamber angle examination with SL-OCT and gonioscope are well consistent.The specificity and sensitivity for SL-OCT in detecting occludable angle is satisfying.SL-OCT can be regard as an objective assistant method for the diagnosis of angle closure glaucoma.

OBJECTIVETo study the protective effects of Shexiang Xingnaonin (SXN) on focal cerebral ischemia/reperfusion injury and its mechanism.

METHODMiddle cerebral artery occlusion (MCAO) was used to make focal cerebral ischemia/reperfusion model by in travascular nylon filament occlusion. The protective effects of SXN at different doses were evaluated by investigating neurological function score, pathomorphology of brain, the ultrastructure of neuron, expression of tumor necrosis factor-alpha, thrombogenesis in vitro, platelet aggregation and lysing effect of blood clot in vitro.

RESULTCompared with model group, SXN (0.08, 0.16 g x kg(-1)) could decrease the neurological score, improve pathomorphology and neuron ultrastructure of brain, inhibit the expression of TNF-alpha, decrease the length, wet weight and dry weight of thromb and inhibit platelet aggregation. And SXN (0.16, 0.32 g x L(-1)) could dissolve blood clot in vitro.

CONCLUSIONSXN has protective effects on focal cerebral ischemia/reperfusion injury. The role of inhibit the expression of TNF-alpha, inhibit thrombogenesis and platelet aggregation might contribute to its neuroprotective effects.

Animals ; Brain ; drug effects ; ultrastructure ; Brain Ischemia ; drug therapy ; genetics ; metabolism ; prevention & control ; Disease Models, Animal ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; Gene Expression ; drug effects ; Humans ; Male ; Platelet Aggregation ; drug effects ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; drug therapy ; genetics ; metabolism ; prevention & control ; Tumor Necrosis Factor-alpha ; genetics ; metabolism