Objective To investigate the effect and mechanism of adenovirus vector mediated SOCS 3 gene transfection in CD4+ Th cell differentiation and expression of inflammatory cytokines of mouse .Methods The CD4+ Th cells were isolated from spleen of C57bl/6 mouse and cultured .Ad‐SOCS3 were transfected into the CD4+ Th cells .PHA was used for culturing with the CD4+ Th cells .RT‐PCR were used to detect the mRNA expression ,and Western blot were used to detect the protein expression of cytokines .Results Compared with the control group ,the gene and protein expression of T‐bet ,IL‐2 ,IFN‐γ,STAT4 and IL‐12Rβ2 in the transfected group were significantly down‐regulated ,the gene and protein expression of SOCS3 ,GATA‐3 ,IL‐4 ,IL‐6 ,IL‐10 and STAT6 were significantly up‐regulated(P<0 .01) .Conclusion The results indicate that SOCS3 gene transfection can up‐regu‐late SOCS3 mRNA and protein expression in the CD4+ Th cells ,down‐regulate the JAK/STAT pathway ,inhibition of Th1 cell dif‐ferentiation ,and down regulation of inflammatory cytokine gene and protein expression ,and indirectly promote Th2 cell differentia‐tion ,and up the corresponding inflammatory cytokine gene and protein expression .

Water soluble extract (WSE) is an important index for the quality evaluation of Astragali Radix (AR). In this study, the WSE of the wild AR from Shanxi province (SX) and the cultivated AR from Gansu Province (GS) were compared. The WSEs of two types of AR were determined according to the appendix of Chinese pharmacopoeia. Then the WSEs were subjected to NMR analysis, and the obtained data were analyzed using HCA, PCA, OPLS-DA, microarray analysis, and Spearman rank analysis. In addition, the Pearson correlation of differential metabolites were also calculated. The results showed that the WSE content of GS-AR (37.80%) was higher than that of SX-AR (32.13%). The main constituent of WSE was sucrose, and other 18 compounds, including amino acids, organic acids, were also detected. Multivariate analysis revealed that SX-AR contained more choline, succinic acid, citric acid, glutamate, taurine and aspartate, while GS samples contained more sucrose, arginine and fumaric acid. In addition, the Pearson correlations between different metabolites of the two types of AR also showed apparent differences. The results suggested that the WSE of two types of AR differs not only in the content, but also in the chemical compositions. Thus, the cultivation way is important to the quality ofAR. This study supplied a new method for the comparison of extract of herbal drugs.

Objective Given the significant differences of ischemia-time tolerance observed in clinical heart transplantation between heart and other solid organs,it is important to make a clinical assessment of the correlation between the cold ischemic time of the donor heart and the survival rate after heart transplantation.With these results,we may standardize the management of clinical heart transplantation by providing a proper heart cold ischemic time frame for reference.Method The clinical data of 131 orthotopic heart transplantation patients in our hospital,from September 2008 to March 2014,were collected and analyzed retrospectively.All donor hearts were preserved in histidinetryptophan-ketoglutarate solution (HTK solution) during cold ischemic time.The cold ischemia time was 103-485 min,with an average of 245.2 ± 120.4 min.According to the cold ischemic time,all patients were divided into three groups:＜ 3 h (n =62); 3-6 h (n =41); ＞6 h (n =28).Result (1) Prolonged cold ischemia time could result in high IABP usage perioperatively (postoperative IABP support rate for the three groups was 3.2％,9.8％ and 14.3％ respectively,P =0.155) and high rejection rate (incidence of rejection was 6.4％,9.8％ and 17.9％ respectively,P =0.245),but there was no statistically significant difference.(2) Three weeks after the transplantation,all EF values of the three groups were reduced within the normal range,with no significant difference.Perioperative overall survival rate was 97.7％ (128/131),while survival rate of the three groups was 97.29％ (72/74),100％ (30/30) and 96.29％ (26/27),respectively (P =0.61).(3) One-year overall survival rate was 89.87％ (71/79),and the one-year survival rate of three groups was 92.2％ (47/51),90.9％ (10/11) and 82.4％ (14/17) respectively (P=0.51).Fifty-two patients were still under 1 year follow-up period.This study aimed to illustrate the effect of different cold ischemic time on perioperative cardiac function,rejection rate,IABP usage postoperatively (intra-aortic balloon pump or intra-aortic balloon counterppulsation) and early/mid-term efficacy after transplantation.Conclusion Cold ischemic time within 6-8 h is clinically safe for heart transplantation,and can provide satisfactory early and medium-term effect.Donor heart with cold ischemia time longer than 6 h may extend the recipient inclusion criteria.But considering the safety of transplantation,these donor hearts may be more applicable for the marginal recipients.This study describes the relationship between cold ischemic time and early and medium-term effect of heart transplantation.However,its long-term effects still require further investigation.

Objective To explore the effects of baicalin in the treatment of a preeclampsia (PE) rat model by detecting the expression of X-linked inhibitor of apoptosis protein (XIAP) and cysteine containing aspartate-9 (Caspase-9) and observing the ultrastructure of mitochondria in trophoblast cells.Methods Forty-eight pregnant Wistar rats were randomly divided into two groups:12 in the control group and 36 in the PE model group.The PE model was established with subcutaneous injection of l-nitro arginine methyl ester with 100 mg/kg per day from the 13th day of pregnancy.Beginning from the 16th day of pregnancy,the PE rats were injected with different doses of baicalin till cesarean section,and divided into three groups:non-intervention PE model group treated with saline (NIP group),low-dose baicalin intervention group (IDB group) at 50 mg/kg per day,and high-dose baicalin intervention group (HDB group) at 100 mg/kg per day.The rat tail artery blood pressure and 24-h urine protein level were measured at pregnant day 10,16 and 20.The levels of XIAP and Caspase-9 in placenta were measured by immunohistochemistry.The ultrastructure of mitochondria of trophoblast cells of the rat placenta was observed under electron microscope.T test,F test and LSD-t test were applied for statistical analysis.Results (1) On pregnant day 10,no significant differences were observed in rat tail artery systolic blood pressure,diastolic blood pressure and 24-h urine protein level between the control group and PE model group (all P＞0.05).On pregnant day 16 and 20,the systolic blood pressure,diastolic blood pressure and 24-h urine protein level of NIP,IDB and HDB groups were significantly higher than those of control group [pregnant day 16:systolic blood pressure:(137.74±5.21),(136.15±4.86),(138.28±4.79) and (110.57±3.79) mmHg (1 mmHg=0.133 kPa),diastolic blood pressure:(89.58 ± 5.50),(88.45 ± 8.59),(89.42 ± 6.29) and (80.28 ± 7.36) mmHg,24-h urine protein:(7.78 ± 0.45),(7.53 ± 0.54),(7.86± 0.57) and (6.45 ± 0.56) mg;pregnant day 20:systolic blood pressure:(145.26 ± 4.67),(131.28 ± 4.34),(130.93 ± 5.33) and (110.40 ± 6.92) mmHg,diastolic blood pressure:(89.87±6.55),(85.34±7.33),(84.64±7.36) and (80.19±7.34) mmHg,24-h urine protein:(11.18±0.42),(9.65±0.54),(9.06±0.56) and (6.31 ±0.45) mg] (all P＜0.01).On pregnant day 20,the systolic and diastolic blood pressure and 24-h urine protein level in IDB and HDB groups were significantly lower than in NIP group (all P＜0.05),but showed no significant differences between IDB and HDB groups (allP＞0.05).(2) Compared with NIP group,the expression of XIAP in control group,IDB and HDB groups were significantly increased(210.39±0.78,180.56±0.82,195.36±0.96 and 192.84± 1.06,all P＜0.01).There was no significant difference in the expression of XIAP between IDB and HDB groups (P=0.66).The expression of Caspase-9 in control group,IDB and HDB groups were significantly decreased compared with NIP group (210.36±0.55,195.53±0.96,198.42± 1.01 and 185.25±0.64,all P＜0.01).There was no significant difference in the expression of Caspase-9 between IDB and HDB groups (P=0.65).Ultrastructure of mitochondria in NIP group showed different degrees of damage,matrix swelling,and mitochondrial cristae bresk or disappearance.In IDB group,mitochondrial matrix swelling was not obvious,and mitochondrial cristae were visible.In HDB group,mitochondrial cristae were neat and clear.Conclusions Baicalin may play an important role in the treatment of preeclampsia by reversing the trophoblast apoptosis and improving the ultrastructure of mitochondria through its regulation of XIAP expression and downregulation of Caspase-9 expression.

Objective To establish and evaluate 3 kinds of minimally-invasive valve implantation model in vivo.Methods A novel tissue engineered heart valve(TEHV) manufactured by branched polyethylene glycol cross-linked acellular porcine valve and a minimally-invasive valve implantation system according to the design of Corevalve revalving system were adopted.After anesthesia,18 adult male goats were randomly divided into 3 groups: the ulrasound-directed orthotropic group (group A,n =6),angiography-directed orthotropic group (group B,n =6) and direct-released heterotopic group (group C,n =6),and all received minimally-invasive valve implantation orthotropically or heterotopically.4 weeks later,the valvular function was evaluated by CTA and/or echocardiography.Results All 3 kinds of caprine model were successfully constructed.The operation success rate of each group was A: 66.7％,B: 50.0％ and C: 100.0％,respectively(multiple x2 analysis,group A and B P ＞0.05; group A and C,group B and C,P ＜0.05).The operation-time of each group was A: (79 ± 18) min,B:(61 ±23) min,C: (45 ± 15) min(one-way ANOVA,P ＜0.05).The survival rate at4 weeks was A: 100％,B: 100％ and C: 83.3％ (multiple x2 analysis,P ＞ 0.05).Echocardiography and CTA proved the short-term function of implanted TEHV was satisfactory.Conclusion All 3 kinds of caprine valve implantation model can be established without cardiopulmonary bypass and blood transfusion.The devices and equipments required in group A is relatively simple,but the procedure cost longer time for it is hard to determine the right position by ultrasound.The application of angiography made the positioning much easier in group B while the procedure had to be performed in specific operation room with angiographic apparatus.Group C did rely on neither special equipments nor complex operation,but the valve leaflets cannot work normally,so this model was only suitable for testing in vivo characteristics such as biocompatiblities.

The aim of the study is to establish a new method of quality evaluation and validate its feasibilities by the simultaneous quantitative assay of four lignanoids in Schisandra chinensis. A new quality evaluation method, quantitative analysis of multi-components by single marker (QAMS), was established and validated with Schisandra chinensis. Four main lignanoids, schisandrin, schisantherin A, deoxyschizandrin and gamma-schizandrin, were selected as analytes and schisandrin as internal reference substance to evaluate the quality. Their contents in 13 different batches of samples, collected from different bathes, were determined by both external standard method and QAMS. The method was evaluated by comparison of the quantitative results between external standard method and QAMS. No significant differences were found in the quantitative results of four lignanoids in 13 batches of S. chinensis determined by external standard method and QAMS. QAMS is feasible for determination of four lignanoids simultaneously when some authentic standard substances were unavailable, and the developed method can be used for quality control of S. chinensis.

AIM: To explore the molecular mechanism in the pathogenesis of dilated cardiomyopathy (DCM) by analyzing the expression of T cell signaling molecules in mice with autoimmune DCM. METHODS: Mouse DCM model was induced by immunizing the animals with adenine nucleotide translocase (ANT) synthetic peptides. P56lck in T cells was detected with real-time fluorescent quantitative PCR in both DCM-group and the sham-immunized controls. At the same time, flow cytometry was used for quantity of Th cell intracellular cytokine IFN-? and IL-4, ELISA for examining the level of serum anti-ANT antibody, immune histochemistry for investigating the expression of CD45 in Th cells. RESULTS: The mRNA expression of P56lck ( 1 369.51 ?874.05 vs 47.93?10.21, P

The cytokine repertoire of ADP/ATP carrier-specific humoral immune responses and the cytokine-dependent anti-ADP/ATP carrier antibody IgG subclasses were examined in a cohort of ADP/ATP carrier-immunized BALB/c mice treated with anti-CD4 monoclonal antibody. Eighteen male BALB/c mice (6-8 weeks old) were randomized into 3 groups: dilated cardiomyopathy (DCM) group, DCM-tolerance (Tol) group and control group. The mice in DCM group were immunized with the peptides derived from human ADP/ATP carrier protein for 6 months and mice in the control group were sham-immunized, while the mice in DCM-Tol group were immunized with ADP/ATP carrier protein and anti-CD4 McAb simultaneously. Serum autoantibody against ADP/ATP carrier and IgG subclasses were measured by ELISA, intracellular cytokines IFN-gamma and IL-4 of Th cells were monitored with flow cytometry, and splenic T cell cytokines IFN-gamma, IL-2, IL-4 and IL-6 were detected by using real-time fluorescent quantitative PCR. The results showed that the autoantibody against ADP/ATP carrier was found in all mice in DCM group, and the antibody level, serum IgG1 and IgG2a subclasses, cytokines in T cells and Th cells were all elevated in DCM group, as compared with those in control group (P<0.01). On the other hand, in DCM-Tol group, the autoantibody level and contents of all the cytokines were significantly different from those in DCM group (P<0.01), and were close to those in control group. And the levels of IgG1, IgG2a, IgG2b and IgG3 were influenced, to varying degrees, by anti-CD4 McAb as compared with those in DCM group. All these four types of IgG subclasses were substantially decreased in DCM-Tol group as compared with DCM group. It is concluded that the treatment with anti-CD4 McAb could prevent the activation of T cells, reverse the abnormal secretion of cytokines and the imbalance between Th1/Th2 cell subsets and abnormal production of autoantibody against ADP/ATP carrier, and eventually avoid myocardial injuries.

Objective Although heart transplantation (HTx) has become a standard therapy for end-stage heart diseases,experience with pediatric HTx is limited in China.In this article,we will try to provide the experience with indications,complications,perioperative management,immunosuppressive therapy,and survival for pediatric HTx based on our clinical work.Method This is a retrospective chart review of the pediatric patients undergoing HTx at Department of Cardiovascular Surgery of Union Hospital from September 2008 to December 2015.We summarized the indications,surgical variables,postoperative complications,and survival for these patients.Result Twenty-three pediatric patients presented for HTx at Union Hospital of Tongji Medical College,of whom 12 were male.The age at the time of transplantation ranged from 3 months to 18years (median 14 years).Patient weight ranged from 5.2 kg to 57.0 kg (median 36.0 kg).Pretransplant diagnosis included cardiomyopathy (16 cases),complex congenital heart disease (5cases) and tumors (2 cases).All recipients received ABO compatible donor hearts.Postoperative complications occurred in 14 patients,including cardiac dysfunction,arrhythmia,pulmonary infection,renal dysfunction,and rejection.Two of them experienced cardiac failure and required extracorporeal membrane oxygenation.The immunosuppression regimen was comprised of prednisone,a calcineurin inhibitor,and mycophenolate.All patients recovered with New York Heart Association (NYHA) Class Ⅰ Ⅱ cardiac function and were discharged.Only one patient suffered sudden death 19 months after transplantation.Conclusion Orthotopic HTx is a promising therapeutic option with satisfactory survival for the pediatric population in China with end-stage heart disease.

Objective To study the clinical effect, safety of stains collapse of traditional Chinese medicine (TCM) combined with tumor necrosis factor-alpha (TNF-α) antagonist-a recombinant human tumor necrosis factor receptor:Fc fusion protein (rhTNF:Fc) and sulfasalazine (SSZ) in treatment of active ankylosing spondylitis (AS). Methods A total of 98 patients of AS of Gansu Provincial Hospital were divided into two groups in random, the western medicine group (rhTNF:Fc and SSZ, n=46) and the combining group (stains collapse of TCM, rhTNF:Fc and SSZ, n=52). The Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), the Bath Ankylosing Spondylitis Functional Index (BASFI), Patient Global Assessment (PGA), Visual Analogue Scale (VAS) of night pain, VAS of total pain, time of morning stiffness, TCM symptom score, distance between occipital and wall, distance between finger and ground,
thoracic activity, Schober test, and erythrocyte sedimentation rate (ESR), C reactive protein (CRP), adverse reactions before and after treatment were observed and the clinical effect was evaluated in two groups. Results A total of 86 patients achieved the study finally, 45 patients in the combining group and 41 patients in the western medicine group. The effective rate of ASAS20 was better in the combining group than that in the western medicine group(χ2=4.814,P=0.028), the Bath ankylosing spondylitis disease activity index (BASDAI) 50 showed no significant difference between the two group (χ2=0.100,P=0.752). The total effective rate in the combining group than that in the western medicine group(χ2=3.873,P=0.049). In comparison with the same group before treatment, the above indicators were improved after treatment (P<0.05). In comparison with the western medicine group after treatment, BASFI, PGA, VAS of night pain, VAS of total pain, time of morning stiffness, TCM symptom score, distance between occipital and wall, distance between finger and ground and Schober test were getting better in the combining group (P<0.05), but BASDAI, thoracic activity, ESR, CRP were not improved in the combining group (P>0.05). The adverse reactions showed no significant difference (P>0.05). Conclusions Stains collapse of TCM combined with rhTNF:Fc was more effective than single method in treatment of AS, it can alleviated the symptoms, body signs and body function, decrease indexes of inflammation, which has dramatic effect and good safety in clinic.