OBJECTIVETo screen the lentiviral vector carrying siRNA and capable of significantly suppressing the ROCK2 gene expression in the corpus cavernosum smooth muscle cells of spontaneously hypertensive rats (SHR).

METHODSWe designed and synthesized 4 siRNA fragments targeting the ROCK2 gene and packaged them into lentiviral vectors. We collected corpus cavernosum smooth muscle cell samples from 5 male SHRs and randomly divided them into groups A (non-transfection control), B (GFP lentiviral transfection), C, D, E, and F (lentiviral transfection with siRNA fragments 1 -4 targeting the ROCK2 gene). Each group consisted of 5 samples and each sample 3 x 10(4) cells. At 48 hours after transfecting MOI = 80 into the SHR corpus cavernosum smooth muscle cells, we detected the expression of GFP under the fluorescent microscope and the mRNA expression of the ROCK2 gene by RT-PCR.

RESULTSThe transfection efficiency of the SHR corpus cavernosum smooth muscle cells was > 50%. Compared with group A, the expression of ROCK2 mRNA in the corpus cavernosum smooth muscle cells showed no remarkable change in group B (P > 0.05) but was inhibited very significantly in C ([43.91 +/- 8.19]%), D ([47.15 +/- 6.64]%), and F ([25.7 +/- 6.03]%) (P < 0.01), and significantly in E ([16.81 +/- 5.94]%, P < 0.05).

CONCLUSIONWe successfully constructed 4 lentiviral vectors carrying siRNA targeting the ROCK2 gene, all of which can significantly suppress the ROCK2 expression in the SHR corpus cavernosum smooth muscle cells, and one has a highly strong inhibitory effect.

Animals ; Gene Silencing ; Genetic Vectors ; Lentivirus ; genetics ; Male ; Myocytes, Smooth Muscle ; Penis ; cytology ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; Random Allocation ; Rats ; Rats, Inbred SHR ; Transfection ; methods ; rho-Associated Kinases ; genetics

Objective To investigate the anti-arrhythmias mechanism of nardostachyos chinensis batal by intracardiac electrophysiologic study. Methods Intracardiac electrophysiologic study were performed in 40 supraventricular tachycardia(SVT)patients before and after oral nardostachys chinensis batal administraton,and the results were compared with 35 SVT patients or normal persons without nardostachys chineusis batal intake. Results The atrial effective,relative,functional refractory periods(AERP,ARRP,AFRP)and ventricular ERP(VERP)were significantly longer in nardostachyos chinensis batal administration group than control(AFRP(358 ± 76)ms vs.(256 ±69)ms;ARRP(365 ±74)ms vs.(287 ±55)ms;VERP(264 ± 36)ms vs.(213 ± 27)ms respectively);but the sinus and atrioventricular node function had no changes. Conclusions The anti-arrhythmia mechanism of nardostachyos chinensis batal were related to the prolonged cardiac refractory periods and the reduced cardiac excitability.

Objective To discuss the significance of abnormal expression of Rb,p53 and PCNA in patients with bladder carcinoma and the correlation with the tumor grade,clinical stage and relapse.Meth- ods Immunohistochemical method was used to detect the Rb,p53 and PCNA gene expression in 60 cases with bladder carcinoma.Results The positive rates of p53,PCNA in 60 cases of bladder cancer is both 40.0 %.There were 27(45 %)cases lack of Rb expression.Multiple altered expression was found in 30 cases (50.0 %).The abnormal expression of Rb,p53 and PCNA was closely related to the tumor grade,clinical stage and relapse.Conclusion The detection of multiple gene is more valuable than that of single gene. Multiple altered expression of Rb,p53 and PCNA and their synergistic action might play an important role in the genesis and development of bladder cancer.

Objective To investigate a new surgical method by using pectoralis minor muscle for its partial transplantation to reconstruct the function of the opposite thumb anastomosed vessels and nerves. Methods The proposed method was evaluated by taking 20 cases of adult cadaveric thoraxes and hands,then compared with the morphology and dimension of both pectoralis minor muscle and palmer muscle to study the feasibility of the new method.Based on the observed data.We selected five suitable cases,in which the opposi- tion function was lost,and then applied the new operation on them with partial transplantation of pectoralis mi- nor muscle anastomosed vessels and nerves according to the results of anatomic study.The follow-up study was conducted to observe the functional recovery of opposition of the thumb.Results The main results can be summarized as follows.First,the anatomical position of pectoralis minor muscle was stable,and every peetoralis minor muscle was provided with self-sustaining artery,vein and nerve.The oppostition process of cadaveric hand succeeded after similar transplantation to clinic operation.Second,follow-up studies conducted 6 - 12 months after the operation showed that all five patients recovered fully.The muscle strength in all five cases re- covered to level four or higher.The shape of palm eminence was satisfactory.Conclusion The surgical method of pectoralis minor muscle transplantation for reconstructing the opposition function of the thumb was based on the clinical and anatomical application.The function of opposition of the thumb reached the satisfacto- ry requirement after the operation.So,the new surgical method can achieve better results than other existing operation methods.

OBJECTIVETo investigate the results of using the modified Jaslow technique to treat the recurent lumbar disc herniation.

METHODSFrom January 2002 to December 2008,62 patients with recurrent lumbar disc herniation were treated with a modified Jaslow technique. There were 42 males and 20 females with an average age of 53.6 years old, ranging from 36 to 70 years. The primary surgical procedures were enlarged fenestration in 20 cases, unilateral semi-laminectomy in 20 cases, bilateral semi-laminectomy in 8 cases and total-laminectomy in 14 cases. The procedures were performed at L3.4 level in 2 cases, L4.5 in 32 cases, L5S1 in 15 cases, L3.4-L4.5 in 3 cases and L4.5-L5S1 in 10 cases respectively. The clinical symptoms included low back pain and radicular pain. Pre-and postoperative JOA score (including subjective symptoms, self-care ability and pain), ratio of disc height and the fusion condition of the involved segments were applied to assess clinical outcome.

RESULTSAll patients were followed up for 1 to 5 years (averaged 3 years). The space height ratio increased from (62.5 +/- 10.4)% to (90.5 +/- 10.3)%, fusion rate was 96.8% (60/62) at the last follow-up. Mean JOA score was (10.42 +/- 2.50) preoperative, improved to (24.26 +/- 2.35) at last follow-up (P < 0.001). The clinic results was excellent in 39 cases, good in 14, fair in 9.

CONCLUSIONThe modified Jaslow technique was a good alternative method for the treantment of recurent lumbar disc herniation with complete decompression, solid interbody fusion and satisfactory clinical result.

Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Intervertebral Disc Displacement ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Spinal Fusion ; methods

Objective To investigate the effect of cell death by HIV-1 infection on gene 90K/Mac-2BP by RNA interference (RNAi) in U937 cell line.Methods We used human monocyte-macrophage cell line U937 as the cell model.Cells were infected by HIV-1 ( R5-tropic) 5 days, and then stained by PE-Annexin V and PerCP-7-AAD.90K/Mac-2BP in U937 cell line was knocked down , and these cells were infected by HIV-1 for 5 days.Then, cells were stained by PE-AnnexinV and PerCP-7-AAD.Apoptosis were examined upon flow cytometry .Results The percentages of Annexin V+cells without 90K/Mac-2BP knock-down were (16.27 ±0.30)% by HIV-1 infection.The percentages of them with 90K/Mac-2BP knock-down were (31.26 ±0.35)%, (25.76 ±0.30)%, (23.69 ±0.33)% respectively.The increase of cell apoptosis rate for HIV-1-infected U937 cells by 90K/Mac-2BP siRNA transfection was significantly greater than that for HIV-1-infected untreated cells (P﹤0.01).Conclusion The apoptosis of HIV-1-infected U937 cells was regulated by the expression of 90K/Mac-2BP.

Objective To observe the muscles and skeleton involvement surrounding sacroiliac joint (SIJ) of axial seronegative spondyloarthropathy (SPA) patients with magnetic resonance imaging (MRI),and to analyze the relationship between them.Methods A prospective study of 38 patients who meet the 2009 axial SpA diagnostic criteria was conducted.We carried out MRI of the SIJ for these patients to evaluate the muscles and skeleton involvement.Those cases were divided into muscles differences between the two groups of image scores,including Spondyloarthritis Research Consortium of Canada (SPARCC) scores and Spondyloarthritis Research Consortium of Canada MRI Sacroiliac Joint Structural Score (SPARCC SSS).The extent of muscles edema in patients with sacroiliac joints was ranked into twelve grades from 0-12,and we did Spearman rank correlation test of muscles edema scope and two indexes.Results We found that 28 cases (73.68％) of the 38 patients had significant muscle involvement by analyzing the STIR sequence,and found erector spinae in 22 cases (57.89 ％)gluteal muscles in 13 cases (34.21 ％),iliacus muscle in 11 cases (28.95 ％),obturator muscle in 5 cases (13.16％),piriform muscle in 5 cases (13.16％) and other 4 cases (10.53％).SPARCC (t =2.28,P =0.03) and SPARCC SSS (t =3.37,P =0.00) were statistically different between the two groups.SPARCC (P =0.00,r =0.67) and SPARCC SSS (P-0.01,r =0.47) were positively correlated with the extent of muscles edema.Conclusions The muscles edema around sacroiliac joint is an important sign of axial SpA magnetic resonance imaging.Patients who had muscles edema tended to have more serious skeleton changes.

Objective To report the clinical result of repairing complex tissue defect in limb caused by traffic accident using both tissue flap transplantation and external fixer. Methods The vascular tissue flap including bone flap and muscular flap to repair the complex tissue defection(soft tissue, bones, joints) in forearm, hand, leg, ankle and foot were applied in 36 cases. At the same time in surgery, injured bones and joints were fixed by external fixer for stabilizing and accelerating cure. Results All 56 eases were applied successfully with transplanted tissue combined with external fixation. The external fixers were moved away about 10-16 weeks(upper extremity) or 6-8 months(lower extremity) later and all the cases recovered satisfactorily. Conclusion It is a reliable method for repairing the complex tissue defection in the limb caused by traffic accident by external fixer of traumarology and microsurgery technique. And it is also characterized as a minor injury method on transplanted bones, enables early exercise for joints, convenient observation of flap circulation and easy nursing.

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Objective To investigate methods and clinical effects of limited open reduction of ankle articular surface and locking compression plate (LCP) placement using minimally invasive plate osteosynthesis (MIPO) in managing tibial Pilon fractures.Methods Twenty-one patients with tibial Pilon fractures treated between August 2009 and August 2011 were involved in the study,including 16 males and 5 females,at age of 21-68 years (average 42.3 years).According to AO/OTA classification,12 patients were with type 43B fractures and nine with type 43C fractures.There were two patients with open fractures (both Gustilo-Anderson type Ⅰ fractures).Limited open reduction of ankle articular surface plus LCP placement using MIPO were performed.Early functional training without weight-bearing was carried out postoperatively.Results Nineteen patients were followed up for 12-24 months (average 14 months).According to Burwell-Chamley' s radiological evaluation system,17 patients obtained anatomical reduction of articular surface and two patients obtained moderate reduction,but they all had bony healing.Ankle function evaluated by Mazur' s criterion were excellent in seven patients,good in 10 and fair in two,with excellence rate of 89％.Conclusion Limited open reduction combined with LCP internal fixation can successfully construct articular anatomic relationship,decrease soft tissue dissection and attain solid fixation in treatment of Pilon fractures and further facilitate functional recovery of the affected extremity in coordination with early functional exercise.