This study aimed to investigate the effects of fusion proteins GnRH-GRP(G3G6)and HSP65-STEAP1(HST1)on dendritic cells(DC)and the sensitization of DCs to B16F10 melanoma. The fusion proteins G3G6 and HST1 were obtained using the previous engineering strains in our laboratory. Group by unsensitized DC(US-DC), the G3G6 fusion protein sensitized DC, the HST1 fusion protein sensitized DC(HST1-DC)and the combined sensitized DC(GH-DC), the mouse bone marrow-derived DCs were sensitized with fusion protein to obtain the fusion protein sensitized DC vaccines. B16F10 melanoma cells were transplanted into C57BL/6J male mice to construct a melanoma model(1×106 cells per mouse), and DC vaccine was injected for treatment. The antitumor efficacy of DC vaccine was explored by in vitro and in vivo experiments. Flow cytometry analysis showed that the fusion protein can effectively stimulate DC into differentiation and maturation; in the animal experiment, the inhibition rate of melanoma treated with G3G6-DC was 35. 75%, that of HST1-DC group and combination group were 34. 03% and 55. 74%. It was initially proved that both G3G6-DC and HST1-DC can effectively inhibit the growth of transplanted tumors of melanoma B16F10 cells in mice, and the combination therapy is superior to the single therapy.

Objective:To expound the effect of Kangxianyixin prescription on cardiocyte apoptosis and expression of Bax and Bcl-2 in rats with dilated cardiomyopathy. Methods:Wistar rats with dilated cardiomyopathy established by furazolidone were treated 8 weeks with Kangxianyixin prescription,to observe cardiocyte apoptosis and to measure Bax and Bcl-2 expression in rats. Results:(1) In comparison of cardiomyocyte apoptosis indexes in rats,when compared with model group,the results showed significant difference in the combined group of high dosage of Kangxianyixin prescription and captopril,the high dosage group,the low dosage group,the captopril group(P

BACKGROUND: Leucine-rich repeats and immunoglobulin-like domains 1 (LRIG1) gene showed low expression in glioma cells. LRIG1 gene overexpression significantly enhanced LRIG1 mRNA and protein expression and inhibited its biological behavior. However, very few researchs are reported from the stand-point of inhibition of LRIG1 gene expression. OBJECTIVE: To construct specific RNA interference plasmids for LRIG1, establish stably transfected human glioma GL15 cell line, and observe its effect on expression of target gene LRIG1. METHODS: Designed and synthesized two shRNAs (named LRIG1-shRNA1 and LRIG1-shRNA2) specific for LRIG1 mRNA according to the GenBank, and one scrambled shRNA sequence as negative control, named pGenesil2-negative shRNA. The shRNA was inserted into pGenesil2 vector and sequenced. The recombinant vectors were transformed into E. coli. Picked up the positive clones and extracted the plasmids, which were transfected into GL15 cells by Metafectine. G418 was applied to select the stably transfected cell clones. Western Blotting was performed to examine the LRIG1 protein level.RESULTS AND CONCLUSION: The recombinant plasmids which contain shRNA were analyzed by restriction endonuclease digestion and DNA sequence, and it was proved that the fragment was inserted into the expected sites. Compared with the negative control group, the level of LRIG1 protein expression in pGenesil2-LRIG1-shRNA1(LRIG11) transfected cells and in pGenesil2-LRIG1-shRNA2(LRIG12) transfected cells was decreased by 47.9% (P < 0.01) and 32.8% (P > 0.05). The results confirmed that RNAi expressing vector specific for LRIG1 gene (pGenesil2-LRIG1-shRNA1) was successfully constructed, and the stable cell clones transfected with the shRNA expression vector showed inhibition of the expression of LRIG1 in glioma cell line GL15.

ObjectiveTo elucidate the effect of portal decompression and FK506 (FK) pretreatment on porcine extended hepatectomy.MethodsForty pigs were equally devided into 4 groups. Group A underwent 80% hepatectomy and portacaval shunt with FK pretreatment, group B did 80% hepatectomy and portacaval shunt, group C did 80% hepatectomy and FK pretreatment and group D did 80% hepatectomy. ResultsThe 5 day survival rate in Group A and B was 80% and 60% respectively, higher than 30% in Group C and 20% in group D ( P 200 mm H 2 O ( P

Objective To investigate the relationship between donor liver warm ischemia and postoperative cytokine expression in liver transplantation from non-heart-beating donors.Methods Rat orthotopic liver transplantation was performed in three groups (HB, NHBD-30 and NHBD-60) according to the non-heart-beating time of 0, 30 or 60 minutes before donor liver was harvested. The serum values of TNF-?, IL-6 and CINC in each group after transplantation were determined.Results Over time of warm ischemia to which the donor liver was exposed, the concentrations of TNF-?, IL-6 and CINC elevated with significant difference among those three groups. TNF-?, IL-6 and CINC reached its peak value at 3 hours, 6 hours and 6 hours after transplantation respectively.ConclusionsKG1 In liver transplantation from non-heart-beating donors, the upregulation of TNF-?, IL-6 and CINC was associated with warm ischemia injury.

Background: Chronic atrophic gastritis (CAG) is a precancerous lesion of gastric cancer.The diagnostic value of serum gastrin-17 (G-17) level for CAG differs substantioulsy, and Helicobacter pylori (Hp) infection may play an important role.Aims: To explore the effect of Hp infection on serum G-17 level, and the diagnostic value of serum G-17 level for CAG under different Hp infection status.Methods: A total of 204 patients with chronic non-atrophic gastritis and 81 patients with CAG from May 2014 to May 2015 at the three different hospitals were enrolled.Gastroscopy was performed, fasting serum G-17 level, postprandial serum G-17 level and Hp-IgG antibody were determined by ELISA.Results: Fasting serum G-17 level was significantly increased in Hp positive group than in Hp negative group (P=0.001), and postprandial serum G-17 level was significantly decreased in CAG group than in non-atrophy group (P=0.002).AUC of fasting serum G-17 level for diagnosing Hp positive and negative CAG were 0.634 (95% CI: 0.537-0.732) and 0.576 (95% CI: 0.478-0.675), respectively, the accuracy were 62.6% and 54.9%, respectively.AUC of postprandial serum G-17 level for diagnosing Hp positive and negative CAG were 0.675 (95% CI: 0.581-0.769) and 0.595 (95% CI: 0.495-0.694), respectively, the accuracy were 61.8% and 53.1%, respectively.Conclusions: Hp infection has impact on serum G-17 level, as a result, the diagnostic value of G-17 level for CAG is different for patients with and without Hp infection.Diagnostic values of fast and postprandial serum G-17 for Hp positive CAG are higher than Hp negative CAG.

Objective To investigate the safety of the laparoscopic pancreaticoduodenectomy (LPD) in 70 years of age or older patients.Methods The retrospective cohort study was conducted.The clinicopathological data of 40 patients (age ≥70 years old) who underwent pancreaticoduodenectomy in the West China Hospital of Sichuan University between January 2012 and December 2016 were collected.Twenty patients undergoing LPD were allocated into the LPD group,and 20 receiving open pancreaticoduodenectomy (OPD) who were selected by random number table during the same period were allocated into the OPD group.Observation indicators included:(1) intraoperative situations;(2) postoperative situations;(3) follow-up situation.Follow-up using outpatient examination and telephone interview was performed to detect the patients' survival after discharge and tumor recurrence and metastasis up to March 2017.Measurement data with normal distribution was represented as x±s,and comparison between groups were evaluated with the t test.Measurement data with skewed distribution were described as median (range) and comparison between groups was analyzed using the nonparametric test.Comparison of count data was analyzed using the chi-square test.Comparison of ranked data was analyzed by non parametric test.Results (1) Intraoperative situations:1 patient in the LPD group was converted to open surgery,with a conversive rate of 5.0％ (1/20).Operative time and volume of intraoperative blood loss were (463 ± 10) minutes,210.5 mL (152.5-300.0 mL) in the LPD group and (332± 25) minutes,420.0 mL (350.1-493.8 mL) in the OPD group,showing statistically significant differences between the 2 groups (t =5.48,Z =-3.98,P＜0.05).Cases with intraoperative blood transfusion and pylorus preservation were respectively 4,14 in the LPD group and 6,10 in the OPD group,showing no statistically significant difference between the 2 groups (x2=0.53,1.67,P＞0.05).The results of intraoperative rapid frozen pathological examination showed negative margin of the 40 patients.(2) Postoperative situations:cases in ICU,cases with postoperative analgesia,time for out-of-bed activity,time to anal exsufflation and time for intake were 17,7,(2.2±0.7)days,(4.2± 0.9)days,(4.8±0.7)days in the LPD group and 6,15,(3.6±0.8)days,(5.7±0.9)days,(7.1 ± 2.7)days in the OPD group,showing statistically significant differences between the 2 groups (x2 =12.34,6.47,t=-6.18,-6.55,-3.65,P＜0.05).Pancreatic fistula,delayed gastric emptying (Grade B),postoperative bleeding (Grade B),biliary fistula,pulmonary infection,intestinal obstruction,wound infection,reoperation and major complication were respectively detected in 2,3,1,1,3,1,0,2,3 patients of the LPD group and 2,4,1,1,4,1,2,3,4 in patients of the OPD group,showing no statistically significant difference between the 2 groups (x2 =0.00,0.17,0.00,0.00,0.17,0.00,2.11,0.23,0.17,P＞0.05).Results of postoperative pathological examination showed that duodenal adenocarcinoma,ampullary carcinoma,lower bile duct carcinoma,pancreatic ductal adenocarcinoma and pancreatic cystic tumor were respectively detected in 8,2,5,3,2 patients of the LPD group and 10,2,4,2,2 patients of the OPD group,showing no statistically significant difference between the 2 groups (x2 =0.53,P＞0.05).Duration of postoperative hospital stay in the LPD and OPD groups were (19± 13) days and (15±7) days,respectively,showing no statistically significant difference between the 2 groups (t =1.28,P＞ 0.05).Results of postoperative oncology showed that tumor diameter,number of lymph node dissected,number of positive lymph nodes,cases with negative margin,cases in T1N0M0,T2N0M0,T3N0M0,T3N1M0,T4N0M0,T4N1M0 of TNM staging were respectively (2.4±0.7)cm,15.4±2.3,2,20,2,7,8,2,1,0 in the LPD group and (2.8±0.9)cm,14.4±2.5,3,20,1,8,5,2,3,1 in the OPD group,with no statistically significant difference between the 2 groups (t =-1.64,1.32,x2 =0.23,0.00,Z =-0.69,P＞ 0.05).(3) Follow-up situation:1 patient died respectively in both groups within the postoperative 30 days.Thirty-eight patients were followed up for 1-26 months,with a median time of 14 months.During follow-up,2 patients had tumor recurrence and 1 died of myocardial infarction in the LPD group;3 had tumor recurrence and 1 died of tumor recurrence in the OPD group.Conclusion LPD in 70 years of age or older patients is not only safe and feasible,but also significantly reduce volume of intraoperative blood loss and demand of analgesia,as well as quickly resume normal diet and activities.

OBJECTIVETo investigate whether the impairment of grafted liver after transplantation was induced by the same inflammatory cells in cold and warm ischemia.

METHODSMale SD rats were divided into two groups randomly, 24 grafted livers in each group were stored for 120 or 240 min at 4 degrees Centigrade Ringer's solution. Also male SD rats were divided into three groups, in which 24 grafted livers in each group were experienced warm ischemia ranged from 90, 120 to 150 min from non-heart-beating donor. The recipients were killed after 1, 3, 6, and 24 hours of transplantation for sample collection.

RESULTSAlong with the prolongation of cold and warm ischemia time, the serum ALT and AST levels were increased gradually after transplantation. Light microscopy showed some necroses in hepatocytes after 3 and 6 hours of transplantation in cold ischemia, and some neutrophilic infiltration in sinusoids. There were a large number of hepatocytes necroses after 3, 6 hours of transplantation in warm ischemia from non-heart-beating donor and a lot of lymphocytic infiltration in sinusoids. The findings in electron microscopy were as the same as those found in light microscopy, and the lymphocytes which infiltrated in sinusoids in warm ischemia were identified as T lymphocytes in electron microscopy.

CONCLUSIONSThe impairment of grafted livers after transplantation seems to be induced by two different inflammatory cells in cold and warm ischemia, that is, neutrophils mediate the cold ischemia-reperfusion, and T lymphocytes mediate the warm ischemia-reperfusion from non-heart-beating donor.

Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Graft Survival ; physiology ; Hepatocytes ; pathology ; ultrastructure ; Liver ; blood supply ; physiopathology ; ultrastructure ; Liver Transplantation ; physiology ; Male ; Neutrophils ; physiology ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; physiopathology ; T-Lymphocytes ; physiology ; Temperature ; Time Factors

The effects of RNAi-mediated gene silencing of LRIG3 expression on cell cycle and survival of human glioma cell line GLI 5 and the possible mechanisms were explored.The plasmids pGenesil2-LRIG3-shRNA l and pGenesil2-LRIG3-shRNA2 were transfected into GL 15 glioma cells respectively by using Metafectine,and the transfected cells that stably suppressed LRIG3 expression were selected by G418.The control cells were transfected with negative control shRNA.The changes in LRIG3 mRNA and protein levels were measured by RT-PCR and Western blot.The apoptosis rate and cell cycle were analyzed by flow cytometry.As compared with the negative shRNA-transfected GL15 cells,LRIG3 mRNA expression in GLI5 cells transfected with pGenesil2-LRIG3-shRNAl and pGenesil2-LRIG3-shRNA2 was silenced by 52.4%,63.8%,and LRIG3 protein expression was re-duced by 50.9% and 67.4% respectively.The LRIG3-specific siRNA transfected cells had higher proliferation rate than control cells.Cell cycle analysis showed that silencing LRIG3 increased the percentage of G2/M phase cells and the proliferation index significantly (P<0.01).Silencing LRIG3 could inhibit the apoptosis of GLl5 cells (P<0.05).These findings suggest that the siRNA targeting LRIG3 gene shows a dramatic inhibitory effect on RNA transcription and protein expression,then promoting the proliferation of GLI5 cells,arresting GLI5 cells in G2/M phase,and suppressing apoptosis ofGL15 cells.

A patient suffered a sustained soft tissue necrosis and infection at the radial interphalangeal joint of left thumb after laser nevus removal. He was treated in the Department of Orthopaedics, No. 920 Hospital of Joint Logistic Support Force of Chinese People's Liberation Army in February 2020. CTA combined with digital technology of Mimics software was used to accurately locate the perforator of posterior tibial artery septal perforator flap at the appropriate part of the calf and the super flap (1.20 cm×0.80 cm×0.46 cm) for the repair was designed. After 1 year of follow-up, the left thumb flap had no swelling with a satisfactory texture and appearance. The sensory recovered to S 3, and the left thumb movement was completely normal. Only a linear scar remained at the donor site of the calf.