HMGB-1, an ubiquitously expressed 25-KD nucleoprotein among mammals, belongs to the HMG family. Recent studies have identified that HMGB-1 is secreted by activated monocytes/macrophages via a non-classical ,vesicle-mediated secretory pathway. Extracellular HMGB-1 is an important proinflammatory cytokine. It participates in the pathogenesis of many diseases such as rheumatoid arthritis, sepsis , acute lung injury, and even leads animals death. Further studies of the mechanism and function of extracellular HMGB-1 may provide a novel strategy for the diagnosis and treatment of these diseases.

OBJECTIVE: To study the effects of ginsenoside Rg1 on the expression of insulin-like growth factor-1 (IGF-1) in the brain of rats after the experimental brain contusion. METHODS: A total of twenty-six Wistar rats were randomly divided into normal control group (n=2), untreated group (n=8) and ginsenoside Rg1 group (n=16). Brain injuries were induced in rats by a mechanical striking device. The brain tissues were extracted at different times after brain injury (6th hour, 12th hour, 2nd day, 6th day), then the expression of IGF-1 in brain tissue was examined by immunohistochemical method. RESULTS: In comparison with the normal control group, the expression of IGF-1 in the brain tissues was increased in the untreated group after the brain contusion (P<0.05). The expression of IGF-1 in brain tissues in ginsenoside Rg1 group was significantly increased as compared with the untreated group (P<0.05). CONCLUSION: Ginsenoside Rg1 enhances the recovery of the contused brain through increasing the expression of IGF-1.

Objective To study the regional cerebral blood flow (rCBF) and correlated factors in patients with hepatolenticular degeneration (HLD).Methods The rCBF of lentiform nucleus,thalamus and other sites in 14 patients with HLD of cerebral type (cerebral type group) and 10 patients with HLD of non-cerebral type (non-cerebral type group) were determined by magnetic resonance-perfusion imaging technology,meanwhile 13 healthy volunteers were selected as control group,and calculated the relative regional cerebral blood flow (rrCBF) for avoiding perfusion time lag.The correlation between the clinical symptom scores,the content of urinary copper,duration and rrCBF in HLD patients were evaluated.Results The rrCBF of cerebral type group in the left and right frontal lobe,temporal lobe,lentiform nucleus,caudate nucleus,thalamus,midbrain,pons and the left hippocampus,cerebellar cortex,dentate nucleus were lower than those of control group (1.91 ±0.35 vs.2.44 ±0.64,1.80 ±0.30 vs.2.37 ±0.65,1.37 ±0.35 vs.2.14 ±0.91,1.58 ±0.52 vs.2.39 ±0.99,1.61 ±0.38 vs.2.59 ±0.74,1.52 ±0.64 vs.2.63 ±0.73,1.88 ±0.32 vs.2.61 ±0.67,1.70 ±0.40 vs.2.35 ±0.50,1.48 ±0.13 vs.2.01 ±0.59,1.46 ±0.38 vs.2.38 ±0.99,1.47 ±0.55 vs.2.02 ±0.72,1.27 ±0.48 vs.1.91 ±0.51,1.24 ±0.38 vs.1.47 ±0.29,1.25 ±0.39 vs.1.53 ±0.37,1.74 ±0.27 vs.2.40 ±0.89,1.79 ±0.50 vs.2.22 ±0.66,2.15 ±0.41 vs.2.64 ± 0.61),and there were significant differences (P ＜ 0.05 or ＜ 0.01).There were no significant differences in the rrCBF of the parietal and occipital lobe,etc between cerebral type group and control group (P ＞ 0.05).The rrCBF of cerebral type group in the left and right lentiform nucleus were lower than those of non-cerebral type group (1.61 ± 0.38 vs.1.94 ± 0.58,1.52 ± 0.64 vs.1.99 ± 0.59),and there were significant differences (P ＜ 0.05).The clinical symptom scores were positively correlated with the rrCBF of the left and right lentiform nucleus in 24 patients with HLD (r =-0.792 and-0.764,P ＜ 0.01),the content of urinary copper and duration had no correlation with the rrCBF of the left and right lentiform nucleus(P ＞ 0.05).Conclusions The rCBF of cerebral type and non-cerebral type HLD is significantly reduced,cerebral type patients have lower rCBF than non-cerebral type patients.The rCBF is correlated with the clinical symptom scores.

Objective To apply RNA interference technique for reducing the expression of MDC1 gene in esophageal carcinoma cell line ECA109, observe the changes in cell cycle and radiosensitivity after radiation, and discuss related mechanisms. Methods Three pairs of effective interference sequences and negative control sequences were synthesized for MDC1 mRNA sequence, and a recombinant plasmid was constructed with the vector pSIH1?H1?copGFP. RT?PCR and Western blot were used to determine the expression levels of MDC1 mRNA and protein. Colony?forming assay was applied to measure radiosensitivity, flow cytometry to determine cell cycle, Western blot to determine the expression of CHK1 and CHK2 proteins, and laser scanning confocal microscope to observe the number of MDC1 blotches inside the nucleus. One?way analysis of variance was used to analyze the differences between groups. Results The pSIH1?H1?copGFP plasmid was constructed successfully and ECA109 cells were infected to obtain ECA109M cells with stable transfection. The expression levels of MDC1 mRNA and protein in ECA109M cells were lower than those in ECA109N and ECA109 cells ( P= 0. 032 and 0. 041, respectively ) . After 5?Gy radiation, ECA109M cells had a lower proportion of G2+M cells than ECA109N and ECA109 cells ( P=0. 026) . After 5?Gy radiation, ECA109, ECA109N, and ECA109M cells had similar expression levels of CHK1 and CHK2 proteins ( P= 0. 345 and 0. 451, respectively ) , and ECA109M cells had a lower expression level of CHK2 T68 protein than ECA109 and ECA109N cells ( P=0. 012) . ECA109 cells had a D0 value of 3. 06 Gy and an SF2 value of 0. 91;the D0 values for ECA109N and ECA109M cells were 2. 90 Gy and 1. 88 Gy, respectively, and the SF2 values for them were 0. 89 and 0. 84, respectively ( P=0. 021 and 0. 037, respectively ) . Conclusions RNA interference can reduce the expression levels of MDC1 protein and cell cycle?related proteins, release cell cycle arrest, and enhance radiosensitivity in esophageal carcinoma ECA109 cells.

Objective To evaluate the effect of ulinastatin on oxidative stress injury to myocardial cells in diabetic rats in vitro.Methods The H9c2 cells were cultured in DMEM culture medium and the cells at the logarithmic growth phase were seeded in 96-well plates (density 1 × 104 cells/ml,200 μl/well) or in 6-well plates (density 1× 105 cells/m1,2 ml/well).The cells were randomly divided into 4 groups (n=18 each) using a random number table:normal control group (group C),high-glucose group (group HG),high-glucose + oxidative stress group (group HG+OS),ulinastatin +high-glucose+oxidative stress group (group U+HG+OS).The cells were cultured in high-glucose DMEM culture medium (25.0 mmol/L) for 48 h in group HG.After the cells were cultured in high-glucose DMEM culture medium for 24 h,H2O2 with the final concentration of 500 μmol/L was added to the high-glucose culture medium,and the cells were continuously cultured for 24 h in HG+OS and U+HG+OS groups.In group U+HG+OS,ulinastatin 400 U/ml was added to the high-glucose culture medium.The cells were collected for determination of cell viability,H9c2 apoptosis,activity of superoxide dismutase (SOD) and contents of malonadehyde (MDA).Apoptosis rate was calculated.The cell culture supernatant was collected for detection of lactate dehydrogenase (LDH) activity.Results Compared with group C,the cell viability and SOD activity were significantly decreased,and the apoptosis rate,MDA content and LDH activity were increased in the other groups.Compared with HG group,the cell viability and SOD activity were significantly decreased,and the apoptosis rate,MDA content and LDH activity were increased in HG+OS and U+HG+OS groups.Compared with group HG+OS,the cell viability and SOD activity were significantly increased,and the apoptosis rate,MDA content and LDH activity were decreased in group U + HG+ OS.Conclusion Ulinastatin can mitigate oxidative stress injury to myocardial cells in diabetic rats,and inhibited cell apoptosis is involved in the mechanism.

Objective To investigate the effects of inhibition of MDC1 protein expression on xenografted tumors in nude mice,and to observe the histopathological and cellular changes in nude mice.Methods Three pairs of effective and control short hairpin RNA targeting MDC1 mRNA were designed and cloned into the pSIH1-H1-copGFP vector.Real-time PCR and Western blot were used to determine the mRNA and protein expression of MDC1.After selection by copGFP reporter gene,cells were divided into negative transfection group (ECA109-N) and MDC1 transfection group (ECA109-M).The transfected cells were injected into nude mice.The mice were divided into ECA109 group,ECA109-N group,and ECA109-M group.Each group was divided into irradiation subgroup and non-irradiation subgroup.The changes in tumor size after irradiation were evaluated in each group.Western blot was used to measure the expression of CHK1,CHK2,and CHK2T68 in xenografted tumors.Flow cytometry was used to analyze the cell cycle distribution and apoptosis of tumor cells in nude mice.The variance analysis was used to compare the mean of multiple groups,and the SNK-q test was used in the two two groups.Results The pMDC1-shRNA plasmid was successfully constructed and used to transfect ECA109 cells.ECA109-M cells were obtained by stable transfection with the recombinant plasmid.All inoculated nude mice survived with visible xenografted tumors at the underside of the paw in about one week.There was no swelling and wound in inoculation sites.There was no significant difference in tumor size between different groups (P＞0.05).The tumor growth in the ECA109 group and the ECA109-N group significantly slowed down after irradiation with a dose of 15 Gy (P＜0.05).Compared with the other two groups,the ECA109-M group had a significant smaller tumor size,significantly slower relative tumor growth,and significantly higher growth inhibition (all P＜0.05).The q value of the ECA109-M group was 1.36.In the ECA109-M group,there were no significant changes in the protein expression of CHK1 and CHK2 after irradiation (P＞ 0.05);however,the phosphorylation of CHK2T68 protein was significantly reduced after irradiation (P＜0.05).There were no significant differences in cell cycle distribution or the proportion of apoptotic cells in tumor tissue between the three groups (P＞0.05).Conclusions Inhibition of MDC1 protein expression by RNA interference can effectively inhibit the growth of xenografted tumors after irradiation in the nude mice by increasing their radiosensitivity.

Objective To investigate the pathological features and prognostic analysis in endoscopic strip-ping microflap excision of vocal cords leukoplakia.Methods A total of 186 cases with vocal cord leukoplakia and treated by endoscopic stripping microflap excision were retrospectively analyzed,calculated the ratios of recurrence and malignant transformation on keratosis without dysplasia,mild dysplasia,moderate dysplasia and severe dysplasia and carcinoma in situ.Then,the association of histopathological classification with recurrence and malignant transformation in vocal cord leukoplakia were evaluated.Results 87 cases of all 186 patients with vocal cord leukoplakia were kera-tosis without dysplasia,17 cases were recurrenced and 3 cases were developed to carcinoma;64 cases were mild dysplasia,15 cases were recurrenced and 7 cases were developed to carcinoma;29 cases were moderate dysplasia, 13 cases were recurrenced and 10 cases were developed to carcinoma;6 cases were severe dysplasia and carcinoma in situ,all 6 cases were recurrenced and 5 cases were developed to carcinoma.Conclusion The histopathological classi-fication is predictive factor for the risk of recurrence and malignant transformation in vocal cord leukoplakia.

Objective To analyze the outcomes and prognostic factors of esophageal carcinoma treated with three-dimensional eonformal radiotherapy (3DCRT). Methods From January 2001 to August 2007, 209 patients with esophageal carcinoma treated with 3DCRT were retrospectively analyzed. The local control rotes, the survival rates and the related prognostic factors were evaluated with SPSS 11.5 software. Results The follow-up rate was 98. 1% by December 2008. The number of patients followed up for 1,3, 4 and 5 years was 209,131,95 and 56, respectively. The 1-, 3- and 4-year local control rates were 74. 9%, 50. 4% and 45. 8%, respectively. The 1-, 3-and 4-year overall survival rates were 64. 6%, 30. 8% and 23.6%, respectively, with a median survival time of 18 months. Univariate analysis showed that the significant prognostic factors included the degree of dysphagia, tumor site, lesion length in barium esophagogram and CT image, the largest diameter of lesion in CT image, T stage, N stage, clinical TNM stage, short term effect, and degree of acute esophagitis. Multivariate analysis revealed that the degree of dysphagia, primary tumor site, clinical stage, and radiotherapy technique (3DCRT or late half course 3DCRT) were independent prognostic factors. Conclusions Three-dimensional conformai radiotherapy is effective and feasible in the treatment of esophageal cancer. The degree of dysphagia, primary tumor site, and clinical stage are independent prognostic factors for survival of patients treated with 3DCRT.

Objective To investigate the effect of pretreatment with dexmedetomidine alone or in combination with sufentanil on myocardial ischemia-reperfusion (I/R) injury in rats.Methods Fifty healthy male Sprague-Dawley rats,weighing 250-300 g,were anesthetized with intraperitoneal pentobarbital sodium 60 mg/kg.Myocardial I/R was induced by occlusion of anterior descending branch of left coronary artery for 30 min followed by 120 min of reperfusion.The rats were then randomly divided into 5 groups (n =10 each):sham operation group (group S),group I/R,dexmedetomidine pretreatment group (group DP),sufentanil pretreatment group (group SP),and dexmedetomidine + sufentanil pretreatment group (group DS).In group S the anterior descending branch was only exposed but not ligated.Dexmedetomidine 0.5μg/kg and sufentanil 0.1μg/kg were injected intraperitoneally 30 min before ischemia in groups DP and SP,respectively.Dexmedetomidine 0.5 μg/kg and sufentanil 0.1 μg/kg were injected intraperitoneally 30 min bbefore ischemia in group DS.Arterial blood samples were collected at 120 min of reperfusion for determination of serum creatine kinase (CK) and lactic dehydrogenase (LDH)concentrations.The rats were sacrificed at 120 min of reperfusion and hearts were removed for microscopic examination.Myocardial infarct size was calculated.The malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in myocardial tissues were measured.Results Compared with group S,the serum CK and LDH concentrations were significantly increased,the myocardial infarct size was enlarged,and SOD activity was decreased in the other groups,MDA content was significantly increased in groups I/R,DP and SP (P ＜ 0.05 or 0.01).Compared with group I/R,the serum CK and LDH concentrations,MDA content and myocardial infarct size were significantly decreased,and SOD activity was increased in groups DP,SP and DS (P ＜ 0.05).Compared with group DS,the serum CK concentration was significantly increased,the myocardial infarct size was enlarged,and MDA content was increased in groups DP and SP,and LDH concentration was significantly increased and SOD activity was decreased in group DP (P ＜ 0.05).The pathological changes were significantly attenuated in groups DP and SP compared with group DS.Conclusion Dexmedetomidine pretreatment can reduce myocardial I/ R injury in rats,dexmedetomidine combined with sufentanil pretreatment provides better efficacy than either alone,and inhibition of lipid peroxidation is involved in the mechanism.

ObjectiveTo analyze the outcomes and prognostic factors of advanced esophageal carcinoma treated by three-dimensional conformal radiotherapy (3DCRT).MethodsFrom Jul 2001 to Dec 2006.375 patients with esophageal carcinoma treated by 3DCRT were retrospectively analyzed of which Ⅰ stage 9,Ⅱ stage 106,Ⅲ stage 158,Ⅳstage 102.The short-term effect,1-,3-,5-year local regional control rates and survival rates were investigated.The local regional control rates and survival rate were calculated by the Kaplan-Meier method. Univariate prognostic factor was analyzed by Logrank method.Multivariate prognostic factor was analyzed using Cox regression model.ResultsThe follow-up rate was 94.7％.The numbers of patients followed-up with 5 years was 191.The 1-,3-and 5-year local control rates were 80.5％,53.7％,44.9％respectively.The 1-, 3-and 5-year survival rates were 67.2％,29.4％,19.0％respectively.Univariate analysis showed the significant prognostic factors included the degree of dysphagia,tumor length,the largest diameter of lesion in CT image,T stage,N stage,clinical TNM stage,grades of acute radiation-induced esophagitis and grades of acute radiation-induced pneumonery ( x2 =46.75,18.52,30.24,42.53,32.71,75.68,7.13,4.64,P =0.000,0.000,0.000,0.000,0.000,0.000,0.008,0.031 ).Multivariate analysis revealed tumor length,clinical TNM stage,chemotherapy and grades of acute radiationinduced esophagitis were independent prognostic factors (x2 =6.70,18.00,4.87,1.1 8,P =0.030,0.000,0.027,0.011 ).Conclusions3DCRT is effective and feasible in treatment of the advanced esophageal carcinoma.Tumor lesion length,clinical TNM stage,chemotherapy and grades of acute radiation-induced esophagitis are independent prognostic factors for survival of patients.