im To study the structure-function relationship of hTNFα . Methods We compared the cytotoxicity, receptor binding ability and toxicity in animal body of wild type(wt)hTNFα with its mutants including R2K-, N30S-, R32W-, L157F-hTNFα , and two multi-site mutants(R32W-L157F-hTNFα and R2K-N30S-R32W-L157F-hTNFα ). Results We found that the two multi-site mutants remained similar cytotoxicity to several human tumor cell lines as wild type hTNFα . However, their cytotoxicity to L929 cells were decreased sharply as compared with those of wt hTNFα . The two multi-site hTNFα mutants had lower binding activity with hTR75 than hTR55. We also found that compared with the wild type, the LD50 of the mutant R32W-L157F-hTNFα was decreased about 300 fold and the dose of mutant R2K-N30S-R32W-L157F-hTNFα resulted in 30％ death was 700 folds lower than LD50 of wt hTNFα . To certify the systematic toxicity of the mutant R2K-N30S-R32W-L157F-hTNFα , we assayed its toxicity to monkeys and found that its systematic toxicity was lower than that of wt hTNFα. Conclusion A 4-site mutants(R2k-N30S-R32W-L157F-hTNFα )of hTNFα is obtained, which the mutant may possess potential application value in clinical therapy.

OBLECTIVE:To promote a sustained and quick development of the high-tech medicine industry in our country.METHODS:The related literature was consulted and the current developmental status of the high-tech medicine in-dustry in our country was analyzed.RESULTS&CONCLUSIONS:The high-tech industry in our country can be pushed on only through rearrange industry mix at the right time,constructing a serial comprehensive development terrace,supporting es-pecially those medicine enterprises that with core technology and independent intellectual property rights,building a virtual strategic league together with the academy of higher learning and scientific research institutes.

Objective To evaluate the effectiveness of mouth care combined with intestinal and endoscopic working channel washing for ERCP related cholangitis.Methods A total of 573 patients diagnosed as having obstructive jaundice were randomized into three groups,190 cases in the control group,192 in the saline group and 191 in the amikacin group.Clinical and laboratory data were collected before ERCP and ERCP related cholangitis were recorded.Results There were no significant differences among the three groups in sex,age,the level of obstruction,the category of obstruction,total bilirubin or WBC counting.The incidences of ERCP related cholangitis were 21.1％ (40/190),13.5％ (26/192) and 4.7％ (9/191)in control group,saline group and amikacin group respectively,which was significantly different (x2 =22.409,P =0.000 ).The incidences of ERCP related cholangitis were 19.5％ ( 65/333 ) and 4.2％(10/240) in patients diagnosed as having hilar duct obstruction and low positioned biliary obstruction respectively (x2 =27.175,P =0.000).There was no significant difference in ERCP related cholangitis between benign and malignant biliary obstruction.Subgroup of hilar duct obstruction showed the incidences of ERCP related cholangitis were 29.7％ (33/111 ),20.5％ (24/117)and 7.6％ (8/105)in the control group,the saline group and the amikacin group,respectively (x2 =16.905,P =0.000).Conclusion The incidence of ERCP related cholangitis is relatively higher in patients with hilar duct obstruction.Mouth care combined with intestinal and endoscopic working channel washing can effectively reduce the incidence of ERCP related cholangitis,especially in the amikacin group.

ObjectiveTo evaluate the feasibility and safety of endoscopic intraductal radiofrequency ablation (RFA) for unresectable malignant biliary strictures.MethodsA total of 12 patients with malignant biliary obstructions,who were unsuitable for surgical resection,were prospectively selected for this study.During ERCP,when successful biliary cannulation achieved,a bipolar radiofrequency probe was introduced into bile duct over a guidewire.RFA was given to the tumor lesion under fluoroscopy,followed by stent placement.The patients were closely observed and followed up after the procedure.ResultsSuccessful RFA was achieved in all patients before stents insertion ( plastic and metal stents in 6 respectively,additional pancreatic stents were given to 3).Mild complications of cholangitis and pancreatitis occurred in 1 patient respectively,which were cured with short-term management.Jaundice was promptly controlled in 7 patients (58.3％ ).During the mean follow-up duration of 3.4 (range 0.5-5.5) months,one patient died of cardiocerebral accident and others survived without obvious disconffort.The 1- month and 3- month stent patency was 100％ (12/12) and 80％ (8/10),respectively.ConclusionIt is technically feasible and safe to perform endoscopic intraductal RFA for the palliation of biliary malignancies.The short-term outcome is satisfying,however,long-term result and the better therapeutic schedule need further evaluation.

Objective To evaluate endoscopic retrograde cholangiopancreatography (ERCP) for diagnosis and management of biliary cast syndrome after orthotropic liver transplantation. Methods A total of 71 consecutive patients with abnormal liver function and MRCP findings after liver transplantation underwent ERCP for diagnosis and management. Their data were retrospectively reviewed. Results A total of 188 sessions of ERCP were carried out on the 71 patients, most of whom were found to have stenosis of anastomotic stoma and/or bile duct. Bile sludge was found and removed in all patients diagnosed within 3 months after liver transplantation, while pigmentoid stones were found and removed in patients diagnosed within 3-6 months and biliary casts in patients diagnosed at more than 6 months. Each patient underwent 2.6 sessions averagely. Biliary casts were formed at an average time of 22. 7 ± 15.6 months after transplantation. PostERCP complications included 2 cases of pancreatitis and 3 cholangitis, with an occurrence rate of 2. 6%(5/188), which were all controlled with conservative treatment. The follow-up data was available in 56 patients showing improvement in liver function after ERCP, among who 42 met the endoscopic criteria of cure,1 0 received second liver transplantation because of progressive sclerosing cholangitis and 4 died from diseases other than liver transplantation. Conclusion Therapeutic ERCP for the biliary cast syndrome after liver transplantation is feasible, safe and effective, and can be performed repeatedly with good short-term effect.

Objective To investigate whether endoscopic nasobiliary drainage (ENBD) after endoscopic stone extraction could reduce post-operative complications.Methods Retrospective analysis of 697 patients with extrahepatic duct stones undergoing ERCP treatment was pedormed.They were assigned to ENBD group (n =538) and control group (n =159).Post-operative complications including pancreatitis,cholangitis and bleeding were observed and compared.Results There were no significant differences in incidence of pancreatitis and bleeding between 2 groups.Incidence of cholangitis in ENBD group was significantly lower than that in control group (1.3％ vs.3.8％,P ＜ 0.05).Conclusion ENBD after endoscopic stone extraction could reduce the incidence of postoperative cholangitis.

Objective To evaluate the effectiveness of mouth care combined with intestinal and endoscopic working channel flushing for ERCP related cholangitis and to investigate the mechanisms.Methods A total of 284 patients who underwent ERCP were divided into the amikacin group (n =96),the saline group (n =95) and the control group (n =93).Specimens were collected before cannulation (Sample 1),the first access to the bile duct (Sample 2) and at the end of ERCP (Sample 3).All the samples underwent bacterial culture and colony counting.Incidence of ERCP related cholangitis and bacterial results were recorded.Results The incidences of ERCP related cholangitis were 7.3％ (7/96),8.4％ (8/95)and 18.3％ (17/93)in amikacin group,saline group and control group respectively,with significant differences among three groups (x2 =6.861,P =0.032).The positive rate of bacterial culture of Sample 1 in the amikacin group,saline group and control group were 6.3％ (6/96),25.3％ (24/95) and 28.0％ (26/93) (x2 =16.833,P =0.000),respectively.The positive rate of bacterial culture of Sample 2 in amikacin group,saline group and control group were 6.3％ (6/96),16.8％ (16/95) and 24.7％ (23/93)(x2 =12.206,P =0.002),respectively.The positive rate of bacterial culture of Sample 3 in amikacin group,saline group and control group were 7.3％ (7/96),17.9％ (17/95) and 33.3％ (31/93) (x2 =20.713,P =0.000),respectively.Conclusion The procedure of ERCP can bring the intestinal bacteria into the bile duct,which could be reduced by mouth care combined with intestinal and endoscopic working channel flushing.And the incidence of ERCP related cholangitis can be effectively reduced.

Background Heat shock proteins (HSPs) are highly conserved proteins that are induced in cells when confronted with a wide variety of proteotoxic stresses.HSP27 has a high degree of similarity with α-crystallin protein.The abnormality of HSP27 structure and expression are closely related to the formation of cataracts.Our previous study showed sodium salieylate has the protective effect on H2O2-induced lens damage.Objective This study was to investigate the roles of MAPK signal pathway in sodium salicylate-induced the expression of HSP27 in human lens epithelial cells (LECs) in vitro.Methods Human LECs were incubated in the fresh media containing sodium salicylate at different concentrations (0-55 mmol/L) for different times (1-5 hours) and allowed to be recovered in fresh medium without sodium salicylate for 1-24 hours with or without pretreatment with P38MAPK inhibitor (SB203580), ERK1/2 inhibitor (PD98059) and JNK/SAPK inhibitor ( SP600125). The expressions of P38MAPK, EBK1/2, JNK/SAPK, phosphorylated P38MAPK, phosphorylated ERK1/2, phosphorylated JNK/SAPK and HSP27 were detected by Western blot. HSP27 mRNA was detected by RT-PCR. The expression of HSP27 was also detected by immunohistochemistry. Results There was only weak expression of HSP27 in normal human LECs.After stimulation of 35-55 mmol/L sodium salicylate was removed and human LECs were cultured again for 6 hours,the expression of HSP27 in LECs were significantly increased ( F= 509. 953,P<0. 01). HSP27 was absent expressed in human LECs in 55 mmol/L sodium salicylate stimulation for 1-5 hours groups, but LECs were re-cultured for 3,6 hours after removed the stimulation, the expression of HSP27 was elevated (F = 452. 534, P<0. 01). Activation of P38 M APK occurred after sodium salicylate stimulation 30 minutes and 1 hour ( F = 865.68, P<0. 01). However, ERK 1/2 was expressed after sodium salicylate was eliminated for 1-6 hours ( F = 388.84, P<0. 01). JNK/SAPK was inactived by sodium salicylate. The expression of HSP27 could be down-regulated with the pretreatment of SB203580 and PD98059. Conclusion Sodium salicylatc can induce the expression of HSP27 in human (LECs) . The effects are mediated,at least in part ,through the activation of P38MAPK and ERK1/2 signaling pathway .

Objective To study the effect of infrasound on the changes of expression of NMDAR1 in hipp- ocampal cells.Methods Eighty-eight male Sprague-Dawley rats were randomized into eleven groups:control group,90 dB/1 d,7 d,14 d,21 d and 28 d infrasound exposed groups;130 dB/1 d,7 d,14 d,21 d and 28 d infra- sound exposed groups.All the animals in the test groups were put in an infrasound field with 8 Hz,90 dB or 130 dB for 2 hours daily.Immunohistochemistry methods were used to detect the changes of intracellular expression of NMDARI in hippocampal cells.Methods The expression of NMDAR1 in hippocampus after the rats were exposed to infrasound of 8 Hz,90 dB SPL showed a procedure from reducing on the 1st day to rising on the 7th and peaked on the 14th day,then to descending on 21st day and returning to the standard level on the 28th day.Exposure to infra- sound of 8 Hz,130 dB SPL induced opposite effects on the expression of NMDAR1 compared with 90 dB SPL,which showed a process of increasing,descending,reaching to the lowest,then ascending and returning to the normal.The lowest expression of NMDAR1 occurred on the 14th day in every observed hippocampal area.Conclusion 8 Hz, 90 dB/130 dB infrasound induced certain reversible reaction in the expression of NMDAR1 of hippoeampal cells in rats,which may disturb their learning and memory function.

Objective To improve the success rate of selective cannulation of major papilla during endoscopic retrograde cholangiopancreatography (ERCP).Methods When cannulation failed with conventional methods,ultra-fine guide wire combined with taped cannulation,precut papillotomy,guide wire pancreatic occupation technique,and percutaneous transhepatic cholangial drainage (PTCD) assisted rendezvous technique was applied to improve the success rate of cannulation of major papilla.Results A total of 5743 patients received ERCP in our hospital during last 5 years,with a success rate of cannulation of major papilla at 98.6% (5664/5743).For 396 patients with difficult cannulation under conventional methods,ultra fine guide-wire technique was applied in 20,pancreatic occupation technique in 67,precut technique in 294 and PTCD assisted rendezvous technique in 15,which achieved a success rate of 80.0% (317/396).Conclusion Application of combined techniques according to the features of papilla can increase success rate of difficult cannulation.