Objective To explore the curative effectiveness of transplantation of bone marrow-derived liver stem cells (BDLSCs) transfected with rat interleukin-10 (IL-10) gene in the treatment of liver fibrosis in rats. Methods β2m-/Thy-1+BDLSCs isolated from Wistar rats were transfected with adenovirus-mediated rat IL-10 gene Wistar rats were subcutaneously injected with CCl4 to induce liver fibrosis, and randomly divided into three groups as follows: (1) the model group, infused with 1 ml normal saline (NS); (2) the BDLSC group, infused with NS containing untreated β2m-/Thy-1+BDLSCs (2×105 cells); (3) the IL-10 group, infused with NS containing β2m-/Thy-1+ BDLSCs transfected with IL-10 gene (2×105 cells). Infusion was done via the portal vein. Rats subcutaneously injected with olive oil served as control (the normal group). The BDLCs labeled with diamidine phenylindole dihydrochloride (DAPI) in the liver were localized. Pathological changes and collagen area in liver tissues were observed. Liver function and blood blotting function were tested. Results β2m-/Thy-1+ BDLSCs labeled with DAPI were observed in liver tissues of rats. Significant pathological changes of liver tissues were observed in the model group. Compared with the model group, pathological changes were alleviated to some extent in the BDLSC group. The morphology of liver tissue in the IL-10 group was mostly close to that in the normal group. Collagen deposition of liver tissues was increased obviously in the model group. However, transplantations of untreated and IL-10-transfected BDLSCs both reduced collagen area. Compared with the BDLSC group, collagen deposition was significantly suppressed in the IL-10 group. Transplantation of IL-10-transfected BDLSCs suppressed obviously the levels of ALT, AST, ALP, TBIL, PT and APTT as compared with the model group (P<0. 05). The levels of ALT, TBIL, PT and APTT in the IL-10 group were significantly reduced to the normal levels as compared with those in the BDLSC group (P<0. 05).Conclusions Transplantation of BDLSCs transfected with rat IL-10 gene was effective in treating liver fibrosis in rats. This combined strategy of IL-10 gene and BDLSCs may represent a feasible, effective and safe therapy form for liver fibrosis.

Objective:To construct a eukaryotic expression vector of leptin gene,and to transfect it into human placental mesenchy-mal stem cells(HPMSCs)and appraise its expression.Methods:Primers were designed and the leptin gene was obtained by RT-PCR from human adipose tissue.The aimed segments were inserted into the eukaryotic expression vector pIRES2-EGFP,plasmid pIRES2-EGFP-LEP was constructed and identified by restricted enzymatic resection,and then transfected it into HPMSCs by liposome.The ex-pression of leptin in the transfected cells was detected by RT-PCR and Western blotting,the multi-differentiation potential of the cells was indentified.Results:The length of specific fragment was 500 bp,the recombinant plasmid pIRES2-EGFP-LEP presented 5.3 kb and 500 bp bands by restriction enzyme digestion.Leptin gene was expressed in transfected HPMSCs and the transfected HPMSCs maintained multi-directional differentiation potential.Conclusion:The eukaryotic expression vector of leptin can be transfected and ex-pressed in HPMSCs.

Objective This retrospective study is to analysis the special medical conditions that most Chinese secondary hospitals are facing with,and to review the safeguards and pitfalls for arterial switch operations,in order to probe intothe feasibility of this procedure for Chinese secondary hospitals and provide our experiences to help other surgeons to avoid pitfalls on complex procedures.Methods Between January 2006 and December 2011,totally 21 newborns and infants with TGA/VSD and TGA/IVS underwent arterial switch operation.There were 15 males and 6 females.In the TGA/VSD group,there were 16 cases,ranging from 30 days to 1 year at surgeries,and weight from 3.4-8.5 kg with average of (5.33 ± 1.42) kg.In the TGA/IVS group,there were 5 cases,ranging from 13 days to 1 month at surgeries,and weight from 3.1-5.5 kg with average of (3.75 ± 1.17)kg.All patients underwent one stage of arterial switch operation.Routine follow-up checking points are set at discharging,3 months,half year and every year after operation.Results The early death rate is 9.5％ (2/21),and the reexploration rate is 9.5％ (2/21).In the TGA/VSD group,average cardiopulmonary bypass time is (151 ± 33) minuntes with the aortic crossclamp time is (1 19 ± 26) minutes.Ventilator support time is 24-159 hours,and the length of ICU stay is 3-17 days.1 case has residual VSD with the diameter less than 2 mm.The pulmonary flow velocity in 2 cases increase mildly to 2.0 m/s and 2.2 m/s,and another 2 cases increase severely to 3.1 m/s and 3.7 m/s.The aortic flow velocity in 3 cases increase to 2.0m/s.ECG instructs no case has myoinfarction signs.In the TGA/IVS group,average cardiopulmonary bypass time is (170 ± 52) minuntes with the aortic crossclamp time is (137 ± 48) minutes.Ventilator support time is 51-144 hours,and the length of ICU stay is 4-14 days;The pulmonary flow velocity in 2 cases increase mildly to 2.0 m/s.The aortic flow velocity in 1 cases increase to 2.0 m/s.ECG instructs no case has myoinfarction signs.Conclusion Unbalanced medical resources distribution causes significant differences between heart centers and the secondary hospitals in China,especially on the complex congenital heart diseases procedures.However,with relatively solid background on correction of the simple congenital heart diseases,the Chinese secondary hospitals can still perform arterial switch operation with satisfactory mortality and morbidity,and provide more prompt medical services for more population.

Aim To observe the effect of antibody NCX-3F10 on the main ion current of rat ventricular myocytes and its effect on arrhythmias induced by ischemia/reperfusion(I/R).Methods ① The whole-cell patch clamp technique was employed to record the Na+/Ca2+ exchange current(INa/Ca) and other major ion currents in rat ventricular myocytes.② The rat models of arrhythmia induced by ischemia/reperfusion were established by ligating the left coronary artery to in vivo and in vitro.Then the effects of antibody on the arrhythmia were observed.③ The IonOptix ion imaging system was used to observe the effect of antibody on calcium transients in single ventricular myocytes.Results ① The antibody NCX-3F10 dose-dependently inhibited INa/Ca from 5 to 40 mg·L-1.The IC50 for outward and inward currents was 11.15 and 11.69 mg·L-1, and the maximum inhibitory rates were 61% and 62%, respectively.The antibody also had an inhibitory effect on calcium current(ICa-L), and had no significant effect on inward rectifier potassium current(IK1), transient outward potassium current(Ito) and sodium current(INa).② In the isolated rat heart group I/R, 100% rats showed ventricular tachycardia, and 88.89% rats had ventricular fibrillation.After administration of antibody NCX-3F10(10 mg·L-1) 5 min before reperfusion, the incidence of ventricular tachycardia decreased to 44.43%(P<0.05), and the duration of ventricular tachycardia and ventricular fibrillation was also shortened remarkably(P<0.05).③ In the anesthetized rats after administration of antibody NCX-3F10(50 μg·kg-1) 5 min before reperfusion, the incidence and duration of ventricular tachycardia,the incidence and duration of ventricular fibrillation, and total number of ventricular premature beats were significantly decreased(P<0.05).④ From 5 to 40 mg·L-1, NCX-3F10 antibody decreased calcium transient amplitude in rat single ventricular myocytes dose-dependently(P<0.05).Conclusions The NCX-3F10 antibody shows significant arrhythmic effects on ischemia-reperfusion induced arrhythmia in rats both in vitro and in vivo, the underlying mechanism of which is related to NCX and L-type calcium current inhibition and calcium overload reduction by the NCX antibody.

AIM: To investigate the effect of zacopride, an inward rectifier potassium channel agonist, on ouabain-induced arrhythmias in adult rats, and to explore the underlying electrophysiological mechanism.METHODS: Using ouabain to establish in vitro and in vivo arrhythmic rat models, the effects of zacopride on ouabain-induced arrhythmias were observed.The technique of whole-cell patch clamp was used to observe the effects of zacopride on inward rectifier potassium current (IK1), resting membrane potential (RMP) and delayed afterdepolarizations (DADs) in single rat ventricular myocyte.RESULTS: Zacopride at 1 μmol/L significantly reduced total number of premature ventricular beats, and the duration and incidence of ventricular tachycardia and ventricular fibrillation induced by ouabain in rat hearts in vitro (P<0.05).In anesthetized rats, zacopride at 15 μg/kg significantly reduced total number of premature ventricular beats, and the duration and incidence of ventricular tachycardia and ventricular fibrillation induced by ouabain (P<0.05).IK1 was significantly inhibited by ouabain (P<0.05), which was partially and even completely reversed by zacopride at 0.1~10 μmol/L.RMP value was significantly reduced by ouabain (P<0.05), and then increased to different levels after treatment with zacopride (0.1~10 μmol/L).Zacopride at 1 μmol/L showed its maximal effect and RMP was restored to normal level.Moreover, zacopride at 1 μmol/L markedly suppressed ouabain-induced DADs in single rat ventricular myocyte.The incidence of DADs decreased from 91.67% to 12.50% after zacopride was applied (P<0.05), and this effect was abolished by 1 μmol/L BaCl2.CONCLUSION: Inward rectifier potassium channel agonist zacopride significantly inhibits ouabain-induced ventricular arrhythmias in adult rats.The mechanism is related to increased RMP level and inhibition of DADs by activation of IK1 channel.

Objective To study the effects of IL-4 in prostate cancer cells glycometabolism and proliferation. Methods We used IL-4 to treat PC3 cells, then tested the changes of LDH-A expression by RT-qPCR, Western Blot, CCK-8 and lactate production assay. Results Our data showed that IL-4 induced LDH-A up-expression in PC3 cells at mRNA and protein levels. Also, IL-4 promoted the proliferation activation and increased lactate production in PC3 cells. Conclusion IL-4 can strengthen the proliferation activation in PC3 cells by up-regulating LDH-A expression.

Objective To study the role of vagusism on diethytlnitrosamine (DEN)-induced hepatocellular carcinoma in rats and to explore its mechanism.Methods The rat vagus nerve was labeled with 1,1'-dioctadecyl-3,3,33 -tetramethylindocarbocyanine perchlorate (DiI) tracer.The rats were divided into three groups:cancer induced rat models (cancer-induced group,n =20),cancer-induced and vagal detachment models (combined group,n =20) and normal rats (negative group,n =20),HE staining was performed on the liver tissue biopsied from the three groups of rats.The pathological grading score was evaluated and statistically analyzed.Immunohistochemistry (IHC) staing was used to determine the expression of vagal neurotransmitters and related receptors in liver tissue.Results The optimal concentration (1 mg/ml) and fixation duration with formalin (4 days) of DiI tracer marker vagal nerve demonstrated superior distribution and density of vagus nerve in the liver tissue.The incidence of hepatocellular carcinoma was significantly higher in the cancer-induced group (75％) than combined group (15％),the difference was statistically significant (P ＜ 0.05).IHC staining results of acetllcholine showed a significant difference between cancerinduced group (9.10) and combined group (2.30) or negative group (2.95),the difference was statistically significant (P ＜ 0.05).The M1 receptor expression was significantly higher in cancer-induced group (6.00) than combined group (2.30) and negative group (2.55),the differences were statistically significant (P ＜ 0.05).There was no significant difference of M2 receptor expression among the cancer-induced group (1.30),combined group (1.25) and negative group (1.35) (P ＞ 0.05).The M3 receptor expression detected with IHC staining shown there were statistical differences among cancer-induced group (8.95),combined group (6.30) and negative group (3.60) (all P ＜ 0.05).Conclusion The vagus nerve may play an important role in the development of DEN-induced hepatocellular carcinoma in rats.

Objective To investigate the effects of CD147/MMP-9 pathway on early left ventricular remodeling Methods 30 healthy eight-week male SHR were divided into 3 groups (n = 10 for each group). SHR group received tail vein injections of normal saline weekly; CD147 group received CD147 of 600 ng·kg-1 weekly; and CD147+DOX group received CD147 of 600 ng/kg weekly and intragastric administration of DOX ( doxycycline ) of 30 mg/kg daily . 10 healthy eight-week male Wistar-Kyoto rats (WKY group) were treated as SHR group. Echocardiography, myocardial sections microscopy examination (HE and VG stain), and Western blot (for assessing levels of MMP-9, TIMP-1, CD147, and collagen I and Ⅲin myocardial tissues) were performed on day 56. Left ventricular weight index (LVWI)was measured and calculated. Collagen volume fractions (CVF) were obtained by image analysis. Results As compared with WKY group , levels of CD147 , MMP-9 , and MMP-9/TIMP-1 were lower but TIMP-1 and collagenⅠand Ⅲ were significantly higher in SHR group. The abundance of CD147 and MMP-9 protein and the ratio of MMP-9/TIMP-1 were obviously increased in CD147 group than in SHR group (P < 0.05). Levels of CD147, MMP-9, and MMP-9/TIMP-1 did no differ between CD147+DOX group and CD147 group. LVWI and contents of collagenⅠand Ⅲ were obviously declined in CD147 group as compare with SHR group. Cardiomyocyte hypertrophy , partial myocardial fibre rupture , myocyte dissolution and fuzzy myocardial fibre boundaries , more abundant of collagen fibers, and higher CVF were found in SHR group. Cardiac fibrosis was significantly improved after CD147 intervention, but the action was suppressed as DOX was administrated simultaneously. Conclusions Early ventricular remodeling may be involved in the inhibition of CD147/MMP-9 pathway in SHR. Input of CD147 to upregulate the pathway can improve the remodeling.

Objective To explore the best cut-point of waist-to-heisht ratio (WHtR) for identifying metabolic syndrome(MS)in adolescents.Methods A cross-sectional study was conducted in 4 507 adolescents aged 13-18 years in Qinhuangdao.Receiver operating characteristic (ROC) curve analysis was employed to determine the optimal WHtR cut-point for detecting MS.Results The prevalences of MS were higher among higher WHtR in both boys and girls.Odds ratios for MS significantly increased from the WHtR≥0.45 in boys(OR =13.85,95%CI 4.08-46.97)and in girls (OR=12.42,95%CI 2.62-58.96,P<0.01).The ROC curve analysis showed that the optimal WHtR cut-point was 0.45 in boys with the sensitivity of 89.4%and specificity of 81.4%.and in girls with sensitivity of 90.3%and specificity of 86.5%.Conclusion There is a significant correlation between WHtR and MS.Optimal WHtR cut-point for predicting MS is 0.45.

Objective To prepare a red blood cells based multifunctional nanoscaled drug delivery system,and to study its in vitro photothermal and photodynamic effects.Methods The indocyanine green (ICG)/doxorubicin (DOX) co-loaded nanoscaled red blood cells (DIRAs) were prepared using an extrusion method.The morphology,particle size,encapsulation efficiency,and stability were determined.The heating related change of particle size was studied using a size and potential tester.The in vitro photothermal effect was studied using an infrared imaging device.The uptake of DIRAs to 4T1 cells was studied using a CLSM examination.The in vitro photodynamic effect was studied using a fluorescence probe and CLSM examination.Results DIRAs were successfully prepared with a uniform and homogeneous size which was about (97.0±20.1) nm.The Zeta potential was about-21.6 mV and the encapsulation efficiency of ICG and DOX were 93.5％ and 95.2％,respectively.The DIRAs had excellent stability within 28 days.This nanoscaled drug delivery system had identical photothermal effect compared to free ICG.The cellular uptake of DOX was significantly improved after the laser irradiation and the photodynamic effect was enhanced.Conclusions The prepared DIRAs have regular shape,suitable particle size,high encapsulation efficiency and high photothermal conversion efficiency.DIRAs can improve the cellular uptake of DOX and enhance the photodynamic efficiency.This biomimetic muhifunctional nano-system could facilitate breast cancer treatment by combining PTT7PDT and chemotherapy.