Objective To observe the transfection efficiency and anti-fibrotic effect of miR-29b transfected by anti-TGF-β Ⅱ R ScFv/Ck/tP fusion protein (new vector) in hepatic stellate cell (HSC),and to provide a new vector in gene therapy for liver fibrosis.Methods The liposome vector,new vector,and lentiviral vector were used as transfection reagents to transfect miR-29b into HSC.Transfection efficiency was observed under fluorescence microscope and flow cytometry.Collagen α1 (Ⅰ) mRNA and protein expression in different groups were analyzed by real-time RT-PCR and Western Blot,respectively.Results Compared to the control,transfection efficiencies in lentiviral vector,new vector,and liposome vector groups were about 70％,58％,and 29％,respectively.Collagen α1 (Ⅰ) mRNA expression in lentiviral vector,new vector,and liposome vector groups was decreased by about 70％,50％,and 38％,respectively ((t =6.316,P ＜0.01 ; t =4.082,P ＜0.01 ; t =3.014,P ＜0.05).Collagen α1(Ⅰ) protein expression in lentiviral vector,new vector,and liposome vector groups was decreased by about 59％,41％,and 27％,respectively (t =4.209,P ＜0.01; t =4.033,P ＜0.01; t =2.842,P ＜0.05).Conclusions The new vector constructed by us has a high transfection efficiency.MiR-29b transfected by the new vector has a good anti-liver fibrosis effect.

Objective To observe the effect of small interfering RNA(siRNA)expression plasmids targeting transforming growth factor p receptor(TαR)Ⅰ gene on the collagen synthesis of hepatic stellate cells(HSCs).Methods Three siRNA expression plasmids were designed and constructed according to TBR Ⅰ sequence.Then the plasmids were transfected into HSC-T6 using 1ipofectamine2000 reagent. The mRNA and protein expressions of TβR Ⅰ were analyzed by reverse transcription polymerase chain reaction(RT-PCR)and Western blot technique, respectively. The cell proliferation was detected using methylthiazo-lyldiphenyl-tetrazolium bromide(MTT)methods. Concentrations of haluronic acid and type Ⅲ pro-collagen in the supernatants were determined by radioimmunoassay. The data were analyzed using least significant difference(LSD).Results Three recombinant plasmids expressing siRNAs were successfully constructed and confirmed by restriction enzyme assay. Compared with the blank control,all the three recombinant plasmids could inhibit the expressions of TβR Ⅰ mRNA,of which plasmid expressing siRNA2 exhibited the strongest inhibitory effect(psiRNA1 group:t=7.354,P＜0.01;psiRNA2 group:t=9.214,P＜0.01;psiRNA3 group:t=5.967,P＜0.01).The expressions of TβR Ⅰ protein were also reduced by all the three recombinant plasmids,of which the plasmid expressing siRNA2 showed the strongest inhibitory effect(psiRNA1 group: t=6.324,P＜0.01;psiRNA2 group:t=8.741,P＜0.01;psiRNA3 group:t=4.128,P＜0.01).The proliferation activity and collagen synthesis of HSCs also decreased in all three HSC groups treated with recombinant plasmids, of which, again, plasmid expressing siRNA2 exhibited the strongest inhibitory effect. However, no significant change was observed in HSCs transfected with non-related siRNA. Conclusion Recombinant plasmids targeting TβR I can inhibit collagen synthesis, which suggests a novel target for gene therapy of liver fibrosis.

Objective:To study heart mitochondrial calcium homeostasis during ischemia- reperfusion under the protection of cardioplegic solution.Methods:Isolated perfused rabbits hearts were subjected to global ischem ia in the presence of cold cardioplegic solution for an hour and reperfused for half an hour.Electron m icroscope was used to observe the change of mitochondrial structure.Heart m itochondria was isolated and Fura2 - AM was used as the indicator of matrix concentration of calcium([Ca2 + ]m) to m easure[Ca2 + ]m,the intake and outtake ability of the calcium when m itochondria was at high calcium concentration liquid or high natrium concentration liquid.Results:[Ca2 + ]m increased significantly but mitochondria was still able to uptake calcium and release calcium of m atrix.There was some electron- dense deposit in mitochondria and the injury of mitochondria could be seen by electron microscope.Conclusion:The reason for mitochondria calcium overload at the beginning of reperfusion include physiological and pathological calcium uptake,which ends after30 min reperfusion under the protection of cardioplegic solution. [

OBJECTIVE:To determine the pharmacokinetics and relative bioavailability of oxetacillin and sulbactam in oxetacillin-sulbactam tablets.METHODS:20healthy male volunteers were randomly divided into2groups.In a crossover de?sign,a single oral dose of two tablets of domestic oxetacillin-sulbactam tablets or imported Tai Ba Meng LBL 500 tablets(one tablet of the two formulations contains oxetacillin250mg,sulbactam250mg)were given,then the drug concentrations in plas?ma were assayed by RP-HPLC.RESULTS:The pharmacokinetic parameters of oxetacillin in domestic and imported tablets were as follows:T max were(1.04?0.26)h and(1.10?0.28)h;C max were(5.16?1.30)mg/L and(5.25?1.21)mg/L;AUC 0～∞ were(13.30?3.12)mg/(h?L)and(13.33?3.53)mg/(h?L)respectively.The pharmacokinetic parameters of sulbactam in domestic and imported tablets were as follows:T max were(0.80?1.0)h and(0.81?0.17)h,C max were(5.73?1.44)mg/L and(5.70?1.46)mg/L,AUC 0～∞ were(13.25?2.41)mg/(h?L)and(13.28?2.26)mg/(h?L)respectively.CONCL_ USION:Domestic oxetacillin-sulbactam tablets and imported Tai Ba Meng LBL 500 tablets were bioequivalent.

Objective To observe the effects of adiponectin on mRNA and protein expressions of connective tissue growth factor (CTGF) in hepatic stellate cells (HSCs) and the levels of procollagen type Ⅲ (PC Ⅲ) and hyaluronic acid (HA).Methods Cultured rat HSCs were treated with different concentrations of adiponectin.CTGF mRNA and protein expressions were analyzed by reverse transcription polymerase chain reaction (RT-PCR) and Western Blot.The levels of PCⅢ and HA were detected by enzymelinked immunosorbent assay (ELISA).Results Compared with the control group,the result of RT-PCR showed that the four groups had different degrees of inhibitory effect,of which group D exhibited the strongest inhibitory effect.The absorbance ratio was 1.54 ±0.18,1.21 ±0.14,0.96 ±0.10,and 0.79 ± 0.08,respectively (t =2.42,P ＜0.05;t =2.73,P ＜0.05;t =3.28,P ＜0.01;t =4.67,P ＜0.01).Western Blot also indicated that four groups had different degrees of inhibitory effect,of which D group exhibited the strongest inhibitory effect.The ratio of integral absorption was 1.54 ± 0.18,1.21 ±0.14,0.96±0.10,and 0.79 ±0.08,respectively (t =2.84,P ＜0.01;t =4.05,P ＜0.01;t =6.25,P ＜0.01;t =9.72,P ＜0.01).The levels of PCⅢ and HA secreted in culture media were also decreased.It was significantly decreased with the concentration of adiponectin increased.Conclusion Adiponectin can inhibit the levels of PC Ⅲ and HA,which may be achieved through reducing CTGF mRNA and protein expressions.

Objective To investigate the effect and mechanism of ischemic postconditioning (IPC) on myocardial perfusion levels of acute ST-segment elevation myocardial infarction (STEMI) patients having underwent primary percutaneous coronary intervention (PCI), and the safety of IPC. Methods One hundred and sixty patients with STEMI were enrolled, and they accepted the primary PCI therapy within the onset of 12 h. The patients were divided into 2 groups according the treatment method:control group (routine PCI group, 82 cases) and IPC group (78 cases). The ST-segment resolution, TIMI myocardial perfusion grade (TMPG), before and after PCI levels of nitrogen monoxidum (NO), endothelin (ET)-1, tissue-type plasminogen activator (t-PA), plasminogen activator inhibitor (PAI)-1, rate of intraoperative complication were observed. The patients were followed up for 6 months, the rate of major adverse cardiac event (MACE) was recorded. Results The rates of ST-segment resolution and TMPG well in IPC group were significantly higher than those in control group:84.62%(66/78) vs. 67.07%(55/82) and 80.77%(63/78) vs. 64.63%(53/82), and the rate of ischemia-reperfusion injury in IPC group was significantly lower than that in control group: 7.69%(6/78) vs. 24.39%(20/82), and there were statistical differences ( P<0.05 or<0.01). The endothelial function and fibrinolysis activity indexes (NO, ET-1, t-PA and PAI-1 levels ) 7 d after PCI in IPC group were significantly better than those in control group: (52.37 ± 3.84) μmol/L vs. (50.95 ±3.85) μmol/L, (75.47 ±3.47) ng/L vs. (76.61 ±3.72) ng/L, (12.96 ±1.25) μg/L vs. (12.52 ±1.23) μg/L, (21.78 ±4.01)μg/L vs. (24.95±7.56)μg/L, and there were statistical differences (P<0.05 or<0.01). There was no statistical difference in the rate of intraoperative complication between 2 groups ( P>0.05). The rate of MACE in IPC group was significantly lower than that in control group:3.85% (3/78) vs. 14.63% (12/82), and there was statistical difference (P<0.05). Conclusions Applying IPC in patients with STEMI having underwent primary PCI is safe and can improve myocardial perfusion levels. The improvement of vessel endothelial function and fibrinolysis activity attained from IPC may be the major mechanism.

Objective To compare the postoperative cognitive function in elderly patients undergoing sevoflurane-based anesthesia versus propofol-based anesthesia.Methods Sixty-two patients of both sexes,aged 65-80 yr,weighing 48-90 kg,of ASA physical status Ⅰ or Ⅱ,were randomly allocated to either sevoflurane-based anesthesia group (group S,n=31) or propofol-based anesthesia group (group P,n =31).At 1 day before operation and 7 days after operation,cognitive function was assessed by MiniMental State Examination,Digit Span Test (forward test and backward test),Digit-Symbol Substitution Test,Trail Making Test A and Word Recognition Test,and the scores were recorded.Results There were no significant differences between the two groups in the scores of each test used for assessment of cognitive function.Compared to group P,Mini-Mental State Examination scores,forward test scores and backward test scores obtained from Digit Span Test,Digit-Symbol Substitution Test scores,and Word Recognition Test scores were significantly decreased at 7 days after operation,and no significant change was found in Trail Making Test A scores in group S.Conclusion Postoperative cognitive function is decreased under sevoflurane-based anesthesia when compared with that under propofol-based anesthesia in the elderly patients.

As the neuron-specific actin binding protein,Drebrin ( developmentally regulated brain protein) can affect spiny and synaptic morphology and function by changing the property of cytoskeleton,and modulate synaptic plasticity. Neural excitability regulates expression and activitiy of Drebrin through a variety of signaling molecules,making Drebrin' s function correlated with that of neurons. Under pathological conditions,the abnormal Drebrin affects synaptic plasticity,which leads to different degrees of cognitive dysfunction,and it is closely relates to the progress of cognitive dysfunction. Extensive studies of physiological and pathological functions of Drebrin in overall and molecular ways will not only contribute to a thorough understanding of cognitive dysfunction,but also develop the new target of therapeutics for cognitive dysfunction.

Objective To evaluate the efficacy of percutaneous CT guided interstitial 125Ⅰ seeds implantation treatment for refractory pelvic malignant tumors and discuss the procedure of technique.Methods Twenty-three patients with refractory pelvic malignant tumors(25 lesions,diameter 3.5-7.0 cm,mean 4.5 cm) showing failure response to full chemotherapy and/or radical radiotherapy after tumors resection were undergone percutaneous CT guided intratumoral 125Ⅰseed implantation.Treatment plan system(TPS)was used to design the distribution and number of 125Ⅰ seeds according to matched peripheral dose(MPD)1-3 days before the procedure.Of which 6 cases received combined internal iliac arterial infusion chemotherapy before or after the 125Ⅰ seed implantation procedure.Results 9 ～ 75(mean 27)125Ⅰ seeds were implanted into a single tumor at first time including 6 patients with intraarterial chemotherapy for 14 cycles(mean 2.3 cycles),showed relief of clinical pain symptoms in 16 of 23 cases,72 h ～ 4 w after the seeds implantation;with the an effective rate of 69.6%.Follow up for 2-34 months(median,21 months),CT or MRI performed 2 months after the seeds implantation showed no CR,but PR in 18 cases,SD in 4 cases,and PD in 1 case,with overall response rate of 78%(18/23),and no serious complication.In addition,20 cases survived,with the longest one of 34 months and the other 3 died.Conclusions Intratumoral 125Ⅰ seeds implantation under CT guidance for pelvic refractory malignant tumors is safe,minimally invasive,and effective.

Objective: To investigate the psychosocial characteristics and mind-physique status of pa-tients with gynecological malignant tumors. Methods: Clinical psycho-rating scale [life event scale (LES) and symptom checklist-90 (SCL-90)] was applied to test life events and mind-physique status of 60 cases of pa-tients with gynecological malignant tumor. The results were compared with those of 40 female patients with benign tumors and 40 normal females. Results: The frequency and tension value of total life events and those of negative life events in the malignant cancer group were much higher than those in the benign tumor group and normal group (P<0.05) and no remarkable differences were found in the frequency of positive life events among the three groups. The frequencies of total life events and negative life events were higher in the be-nign tumor group than those in the normal group (P<0.05) but there were no significant differences in the fre-quency of positive life events, total tension value or positive tension value among the three groups. No signifi-cant differences were found in the indices of SCL-90, positive scores and scorns of other factors among the three groups (P>0.05). The scores of somatization, obsession, interpersonal relationship sensitivity, anxiety, hostility, phobia, and paranoia were higher in the malignant tumor group than in the benign tumor group and normal group (P<0.05). The average positive scores and the depression scores in the benign tumor group were higher than those in the normal group (P<0.05), but there was no statistical significance in other indices between the benign tumor group and the normal group (P>0.05). Conclusion: Negative psychosocial factors are closely correlated with the occurrence and growth of gynecological malignant tumors. Active psychological intervention should be performed as eady as possible.