Objective:To evaluate the preventive effect of proximal splenic vein ligation after splenectomy on the splenic vein originated portal vein thrombosis (PVT) in portal hypertension.Methods:The clinical data of 94 patients of portal hypertension who had received splenectomy were retrospectively analysed. The proximal splenic vein was ligated in 36 cases during pericardial devascularization and coronary renal shunt with splenectomy. The other 58 cases who had received pericardial devascularization without proximal splenic vein ligation served as control. All of the patients in both groups were given heparin infusion postoperatively through the catheter which was placed in the right gastroepiploic vein during operation. CT portal veinography was performed at the 7th-14th postoperative day for detection of PVT.Results:None of the PVT occurred in the splenic vein ligation group. In the control group, PVT occurred in 22 cases(38%) and splenic vein thrombosis occurred in all the 58 cases (100%). PVT incidence between the two groups is significantly different (0 vs. 38%, χ 2=17.828, P<0.05). Conclusions:Ligation of the proximal splenic vein during splenectomy can effectively prevent the postoperative splenic vein originated PVT in portal hypertension.

A special medical device for the treatment of congenital megacolon is designed.This device is made of avirulent and high-intensity plastics.The resultant which formed by up and down curve arm detained lateral anus can spur the up and down leaf of the ring clamp device.Under the continuous pressurized condition,it can clamp the rectum and the downward pull-through colon,then the colon becames necrosis,so that the aim of confluence is achieved.This device is disposable.

Objective:To study the effects of PRF and recombinant hPDGF-AB,TGF-β1 and VEGF on the proliferation and adhe-sion of rat adipose tissue-derived stem cells(ADSCs)in vitro.Methods:ADSCs were cultured with PRF membrane and various do-ses of PDGF-AB,TGF-β1 and VEGF,cell adhesion was examined by adhesion assay after 2 culture,cell proliferation was examined by CCK-8 kit after 1 -7 d culture.Results:Cell adhesion assay showed that the adhesive numbers of rat ADSCs in PRF group were significantly higher than those in the negative group(P <0.05).The adhesive numbers of the ADSCs treated by PDGF-AB showed no significantly difference among different concentration groups(P >0.05).The adhesive numbers of the ADSCs treated by VEGF or TGF-β1 at different concentrations showed significant difference(P <0.05).CCK-8 kit assay showed that at different time points, the A values of ADSCs in PRF group were significantly higher than those of the negative control group(P <0.05).The A values of ADSCs in VEGF or PDGF-AB groups at different concentrations showed significant difference(P <0.05).The A values of rat AD-SCs in TGF-β1 group at different concentrations were lower than those in the negative control group(P <0.05).Conclusion:PRF as a combination of growth factors may stimulate the proliferation and adhesion of rat ADSCs in vitro.PDGF-AB and VEGF may stim-ulate the proliferation of rat ADSCs.TGF-β1 and VEGF may stimulate the adhesion of rat ADSCs in a dose-response manner to some degree.

Objective To compare the normal anatomy of the anterior cruciate ligament (ACL) of fresh frozen cadaveric knee specimen in oblique coronal thin-slice section with oblique coronal magnetic resonance imaging. Methods One fresh cadaveric knee specimen was scanned with MR T1-weighted spinecho sequence.then the specimen was frozen and sliced with a band saw along the oblique coronal plane into 1.0-mm-thick sections that corresponded to the MR images,MR images including oblique coronal T1-weighted and T2-weighted images of 50 normal the knee joints were retrospectively reviewed to observe the MR imaging features of the cruciate ligament. Results Anteromedial and posterolateral bundles of ACL were clearly depicted on both anatomic slices and MR images.The anteromedial bundles originated from the posteromedial aspect of the lateral femoral condyle,coursing through the lateral intercondylar notch in an anterior,inferior,and medial direction,and inserted on the anteromedial aspect of the intercondylar eminence. The posterolateral bundles originated from the anteromedial aspect of the lateral femoral condyle,passing laterally and inferiorly through the lateral intercondylar notch,and inserted on the posterolateral side of the intercondylar eminence.The full length of ACL of all 50 individuals was showed on MR images.MRI clearly differenitated the anteromedial and posterolateral bundles of ACL and depicted the full length of the bundles.similar to the findings on sectional anatomy.Conclusion Oblique coronal MR imaging is the best way to demonstrate ACL and should be used for clinically suspected injury of ACL.

OBJECTIVEThe objective of this study is to investigate the effect of epidermal growth factor receptor (EGFR) monoclonal antibody MAb225 on repair of DNA double strand break (DNA-DSB) after radiation in tongue squamous cell carcinoma cell.

METHODSThe single cell gel electrophoresis (SCGE) was performed to estimate the repair of DNA-DSB induced by radiation in human tongue carcinoma cells Tca8113 treated with or without MAb225. Expression of Ku70 and Ku80 were detected by semiquantitative reverse transcription polymerase chain reaction and Western bolt.

RESULTSComet tail moment of MAb225 treated cell was significantly higher than untreated cell (P < 0.05). The expression of Ku70 and Ku80 were inhibited by MAb225.

CONCLUSIONMAb225 can inhibit repair of DNA-DSB induced by down-regulated expression of Ku70 and Ku80.

Objective To investigate the changes and significance of ubiquitination of Foxp3 pro-tein during the acute phage of Kawasaki disease ( KD) .Methods Forty-eight children with KD and twenty-eight age-matched healthy children were recruited in this study.Co-immunoprecipitation and Western blot assays were performed to determine the poly-ubiquitination status of Foxp3 in CD4+T cells.The percentages of CD4+CD25high Foxp3+regulatory T cells ( Treg) and the levels of IL-10, transforming growth factor-β( TGF-β) , cytotoxic T lymphocyte-associated protein-4 ( CTLA4 ) , STIP1 homology and U-Box containing protein 1 (STUB1), heat shock protein 70 (HSP70), ubiquitin-specific-processing protease 7 (USP7) and pSTAT3 were analyzed by flow cytometry analysis.Quantitative real-time PCR was used to evaluate the tran-scription levels of TLR1-10, IL-1R1, IL-1RAP, IL-6Rα, gp130, TNFR1, MyD88 and RIP1 in CD4+T cells.Plasma concentrations of IL-1β, IL-6 and TNF-αwere measured by enzyme-linked immunosorbent as-say.Results (1) The percentages of Treg cells and the levels of IL-10, TGF-β, CTLA4 and forkhead box P3 (Foxp3) in patients with acute KD were lower than those of healthy subjects (P<0.05), while the poly-ubiquitination of Foxp3 protein was significantly enhanced in patients with acute KD [(0.52 ±0.19) vs (0.08±0.02),P<0.05].Meanwhile, the four former items in KD patients with coronary artery lesions (KD-CAL+) were lower than those of KD patients without coronary artery lesions (KD-CAL-) (P<0.05), while the polyubiquitination level of Foxp3 protein in KD-CAL+group was much higher than that of KD-CAL-group [(0.70±0.28) vs (0.43±0.17), P<0.05].The levels of Treg cells, IL-10, TGF-βand CTLA4 in patients with KD were increased and the ubiquitination of Foxp3 protein was inhibited [(0.24±0.10) vs (0.52±0.19), P<0.05] upon the treatment with IVIG.(2) The levels of STUB1 and HSP70 in CD4+T cells were significantly elevated during acute KD, while the levels of USP7 were decreased (P<0.05).The ratios of STUB1/USP7 in patients with acute KD were much higher than those of the control group [(2.65± 0.92) vs (1.09±0.37), P<0.05], but were significantly decreased after IVIG therapy [(1.46±0.53) vs (2.65±0.92), P<0.05].A negative correlation was found between STUB1/USP7 ratio and Foxp3 level during acute KD (r=-0.56, P<0.05).Moreover, KD patients with CAL+showed higher levels of STUB1 and HSP70 and higher ratios of STUB1/USP7 (P<0.05), but lower levels of USP7 as compared with those of KD-CAL-group (P<0.05).(3) The plasma concentrations of inflammatory cytokines (IL-1β, IL-6 and TNF-α), the levels of surface receptors ( IL-1R1/IL-1RAP/TLR4, IL-6Rα/gp130 and TNFR1) and its downstream molecules ( MyD88, pSTAT3 and RIP1) in CD4+T cells were up-regulated during acute KD ( P<0.05), especially in patients with CAL+, but were down-regulated upon the IVIG therapy (P<0.05).No significant differences with other TLRs were found among the groups (P>0.05).Conclusion Hyper-ubiq-uitination of Foxp3 protein might be involved in the immune dysfunction during Kawasaki disease.

Objective To investigate the role of IL-35-producing regulatory B cells(IL-35 + Breg)in immunological pathogenesis of Kawasaki disease (KD).Methods Thirty-two children with KD and 28 age-matched healthy children were allowed to participate in this study.Flow cytometry was performed to evaluate the proportions of IL-35 + Breg as well as requlatory T cells (Treg)and expression levels of associated molecules such as programmed death-ligand 1 (PD-LI),CD169,programmed death 1 (PD-1),CD43,IL-12p35,epstein-Barr virus induced 3 (IL-27 EBI3).IL-12 receptor beta 2 (IL-12 Rβ2),IL-27 receptor alpha (IL-27 Rα),phosphated signal transducer and activator of transcription 1 (pSTAT1) and phosphated signal transducer and activator of transcription 3 (pSTAT3).Transcription levels of the Src homology 2 domain-containing protein tyrosine phosphatase 2 (SHP-2),phosphatase and tensin homolog (PTEN),vav1 guanine nucleotide exchange factor(Vav) in CD19 + B cells were determined by using quantitative real-time PCR.Plasma concentrations of IL-35,tumor necrosis factor α(TNF-α) and IL-12 were measured by adopting enzyme-linked immunosorbent assay.Results (1) The proportions of IL-35 +Breg and its expressions of IL-12p35,IL-27EBI3 and IL-10 in patients with acute KD were lower than those of healthy controls [IL-35 + Breg:(5.79 ± 2.60) ％ vs (12.65 ± 5.34) ％;F =19.23,9.70,14.30.7.08;all P ＜ 0.05],but they were significantly increased after intravenous immune globulin (IVIG) treatment [IL-35 + Breg:(10.52 ± 4.95) ％;all P ＜ 0.05].(2) The proportions of Treg and its transcriptional levels of IL-12p35 and IL-27 EBI3 were down-regulated during acute KD [Treg:(4.12 ± 1.51) ％ vs (8.06 ± 3.32) ％;F =19.70,17.69,38.22;all P ＜ 0.05],but were increased after therapy [Treg:(7.39 ± 2.85) ％;P ＜ 0.05].A positive correlation was found between the proportions of Treg and IL-35 + Breg during acute KD (r =0.69,P ＜ 0.05).Meanwhile,plasma concentrations of IL-35 and expression levels of IL-12Rβ2,IL-27Rα,pSTAT1 and pSTAT3 in CD19 + B cells were significantly down-regulated in children with acute KD,but they were increased after treatment(F =8.09,7.54,7.69,5.89,12.59,all P ＜ 0.05).(3) Compared with healthy controls,expressions of PD-L1 and CD169 on CD14 + cells and plasma concentrations of TNF-α and IL-12 were elevated during acute KD (F =24.94,16.53,34.71,19.51;all P ＜ 0.05).Expression levels of PD-1,CD43 and its downstream molecules (SHP-2,PTEN,Vav) in CD19 + B cells were down-regulated during acute KD (F =6.43,5.57,19.52,10.37,11.37;all P ＜ 0.05),and restored remarkably after therapy (all P ＜ 0.05).Conclusion Insufficiency of IL-35 + Breg and its expression of IL-35 may be the important factors contributing to immunological dysfunction in KD.

Objective To investigate the changes and significances of inducible IL-35-producing regulatory T cells(iTR35) in immunological pathogcnesis of Kawasaki disease (KD).Methods Forty-eight children with KD and 32 age-matched healthy children (healthy control group) consented to participate in this study.Flow cytometry was performed to evaluate the proportions of CD4+ FOXP3-IL-12p35+IL-27EBI3+iTR35 and CD4+CD25high FOXP3+regulatory T cells (Treg),and expression levels of associated molecules such as programmed death-ligand 1 (PD-L1),CD169,programmed death 1 (PD-1),CD43,IL-12p35,Epstein-Barr virus induced 3 (IL-27EBI3),glycoprotein 130(gp130),IL-12 receptor beta 2 (IL-12Rβ2),phosphated signal transducer and activator of transcription 1 (pSTAT1) and phosphated signal transducer and activator of transcription 4 (pSTAT4).Transcription levels of the Sre homology 2 domain-containing protein tyrosine phosphatase 2 (SHP-2),phosphatase and tensin homolog (PTEN),Vavl guanine nucleotide exchange factor(Vav) in CD4+T cells were determined by quantitative real-time PCR.Plasma concentrations of IL-35,IL-10,TNF-α and IL-12 were measured by enzyme-linked immunosorbent assay.Results (1) The proportions of iTR35 and its expressions of IL-12p35 and IL-27EBI3 in patients with acute KD dccreased remarkably[iTR35:(0.72±0.26) ‰ vs (1.65±0.43) ‰,P＜0.05],and restored after treatment [iTR35:(1.58±0.63) ‰ vs (0.72±0.26) ‰,P＜0.05].(2) The proportions of Treg and transcriptional levels of IL-12p35 and IL-27EBI3 were down-regulated during acute phase of KD [Treg:(3.26±1.21) ％ vs (7.26±2.86) ％,P＜0.05],and increased to some extent after therapy [Treg:(5.89±2.60)％ vs (3.26±1.21)％,P＜0.05].Meanwhile,plasma concentrations of IL-35 and IL-10,and expressions of gp130,IL-12Rβ2,pSTAT1 and pSTAT4 in iTR35 of patients with acute KD were found lower than those of the healthy control group (all P＜0.05),and increased after treatment (P＜0.05).Additionally,positive correlations were found between plasma concentrations of IL-35 and the proportion of iTR35 or its expressions of IL-12p35 and IL-27EBI3,respectively.(3) Expressions of PD-L1 and CD169 on CD14 + cells and plasma concentrations of TNF-α and IL-12 were elevated significantly during acute KD(all P＜0.05),as well as expression levels of the ligands (PD-1 and CD43) and its downstream molecules (SHP-2,PTEN,Vav) in CD4 + T cells were found to be lower in patients with acute KD (P＜0.05),and restored remarkably after therapy.Conclusion Insufficiency of iTR35 and its expression of IL-35 might be one of the important factors contributing to immunological dysfunction in KD.

Objective To investigate the liver repair effects of Pim-3 gene in rat with fulminant hepatic failure (FHF). Methods Thirty-two rats were divided into four groups (eight for each group). Three groups of rats were pretreated with Ringer's solution, vector plasmid or Pim-3 gene recombinant plasmid respectively and, one day later, received intraperitoneal injections with lipopolysacchride (LPS) and D-galactosamine (D-GalN). The fourth group served as normal control.Eight hours after the LPS/D-GalN injection, the liver tissues and blood samples were collected. The contents of serum transaminase was tested by automatic blood biochemistry meter. The morphological changes were observed by light microscopy using hematoxylin and eosin (HE) staining. Tumor necrosis factor (TNF)-α and interleukin (IL)-1β gene expression was detected by reverse transcriptionpolymerase chain reaction(RT-PCR). The serum levels of TNF-α and IL-1β were measured by enzyme linked immunosorbent assay (ELISA), and cell apoptosis by TdT-mediated dUTP nick end labeling (TUNEL) assay. Comparisons between groups were done by analysis of variance. ResultsThe over expressions of Pim-3 gene and reporter gene, green fluorescent protein (GFP) were induced by injection with recombinant plasmid solution.In comparison with the rats retreated with Ringer' s solution or vector plasmid, those pretreated with recombinant plasmid had a lower mortality and lower serum transaminase levels. The injection of recombinant plasmid significantly reduced hemorrhage, necrosis and inflammatory infiltration in the liver. Liver apoptotic index (AI) was dramatically lower in rats treated with recombinant vectors compared to the rats treated with Ringer's solution or vector plasmids [(10. 2±6.9)％ vs (83. 1±12.6) ％ and (79.9±13.4) ％ respectively, P＜0. 01]. In addition, the expression of exogenous Pim-3 gene remarkable inhibited the transcriptions and expressions of TNF-α and IL-1β. ConclusionsPim-3 gene can protect rats from LPS/D-GalN-induced FHF possibly by inhibiting expressions and secretions of inflammatory cytokines, such as TNF-α and IL-13, in liver tissues.

Objective To design and create a C/S-J type of biliary self-releasing stent,and to study its safety and efficacy in preventing post-ERCP complications.Methods 118 patients with common bile duct stones treated in our hospital were enrolled into this study from October 2013 to May 2015.These patients were randomly divided into two groups:the experimental group who underwent ERCP + EST + C/S-J type of self-releasing biliary stent drainage,while the control group underwent ERCP + EST + ENBD.The incidences of post-ERCP acute pancreatitis (PEP) and cholangitis in the two groups and the time the self-releasing stent was dislodged from the biliary system in the experimental group were recorded.Results The incidence of PEP was 6.4％ (5/78) and 7.5％ (3/40) in the experimental and the control group,respectively (P ＞ 0.05).There were no patients who developed postoperative acute cholangitis in the two groups.The stents were dislodged from the biliary system on the first day after the procedure in 2 patients in the experimental group without any complications.One stent failed in self-releasing but was removed successfully with endoscopy 3 months later.In the other 75 patients,the stents were successfully dislodged and were excreted outside the patient's body through the intestinal tract (mean 11.4,range 9 ～ 14) days,without any complications.Conclusion The C/S-J type of biliary self-releasing stents is safe and efficacious in preventing post-ERCP pancreatitis and cholangitis.