Trojan peptides,also named cell-penetrating peptides or protein transduction domains,are a class of small cationic peptides that often contains less than 30 amino acids.They can deliver a wide range of "cargos" such as peptides,proteins and nucleic acids efficiently through the cellular membrane.This review mainly focuses on the recent progress on utilizing Trojan peptides to deliver plasmid DNA and siRNA into cells in vitro and in vivo,and also highlights the implications of this technology in both gene function study and therapeutic potential.

Objective To investigate the effect of pioglitazone on the activation of pancreatic apoptosis in the pathogenesis of rats with acute necrotizing pancreatitis.Methods Eighty Sprague-Dawley (SD) rats were randomly divided into four groups,including acute necrotizing pancreatitis (ANP),sham operation (SO),solvent control (Solvent),pioglitazone intervention (pioglitazone) group,with 20 rats in each group.ANP model was induced by retrograde injection of 4％ sodium taurocholate (1ml/kg body weight) into the biliary-pancreatic duct.The rats in pioglitazone group were injected pioglitazone (40 mg/kg body weight) into the ANP abdominalcavity 30 min before mldel induction.The rats were sacrificed at 1 h,3 h,6 h,and 12 h after ANP model induction.The pancreatic tissues were harvested.Routine HE staining was used to evaluate pancreatic pathological damage.The apoptosis was determined by TUNEL method.The expression of PPARγ was determined by using immunohistochemistry and Western-blot methods.The activity of caspase3 in pancreatic tissues was detected by using spectrophotometry.Results The pancreatic pathological damage was attenuated in rats in pioglitazone group compared with that in rats of ANP group,and the difference between the two groups was statistically significant (P ＜ 0.05).The PPARγ expression of pioglitazone group was 1.34 ± 0.09,which was significantly higher than that in ANP group (0.75 ± 0.05),and the difference between the two groups was statistically significant (P ＜ 0.05).The apoptotic index in pioglitazone group at 3 h was 8.35 ± 0.95,which was significantly higher than that in ANP group at 3 h (4.37 ± 1.22) ; the caspase3 activity was 9.24 ± 1.78,which was significantly higher than that in ANP group (5.04 ± 0.86),and the difference between the two groups was statistically significant (P ＜0.05).Conclusions Pioglitazone intervention attenuates pancreatic inflammation,increases PPARγ expression and caspase3 activity and induces apoptosis in pancreas of rats with acute necrotizing pancreatitis.

Objective To investigate the effects of pioglitazone pre-treating on pancreatic acinar cell (AR42J cells) apoptosis induced by caerulein.Methods AR42J cells were divided into blank control group (with normal culture),pioglitazone group (40 μmol/L),caerulein control group (1 × 10-8 mol/L),pioglitazone+ caerulein group (40 μmol/L pioglitazone + 1 × 10-8 mol/L caerulein) and pioglitazone + GW9662+caerulein group (40 μmol/L pioglitazone+ 5 μmol/L GW9662 + 1 × 10-8 mol/L caerulein).Pioglitazone and GW9662 were added 30 minutes earlier than caerulein.Cell proliferation rate of each group was determined by MTT assay at three,six,12 and 24 hour.The cell apoptosis rate was detected by flow cytometry with Annexin Ⅴ/PI staining and terminal dexynucleotidyl transferase-mediated deoxyuridine triphosphate (dUTP) nick end labeling (TUNEL) staining.The activity of Caspase 3,8 and 9 of each group was measured.Mitochondrial membrane potential (MMP) was detected by flow cytometry with JC-1 staining.Single factor analysis of variance and LSD test were performed for data analysis.Results At six,12 and 24 hour,the cell proliferation rate of pioglitazone group and pioglitazone + caerulein group was 0.19±0.02,0.22±0.02,0.36±0.02 and 0.20±0.04,0.23±0.02,0.38±0.02,respectively,which were significantly lower than those of blank control group (0.25 ±0.04,0.28 ± 0.03 and 0.46±0.02) and caerulein group (0.23±0.02,0.29±0.01 and 0.46±0.05,t lgroup=-3.16,-4.61 and-6.25,tcaerulein group =-1.58,-4.61 and-6.15,all P＜0.05).And the cell proliferation rates of pioglitazone+GW9662+caerulein group at six,12 and 24 hour (0.23±0.02,0.27±0.02 and 0.45±0.01) were significantly higher than those of pioglitazone+caerulein group (t=2.25、3.87、4.56,all P＜0.05).There was no significant difference in cell apoptosis rate detected by flow cytometry with Annexin Ⅴ/PI staining between pioglitazone group ((11.80 ± 0.47) ％,(9.62 ± 2.63) ％ and (14.92 ± 2.52) ％) and pioglitazone+caerulein group ((8.78±0.47)％,(11.89±2.80)％ and (14.25±2.67)％,all P＞0.05),but cell apoptosis of these two groups were higher than those of control group ((5.52± 0.64)％,(5.30±0.97)％ and (5.47±0.88)％) and caerulein group ((5.98±1.21)％,(7.47± 0.58) ％ and (8.11 ± 1.32) ％) respectively,and the differences were statistically significant (t l group =9.81,4.45 and 10.74,tcaerulein group =4.38,7.62 and 6.98,all P ＜0.05).There was no significant difference in apoptosis rate between pioglitazone+GW9662+caerulein group ((5.82±0.26) ％,(6.05± 0.83) ％ and (9.23±0.90)％) and caerulein group; while significantly higher when compared with those of pioglitazone+ caerulein group (t=-4.63,-10.07 and-5.70,all P＜0.05).At 12 hour,the apoptosis rate detected by TUNEL staining of pioglitazone group ((3.93 ± 0.40)％) was significantly higher than that of control group ((2.73 ±0.68) ％),the apoptosis rate of pioglitazone+ caerulein group ((8.43 ± 1.65)％) was significantly higher than that of caerulein group ((2.80 ± 0.56)％),the apoptosis rate of pioglitazone+GW9662+caerulein group ((3.87±0.35)％) was lower than that of pioglitazone+ caerulein group (t=7.93,8.92,-5.35,all P＜0.05).At 24 hour,the activity of Caspase 3,8 and 9 of pioglitazone+ caerulein group (1.28 ± 0.05,1.38 ± 0.04 and 1.53 ± 0.09) significantly increased compared with those of caerulein group (1.12±0.88,1.22±0.02 and 0.53±0.07,t=3.20,8.62 and 1.29,all P＜0.05).After treated with GW9662,part of activity of Caspase enzymes recovered.The number of cells with potential change of mitochondrial membrane in pioglitazone group and pioglitazone + caerulein group was more than that of caerulein group (28.50±0.91)％ and (28.20±2.56)％ vs (15.00±3.67)％) and part of membrane potential recovered after GW9662 added ((20.67 ± 2.20) ％).Conclusions Pioglitazone might promote AR42J cell apoptosis through the activation of caspases enzymes and changing membrane potential.And the antagonist GW9662 would partially inhibit the apoptosis induced by pioglitazone.

Objective To investigate the serum leptin levels of chronic hemodialysis patients and its relation to their nutritional status.Methods Twenty-nine maintenance hemodialysis patients were included in the study.TSF(triceps skin fold),BMI(body mass index) and FAT%(content of fat),lymphocyte count,serum albumin,globulin,total iron binding capacity,BUN,creatinine,cholesterol,triglyceride and leptin were measured.Malnutrition-inflammation score(MIS) was used to assess the patients nutritinal status.Results Levels of leptin were positively correlated with BMI,FAT%,TSF and MIS(P

Objective To investigate the effect of leflunomide and prednisone in treating refractory nephropathy syndrome.Methods The 60 refractory nephropathy patients in our hospital from Oct,2000 to May,2003 were divided into two groups at random.Paients in test group received leflunomide and prednisone.Patients in control group received mycophenolate mofetil and prednisone.Clinical data were observed in the 2nd,4th,6th,8th,12th,16th,20th,24th and 28th week.Results After receiving leflunomide therapy in the test group,the proteinuria was decreased significantly(P0.05),but rate of showing efficacy in 12th week in Leflunomide group was higher than mycophenolate mofetil group(P

Objective To investigate the efficacy and safety of mycophenolate mofetil combined with methylprednisolone and lamivudine on the treatment of hepatitis B virus associated glomerulonephritis(HBV-GN).Methods Twenty-four patients with hepatitis B virus associated glomerulonephritis were confirmed by renal biopsy and immunohistochemistry,these participant patients were admitted to the Third Affiliated Hospital of Sun Yat-Sen University from Jan,1999 to Jan,2004.They were treated by MMF combined with methylprednisolone and lamivudine.The initial dosage of MMF was 1.0~1.5 g/d.Methylprednisolone at the dosage of 0.4 mg/(kg?d)was used at the beginning of the combined treatment.Lamivudine was in the dosage of 0.1 g/d.The duration of the treatment was six months.Regular test was conducted every two weeks.Results Nine cases had fully remission,11 cases had partial remission and 4 cases had no efficiency;no patient deterioration.Renal and hepatic function remained stable,blood cell didn't decrease and the reproduction of HBV didn't increase during the treatment.Conclusion MMF combined with methylprednisolone and lamivudine is an effective and safe method for HBV-GN.

Objective To explore the relationship between poor sleep quality and stoke.Methods A total of 738 stroke patients in Xuzhou city in 2013 were selected as the case group and age-and sex-matched healthy non-stroke subjects (n =738)as control group.The writer-designed general situation questionnaire and the Pittsburgh Sleep Quality Index(PSQI)analyses were conducted for a face-to-face investigation.Results No significant difference in mean age(66.1±10.9 and 65.8 ± 10.6,t =0.60,P =0.58)and in sex (50％ vs.50 ％) was found between two groups.There were statistically significant differences between case and control groups in baseline values of BMI(t=2.40,P =0.02),histories of hypertension(x2 =174.30,P =0.00),diabetes mellitus (x2 =27.20,P =0.00),coronary heart disease(x2 =115.60,P =0.00),smoking(x2 =6.10,P =0.01),drinking (x2 =7.30,P =0.01)and living stress(x2 =11.40,P =0.01).The PSQI sub-scores and PSQI total scores were higher in case group than in control group.The rate of poor sleep quality was higher in case group(279 cases,37.8 ％) than in control group(136 cases,18.4 ％) (x2 =6.10,P =0.01).Multivariate logistic analysis showed that,after adjusting for confounding factors of BMI,histories of hypertension,diabetes,coronary heart disease.smoking,drinking and living stress,the poor sleep quality in total male plus female was independent predictor variables for stroke[odds ratio(95 ％ CI) of 2.3 (1.8-3.0)],no matter their sex,with odds ratio (95 ％ CI) in male (2.5,1.7-3.7) or in female (2.2,1.5-3.2),respectively,but there was no significance difference in the odds ratio between male and female in case group(x2 =0.04,P=0.85).The risk stroke was 2.3 folds higher in poor sleep quality versus control in male plus female,with pure male or female of 2.5 or 2.2 folds.There was no significance difference between male and female in case group versus.the control(x2 =0.04,P=0.85).Conclusions Poor sleep quality is associated with the occurrence of stroke and may be a risk factor for stroke.

Objective To study the immune protective effects of liver graft on intestinal graft in simultaneous liver/small bowel transplantation. Method Rat models(Wistar to SD) of liver/small bowel transplantation(LSBTx),single liver transplantation(LTx)and single small bowel transplantation(SBTx) were established respectively. Twelve recipients from each group were used to determine survival time. Six recipients from each group were sacrificed for liver and/or small bowel biopsy at posttransplantation day 5,7 and 14,respectively. Rejections were detected by histopathology and mRNA expression of IL-2,IL-4,perforin and granzyme B in grafts by semi-quantitative reverse transcript PCR. Result The survival time of LSBTx recipients was (27.83?4.47) d,much longer than SBTx recipients (11.58?3.26) d,but was similar to LTx recipients (28.92?2.39) d. Allograft rejections of intestinal graft in LSBTx group were milder than that in STBx group,accompanied by down-regulated mRNA expression of perforin and granzyme B. Conclusion Simultaneous liver/small bowel transplantation in rats provides immune protection on intestinal graft.

Objective: To investigate the effects of chemokine like factor 1 (CKLF1) on proliferation and metabolism of chondrocytes. Methods: Cell culture, 3H TdR and 3H Proline incorporation methods were used. Chondrocytes were harvested from rabbit articular cartilage. First passage cells were seeded on 96 well plates. After synchronization,the medium was replaced by DMEM containing 5% FCS with various concentration of CKLF1 conditioned medium. The synthesis of mucopolysaccharide was detected by Saffraan O staining. The transcription of inducible nitricoxide synthase (iNOS) was detected by semi quantitative RT-PCR. Results: CKLF1 inhibited the DNA, collagen and mucopolysaccharide synthesis significantly, meanwhile, stimulated the transcription of iNOS. Conclusion: CKLF1 inhibits the proliferation and matrix synthesis of chondrocytes, which might be an important factor resulting in cartilage destructive lesions. CKLF1 may exert its effects on chondrocytes through iNOS pathway.

Objective: To characterize the expression of programmed cell death 5 (PDCD5) in normal and osteoarthritic human cartilage. Methods: Articular cartilage specimens were obtained from 20 patients with osteoarthritis and 10 with femoral neck (normal cartilage) at the time of arthroplasty. Expression of PDCD5 was detected by flow cytometry, immunofluorescence, RT-PCR and immunohistochemical analysis. Results: PDCD5 expression in osteoarthritis cartilage was significantly higher than in normal cartilage especially in the nucleus. PDCD5 positive chondrocytes were mainly observed in the superficial and deep zone of osteoarthritis tissue sections,and in contrast, in the superficial and middle regions of normal controls. Conclusion: Since apoptotic chondrocyte death occurs more frequently in osteoarthritis compared to normal cartilage and PDCD5 is an apoptosis related protein, the different expression patterns of PDCD5 in osteoarthritis and normal cartilage suggest that it might be involved in the pathogenesis of osteoarthritis.