Objective To construct the lentiviral overexpression vectors of mouse neuromedin B(NMB) and neuromedin B receptor(NMBR) genes and express them in RAW264.7 cells, so as to lay a foundation for further study on the effects of mouse NMB and NMBR gene overexpression on the proliferation and apoptosis of RAW264.7 cells. Methods The coding sequences(CDSs) of mouse NMB and NMBR genes were amplified by RT-PCR and inserted into vector pCD513B-1 to construct recombinant plasmids pCD513B-1-NMB and pCD513B-1-NMBR. Mouse NMB and NMBR gene overexpression lentiviruses were obtained through packaging by HEK-293T cells, and the virus titers were detected by double dilution method. After infection with lentivirus for 48 h, RAW264.7 cells were detected for the expression of NMB and NMBR mRNA by qPCR using the uninfected cells as control. Results The recombinant plasmids were constructed correctly as identified by double enzyme digestion. The virus titers of mouse NMB and NMBR gene overexpression lentiviruses were 6 × 106and 8 ×106TU/mL, respectively. The mRNA expression levels of mouse NMB and NMBR genes in RAW264.7 cells transfected with two lentiviruses were significantly higher than those in the control group(t = 24. 158 and 14. 958, respectively, each P <0. 01). Conclusion Mouse NMB and NMBR gene overexpression vectors were successfully constructed, which can significantly increase the expression of NMB and NMBR genes in RAW264.7 cells.

Surgical treatment is the only cure treatment for patients with inferior vena cava tumor thrombus in hepatic segment and upper hepatic segment.The accurate diagnosis of tumor thrombus is very important.In preoperative imaging examination,the abdominal enhanced CT scan and the inferior vena cava MRI scan were the best methods for the diagnosis and evaluation of the tumor thrombus in hepatic segment and upper hepatic segment.Compared with the tumor thrombus below the liver,the tumor thrombus in hepatic segment or above hepatic segment extend widely,and the operation are more difficult.For simple inferior vena cava tumor thrombus (the top of the thrombus has reached the level of hepatic vein),Retroperitoneal approach combined with transperitoneal approach should be used.Open surgery is the standard procedure for other tumor thrombus in hepatic segment and upper hepatic segment.In addition to exposure of inferior vena cava below the hepatic vein,the liver and the first hepatic hilum should be exposed.For tumor thrombus in the atrium,after the longitudinal incision of diaphragm,we use Milking technology to squeeze thrombus into inferior vena cava.Then we use catheterization technology to remove thrombus.For difficult atrial tumor thrombus,an extracorporeal circulation should be performed.The median incision in the chest should be performed to open the chest and open the pericardium and remove the tumor thrombus.Patients with tumor thrombus in hepatic segment or upper hepatic segment should be diagnosed as early as possible and they need actively treated by operation.

OBJECTIVE: To investigate the protective effect and probable mechanism of 20-Hydroxyecdysone on SH-SY5Y cells injured by MPP+. METHODS: MPP+ induced SH-SY5Y cells injury model was established. 20-Hydroxyecdysone was added into the culture to test its protective effect. The morphological changes of cells were observed. Cell viability was detected by method of MTT. The contents of LDH, MDA and activity of SOD were inspected by kits. Apoptosis rate of cells was detected by flow cytometry. RESULTS: Compared with MPP+ group, 20-Hydroxyecdysone group (50, 100, 150 μmol·L-1) alleviated the damage of SH-SY5Y cells induced by MPP+, significantly improved cell survival rate, reduced the release of LDH, increased the activity of SOD, decreased the contents of MDA and reduced the apoptosis of cells. CONCLUSION: 20-Hydroxyecdysone has a significant protective effect on SH-SY5Y cells injured by MPP+. The probable mechanism may be anti-oxidation and anti-apoptosis.

Objective To determine the effect of ABCC4 gene silencing on cell proliferation and cell cycle in human pancreatic cancer cell lines PANC1 and BxPC-3.Methods PANC1 and BxPC-3 pancreatic cancer cells were transfected with a lentivirus expressing an ABCCA short hairpin RNA (shRNA).ABCC4 mRNA and protein expression of transfected cells was determined by RT-PCR and Western blot,colony formation ability was measured by colony assay,and cell cycle change was investigated by the flow cytometric analysis.Results Lentivirus expressing an ABCC4 short hairpin RNA (shRNA) was successfully established.After transfection with shRNA lentivirus,ABCC4 mRNA and protein expression were significantly inhibited (0.28 ±0.01 vs 1.00 ±0.03,0.22 ±0.02 vs 1.00 ±0.03,P ＜0.05).Colony formation ability was significantly decreased (4 vs 65,P ＜0.05),and cell cycle was significantly blocked at G1 phase [(54.98 ±1.78) ％ vs (42.93 ± 0.88) ％,(68.55 ± 0.75) ％ vs (54.76 ± 0.29) ％].Conclusions ABCC4 gene silencing can significantly inhibit cell proliferation of human pancreatic cancer cell lines PANC1 and BxPC-3,and block the cells at G1 phase.

Objective To investigate the level change and correlation of serum IL-18 and soluble Fas/Fas ligand(sFas/sFasL) in patients with chronic obstructive pulmonary disease (COPD) to evaluate their roles in the pathogenesis of COPD. Methods The levels of serum IL-18 and sFas/sFasL in 36 patients with COPD of acute aggravation stage and 20 healthy control subjects were detected by ELISA. Results The levels of serum sFasL and IL-18 in patients with COPD were significantly lower than those in healthy control subjects, while there was not significant differene in serum sFas level between the two groups. Conclusion The serum sFasL and IL-18 level decreass in patients with COPD of acute aggravation stage indicated that cell apoptosis level and immune function reduced in the patients.

Objective To study the relationship between the dose indicator of computed radiography (CR) and the entrance surface dose (ESD),and to build a model for estimating ESD based on this relationship.Methods Taking Kodak CR system as the research object,a theoretical model for estimating the ESD was established according to theoretical derivations,and the key parameters in the model were determined through experiments in a CR system.Further experiments in another CR system were also conducted to verify the model.Results The ESDs were not only dependent on the dose indicator provided by the CR system,but also influenced by other factors,such as tube potential,patient's body thickness and energy response of the imaging plate.The estimation results of the model agreed well with the experiment results,and the relative deviation was confirmed within 20％.Conclusions The dose indicator based model can provide a relatively fast and easy way for evaluating the doses of patients undergoing X-ray diagnoses with the CR system.

Objective To summarize the experience of surgical management of postoperative intrahepatic duct stones and strictures with choledochoscope.Methods The clinical data of 1241 patients with intrahepatic duct stones and strictures who were admitted to the Shenzhen People's Hospital from January 2000 to December 2011 were retrospectively analyzed.All patients were divided into postoperative choledochoscopy group (652 patients were treated by choledochoscope through T-tube fistula) and conservative treatment group (589 patients were treated by cholangic and antibiotic drugs) according to treatment plan.The clinical data including stricture alleviation,incidence of complications and treatment efficacies of the 2 groups were compared using the chi-square test.Results In the postoperative choledochoscopy group,the intrahepatic duct stones of 630 patients were completely removed,with the removal rate of 96.63％ (630/652),which was significandy higher than 76.23％ (449/589) of the conservative treatment group (x2=113.407,P ＜ 0.05).A total of 1237 intrahepatic duct strictures were detected in the postoperative choledochoscopy group,including 698 cases of membranous stricture (the length of strictured bile duct ＜ 2 mm),529 cases of spool-shaped stricture (2 mm ≤ the length of strictured bile duct ≤5 mm),8 cases of long tube-shaped stricture (the length of strictured bile duct ＞ 5 mm),2 cases of crackshaped stricture.The alleviation rate of the stricture was 99.35％ (1229/1237).The conditions of 6 cases of long tube-shaped stricture and 2 cases of crack-shaped stricture were not alleviated.There were 986 cases of intrahepatic duct strictures in the conservative treatment group,and the stricture alleviation rate was 80.43％ (793/986).There was a significant difference in the stricture alleviation rate between the 2 groups (x2=238.994,P ＜ 0.05).Nine patients in the postoperative choledochoscopy group had severe complications.Five patients had intrahepatic duct hemorrhage with a volume of blood loss more than 1500 ml,and they were treated by balloon dilatation hemostasis.One patient with repeated intrahepatic duct bleeding received hepatectomy.Two patients with severe intrahepatic cholangitis received choledochoscopy.Two patients had incomplete formation of sinus and they received implantation of biliary stents under duodenoscope.Conclusion Standard surgical management with choledochoscope can effectively alleviate postoperative intrahepatic duct stones and strictures.

Objective To determ ine the norm al reference values of 33 elem ents, Ag, Al, As, Au, B , B a, Be, Ca, Cd, Co, Cr, Cs, Cu, Fe, G a, H g, Li, Mg, Mn, Mo, N i, Pb, Rb, Sb, Se, Sr, Th, Ti, Tl, U , V , Zn and Zr, in the blood and urine sam ples from the general population in Sanm en County of Zhejiang province, a typical coastal area of eastern China. Methods The 33 elem ents in 272 blood and 300 urine sam ples w ere determ ined by inductively coupled plasm a-m ass spectrom etry (ICP-MS ). The norm ality test of data w as conducted using SPSS 17.0 Statistics.The data w as com pared w ith other reports. Results The norm al reference values of the 33 elem ents in the blood and urine sam ples from the general population in Sanm en County w ere obtained, w hich of som e elem ents w ere found to be sim ilar w ith other reports, such as Co, Cu, Mn and Sr, w hile As, Cd, H g and Pb w ere generally found to be higher than those previously reported. There w as a w ide variation betw een the reports from different countries in blood B a. Conclusion The norm al reference values of the 33 elem ents in the blood and urine sam ples from the general population in Sanm en County are established, and successfully applied to tw o poisoning cases.

Background Bevacizumab has been widely used in the treatment of new blood vessel disease in ophthalmology.The investigation of the pharmacokinetics and safety after intracameral injection of bevacizumab can offer the basis for the management of iris neovascularization and neovascular glaucoma.Objective The present study was to observe the distribution of bevacizumab(avastin)in eye tissue and toxic effects following the injection of anterior chamber.Methods Twenty-four New Zealand albino rabbits were divided into two groups randomly.0.05 ml (1.25mg)of Bevacizumab was intracamerally injected into the left eyes in the experimental group,and a balanced salt solution of 0.05 ml was injected in the same way into the left eyes of the control group.The anterior segment of eyes and ocular fundus were examined by slit-lamp microscope and direct ophthalmoscope after injection.Intraocular pressure was measured and corneal endothelial microscopy was performed before and after the injections.Five rabbits of the two groups were sacrificed on the first day,the fourth day,the seventh day,the fourteenth day,and the thirtieth day after injection,and the eyeballs were enucleated for histopathological examination.The ultrastructure of eye tissue was observed under the transmission electron microscope on the fourth day and the thirtieth day,and then immunofluorescence staining were performed to assess the distribution of bevacizumab in the eye tissues.This experiment complied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission(Version 1988).Results No abnormality in the cornea,lens,vitreous and retina was observed after the injection of bevacizumab under the slit lamp microscope and direct ophthalmoscope.No significant differences were found in intraocular pressure and corneal endothelial cell density in the bevacizumab group compared with the control group before injection and 2 hours,1 day,7 days,14 days,30 days after injection(P =0.760,P =0.956).No histopathological and ultrastructural changes of the cornea,lens,chamber angle,iris,ciliary body and retina were seen after the injection in the experimental group and control group under the light microscope and transmission electron microscope.Bevacizumab was distributed in the anterior chamber angle,iris,ciliary body,choroid and retina in injected eyes and fellow eyes after intracameral injection with red fluorescence and presented the dynamic changes with the lapse of time.The immunofluorescence response of eye tissue to bevacizumab was weaker in the fellow eyes compared with injected eyes.Bevacizumab was mainly distributed in the vessel wall and lumen.Conclusions Bevacizumab can quickly distribute in the vascular tissue of the anterior chamber angle,iris,ciliary body,choroid and retina in injected eyes after intracameral injection without obvious toxic effects to eye tissue.Bevacizumab administered intracamerally may be a new strategy or a joint strategy for iris neovascularisation.