Objective To investigate the inhibitory effects of hepatitis B virus (HBV) S gene-specific antisense locked nucleic acid (LNA) on HBV replication and expression in HepG_22.2.15 cells,and to screen the effective short sequence of LNA.Methods Four different lengths of short sequence of antisense locked nucleic acid which were complementary to the initiator of HBV S gene were designed,synthesized and transfected by cationic liposomes into HepG_22.2.15 cells.The HBsAg and HBV DNA of supematant was tested by enzyme linked immunoadsorbent assay(ELISA) and real-time fluorescent quantitative PCR(FQ-PCR) at 24,48 and 72 hours after treatment.LNA's cyto-toxicity on cell was evaluated by MTT method.Results Four different lengths of short sequence of LNA inhibi-ted the expression of HBsAg and the replication of HBV DNA with the inhibition rates of 46.58%,54.38%,72.43% ,69.92% and 27.09% ,28.77% ,34.71% ,32.68% respectively after 72 hours.There's no obvious tox-icity on cell.Conclusion Antisense LNA that targeting at HBV S gene has strong inhibition on HBV in vitro,and the optimal length of LNA sequence might be in the range of 15 base to 25 base.It has a therapeutic potential in the treatment of patients infected with HBV.

Objective To investigate the expression and biological function of miRNA-449 a in lung cancer . Methods A case-control study was conducted in 58 patients diagnosed with lung cancer ( carcinoma and adeno-carcinoma) and normal tissue closely adjacent to tumor.MiRNA-449a simulation was designed and synthesized, was dissolved into two different concentrations as 10 and 20 mg/mL.The expression of miRNA-449a in lung cancer tissues and matched normal tissues were detected by Real time PCR .The expression of luciferase gene was detected by chemiluminescence technique.MiRNA-449a mimics on cell apoptosis was evaluated by MTT assay . Results The mean tissues expression levels of miRNA-449 a in squamous carcinoma group and adenocarcinoma group were 1.48 ±1.63 and 1.52 ±1.54 respectively, and were significantly lower than in control group (2.74 ± 1.55 ) ( P<0.01 ) .The average intensity of fluorescent protein in 10 mg/mL group and 20 mg/mL group were 2 115 ±168 and 1 352 ±159 respectively , and were significantly lower than that in control group ( 4 975 ±115 ) ( P<0.01 ) .Conclusions MiRNA-449 a was down-regulated expression in lung cancer and induced apoptosis .

Objective To evaluate the curative effects of conversion from mycophenolate mofetil (MMF) to enteric-coated mycophenolate sodium (EC-MPS) on MMF-related chronic diarrhea in the renal transplant (RT) recipients of long-term stage following transplantation.Methods Twenty-six RT recipients with persistent and severe diarrhea,diagnosed as MMF-related diarrhea after a systemic examining procedure including macroscopic and microscopic examinations of the upper and lower gastrointestinal tracks,serology of the blood for CMV and culture of stool for the bacteria.In all the 26 recipients,the dosage of MMF was reduced to 250 mg,twice every day for 2 weeks.Those without significant improvement at the end of this period were shifted to EC-MPS at a dose of 180 mg,twice every day for 2 weeks.The EC-MPS dose was increased to 360 mg,twice every day if the symptoms were improved significantly at the third week after conversion,or EC-MPS was withdrawn if the diarrhea still existed.The dosage of EC-MPS would be reduced to 180 mg,twice every day if the diarrhea recurred in the next 3 months. The clinical symptoms,biological parameters and renal function were observed for 3 months after the conversion.Results ( 1 ) All the 26 recipients were switched to EC-MPS because of the persistent existence of diarrhea after reduction of MMF.After conversion,the diarrhea disappeared completely in 19 out of the 26 recipients in 2 weeks and 2 patients also showed significantly improvement of diarrhea with the total efficiency being 80.8％ (21/26).In the rest 5 cases,EC-MPS was withdrawn at the second week; (2) The disturbance of internal environment was improved significantly following the EC-MPS conversion.Serum potassium,sodium and TCO2 were elevated to normal level.The benefit was predominantly observed in the recipients with moderate to severe proteinuria.The 24-h urinary protein secretion was significantly reduced from 0.76±0.48 to 0.46±0.53 (g/24 h) (P＜0.05) at the third month.Conclusion In RT recipients with MMF-related chronic diarrhea after long-term stage following renal transplantation,switching MMF to EC-MPS can significantly alleviate the diarrhea and rectify the imbalance of internal environment of the recipients.

Objective To explore the effects of cyclosporin A (CsA) and tacrolimus (Tac) on biological behaviors of lung cancer A549 cells in nude mice.Methods Thirty-six models of transplanted tumor in Balb/c mice were established by using lung cancer A549 cells and divided into three groups:control group,without given any immunosuppressant; CsA group,intraperitoneally given CsA; Tac group,intraperitoneally given Tac.The transplanted tumor growth curve was drawn according to the transplanted tumor volume,and the influencing ratio was calculated according to the final tumor weight.The changes in cell migration ability were observed by using Transwell system.Terminal deoxynucleotidyl transferase mediated UTP nick end labeling (TUNEL) assay was used to examine the apoptosis index of the transplanted tumor.Quantitative RT-PCR was used to detect the expression levels of Bcl-2 mRNA and Bax mRNA.Results The growth of transplanted tumor in CsA and Tac groups was faster than in control group.Final tumor volume and final tumor weight in CsA and Tac groups were greater than those in control group.The influencing ratio in CsA and Tac groups was 19％ (P＜0.05) and 25％ (P＜0.05),respectively.The migration ability was greater in CsA and Tac groups than in control group (P＜0.01).The apoptosis index of the transplanted tumor in CsA and Tac groups was lower than in control group (P＜0.05).The expression level of Bcl-2 mRNA was higher in CsA and Tac groups than in control group (P＜0.05),and that of Bax mRNA was lower in CsA and Tac groups than in control group (P＜0.05).Conclusion Both CsA and Tac can promote the growth of transplanted tumor in nuce mice bearing 549 cells and enhance the invasion forces,which is probably related with the apoptosis induction of tumor cells.

Objective To summarize the clinical experience of combined liver and kidney transplantation (CLKT). Methods CLKT was performed on 22 patients. The orthotopic liver transplantation (LT) was preceded with the classic fashion in 10 patients and piggyback fashion in 12 patients. The renal allograft was implanted to the iliac fossa routinely. After operation, the patients received an induction therapy with anti-CD25 monoclonal antibody or antithymocyte globulin ( ATG) and a maintenance therapy with tacrolimus (Tac), mycophenolate mofetil and prednisone. Results The CLKT was successfully performed on all 22 patients, and the graft function was restored well postoperation. During the perioperative period, an acute rejection episode of liver occurred in one patient and acute renal allograft rejection episode in 2 patients. The Tac toxicity occurred in one patient. The hemorrhage of digestive tract occurred in one recipient and the hemorrhage of peritoneal cavity in one patient. The pleural effusion occurred in 6 recipients. The pneumonia occurred in 2 cases and the peritoneal infection in one patient During a follow-up period of 6 months to 7 years 11 months, three patients died because of cytomegalovirus pneumonia in 2 patients and acute myocardial infarction in, one patient, The 1-, 3-, 5-year survival rate of recipients was 86,4 %, 81.3 %, 72.7 % respectively. Conclusion The CLKT is an effective method for treatment of patients with end-stage liver djsease and chronic renal failure.

Objective To investigate a possible association of donor-recipient compatibility for ABO blood group alleles with acute rejection (AR) in renal transplantation. Methods A study comprising 87 pairs of donor and recipient was performed. The ABO genotype A1, A2, O1, O2, and B alleles of renal transplanted recipients and their respective donors were assessed by PCR amplification with sequence-specific primers (PCR-SSP). Accordingly, recipients were divided into donor-recipient ABO genotype matched and mismatched groups. Results The PCR-SSP based types of all cases showed total concordance with their serologically assigned ABO groups. Fifty pairs (57. 5%) were matched for ABO genotype among the 87 pairs of donor and recipient while 37 (42. 5%) were mismatched, including 1 allele mismatch in 31 pairs (83.8%), 2 alleles mismatches in 6 pairs (16. 2%).The incidence of AR was 12.0% (6 cases) and 29. 7% (11 cases) for ABO genotype matched and mismatched transplant patients, respectively ( P ＜ 0.05). After high dose methylprednisolone (MP)treatment, all cases exepienced reversion of AR except a A2O1 recipient receiving kidney from a A1O1enced 4 AR episodes within 3-10 months, and the period of AR was gradually shortened. After high dose MP was administered empirically, even though short-term improvement of renal function was observed, the serum creatinine continued to increase progressively with decreased efficacy of high dose MP. One year after operation the serum creatinine rose to 441 μmol/L. Conclusions Simultaneous definition of the ABO genotype and HLA is highly feasible. The A2 patient is suitable for receiving kidneys from blood group O donors. DNA mismatch for ABO genotype of renal transplant recipients and their respective donors is an independent risk factor for AR. Genotyping of ABO blood group is conducive to prevent AR.

Objective To assess the change of the CD4~+ CD25~+ Treg after the transplant nephrectomy in recipients suffering from chronic allograft nephropathy (CAN) with normal PRA level.Methods Thirty recipients suffering from CAN,aged 20-55 years old,with norlTlal PRA level,were divided into two groups:removal group (n=17) and preserving group(n=13).CD4~+ CD25~(high)/CD4~+,CTLA-4 and Foxp3 in peripheral blood were tested at two time ends:O month and 2 months after the transplant nephrectomy operation.Results (1) Ratio of CD4~+ CD25~(high)/CD4~+ at the Oth and 2nd month in the removal group was(1.47±0.19)%,(1.08±0.16)%,and that was(1.44±0.25)% and (1.77±0.24)%,at the same time in the preserving group (P<0.01).(2) The expression of CTLA-4 at the Oth and 2nd month in the removal group was (76.82±5.31)% and (72.56±4.99)%,and that was (76.20±4.22)% and (75.24±4.26)% in the preserving group (P>0.05).(3)The expression of Foxp3 was (79.77±1.59)% and (69.07±4.37)% in the removal group,and that was (79.56±1.75)% and (79.09±2.05)% in the preserving group.The expression rate of Foxp3 at the 2nd month in the removal group was significantly lower than in the preserving group (P<0.01).Conclusion Remoral of the graft can reduce the ratio of CD4~+ CD25~(high)/CD4~+ and the expression of Foxp3,suggesting that the removal of the renal graft may inhibit the activity of CD4~+ CD25~+ Treg.

Objective To analyze the MP infection status in children with respiratory tract disease and the correlation with gen‐der ,age ,season and clinical conditions .Methods To investigate the clinical data retrospectively of 655 children with respiratory tract infection from January to December 2013 .Results The positive rate of MP antibody was 48 .09% with a higher incidence in girls than boys ,and those above 3 were more susceptible to it .Winter and spring were the peak seasons .The older group had a higher positive rate of MP antibody ,together with high morbidity of MPP and LP ,the differences were statistically significant (P<0 .05) ,while the younger group was inclined to get higher WBC count and percentage of increasing CRP and neutrophils ,the differ‐ences were statistically significant(P<0 .05) .Conclusion MP is a common pathogenic bacteria causes respiratory tract infection in children above 3 years ,especially in winter and spring .The antibody positive rate rise with age and the infected children are more likely to have pneumonia meanwhile the younger group has higher WBC count ,in which more cases get higher level of CRP and neutrophils .

Objective:To investigate the relationship between vulnerability of mouse coronary artery plaque and downstream myocardial perfusion and myocardial strain.Methods:Thirteen ApoE knockout mice with stable coronary plaques (stable plaque group)and 13 ApoE knockout mice with vulnerable coronary plaques(vulnerable plaque group) were selected as the experimental group, and 15 sex- and age-matched C57BL/6 mice with the same genetic background as ApoE mice were chosed as the control group. Myocardial contrast echocardiography (MCE) was carried out to quantify regional myocardial perfusion at rest and during adenosine stress using a Vevo 2100 system (Visual sonics). Replenishment curves of myocardial contrast were obtained, and rates of signal rise (β) and plateau intensity (A) were recorded. MBF was estimated by the product of A and β. Speckle tracking imaging combined with adenosine stress test was used to evaluate the longitudinal strain of left ventricular myocardium in mice. The vulnerability of the plaque was assessed by histopathology in serial tissue sections of proximal and middle left coronary artery according to the previously reported method.Results:There were no significant differences in body weight, heart rate, left ventricular end diastolic volume, left ventricular end systolic volume, left ventricular mass and ejection fraction among the three groups( P>0.05). The levels of serum triglyceride, total cholesterol, high density lipoprotein and low density lipoprotein in stable plaque group and vulnerable plaque group were significantly increased when compared with those in control group (all P<0.05). The pathological results showed that the coronary luminal stenosis rates in the stable plaque group and the vulnerable plaque group were (74.3±4.9)% and (75.5±7.1)% respectively, with no significant difference between the two groups( P>0.05). MBF of the middle anterior septum and left ventricular posterior wall in the experimental groups were significantly decreased when compared with that in the control group both in the resting status and during adenosine stress(all P<0.05). There were no significant differences in the MCE parameters between the stable plaque group and the vulnerable plaque group at rest( P>0.05). However, during adenosine stress, MBF of the vulnerable plaque group was decreased more significantly than that of the stable plaque group ( P<0.05). Compared with the control group, the values of longitudinal strain of the left ventricle in both experimental groups were decreased during resting status, without statistical significance (all P>0.05), but decreased significantly during adenosine stress and with more decrease in the vulnerable plaque group (all P<0.05). Conclusions:For the same degree of coronary artery stenosis in mice, the coronary artery vulnerable plaque group has less downstream myocardial perfusion and myocardial strain than the stable plaque group during adenosine stress. That is, the plaque vulnerability can affect the downstream myocardial perfusion and myocardial strain in the mouse model.

Objective To explore the impact of induction therapy with anti-lymphocyte agents on long-term survival of kidney transplantation.Methods 271 recipients of first cadaveric kidney transplants were treated with tacrolimus,mycophenolate mofetil and prednisone.110 patients of them received induction therapy with anti-thymocyte globulin(ATG group),88 patients received Basiliximab(Bax group),and the remaining 73 patients did not receive induction therapy(control group).The data of AR,DGF,CMV infection,and 1- 3- 5-year patient/allograft survival rate in three groups were retrospectively during a follow-up period of 1 to 5 years postoperatively.Results Within 6 months after operation,the incidence of AR in control group,ATG group and Bax group was 17.8 %(13/73),9.1 %(10/110)and 10.2 %(9/88)respectively.The incidence of AR in ATG group and Bax group was significantly lower than in control group (P<0.05).There was no significant difference in incidence of DGF and CMV infection among three groups.The 1-,3- and 5-year allograft survival rate postoperation in ATG group and Bax group was 95.5 %,90.9 %,87.3 % and 93.2 %,87.5 %,83.8 % respectively,which was significantly higher than in control group(87.7 %,80.8 % and 75.3 %,P<0.05).Conclusion Induction therapy with anti-lymphocyte agents may reduce the early incidence of AR and prolong long-term allograft survival significantly.