Objective To study the bacterial types and their drug resistance in intra-abdominal infections after pancreatic surgery,and to evaluate the appropriate treatment measures.Methods 113 patients who underwent pancreatic surgery from Jan 2012 to Dec 2012 in our hospital were included into this study.The drainage liquid from the surgical sites were collected for bacterial culture and drug susceptibility tests.Results The incidence of intra-abdominal infections was 39.8％ (45/113).There were 54 pathogenic strains of bacteria isolated,including 49 strains of gram-negative bacteria (90.7％),4 strains of gram-positive bacteria (7.4％),and 1 strain of fungus (1.9％).The top three pathogens were Pseudomonas aeruginosa (50.0％),Acinetobacter baumannii (14.8％) and Singular deformation bacteria (1 1.1％).Most gram-negative bacteria were sensitive to Polymyxin B and Aminoglycoside antibiotics (＞ 70％),but they were resistant to Imipenem and Cephalosporin which were commonly administered.Pancreatic fistula was closely related to intra-abdominal infections.Concluusions A gram-negative bacteria,Pseudomonas aeruginosa,was the predominant organism in intra-abdominal infections after pancreatic surgery in our hospital.The situation of drug-resistance was still severe.More effective measures should be taken to prevent growth of resistant strains such as using antibiotics according to drug sensitivity and avoiding empirical single use of broad-spectrum antibiotics.Pancreatic fistula commonly led to intra-abdominal infections.

Objective To study the impact of lymph node metastasis on prognosis of patients with pancreatic cancer and to evaluate predictors of postoperative survival of these patients.Methods The clinical data on patients with pancreatic cancer who underwent pancreatic cancer radical surgery in our hospital from January 2002 to December 2013 were reviewed and analyzed.Data on lymph node metastasis,number of lymph node dissection,number of positive lymph nodes and positive lymph ratio were analyzed.Results Of 101 patients,the 6-month,1-year and 2-year survival rates were 84.2％,56.6％ and 28.5％,respectively.The median survival was 13.8 months.Univariate and multivariate analyses showed lymph node metastasis,a positive lymph node ratio,number of lymph node dissection and positive lymph nodes were independent influential factors of prognosis.Results of subgroup analysis showed the number of lymph node dissection was a prognostic factor for pNO patients,while a positive lymph ratio had no impact on survival of pN1 patients.In the subgroup of patients with pancreatic head cancer,lymph node metastasis was associated with prognosis but not in the subgroup of patients with pancreatic body and tail carcinoma.Conclusions For patients with pancreatic head cancer,lymph node metastasis was closely correlated with prognosis.In addition,factors including lymph node metastasis,number of lymph node dissection,a positive lymph node ratio and number of positive lymph nodes were independent influential factors of prognosis for patients with pancreatic head cancer.However,for pN1 patients,a positive lymph node ratio has no influence on prognosis.

Objective To analyze the risk factors of pancreatic fistula after pancreaticoduodenectomy,in order to provide evidence to reduce post-operative complication in clinical practice.Methods The clinical data of 352 patients with malignancy who received pancreaticoduodenectomy at the Shanghai Renji Hospital from September 2009 to September 2012 were retrospectively analyzed.The patients were divided into pancreatic fistula group and non-pancreatic fistula group.Peri-operative risk factors of pancreatic fistula after pancreaticoduodenectomy were analyzed by univariate and multivariate logistic regression analysis.Results Forty-nine cases of pancreatic fistula occurred,and the incidence rate of pancreatic fistula was 13.9％ (49/352).Univariate and multivariate logistic regression analysis showed sex,age,history of diabetes,operation time,intra-operative blood loss,vessel reconstruction,pancreatic tube placement,anastomosis time,type of digestive tract reconstruction were not risk factors of pancreatic fistula; however,brittle pancreatic tissue,diameter of pancreatic duct ＜3 mm,pre-operative total bilirubin level ＞ 171 μmol/l,duration of preoperative jaundice ＞ 8 weeks,pre-operative albumin level ＜30 g/L were the independent risk factors of pancreatic fistula (P ＜ 0.05).Conclusions Brittle pancreatic tissue,small pancreatic duct,high level of serum bilirubin,long duration of preoperative jaundice,low level of serum albumin are the independent risk factors of pancreatic fistula after pancreaticoduodenectomy.

Objective To establish the HeLa cell line that can stably express EYFP fluorescent protein as the model for anion channel blocker (halide ion) screening ,which lays the foundation for high throughput screening of anion channel blocker (halide ion) .Methods Through gene recombination technology ,a new lentivirus vector which can express mutant protein YFP (EYFP‐H148Q/I152L) and puromycin resistance ,was built .The mixture of lentivirus vector and packaging plasmid was transfected into 293T cells to produce lentivirus particles . After infection of HeLa cells by the lentivirus particles ,puromycin was used to screen the cells as YFP‐positive HeLa cell line .Then cell amplification was carried out after purification and efficiency of EYFP‐H148Q/I152L was further detected by Real‐time quantitative PCR (RT‐PCR) and Western blot .We then verified the activity of EYFP‐HeLa transfected cell line as a screening model of anion channel blocker .Results Gene sequencing verified that EYFP‐H148Q/I152L was successfully inserted into lentivirus vectors .RT‐PCR and Western blot results showed that the target gene was overexpressed in HeLa cells . The specific yellow fluorescence of EYFP of HeLa cells could be observed under fluorescence microscope with the efficiency of nearly 100% . I- (low permeability ) solution stimulated the opening of anion (halogen) channels ,and the yellow fluorescence was quenched by I - flow into cells . Conclusion The EYFP‐HeLa cell line can stably express EYFP yellow fluorescent protein and is sensitive to the internal flow of I - .Therefore ,it can be used as an ideal screening model of anion channel blocker (halide ion) .

A series of novel self-assembled polymeric micelles based on carboxymethyl chitosan bearing long chain alkyl chains (N-octyl-O, N-carboxymethyl chitosan, OCC) was synthesized. PTX loaded OCC polymeric micelles (PTX-OCC) were prepared by dialysis method. The effects of the degree of substitutions (DS) of octyl groups on the solubilizing abilities of OCC for paclitaxel were studied. The PTX-OCC were characterized using drug loading content, drug encapsulation efficiency, dynamic light scattering, zeta potential and transmission electron microscopy (TEM). Take PTX injection (PTX-INJ) as control, the safety of PTX-OCC including hemolysis, hypersensitiveness in guinea pigs and acute toxicity in mice were also evaluated. OCC showed excellent loading capacities for paclitaxel with the DS of octyl groups in the range of 37.9% - 58.6%. Drug loading contents were up to 24.9% - 34.4% with drug encapsulation efficiency 56.3% - 89.3%, which both increased with the increasing of DS of octyl groups. The mean size of PTX-OCC was 186.4 - 201.1 nm which decreased with the increasing of DS of octyl groups. The zeta potential was -47.5 to -50.9 mV, which had no obvious relation with the DS of octyl groups. The TEM images showed a spherical shape. No burst release phenomena were observed and drug cumulative release was in the range of 60% -95% in 15 days. PTX-OCC with higher DS of octyl groups showed stronger sustained releasing ability. In terms of the induction of membrane damage and hypersensitiveness, PTX-OCC was superior to PTX-INJ. The LD50 and its 95% confidence interval of PTX-OCC were 134.4 (125.0 - 144.6) mg x kg(-1), which was 2.7 fold of PTX-INJ. The present PTX-OCC could be potentially useful as safety carriers for intravenous delivery.
Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacology ; toxicity ; Chitosan ; analogs & derivatives ; chemistry ; Delayed-Action Preparations ; Drug Carriers ; Drug Delivery Systems ; Female ; Guinea Pigs ; Hemolysis ; drug effects ; Humans ; Hypersensitivity, Immediate ; chemically induced ; Male ; Mice ; Micelles ; Nanoparticles ; Paclitaxel ; administration & dosage ; pharmacology ; toxicity ; Particle Size ; Polymers

OBJECTIVETo study the inhibitory effect of paeoniflorin (PAE) on TNF-α-induced TNF receptor type I (TNFR1)-mediated signaling pathway in mouse renal arterial endothelial cells (AECs) and to explore its underlying molecular mechanisms.

METHODSMouse AECs were cultured in vitro and then they were treated by different concentrations PAE or TNF-α for various time periods. Expression levels of intercellular cell adhesion molecule-1 (ICAM-1) were detected in the normal group (cultured by serum-free culture media), the TNF-α group (cultured by 2-h serum-free culture media plus 6-h TNF-α 30 ng/mL), the low dose PAE group (cultured by 2-h PAE 0.8 μmo/L plus 6-h TNF-α 30 ng/mL), the middle dose PAE group (cultured by 2-h PAE 8 μmol/L plus 6-h TNF-α 30 ng/mL), the high dose PAE group (cultured by 2-h PAE 80 μmol/L plus 6-h TNF-α 30 ng/mL) with Western blot analysis. Nuclear translocation of transcription factor NF-κB (NE-κB) was detected in the normal group (cultured by serum-free culture media), the TNF-α group (cultured by 2-h serum-free culture media plus 45-mm TNF-α 30 ng/mL), and the high dose PAE group (cultured by 2-h PAE 80 μmol/L plus 45-min TNF-α 30 ng/mL) by immunofluorescent staining. Expression levels of the phosphorylation of extracellular signal-regulated (protein) kinase (ph-ERK) and p38 (ph- p38) were detected in the normal group (cultured by serum-free culture media) and the high dose PAE group (2-h PAE 80 μmol/L culture) by Western blot. NF-κB inhibitor-α (IκBα) protein expressions were detected in the normal group (cultured by serum-free culture media), the TNF-α group (cultured by 2-h serum-free culture media plus 30-min TNF-α 30 ng/mL), the high dose PAE group (cultured by 2-h PAE 80 μmol/L plus 30-min TNF-α 30 ng/mL), the p38 inhibitor group (SB group, pretreatment with SB238025 25 μmol/L for 30 min, then treated by PAE 80 μmol/L for 2 h, and finally treated by TNF-α 30 ng/mL for 30 min), the ERK inhibitor group (PD group, treated by PD98059 50 μmol/L for 30 min, then treated by PAE 80 μmol/L for 2 h, and finally treated by TNF-α 30 ng/mL for 30 min) by Western blot.

RESULTSCompared with the normal group, ICAM-1 protein expression levels obviously increased (P < 0.01). Compared with the TNFα group, ICAM-1 protein expression levels were obviously inhibited in the high dose PAE group (P < 0.05). Protein expression levels of ph-p38 and ph-ERK were obviously higher in the hIgh dose PAE group (P < 0.05). Compared with the normal group, IκBα protein expression levels obviously decreased in the TNF-α group (P < 0.01). Compared with the TNFα group, TNF-α-induced IκBα degradation could be significantly inhibited in the high dose PAE group (P < 0.01); the inhibition of PAE on IκBα degradation could be significantly inhibited in the SB group (P < 0.05). NF-κB/p65 signal was mainly located in cytoplasm in the normal group. NF-κB/p65 was translocated from cytoplasm to nucleus after stimulated by 45 min TNF-α in the TNF-α group, while it could be significantly inhibited in the high dose PAE group.

CONCLUSIONSPAE inhibited TNF-α-induced expression of lCAM-1. Its action might be associated with inhibiting TNFR1/NF-κB signaling pathway. p38 participated and mediated these actions.

Animals ; Cells, Cultured ; Endothelial Cells ; cytology ; drug effects ; Glucosides ; pharmacology ; Intercellular Adhesion Molecule-1 ; metabolism ; Mice ; Monoterpenes ; pharmacology ; NF-kappa B ; metabolism ; Receptors, Tumor Necrosis Factor ; metabolism ; Signal Transduction ; drug effects ; Tumor Necrosis Factor-alpha ; pharmacology

Objective To investigate the relationship between PPAR coactivator 1 (PGC-1), nuclear respiratory factor (NRF), mitochondrial transcription factor A (mtTFA) expressions of vascular smooth muscle cells (VSMC) and development of atherosclerosis in a rabbit model.Methods Atherosclerotic model was established by feeding the rabbits with high-fat diet for 4, 8 and 12 weeks ( n = 10 each).Another 8 rabbits fed with normal diet served as normal controls.Intima-media ratio, mRNA and protein expressions of PGC-1, NRF, mtTFA and SMemb, a marker for synthetic VSMC, were detected on aorta specimens.Results With the blood lipid increased, the intima-media ratio rose from (0.031 ± 0.010) μm up to (0.814±0.258) μm during 12 weeks.Increasing SMemb means that synthetic VSMC grew more and more.The expressions of PGC-1 became significant after 4 weeks (P < 0.01 ), while that of NRF-1 and mtTFA rose significantly after 8 weeks (P < 0.01 ).conclusions The PGC-NRF-mtTFA pathway might play a critical role in VSMC proliferation and development of atherosclerosis.

Objective To investigate the prognostic factors of survival for patients with duodenal papilla carcinoma (DPC) after pancreaticoduodenectomy.Methods 98 DPC patients undergoing pancreaticoduodenectomy with follow-up from Jan 2010 to Dec 2017 at Renji Hospital,School of Medicine,Shanghai Jiao Tong University were analyzed retrospectively.Results 80 Cases were followed up.The 1,3,and 5 year survival rates of 80 patients were 89.9％,72.4％,and 66.6％,respectively.Univariate analysis showed tumor size,T stage,N stage,TNM stage,tissue differentiation degree were related to postoperative survival(all P ＜0.05).Multivariate analysis showed that tumor size,N stage,TNM stage,and tissue differentiation degree were independent factors influencing postoperative prognosis(all P ＜ 0.05).Conclusions Tumor size,N stage,TNM stage and tumor tissue differentiation degree were independent factors influencing the prognosis of DPC after pancreaticoduodenectomy,suggesting that early diagnosis,early treatment and radical resection were the key to improve the postoperative prognosis of DPC.

Objective:To evaluate the effect of prophylactic octreotide administration on pancreaticoduodenectomy (PD)associated postoperative pancreatic fistula (POPF), total complications, peri-operative death and postoperative in-hospital days.Methods:From January 2020 to August 2021, 148 patients who underwent PD in the Department of Biliary-Pancreatic Surgery in Ren Ji Hospital affiliated with School of Medicine of Shanghai Jiao Tong University were recruited into this single-center randomized control double-blinded clinical trial. Patients were randomly assigned into octreotide group ( n=74) and control group ( n=74). Octreotide group was subcutaneously injected with 0.1 mg (1 ml) octreotide after preoperative anesthesia, and was subcutaneously injected with the same dose every 8 hours for 5 days, with a total of 16 doses. Control group was injected with 1 ml normal saline in the same way, and relevant clinical data and indicators of the two groups were recorded. The primary endpoint was clinically relevant pancreatic fistula, and the secondary endpoints were total complications, perioperative death and postoperative in-hospital days. Univariate and multivariate logistic regression analysis were used to screen the risk factors of clinically related POPF after PD. Results:120 patients were finally enrolled, including 61 in octreotide group and 59 in control group. There were no significant differences on age, gender ratio, body mass index, preoperative surgery rate of jaundice reduction, preoperative major biochemical indicators, operation time, intraoperative blood loss, pancreatic duct diameter, pancreatic texture and pathological type composition ratio. The total incidence of clinical relevant POPF was 8.3%, and there were no significant differences on biochemical leakage (4.9% vs 8.5%, P=0.435), grade B fistula (4.9% vs 8.5%, P=0.435) and grade C fistula (1.6% vs 1.7%, P=0.981). The total complication incidence (24.5% vs 28.8%, P=0.601), perioperative mortality (0 vs 3.3%, P=0.147) and postoperative in-hospital days (20.6±11.1 d vs 19.5±12.2 d, P=0.633) were not significantly different between two groups. Univariate analysis showed that preoperative serum albumin level <30 g/L( P<0.001) and pathological type of pancreatic ductal adenocarcinoma ( P=0.036) were independent risk factors for POPF after PD, while multivariate analysis found no statistically significant risk factors. Conclusions:Octreotide can neither reduce the incidences of POPF, total complications and postoperative mortality, nor shorten postoperative in-hospital days. However, for patients with preoperative hypoproteinemia and (or) the pathological type of pancreatic duct adenocarcinoma, the prophylactic use of octreotide during PD and after PD may reduce the occurrence of POPF.

During the progression of heart failure(HF),abnormal transduction of the transforming growth factor-β(TGF-β)/Smads signaling pathway is important mechanism of myocardial fibrosis(MF)in HF.TGF-β,a key factor in MF,is in an overexpression state in the process of MF in HF,and Smads is a major effector downstream of TGF-β.The TGF-β/Smads pathway induces abnormal proliferation of myofibroblasts,aggravates myocardial extracellular matrix deposition,and reduces the ability of the cardiac tissues to resist fibrosis,which plays a complex role in the pathogenesis of MF in HF.Traditional Chinese medicine(TCM)has the efficacy of unequivocal inhibiting myocardial collagen deposition,anti-MF,protecting the myocardium and improving cardiac function in the prevention and treatment of MF in HF and so on,and the TGF-β/Smads pathway is one of the key pathways through which TCM monomers,TCM combinations,and proprietary medicines can exert their cardioprotective effects on the HF.This paper reviews the existing experimental research results of TCM intervening in the TGF-β/Smads pathway for the treatment of MF in HF over the past 10 years,with a view to providing theoretical basis for the prevention and treatment of HF MF well as the development and of new drugs.