Objective To explore the protectional function of CpG ODN 1826 against radiation pulmonary fibrosis in rats.Methods The rat left lung was exposed to 20 Gy of 6 MV X-rays for establishing a radiation pulmonary fibrosis model.SD rats were randomly divided into control group,irradiated group and intervention group,with 30 rats in each group.CpG ODN 1826 was intraperitoneally injected into rats at 0,1,2,5 and 7 d post-irradiation.The rats were terminated at 5,15,30 and 90 d post-irradiation,and the lung indexes were recorded.Paraffin sections of the radiated lung were conducted with HE staining and Masson staining,the pulmonary fibrosis scores were recorded.The serum concentrations of TGF-β1 and hydroxyproline (Hyp) were measured.Results The radiation pulmonary fibrosis rat model was successfully established.The lung indexes of the control group were lower than those of the irradiated and intervention groups at 5 d post-irradiation (t =3.046,2.252,P ＜ 0.05).The lung indexes of the intervention group were lower than those of the irradiated group (t =4.120,5.226,5.719,P ＜ 0.05).Pulmonary fibrosis scores of intervention group were lower than those of irradiated group (t =3.212,4.959,P ＜ 0.05).The serum concentrations of TGF-β1 of irradiated group were higher than those of the intervention group (t =4.138,5.924,4.138,5.924,P ＜ 0.05).The Hyp in the lung of irradiated group was higher than that of intervention group (t =7.527,8.416,P ＜ 0.05).Conclusions CpG ODN1826 will not worse the radiation pulmonary fibrosis,on the contrary,it could reduce the serum concentrations of TGF-β1 and the lung content of Hyp in radiation pulmonary fibrosis,and protects rat against radiation pulmonary fibrosis.

Objective To investigate the effect of CaMK Ⅱ expression on apoptosis of rat hepatocytes BRL-3A.Methods Rat BRL-3A cells were stable passage were cultured.The CaMK Ⅱ γ protein (LV-CaMK Ⅱ γ group) and CaMK Ⅱ γshRNA (shRNA group) lentiviral expression systems were constructed.The corresponding blank vectors (LV-NC group and shRNA-NC group) and normal saline (CON group) were perfused into the control groups.The expression levels of CaMK Ⅱ,Cyt C and MF proteins were detected by Western blotting,and the apoptosis rate of BRL-3A cells was measured by Tunel method.Results The protein expression of CaMK Ⅱ,Cyt C and AIF in LV-CaMK Ⅱ γ group was significantly higher than that in CON group (P＜0.05).The protein expression of CaMK Ⅱ,Cyt C and AIF in shRNA group was significantly lower than that in CON group (P＜ 0.05).There was no significant difference among CON group,LV-NC group and shRNA-NC group (P＞0.05).At the same time point,the apoptosis rate of hepatocytes in LV-CaMK Ⅱ γ group was significantly higher than that in CON group (P＜0.05).At the same time point,the apoptosis rate of hepatocytes in shRNA group was significantly higher than in CON group (P＜0.05).There was no significant difference in the apoptosis of hepatocytes among CON group,LV-NC group and shRNA-NC group (P＞0.05).Conclusion The specific CaMK Ⅱ signaling pathway can inhibit the apoptosis of BRL-3A cells,while the enhanced CaMK Ⅱ signaling pathway promotes the apoptosis of BRL-3A cells.

BACKGROUND:The proteome is a highlight technology in medical research fields lately, and has been reported to be applied in basic research fields related to liver transplantation. However, it has not been heard that the proteome has been used in research related to reduced-size liver transplantation. OBJECTIVE: To study expression of hepatic differential proteins related to signal transduction using proteomics after reduced-size liver transplantation in rats. METHODS:On the basis of successful establishment of rat models of reduced-size liver transplantation, transplanted liver tissues were obtained at 1, 3 and 7 days after transplantation. Postoperative liver tissue and normal donor, receptor liver tissues were subjected to solid pH gradient two-dimensional gel electrophoresis. Two-dimensional gel electrophoresis patterns were set up. Differentialy expressed protein spots were identified using tandem mass spectrometry analysis and database. RESULTS AND CONCLUSION:Seventy-two differential protein stains were found taking 10 times measure. Finaly, 32 proteins with clear functions were identified. Of them, four proteins participated in signal transduction, and they distributed at 3 and 7 days after liver transplantation, accounting for 6%. Results verified that on the basis of successful and stable establishment of rat models of reduced-size liver transplantation, proteomics technology was utilized to study differential proteins involving in signal transduction after reduced-size liver transplantation, and this study provides data for further deep investigation of regulating MicroRNA of these proteins.

Abstract BACKGROUND: Looking for the early diagnosis of acute rejection indicators after liver transplantation can assess the risk after liver transplantation quickly and effectively, and T lymphocytes play the significant role in acute rejection. OBJECTIVE:To observe the relationship between acute rejection and variation of expression of T cel subset in blood after liver transplantation in rhesus monkey. METHODS: The sixteen liver transplant models in rhesus monkey which were constructed successfuly by the method of “double-cuff and one support tube” were divided into two groups randomly: experiment group (no treated by immunosuppressant in perioperative period) and control group (treated by immunosuppressant in perioperative period). Then the blood specimen and liver tissue respectively were colected at 6, 12, 24 and 72 hours after operation. The levels of alanine transferase, aspartate aminotransferase, and total bilirubin were detected with the fuly automatic biochemical analyser. The levels of CD4+/CD8+were tested by flow cytometry. The liver tissue in rhesus monkey after liver transplantation was detected by hematoxylin-eosin staining. The degree of acute rejection was evaluated by Banff Score System. RESULTS AND CONCLUSION: Acute rejection appeared in the experiment group at 12, 24, and 72 hours after liver transplantation. Levels of alanine transferase, aspartate aminotransferase, and total bilirubin were significantly higher in the experimental group than in the control group at 24 and 72 hours after transplantation (P < 0.05). The expression of CD4+/CD8+of the experiment group and control group began to rise at 6 hours after surgery, but the experiment group increased the most obvious. CD4+/CD8+ expression was significantly greater in the experimental group than in the control group at 24 and 72 hours after transplantation (P < 0.05). Morphological pathology was severer, and Banff score was higher in the experiment group than in the control group at 72 hours (P < 0.05). These data suggested that the variation of expression of CD4+/CD8+was earlier than the change of liver tissue pathology and the change of liver function in the early acute rejection after liver transplantation. The rise of level of CD4+/CD8+ after liver transplantation indicated the increase of celular immunity in body, which had an important role in the early diagnosis of acute rejection after liver transplantation.

BACKGROUND:At present, the proteome is a mature technology that has been applied in basic research fields related to liver transplantation. But, it has been not reported in research related to reduced-size liver transplantation.
OBJECTIVE:To explore the expression of differential proteins related to hepatic energy metabolism fol owing reduce-size liver transplantation in rats by using by proteomic technology.
METHODS:The improved model of reduced-size liver transplantation was used in this experiment. The donor was health female Lewis rats and the recipient was male Wistar rats for liver transplantation. The difference between the donor and the recipient was about 20 g. The weight of donor liver/the weight of recipient donor was approximately equal to 50%. The donor liver tissue was harvested and trimmed to the required size. The portal vein and infrahepatic vena cava were cannulated, and the biliary tract was implanted into the donor bile duct for transplantation. Then the donor was transplanted into the recipient after the removal of original liver tissue. Hepatic specimens were harvested by 1, 3 and 7 days after reduced-size liver transplantation. Then, the harvested specimens were compared with the normal donor and recipient liver tissue that were previously harvested and frozen, to generate two-dimensional gel electrophoresis profile using proteome technology. Then tandem mass spectrometry and databases analysis were performed after two-dimensional electrophoresis for identifying differential protein stains.
RESULTS AND CONCLUSION:In this experiment, 72 differential protein stains with over lo-fold changes were selected. After identification, 32 proteins showed clear functions, and among them three differential proteins (ATP synthase beta subunit, electron-transferring flavoprotein beta peptide and proton-transferring ATP synthase) were involved in the process of cel energy metabolism. The proteins were distributed on 1 and 7 days after reduce-size liver transplantation, accounting for 6%.

BACKGROUND: Various factors contribute to the establishment of liver transplantation models in rhesus monkey, the rate of successful operation and long-term survival are very low. OBJECTIVE: To analyze the cause of early death following liver transplantation in rhesus monkey. METHODS: Liver transplantation models were fabricated with the classical and modified methods in rhesus monkeys. Operation of donor was performed quickly by a big crucial incision of abdomen. The improved double-cuff of the portal vein and inferior vena cava were finished, in addition to stay pipe of biliary tract in the process of repairing donor liver. Operation of the receptor was performed by classical orthotopic liver transplantation. RESULTS AND CONCLUSION: A total of 25 pairs of rhesus monkeys were successfully for establishing liver transplantation models. Seven rhesus monkeys died within early stage of post-operation, including six out of nine monkeys died by using the classical approach and one out of sixteen monkeys died by using the improved approach. There were five of seven monkeys died of intra-abdominal hemorrhage, one died of primary graft nonfunction and one died of respiratory failure. Results indicated that, the major death cause after classical orthotopic liver transplantation in rhesus monkey is abdominal hemorrhage. The improved methods of liver transplantation apparently reduce the hemorrhage and raise early survival rate following liver transplantation.

Objective To explore the effect of nano-hydroxyapatite/collagen scaffolds incorporating ADM-PLGA microspheres in repairing large bone defects of rabbit femoral condyle. Methods Animal models of bone defects were induced in 21 New Zealand white rabbits by drilling holes in bilateral femoral lateral condyles , and the rabbits were equally divided into 3 groups:group A as the control group with the defects untreated , group B treated by filling with nano-hydroxyapatite/collagen scaffolds (NHAC), and group C treated by filling with the nano-hydroxyapatite/collagen scaffolds incorporating ADM-PLGA microspheres (ADM-PLGA-NHAC). At week 12 after implanting , the rabbits were all sacrificed for the implanted scaffolds , which were then examined by X-ray , and Micro-CT 3D reconstruction and in histology for evaluation of the new bone formation. Results X-ray, Micro-CT and the measurement and analysis of BMD indicated thatthere was no significant differencein the new bone formation between group B and group C (P > 0.05). The histological examination revealed that. 12 weeks after operation an evident number of new born bones were seen on the implanted scaffolds in groups B and C , while very few were seen scattering in group A. Conclusion The nano-hydroxyapatite/collagen scaffolds incorporating ADM-PLGA microspheres is effective in repairing bone defect without influencing the prosthetic process.

Objective To explore the role of pyruvate kinase M2 (PKM2) siRNA in the radiosensitivity of human lung cancer A549 cells.Methods PKM2 siRNA was synthesized according to the coding sequence of PKM2 mRNA and then was transferred into A549 cells with lipofectamine.The expressions of PKM2 gene and protein was detected by RT-PCR and Western blot,respectively.The experiments were divided into PKM2 siRNA interference group,siRNA negative control group,and blank control group.The cells of each group were exposure to 6 MV X-rays in different dose.Radiosensitivity was evaluated by colony formation assay.Flow cytometry was applied to analyze cell cycle distribution and apoptosis.Data are representative of three independent experiments.Results Ccompared with blank control cells,the expressions of PKM2 gene and protein in the PKM2 siRNA transferred A549 cell was efficiently diminished (t =20.91,47.00,P ＜0.01) with inhibition rates of (70.27 ± 1.38)％ and (70.42 ± 1.18) ％,respectively.Compared with control,PKM2 siRNA transfection significantly decreased the D0,Dq,N and SF2 values (t =43.82,28.44,15.60,29.63,P ＜ 0.01) and hence yield a sensitization enhancement ratio (SER) of 1.27.In addition,the percentage of G2/M phase cells in the siRNA group and irradiated group were both significantly higher than that of the blank control group (t =8.35,27.87,P ＜ 0.01).The combined treatments of PKM2 siRNA interference and irradiation arrested more cells in the G2/M phase compared to either treatment alone.The apoptosis rate of siRNA group was not dramatically different from that of blank control group.The apoptosis rate of irradiation group was higher than that of blank control group (t =23.99,P ＜ 0.01),and the combined treatments of siRNA and irradiation enhanced the apoptotic rate compared to either treatment alone (t=9.42,65.21,P ＜ 0.01).Conclusions Specific blockage of PKM2 expression by gene silencing could enhance the sensitivity of human lung cancer A549 cells to radiotherapy in vitro,which may due to the cell cycle arrest and apoptosis induction after irradiation.

Objective To compare the effects of smoking and non smoking on postoperative pain of laparoscopic cholecystectomy. Methods Sixty patients having underwent selective laparoscopic cholecystectom were divided into smoking group and non smoking group by random digits table with 30 cases each. In smoking group, 14 cases quitted smoking within 1 week before operation. The Fagerstrom test of nicotine dependence (FTND) was evaluated before operation in smoking group, and FTND ≥ 6 scores was in 11 cases. The visual analog score (VAS), Bruggrmarm comfort score (BCS), sedation-agitation score (SAS), immediately, 15 min, and 30 min after entering postanesthesia care unit (PACU) and leaving PACU was evaluated. The operation time, anesthesia time, wake up time, extubation time, PACU time, using rate of remedial measures and untoward reaction were recorded. Results There were no statistical differences in operation time, anesthesia time, wake up time, extubation time, SAS and incidence of untoward reaction between 2 groups (P>0.05). The PACU time and using rate of remedial measures in smoking group were significantly higher than those in non smoking group:(39.7 ± 5.1) min vs. (31.3 ± 6.1) min and 30.0% (9/30) vs. 0, and there were statistical differences (P<0.05). The VAS immediately, 15 min and 30 min after entering PACU and leaving PACU in smoking group was significantly higher than that in non smoking group: (2.90 ± 0.85) scores vs. (1.00 ± 0.83) scores, (2.70 ± 0.47) scores vs. (0.73 ± 0.69) scores, (2.60 ± 0.56) scores vs. (1.13 ± 0.73) scores, (2.23 ± 0.57) scores vs. (1.13 ± 0.73) scores; and the BCS was significantly lower than that in non smoking group:(1.80 ± 0.61) scores vs. (2.90 ± 0.99) scores, (1.90 ± 0.31) scores vs. (2.87 ± 1.00) scores, (2.10 ± 0.31) scores vs. (2.47 ± 0.82) scores, (2.17 ± 0.38) scores vs. (2.47 ± 0.82) scores, and there were statistical differences (P<0.05). The VAS immediately after entering PACU in patients of FTND ≥ 6 scores was significantly higher than that in patients of FTND<6 scores:(3.6 ± 0.7) scores vs. (2.5 ± 0.7) scores, the BCS was significantly lower than that in patients of FTND <6 scores:(1.5 ± 0.5) scores vs. (2.0 ± 0.6) scores, and there were statistical differences (P<0.05). The VAS immediately after entering PACU in patients of non- quit smoking was significantly higher than that in patients of quit smoking: (3.4 ± 0.7) scores vs. (2.4 ± 0.6) scores, and there were statistical differences (P<0.05). Conclusions Smokers have more severe postoperative pain in laparoscopic cholecystectomy and higher postoperative opioid requirement than nonsmokers. Quit smoking before surgery will reduce postoperative pain and related complications. Appropriate increase of analgesic drugs can prevent postoperative pain in patients with smoking.

BACKGROUND:Nuclear factor-κB as an important nuclear transcription factor, is a converge point for various signal transduction pathways, and participate in the regulation of reactive substances gene expression such as the immune cel proliferation, differentiation and apoptosis. The nuclear factor-κB plays an important role in humoral and cel ular immune. OBJECTIVE:To investigate the relationship between the nuclear factor-κB p65 protein expression and acute rejection in transplanted liver tissue of rhesus monkey. METHODS:The rhesus monkey recipients were randomly divided into two groups:acute rejection group and control group. The acute rejection group did not received anti-rejection treatment after liver transplantation, and the control group was given anti-rejection treatment during and after liver transplantation. The blood samples were col ected at 6, 12, 24 and 72 hours after transplantation, and the automatic biochemical analyzer was used to detect the levels of alanine aminotransferase and total bilirubin. Hematoxylin-eosin staining of transplanted liver tissue was performed to observe the morphological structure and rejection. The degree of rejection was evaluated according to the Banff scoring system, and the expression of nuclear factor-κB p65 in the liver tissue was detected with Western blot. RESULTS AND CONCLUSION:When the acute rejection occurred after liver transplantation, the liver function change was observed after liver histopathological examination, the expression of nuclear factor-κB p65 in the liver tissue was up-regulated, and the degree of acute rejection was increased. In the early stage of acute rejection, the liver function and pathology were changed slightly, while the expression of nuclear factor-κB p65 was significantly increased. The results indicate that the detection of nuclear factor-κB p65 in the transplanted liver tissue has great significance for the early diagnosis of acute rejection after liver transplantation, and the nuclear factor-κB may be the new target for control ing the acute rejection.