How to reduce immune response is an unprecedented challenge for rAAV gene medicine. Recent studies suggested that lowering dosage of the vector used could reduce immune response caused by rAAV gene medicine. Nevertheless, it would also decrease the transgene expression, leading to failure of gene treatment. It is therefore important to take appropriate steps to maintain high gene expression level and pharmacodynamic, while the dosage of rAAV used is reduced. Here, steps to enhancing gene therapy, such as optimization of the administration, reconstruction of the viral vector and selection of the promoter, are discussed in order to achieve maximum outcome.

0.05). The scaffolds produced no effect on the cell proliferation and had a low cytotoxicity, with chondrocytes tiled and lots of microspikes and matrixes secreted on the surface. Conclusion: The PCL-b-PLLA scaffold has a good compatibility with cells and is suitable for cartilage tissue engineering.

[Objective] To observe the clinical effects of Zhishang Jiaonang on soft tissue injury of chest wall.[Method] A randomized,positive-controlled trial was adopted,120 cases was divide into trial group(n=90) and the control group(n=30).[Result] 60 cases were cured(66.7%),29 better(32.2%),1 had no effect(1.1%) in trial group;14 cases were cured(46.7%),15 better(50.0%),1 had no effect(3.3%) in control group. With marked difference after statistical dealing,P

Objective To investigate the effects of different doses of dexmedetomidine pretreat-ment on cardiac toxicity of bupivacaine in rats.Methods Forty eight adult male SD rats weighing 250-300 g were randomly divided into 4 groups (n=12):saline control group (group C),dexmedetomi-dine 5 μg/kg group(group D5),dexmedetomidine 10 μg/kg group(group D10)and dexmedetomidine 1 5 μg/kg group(group D1 5 ).A Ⅱ-lead electrocardiogram(ECG)was continuously monitored,the femoral artery was cannulated for direct measurement of MAP and the femoral vein was cannulated for infusion of drugs.Groups D5,D10 and D1 5 were received infusion of dexmedetomidine 5,10 and 1 5μg/kg respectively 1 5 minutes before administration of bupivacaine,while the equal volume of saline was given in group C,then all rats received infusion 0.75% bupivacaine at the rate of 2 mg·kg-1· min-1 until asystole occurred.The doses of bupivacaine and the times of bupivacaine-induced convul-sions,arrhythmia and asystole were recorded respectively,and the myocardial concentration of bupiv-acaine was observed.Results Compared with group C,the doses of bupivacaine and the times of bupivacaine-induced convulsions,arrhythmia and asystole were all increased in groups D5,D10 and D1 5 (P <0.05).Compared with group D5,the above parameters were increased in groups D10 and D1 5 (P <0.05 ).There was no statistical significance of the above parameters between groups D10 and group D1 5.Conclusion Dexmedetomidine pretreatment can raise the threshold toxic dose of bupi-vacaine,delay the time of occurrence of cardiotoxicity of bupivacaine,so that to prevent the cardiac toxicities of bupivacaine in rats,and it produces a dose-dependent protective effect within a certain dose range.

Objective To investigate the role of Rho/Rock signaling pathways in ventilator-in-duced lung injury in rats.Methods Ninety-six male SD rats,weighing 300-350 g,were equally and randomly divided into four groups using a random number table (n =24 each):control group (group C),fasudil group (group F),high tidal volume group (group H)and high tidal volume + fasudil group (group HF).Rats in group C and group F received no mechanical ventilation,rats in group H and group HF were intubated and mechanically ventilated (VT = 40 ml/kg,RR = 40 beats per minutes,FiO 2 =40%)for 4 h.The animals in group F and group HF were given intraperitoneal in-jection of fasudil 10 mg/kg at the time 1 h before mechanically ventilated.Six rats were chosen in each group at the time before ventilation (T0 )and at 4,8,24 h after ventilation (T1-T3 ),and blood sam-ples were taken for determination of the levels of serum tumor necrosis factor-α (TNF-α),IL-6 and IL-10,lungs were removed,bronchoalveolar lavage fluid (BALF)was collected to examine protein content,wet/drying (W/D)ratio was determined,which were then stained with haematoxylin and e-osin and examined under microscope,the pathological changes of lungs were scored.Myeloperoxidase (MPO)activity in lung tissue was determined by spectrophptometry.Protein and gene expression of RhoA and Rock2 in the lung tissue were detected by Western blot and reverse transcription PCR (RT-PCR).Results Compared with group C,the serum TNF-α,IL-6 and IL-10,BALF protein content, W/D ratio,the pathological scores,MPO activity,the expression of RhoA,Rock2 protein and mR-NA were significantly increased in group H and HF at T1-T3 (P <0.05 ).Compared with group H, the serum TNF-αand IL-6,BALF protein content,W/D ratio,the pathological scores,MPO activi-ty,the expression of RhoA,Rock2 protein and mRNA were significantly decreased,and the serum IL-10 was significantly increased in group HF at T1-T3 (P <0.05).Conclusion Fasudil can attenuate ventilator-induced lung injury in rats,and the mechanism may be associated with inhibition of the Rho/Rock signaling pathways reducing the inflammatory response.

Objective To investigate the effects of penehyclidine hydrochloride(PHC)precon-ditioning on hepatic ischemia-reperfusion injury (HIRI)in rats.Methods Fifty-four male Sprague-Dawley rats,weighing 230-250 g,were randomly divided into three groups (n =18 each):sham op-eration group (group S),HIRI group (group HR)and penehyclidine hydrochloride group (group PHC).In group S,the hepatoduodenal ligaments of rats were only pulled and separated,then abdo-mens were closed.In group HR,an atraumatic vascular clip was placed on the vessels blocking the portal venous blood supply to the median and left lateral lobes of the liver for 45 minutes,which re-sulted in approximately 70% rat liver ischemia injury.In group PHC,the rats were treated with 0.45 mg/kg penehyclidine hydrochloride at 30 minutes before the portal venous and hepatic arterial were blocked like group HR.Animals were killed at 2 h (T1 ,n =6),4 h (T2 ,n =6),24 h (T3 ,n =6) after HIRI or sham surgery.Liver tissues and blood samples were taken for analysis.The serum con-centration of ALT and AST were measured as the markers of hepatic functional damage.The TNF-a and IL-1βconcentration were measured by the enzyme linked immunosorbent assay (ELISA)tech-nique.The endothelial nitric oxide synthase (eNOS ) and hypoxia-inducible factor (HIF-1α) expression were measured by immunohistochemical staining.Results Compared with group S,the expressions of ALT,AST,TNF-αand IL-1βin groups HR and PHC increased at T1-T3 (P <0.05), which were similar to the change trend of HIR-1αand eNOS expressions (P <0.05).Compared with group HR,the expression of ALT,AST,TNF-αand IL-1βin group PHC decreased at T1-T3 (P <0.05).Nevertheless expression of HIF-1αand eNOS of groups HR and HPC were increased than that of group S,and experssion of HIF-1αand eNOS of group HPC were increased than that of group HR at T1 and T2 (P < 0.05 ).Conclusion PHC preconditioning can protect the liver from HIRI.The mechanism may be associated with the up-regulation of eNOS and HIF-1α,as well as reducing the in-flammatory response.

A safe and effective targeting viral vector is the key factor for successful clinical gene therapy. microRNA, a class of small, single-stranded endogenous RNAs, act as post-transcriptional regulators of gene expression. The discovery of these kind regulatory elements provides a new approach to regulate gene expression more accurately. In this review, we elucidated the principle of microRNA in regulation of targeting viral vector. The applications of microRNA in the fields of elimination contamination from replication competent virus, reduction of transgene-specific immunity, promotion of cancer-targeted gene therapy and development of live attenuated vaccines were also discussed.
Gene Expression Regulation ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; MicroRNAs ; genetics ; Viruses ; genetics ; metabolism

Objective To investigate the effect of stellate ganglion block (SGB) on the postoperative cognitive function in aged rats.Methods Sixty-four male Sprague-Dawley rats,aged 18-20 months,weighing 500-700 g,were randomly divided into 4 groups (n =16 each):control group (group C),operation group (group O),normal saline + operation group (group NS) and SGB + operation group (group SGB).Group SGB received right SGB with 0.25％ bupivacaine 0.15 ml,while the equal volume of normal saline was given in group NS.Groups O,SGB and NS underwent 30 min of exploratory laparetomy starting from 15 min after the end of administration.Morris water-maze test was performed at days 1-6 after operation in 10 rats chosen from each group.The escape latency and frequency of crossing the original platform were recorded.Two rats were chosen from each group at 1,2 and 3 d after operation and sacrificed and the hippocampi were removed for microscopic examination.Results Compared with group C,the escape latency wag significantly prolonged on 1-5 days after operation in groups O and NS,the escape latency wag significantly prolonged on 1 and 2 days after operation in group SGB,and the frequency of crossing the original platform was decreased in groups O,NS and SGB (P ＜ 0.05).Compared with group NS,the escape latency wag significantly shortened,and the frequency of crossing the original platform was increased on 1-5 days after operation in group SGB (P ＜ 0.05).The number of hippocampal neurons was significantly larger at 2 and 3 d after operation in group SGB than in group NS.Conclusion SGB can improve the postoperative cognitive function in aged rats.

Objective To evaluate the effect of stellate ganglion block (SGB) on cellular immune function in diabetic rats.Methods Healthy male Sprague-Dawley rats,aged 3 months,weighing 240-280 g,were used in this study.Diabetes mellitus was induced by intraperitoneal 1％ streptozotocin 60 mg/kg and confirmed by blood glucose ≥ 16.7 mmol/L 3 days later.Forty-eight rats with diabetes mellitus were randomly divided into 2 groups (n=24 each) using a random number table:diabetes mellitus group (group DM) and group SGB.Another 24 healthy rats,aged 3 months,were selected and served as control group (group C).At 1 week after successful establishment of the model,unilateral transection of cervical sympathetic trunk (TCST) was performed in group SGB,while the right cervical sympathetic trunk was only exposed in C and DM groups.Before TCST (T0) and on 1,3,7 days after TCST (T1-3),6 rats were randomly selected from each group,and blood samples were collected from the inferior vena cava for determination of the blood glucose,plasma norepinephrine (NE) concentrations (by enzyme-linked immunosorbent assay),and levels of T lymphocyte subsets CD3+,CD4+ and CD8+ in whole blood (using FACSCalibur flow cytometer).C D4+/CD8+ratio was calculated.The rats were weighed before sacrifice,and the rats were sacrificed to obtain the thymus which was weighed.The thymus index (thymus weight/body weight) was calculated.Results Compared with group C,the blood glucose was significantly increased,and the levels of CD3+ and CD4+ in whole blood,CD4+/CD8+ ratio,and thymus index were significantly decreased at T0-3 (P＜0.05),and no significant change was found in CD8+ levels in DM and SGB groups (P＞0.05),the plasma NE concentrations were significantly decreased at T1-3 in group SGB (P＜0.05),and no significant change was found in plasma NE concentrations in group DM (P＞0.05).Compared with group DM,the blood glucose and plasma NE concentrations were significantly decreased,and the levels of CD3+ and CD4+ in whole blood,CD4+/CD8+ ratio,and thymus index were significantly increased at T1-3 (P＜0.05),and no significant change was found in CD8+ levels in group SGB (P＞0.05).Conclusion SGB can improve the cellular immune function in diabetic rats.