Objective To investigate the protective effects and the undedying mechanism of recombinant human tumor necrosis factor receptor: Fc fusion protein (Yisaipu, rhu TNFR: Fc) on the lipopolysaccharide (LPS) induced acute liver injury of rats. Method Totally48 SD rats were randondy divided into four groups , in-cluding control gronp (n = 12), Yisaipu group(n = 12), LPS gronp(n = 12) and Yisaipu + IPS group(n = 12). The models of acute liver injury were produced by injection of LPS intravenously. Being fasted for 12 h, the rats were anaesthetized (60 mg/kg pentobarbital sodium, i.p.) and cannulated into carotid arteries. The cannula was connected with the multi-channel creature signal analysis system. The rata in control group and LPS group were injected with normal saline or LPS in dose of 5 mg/kg through rats' sublingual vein respectively. While the rats in Yisaipu group and Yisaipu + LPS group was pretreated with Yisaipu in dose of 0.4 mg/kg subcutaneously 24 h be-fore normal saline or LPS infusion. Six rats of each goup were randomly selected and mean arterial pressure (MAP) were monitored for 6 h via multi-channel creature signal analysis system, and rats' survival rate was calcu-lated. The rats whose MAP less than 10 mmHg were considered to die and the alive rats during period of observa-tion sacrificed by exsanguination. The liver tissue at the same site was removed, fixing in 10% formalin or stored at -80 ℃. To detect serum TNF-α, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) level, 0.2 mL blood samples were collected from the carotid artery 2 and 3 h after the injection of saline or LPS. The serum was collected from centrifuged blood samples and stored at -80 ℃. Enzyme-linked immunosorbent assay (ELISA) and flow cytometry were used to assess serum TNF-α level and bioactivity respectively. We also measured the serum ALT and AST levels, the myeloperoxiase (MPO) and superexide dismutase (SOD) activity, and malon-dialdehyde (MDA) content in liver tissue The pathology of hepatic tissue was evaluated by HE staining. Statistical-ly,the data of TNF-α level and bioactivity, ALT and AST release, and MDA content were analyzed by ANOVA, and rat survival rate were analyzed by Chi-square Tests. Results The rats in control group and Yisaipu group were all survived. Rat survival rate was significantly higher in Yisaipu + LPS group (67%) than in LPS group (17%) (P < 0.05). Serum TNF-α bioactivity was significantly lower in Yisaipu + LPS group than in LPS group [(7.3±2.8)% vs.(51.3±6.4)%, P <0.05]. Compared with IPS group, Yisaipu pretreatment decreased MDA content [(1.40±0.10)vs. (2.81±0.11) nmol/mgprot, P <0.05]and MPO acticity [(0.38±0.04) vs. (0.54±0.02) U/g, P <0.05]in hepatic tissue, while SOD activity [(188.4±20.2) vs. (142.5 ± 18.3) U/mgprot, P <0.05]was increased. The serum AST level, ALT level and the pathology in the liver were also ameliorated correspondingly. Conclusions These data suggest that Yisaipu could protect rats from LPsinduced a-cute liver injury by inhibiting TNF-α bioactivity and by enhancing anti-oxidation.

Objective To investigate the protective effects of recombinant human tumor necrosis factor receptor:Fc fusion protein (rhuTNFR:Ice)on the acute renal injury induced by lipepelysaccharide(LPS)in rats.Methods Models ofacute renal injury in rats were constructed by intravenous injection of LPS 10 m/kg.Forty-eight SD rats weighing 180～240g were randomly divided into 4 groups with 12 animals each group,including control group,rhuTNFR:Fc group,LPS groupand rhuTNFR:Fc+LPS group.Mean arterial pressure(MAP)wKs continuously monitored for 6 h.The levels of blood tLrea nitrogen(BUN)。Creatinine(Cr),TNF-αas well as TNF-α bioactivity were assessed.The myeloperoxinse(MPO)and superoxide dismutase(SOD)activity,content ofmalondialdehyde(MDA)were also measured.Pathologic changes of lung tissue in each group were observed by HE staining.Results Compared with LPS group,the status of hypotension and pathological manifestation in kidneys were ameliorated,and MPO activity significantly decreased in rhuTNFR:Fc+LPS group(P<0.05).Conclusion These data suggest that rhuTNFR:Fc can ablate the rise in serum TNF-α bioactivity that occurs in response to LPS,and rhuTNFR:Fc could in part protect rats from the acute renal injury induced by LPS.

Objective To investigate the role of calcitonin gene-related peptide(CGRP)and IP3 signal pathway in ischemic preconditioning(IPC)in the isolated perfused hearts of rats with type 1 diabetes mellitus. Methods Type 1 diabetes mellitus rat models were established in 80 SD rats,and were randomly divided into 4 week(D-4w)group and 8 week (D-8w)group.These two groups were randomly subdivided into model control(D-Cont)group,type 1 diabetes mellitus ischemia-reperfusion(IR)group,IPC group,CGRP(IPC+CGRP)group and IP3 inhibitor wortmanin(IPC+WMN) group.Another 16 rats were served as normal control(N-Cont)group.In vitro perfusion models of isolated hearts were established by Langendorff methods,and CGRP or wortmanin(WMN)were administered during perfusion.The left ventricle function of isolated heart in each group was monitored by multichannel biosignal analysis system,and coronary artery flow was recorded.The serum CGRP levels were detected by ELISA.The activity of lactate dehydrogenase(LDH)and creatine kinase(CK)in effluent of coronary artery was detected by biochemical method.The size of myocardial infarction was determined by NBT staining,and apoptosis of cadiocytes was detected by TUNEL method. Results Compared with N- Cont group,the CGRP level in serum of rats with type 1 diabetes mellitus decreased with time,the basic left ventricle function decreased,while the activity of LDH and CK in effluent of coronary artery,size of myocardial infarction and cardiomyocyte apoptosis index increased(P<0.05).Compared with N-Cont group,the left ventricle function was significantly lower in IR group,and more severe myocardial damage was observed.IPC improved myocardial damage of D-4w IR group,while had no protection on D-8w IR group.Compared with IPC group,the left ventricle function was significantly improved in IPC+CGRP group.IPC+WMN blocked the myocardial protection of D-4w group from IPC.Conclusion CGRP and IP3 signal pathway are involved in the protection provided by IPC in isolated hearts of rats with type 1 diabetes mellitus.

Objective·To observe mitochondria permeability transition pore (mPTP) opening and apoptosis of H9c2 myocardial cell stimulated by lipopolysaccharide (LPS),and to explore the anti-apoptotic effect of combined application of cyclosporine A (CsA) and ryanodine (Rya).Methods·The H9c2 cells were divided into Control group,LPS group,LPS+CsA group,LPS+Rya group,and LPS+CsA+Rya group.The mPTP opening state,Ca2+ concentration within cell and mitochondrial,mitochondrial membrane potential (AΦm),cell apoptosis,expression of Bax and Bcl-2 at mRNA and protein levels,and activity of caspase 3 were determined respectively.Results·mPTP opened after being stimulated by LPS for 24 h,which increased the fluorescence intensity for Ca2+in cytosolic and mitochondria by 298％ and 231％ respectively,induced about 1/3 cell apoptosis,improved the activity of caspase 3 approximately twice,and enhanced expression ofBax mRNA (P=0.008).The combined use of CsA and Rya effectively inhibited mPTP opening,increased the enhancement of fluorescence intensity for Ca2+in both cytosolic and mitochondria,maintained normal AΦrn,reduced LPS-induced apoptosis,inhibited the activity of caspase 3,and decreased Bax mRNA expression level induced by LPS in the myocardial cells.Conclusion·mPTP plays an important role in in LPS-induced myocardial apoptosis,whereas the combination of CsA and Rya can alleviate it effectively.

Objective To analyze the imaging features and to enhance the understanding of pancreatic and renal involvement in yon Hippel-Lindau disease (VHLD). Methods CT and MRI appearances and clinical data of six patients with pancreatic and renal involvement in VHLD were studied retrospectively.Six patients underwent CT scanning, and two of them also had MRI.Results Pancreatic cysts found in all six patients varied from several small cysts to cystic replacement of the entire gland.Calcifications were detected in four patients. Multiple bilateral renal lesions were detected in six patients.The renal lesions were classified as cystic, cystic with solid components and solid.Multiple combined renal lesions were found in five patients, and multiple simple cysts in one patient.Unilateral nephrectomy was performed in two patients, and the renal masses were diagnosed as clear cell carcinoma by pathology.Bile carcinoid was found in one patient, and retroperitoneal metastasis in another.Conclusion Multiple pancreatic cysts and/or multiple and bilateral combined renal lesions are highly suggestive of VHLD.

Objective To discover radioresistance-associated proteins by performing comparative proteomic analysis on nasopharyngeal carcinoma cell lines.Methods The total proteins were extracted from radioresistant human nasopharyngeal carcinoma cell line CNE-2R and its parental cell line CNE-2,respectively.These proteins were separated by high quality two-dimensional polyacrylamide gel electrophoresis (2-DE) and then the 2-DE profiles were screened for differentially expressed protein spots by the Image Master 5.0 software.Those spots were identified by a matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry.Results 32 significantly differentially expressed protein spots were screened in two different radiosensitivity cell lines and 11 proteins were identified by tandem mass spectrometry,among which 3 proteins were up-regulated in radioresistant human nasopharyngcal carcinoma cell line CNE-2R and the other 8 proteins were down-regulated.Conclusions The differentially expressed proteins of nasopharyegeal carcinoma cells with different radiosensitivity were mainly involved in apoptosis regulation,DNA damage and repair,cell cycle regulation,RNA transcription,cell signaling,cytoskeleton formation and radiation stress responses.

Objective To discover radioresistance associated molecular biomarkers and its mechanism in nasopharyngeal carcinoma by protein-protein interaction network analysis.Methods Whole genome expression microarray was applied to screen out differentially expressed genes in two cell lines CNE- 2R and CNE-2 with different radiosensitivity.Four differentially expressed genes were randomly selected for further verification by the semi-quantitative RT-PCR analysis with self-designed primers. The common differentially expressed genes from two experiments were analyzed with the SNOW online database in order to find out the central node related to the biomarkers of nasopharyngeal carcinoma radioresistance. The expression of STAT1 in CNE-2R and CNE-2 cells was measured by Western blot.Results Compared with CNE-2 cells,374 genes in CNE-2R cells were differentially expressed while 197 genes showed significant differences.Four randomly selected differentially expressed genes were verified by RT-PCR and had same change trend in consistent with the results of chip assay. Analysis with the SNOW database demonstrated that those 197 genes could form a complicated interaction network where STAT1 and JUN might be two key nodes.Indeed,the STAT1-α expression in CNE-2R was higher than that in CNE-2 (t =4.96,P ＜ 0.05).Conclusions The key nodes of STAT1 and JUN may be the molecular biomarkers leading to radioresistance in nasopharyngeal carcinoma,and STAT1-α might have close relationship with radioresistance.

Malic acid is a dicarboxylic acid that is widely used in food, pharmaceutical and chemical industries. We studied the effects of overexpression of carboxylation pathway genes and inactivation of malic enzymes on the aerobic production of malic acid. Over expression of phosphoenolpyruvate (PEP) carboxylase (ppc) generated strain E21, which increased malic acid production from 0.57 g/L to 3.83 g/L. Then pyc gene from Coryenbacterium glutamicus and pck gene from Actinobacillus succinogenes were overexpressed in E21 separately. The resulting strains E21 (pTrcpyc) and E21 (pTrc-A-pck) produced 6.04 and 5.01 g/L malate with a yield of 0.79 and 0.65 mol/mol glucose, respectively. Deleting two malic enzymes (encoded by maeA and maeB) also led to an increase of 36% in malic acid production with a production of 5.21 g/L. However, the combination of malic enzymes deletion and pyc overexpression could not further increase the yield of malic acid. After optimization of fermentation conditions, strain E21 (pTrcpyc) produced 12.45 g/L malic acid with a yield of 0.84 mol/mol which is 63.2% of the theoretical yield.