Objective To study the influence of microenvironment created by dissoluble cytokines derived from hepatocellular carcinoma cells (HCCs) on the functional status of mitochondria in dendritic cells (DCs) in order to investigate the mechanisms of tumor immune escape.Methods CD14+ monocytes,which were isolated with immune magnetic beads from fresh peripheral blood of healthy human,were then cultured in RPMI1640/10% FBS supplemented with 100 ng/ml rhGM-CSF and 100 ng/ml rhIL-4 for 7 d to develop into immature DCs (imDCs).Maturation was induced by addition of 20 ng/ml rhTNF-? to imDCs for another 3 d of culture.Acquired imDCs and mDCs were respectively co-cultured with human HCCs for 48 h in Transwell chamber.These treated cells were investigated for mitochondrial membrane potential,enzyme activity and intracellular Ca2+ concentration ([Ca2+]in) in the cytoplasma by immune fluorescence and MTT assay.Those untreated cells served as control.Results After co-cultured with HCCs,flow cytometry showed that the mitochondrial membrane potentials of imDCs and mDCs were respectively decreased from (659.991?17.052)and(473.741?11.676)to(482.681?7.935)and(407.189?5.051)(P

Objective To study the influence of microenvironment created by dissoluble cytokines derived from hepatocellular carcinoma cells (HCCs) on the expression of IL-12 of dendritic cells (DCs) at different differentiating stage. Methods The CD14+ monocytes,isolated with immune magnetic beads from fresh peripheral blood of healthy human,were cultured in RPMI 1640/10% FBS supplemented with 100 ng/ml rhGM-CSF and 100 ng/ml rhIL-4 for 7 d to develop into immature DCs (imDCs). Maturation was induced by addition of 20 ng/ml rhTNF-? to imDCs for another 3 days' culture. Acquired imDCs and mDCs were respectively co-cultured with human HCC Bel7402 cells for 48 h in Transwell chamber. The concentrations of IL-10,IL-12,TGF-?1 and VEGF in culture supernatant were investigated by ELISA,imDCs and mDCs cultured alone served as control. Results After co-cultured with Bel7402 cells,IL-12 were decreased from (115.076?8.129) pg/ml to (17.599?1.757) pg/ml in imDCs,and from (258.346?3.609) pg/ml to (6.787?1.123) pg/ml in mDCs (P

Objective To observe the effect of Xingnaojing injection in myocardial mitochondrial oxidative stress injury in sepsis mice. Methods The septic model were set up by receiving endotoxin through interaperitoneal injection. After Xingnaojing injection by gastric tube , seventy mice were randomly divided into 7 groups in ten of each group including group LPS-6 h , group LPS-24 h , group LPS-48 h; group XNJ-6 h , group XNJ-24 h, group XNJ-48 h and control group. The myocardial mitochondrial changes , the semi-quantitative scores and the level of SOD、MDA、NO、iNOS in sepsis mice were observed. Results Xingnaojing injection could improve the myocardial mitochondrial changes , reduce the semi-quantitative scores , significantly reduce the level of MDA、NO、iNOS and elevate the level of SOD. Conclusion Xingnaojing injection could significantly reduce the myocardial mitochondrial oxidative stress level and improve the ability of clearing oxygen radicals , thereby protect the myocardial mitochondrion in sepsis mice.

Objective To investigate the clinical efficacy of laparoscopic hepatectomy.Methods The clinical data of 258 patients who received laparoscopic hepatectomy at the First Affiliated Hospital of Fujian Medical University from March 2010 to January 2013 were retrospectively analyzed.There were 196 patients with primary hepatic cancer,45 with hepatic hemangioma,13 with hepatic focal nodular hyperplasia,2 with hepatic metastatic cancer,1 with carcinoma of gallbladder and 1 with hepatic hamartoma.All patients were followed up via phone call or out-patient examination till March 2013.Results A total of 142 patients received single hepatic segmentectomy,98 received multiple hepatic segmentectomy,18 received multiple lesions resection.Fifty-one patients received hepatic tumorectomy + cholecystectomy.All the operations were successfully done under laparoscope without conversion to the open surgery.The mean tumor diameter and the operation time were (5 ± 3) cm (range,1.0-11.5 cm) and (113 ± 56) minutes (range,50-310 minutes),respectively.Intraoperative hepatic portal occlusion was performed on 122 patients,and the time for hepatic portal occlusion was (15 ± 7)minutes.The volume of intraoperative blood loss was (211 ± 195)mL (range,10-650 mL),and blood transfusion was not needed.The capsule of the tumor was complete.The distance between the resection margin and the malignant tumor was above 1.5 cm,and there was no residual tumor in the resection margin.The hepatic function was back to the normal level in 1 week after the operation,and no patient had hepatic failure.The duration of postoperative hospital stay was (7.2 ± 1.3)days (range,5-10 days).One patient was complicated with bile leakage,6 with slight peritoneal effusion,and other patients had no postoperative complications.The rate of follow-up was 91.47％ (236/258),and the time of follow-up was (16 ± 10) months.A total of 199 patients with malignant hepatic tumors were followed up.During the follow-up,180 patients had tumor-free survival; 18 patients had postoperative tumor recurrence; 1 patient had omental metastasis and received surgical resection.Thirty-seven patients with benign hepatic tumor survived without complication during the follow-up.Conclusion Laparoscopic hepatectomy is effective for the treatment of hepatic tumors.Multiple hepatic inflow occlusion under laparoscope in a short time may improve the safety of surgery,without prolonging the recovery time of patients.

LCRG1(laryngeal carcinoma related gene1,LCRG1),a new candidate tumor suppressor gene of laryngeal carcinoma.However,it is known little about the possible regulatory mechanisms of LCRG1 gene expression.Restriction endonuclease digestion was used to obtain a set of the 5',or 3'deletion mutants from the region(-169～+127) of the LCRG1 gene.It has been found that the minimal promoter of the LCRG1 gene is mapped at the region from-169～-57.Linker scanning mutational analysis in the region(-169～+127) of the LCRG1 gene was used to identify the crucial cis-elements within the promoter region,The key cis-elements are within the region from-137～-122.SP1,E2F1/DP1,EKLF and ZF9 transcription factor binding site sites were predicted in the region by bioinformatics analysis.Co-transfection with each of a panel of the expression plasmids of the known transcription factors with the relevant reporter construct indicates Sp1 is potent transcription factor for enhancement of the promoter activity,SP1 can also up-regulate the endogenous expression of LCRG1 gene.Electrophoretic mobility shift assay(EMSA) was applied to verify that the key cis-elements of LCRG1 gene exist sequence of Sp1 binding sites.The findings,which showed that the key cis-elements within the region from 137～-122 play an important role in expression of the LCRG1 gene,provide a novel evidence for further study of the function of LCRG1 gene.

Objective Mitochondrial dysfunction, cell energy metabolism, and oxidative stress play important roles in sepsis-induced acute brain injury.This study was to investigate the effects of Xingnaojing Injection (XNJ) on brain mitochondrial oxidative stress in rats with lipopolysaccharide (LPS)-induced sepsis.Methods Totally, 252 male SD rats were randomly divided into a normal control group, 3 LPS-induced sepsis model groups (LPS 6, 24, and 48 h), and 3 XNJ treatment groups (XNJ 6, 24, and 48 h), with 36 in each group.After treatment, the mitochondrial membrane potential (MMP) was monitored by flow cytometry and the levels of manganese superoxide dismutase (Mn-SOD), malondialdehyde (MDA), nitric oxide (NO), and nitric oxide synthase (NOS) were determined by chromatometry.Results The MMP was significantly increased in the XNJ 6 h group as compared with the LPS 6 h group (0.80±0.11 vs 0.54±0.19, P<0.05).In the LPS and XNJ groups, the levels of MDA and NOS reached the peak at 6 hours and then dropped gradually, while those of NO and Mn-SOD rose to the peak at 24 hours followed by a gradual fall.Statistically significant differences were observed in the levels of MDA, NOS and NO between the LPS 6h and XNJ 6 h groups (P<0.05), as well as in those of NOS, NO and Mn-SOD between the LPS 24 h and XNJ 24 h groups (P<0.05).Conclusion Xingnaojing Injection can elevate the level of the brain mitochondrial membrane potential, improve anti-oxidation indexes in the mitochondria, and protect brain mitochondria in sepsis rats.

Objective: To explore influence of coping style on sleep quality and blood pressure in community male population with high normal value blood pressure. Methods: The Pittsburgh sleep quality index (PSQI) and coping style questionnaire (CSQ) were used to assess 120 men with high normal blood pressure in community. With PSQI>7 scores as criterion for judging sleep quality disorders, the subjects were divided into sleep disorder group (n=51) and normal sleep group (n=69), and sleep disorder group received psychological intervention. Results: Sleep disorders existed in 42.5% male population with high normal blood pressure. Compared with normal sleep group, there was significant increase in PSQI [（6.43±2.59）scores vs. (8.33±3.14）scores] and diastolic blood pressure [(81.00±8.91) mmHg vs. (88.00±5.69) mmHg] and significant decrease in factor scores of “problem solving” [(0.76±0.21) scores vs. (0.61±0.18) scores] and “asking for help” [(0.52±0.26) scores vs. (0.41±0.11) scores] in sleep disorder group, P<0.05 all; Compared with before intervention there were significant increase in scores of “problem solving” [(0.61±0.18) scores vs. (0.71±0.12) scores]and “asking for help” [(0.41±0.11) scores vs. ( 0.51±0.13) scores]and significant decrease in PSQI score [(8.33±3.14) scores vs. (7.41±2.37) scores] and diastolic blood pressure [(88±5.69)mmHg vs. (80± 4.17)mmHg] after psychological intervention 12 weeks in sleep disorder group, P<0.05 all. Conclusion: Psychological intervention may improve sleep quality and reduce blood pressure in community male population with high normal value blood pressure.

ObjectiveTo investigate the effects of autophagy on exocrine function of pancreas in rats with acute sepsis, and to determine whether the mitochondrial coenzyme Q (Mito Q) can prevent exocrine dysfunction of pancreas mediated by autophagy.Methods ExperimentⅠ: 30 Sprague-Dawley (SD) rats were randomly divided into three groups, with 10 rats in each group. All the rats were given lipopolysaccharide (LPS, 10 mg/kg) intraperitoneally, and Wortmannin (2 mg/kg), the specific inhibitor of autophagy (LPS+ Wortmannin group), Mito Q (6.5μmol/kg, LPS+Mito Q group), or the same volume of normal saline (LPS group) was respectively injected via the tail vein 1 hour later. Survival rate was assessed within 12 hours after LPS injection. ExperimentⅡ: another 100 male SD rats were randomly divided into ten groups with 10 rats in each group: namely control 4, 6 and 12 hours groups, LPS 4, 6 and 12 hours groups, and LPS+ Wortmannin 4 hours group, Wortmannin 4 hours group, LPS+ Mito Q 6 hours group, and Mito Q 6 hours group. The protocols of model reproduction and drug administration were the same as in the experimentⅠ. Blood samples were collected at each time point, and the amylase content was determined with the velocity method. The levels of reactive oxygen species (ROS) in the pancreases were measured with enzyme-linked immunosorbent assay (ELISA). The expression of the autophagy-related protein LC3 was determined with Western Blot. The pathological changes in the pancreas were observed with microscopy.Results① The survival time in the LPS+ Wortmannin group was significantly shorter than that in the LPS group (hours: 7.50±0.64 vs. 11.90±0.13,χ2= 19.847,P= 0.001). There was no significant difference in the survival time between LPS+ Mito Q and LPS groups (hours: 11.60±0.24 vs. 11.90±0.13,χ2= 1.055,P= 0.137).② The serum amylase in the LPS 6 hours, LPS+ Wortmannin 4 hours, and LPS+ Mito Q 6 hours groups were significantly higher than those in the control group at the same time points (U/L:2 881.00±550.12 vs. 2 099.20±249.57, 3 672.00±779.24 vs. 2 081.36±245.18, 2 975.20±687.03 vs. 2 099.20± 249.57, allP< 0.05), and were significantly lowered in LPS 12 hours group (U/L: 794.00±218.71 vs. 2 086.80±261.75, P< 0.01). The pancreatic ROS in the LPS 6 hours and 12 hours groups, LPS+ Wortmannin 4 hours group, and LPS+Mito Q 6 hours group were significantly higher than those of the control group at the same time points (kU/L: 3.18±1.06 vs. 1.78±0.37, 3.63±1.08 vs. 1.85±0.41, 3.14±0.98 vs. 1.65±0.34, 3.17±1.03 vs. 1.78±0.37, allP< 0.05). The serum amylase and pancreatic ROS in LPS+ Wortmannin 4 hours group were significantly higher than those of the LPS group at the same time points (U/L: 3 672.00±779.24 vs. 2 432.20±442.85, kU/L: 3.14±0.98 vs. 1.87±0.42, both P< 0.05), but there were no differences in above two parameters between LPS+ Mito Q 6 hours group and LPS group (U/L: 2 975.20±687.03 vs. 2 881.00±550.12, kU/L: 3.17±1.03 vs. 3.18±1.06, bothP> 0.05). Light microscopy showed that obvious pathological changes were found in the pancreas in the LPS 6 hours and 12 hours groups, LPS+Wortmannin 4 hours group, and LPS+ Mito Q 6 hours group. Electron microscopy showed that the number of autophagic vacuoles increased 6 hours after LPS administration. There was no difference at any time point in the number of autophagic vacuoles between LPS+ Mito Q 6 hours group and LPS 6 hours group, and the autophagic vacuoles were not found after Wortmannin intervention. It was demonstrated by Western Blot that the levels of LC3 protein in the LPS 6 hours and 12 hours groups, and LPS+ Mito Q 6 hours group were significantly higher than those of the control group at the same time points (A value: 0.34±0.02 vs. 0.17±0.02, 0.37±0.03 vs. 0.18±0.04, 0.36±0.02 vs. 0.17±0.02, allP< 0.05), but there were no differences between LPS 12 hours group or LPS+ Mito Q 6 hours group and LPS 6 hours group (bothP> 0.05).Conclusions Autophagy prevents exocrine dysfunction of pancreas in septic rats, and the autophagic capacity or autophagosome-formation rate may determine the development of exocrine pancreatic dysfunction. The mitochondria-targeted antioxidant Mito Q does not prevent exocrine dysfunction of pancreas.

Objective To investigate the effects of autophagy on cardiac function and to determine whether the mitochondrial coenzyme Q (MitoQ) prevents cardiac dysfunction,mediated by autophagy,in rats with acute sepsis.Methods Forty-five Sprague Dawley (SD) rats were randomly divided into 9 groups (n =5,each group):control group,4 h lipopolysaccharide(LPS) group,6 h LPS group,12 h LPS group,4 h LPS + Wortmannin group,4 h LPS + MitoQ group,6 h LPS + MitoQ group,MitoQ group and Wortmannin group.Rats in LPS + Wortmannin group and LPS + MitoQ group were intraperitoneally given LPS(10 mg/kg) and followed by an injection of Wortmannin(2 mg/kg) and MitoQ (6.5 μmol/kg) via tail vein 1 hour later,respectively.Rats in each group were given the same amount of normal sodium in addition to different intervention drugs.The cardiac function parameters were measured by a BL-420E + biosignal collection system.Blood samples from abdominal aorta were taken at each time point,and creatine kinase MB isoenzyme (CK-MB) content was detected by using the velocity method.The content of reactive oxygen species (ROS) in isolated myocardial tissues in rats was measured by enzyme-linked immunoadsorbent assay(ELISA).The protein expression of microtubule-associated protein 1 light chain 3 (LC3) was detected by Western blot method.The pathological changes of myocardial tissue were observed by light and electronic microscopy.Results Compared with the control group,the left ventricular systolic pressure(LVSP),the rate of the rise in left ventricular pressure (± dp/dt max) were significantly decreased in 6 h LPS group,6 h LPS + MitoQ group and 4 h LPS + Wortmannin group(P ＜0.05),left ventricular end-diastolic pressure(LVEDP) was significantly increased in these 3 groups(P ＜0.05).The contents of CKMB and ROS in 6 h LPS group,6 h LPS =MitoQ group and 4 h LPS + Wortmannin group were higher than those in the control group(P ＜ 0.05).Electron microscopy showed that the number of autophagic vacuoles increased 6 h after LPS was administered,but did not increase significantly thereafter to 12 h.There was no difference at any time point in the number of autophagic vacuoles in the group given MitoQ and LPS.Immunoblotting demonstrated that the levels of LC3Ⅱ protein in the LPS 6 h group and LPS + MitoQ 6 h group were higher than those in the control group(P ＜0.05),but there was no difference between the LPS 12 h and LPS 6 h groups (P ＞ 0.05).Conclusions The mitochondria-targeted antioxidant MitoQ does not prevent cardiac dysfunction.However,autophagy prevents cardiac dysfunction,and the autophagic capacity or autophagosome-formation rate may determine whether cardiac dysfunction develops.

Objective To study the quality control standard of Gushuling capsule. Methods Fructus Psoraleae and Fructus Ligustri Lucidi in Gushuling capsule were identified by TLC; The content of ieariin was determined by HPLC with acetonitrile-water (30: 70) as mobile phase and the wavelength at 270 nm. Results TLC identification was positive; the linear range of icariin was 0.0968～0.4840 ?g(r=0.9999), the average recovery was 97.27%(RSD=1.88%, n=5). Conclusion This method can be used as a standard of the quality control for Gushuling capsule.