Objective To induce highly active hepatocellular carcinoma (HCC) specific cytotoxic T-lymphocyte (H-S-CTL) lines by co-culture of lymphocytes and hepatocellular carcinoma cells. Methods H-S-CTL were established by co-culture of autologous HCC cells and lymphocytes, under stimulation of IL-1,IL-2,IL-4 and IL-6.The cytotoxic activity of H-S-CTL for target cells were detected by 51Cr release assay and indirect immunofluorescence assay. Results Compared with auto-LAK, H-S-CTL showed much higher antitumor activity of significantly increased CD3+, CD4+ and CD8+ cells. Conclusions H-S-CTL lines with massive proliferations and high antitumor activity can be successfully obtained by ex vivo co-culture under the stimulation of interlukins.

Objective To investigate the effects of propofol and sevoflurane on adriamycin chemotherapy-induced myocardial damage in rats.Methods Twenty-four healthy male Wistar rats, aged 3 months, weighing 212-270 g, were randomly divided into 4 groups (n =6 each) using a random number table: control group (group C) , adriamycin group (group ADM) , adriamycin-propofol group (group ADM-Pro) , and adriamycin-sevoflurane group (group ADM-Sevo).The equal volume of normal saline was injected intraperitoneally in group C.In group ADM, adriamycin 2.5 mg/kg was injected intraperitoneally once every other day for 6 times (11 days in total).In group ADM-Pro, adriamycin 2.5 mg/kg was injected intraperitoneally once every other day for 6 times, and at 3 days after the last administration of adriamycin, propofol anesthesia was performed as follows : the total amount of propofol injected intraperitoneally was 200 mg/kg, the initial dose was 100 mg/kg, and 1 h later the remaining amount was added, and the duration of anesthesia was about 2 h.In group ADM-Sevo, adriamycin 2.5 mg/kg was injected intraperitoneally once every other day for 6 times, and at 3 days after the last administration of adriamycin, sevoflurane anesthesia was performed as follows: 1.5％-3.0％ sevoflurane was inhaled for 2 h.After the end of anesthesia, blood samples were collected for determination of serum cardiac troponin (cTnI) concentrations.Myocardial specimens were collected for detection of caspase-3 expression by immuno-histochemistry.Results Compared with group C, the serum cTnI concentration was significantly increased, and the caspase-3 expression was up-regulated in ADM, ADM-Pro and ADM-Sevo groups (P＜ 0.05).Compared with group ADM, the serum cTnI concentration was significantly decreased, and the caspase-3 expression was down-regulated in ADM-Pro and ADM-Sevo groups (P＜0.05).There was no significant difference in the parameters mentioned above between group ADM-Pro and group ADM-Sevo (P＞0.05).Conclusion Propofol and sevoflurane both can mitigate adriamycin chemotherapy-induced myocardial damage in rats, without significant difference in the efficacy.

Objective To study the relationship between the level of fluoride in urine and the prevalence of dental caries in children before and after the defluoridation, and to provide a basis for assessment of the effects of defluoridation projects and for control of dental caries.Methods Wamiao Village, in Jiangsu Province, a formerly severe endemic fluorosis area, was selected to carry out the study.All children aged 8-13 years old before the defuoridation were investigated from September to November in 2002, and children 8-10 years old who were born after the defluoridation were investigated from September to November in 2013.Urinary fluoride level of the children and the prevalence of dental caries were determined, and their relationships were analyzed.The urina sanguinis samples of children were collected in the morning, and the urine fluoride level was tested using the fluoride ion selective electrode.Dental caries diagnoses was referenced to Dental Caries.Results Totally children's urine samples were 236 and 68 respectively before and after defluoridation.Urinary fluoride level of the children was significantly decreased from (3.53 ± 1.81)mg/L (before defluoridation) to (1.39 ± 0.66)mg/L (after defluoridation, t =9.506, P ＜ 0.01);the prevalence of dental caries was increased from 52.73％ (29/55, before defluoridation) to 63.24％ (43/68, after defluoridation), however, the difference was not significant (x2 =1.383, P ＞ 0.05).The DMFT increased from 1.18 (before defluoridation) to 1.68 (after defluoridation), and the epidemic levels of dental caries were all at lower levels.The relationship between urine fluoride level and the prevalence of dental caries as well as the DMFT before defluoridation was a U-shape dose-response curve;which was gone after defluoridation.Conclusions The urinary fluoride level is significantly decreased after defluoridation for 10 years, the prevalence of dental caries is increased but not significantly.The results of this study indicate that the measure of fluoridation to prevent dental caries needs to be further validated.

Objective:To study the mechanism of the effectiveness loss of methotrexate(MTX)in the treatment of rheumatoid arthri-tis.Methods:The culture system of rat whole bone marrow cells(WBMCs)and tartrateresistant acid phosphatase(TRAP)staining were utilized to evaluate osteoclastogenesis.The mRNA expression of osteoclastogenesis factors in the WBMCs culture system was examined by semi-quantitative RT-PCR.Results:Deoxyadenosine(dAdo)decreased MTX-induced suppression of osteoclastogenesis.The recov-ery effect of dAdo on MTX was partially prevented by caffeine.MTX significantly reduced mRNA expression of receptor activator of nu-clear factor kappa-B ligand(RANKL),dAdo partially recovered RANKL mRNA expression and inhibited osteoprotegerin(OPG)expres-sion.Conclusion:The accumulation of dAdo may induce the effectiveness loss of methotrexate in rheumatoid arthritis treatment.Com-bination of MTX and caffeine can be a potential therapeutic strategy.

ObjectiveTo investigate the association between glucokinase regulatory protein (GCKR) gene rs780094 (C＞T) polymorphism and non-alcoholic fatty liver disease (NAFLD) in the Han population in Qingdao, China. MethodsA case-control study was conducted in 180 patients with NAFLD and 172 healthy controls. Blood samples were collected from all the subjects and were genotyped for rs780094 by polymerase chain reaction and DNA sequencing method. Genotype and allele frequencies were compared between the control and case groups. ResultsNo significant differences between the two groups were observed in both genotype and allele frequencies (both P＞0.05). The variant GCKR rs780094 T allele did not significantly increase the risk of NAFLD (OR=1.068, 95% CI: 0.748-1.525, P=0.719) compared with the C allele. Logistic regression model analysis adjusted for confounding factors exhibited that the TT + CT genotype of GCKR rs780094 did not significantly increase the risk of NAFLD relative to the CC genotype (P＞0.05). ConclusionGCKR rs780094 T allele may be not associated with the increased risk of NAFLD in the Han population in Qingdao.

Objective To investigate the effects of preconditioning with different doses of levothyroxine sodium on myocardial ischemia-reperfusion (I/R) injury in immature rats. Methods Forty-eight female immature Wistar rats, aged 35 days, weighing 120-140 g, were randomly allocated into 6 groups ( n = 8 each): control group (group C), I/R group, and preconditioning with levothyroxine sodium 10, 20, 40 and 80 μg/100 g groups (groups LS1-4 ) . The rats received levothyroxine sodium 10, 20, 40 and 80 μg/100 g through a gastric tube every day for 7 days in groups LS1-4 , respectively. Venous blood samples were taken on 8th day for determination of serum thyroid hormone levels. The hearts were removed from the animals and perfused in a Langendorff apparatus with K-H solution saturated with 95% O2-5% CO2 at 37 ℃. The hearts were continuously perfused for 80 min in group C. After 30 min of equilibration, the isolated hearts were subjected to 20 min of ischemia followed by 30 min of reperfusion in I/R and LS1-4 groups. HR, SP and ± dp/dtmax were recorded at 20 min of perfusion and 30 min of reperfusion. The recovery rates of HR, SP and ± dp/dtmax were calculated at 30 min of reperfusion. The coronary effluent was collected at 10 min of perfusion and 15 min of reperfusion for determination of creatine kinase (CKMB) activity. The samples of ventricular myocardial tissues were taken at 30 min of reperfusion to detect the expression of heat shock protein 70 (HSP70), thyroid hormone receptor (TR) mRNA (TRa, , TRoj and TRft ) and myosin heavy chain (MHC) mRNA. Results Compared with group C, the recovery rates of HR, SP and. ± dp/dtmax were significantly decreased, the CK-MB activity was significantly increased, and MHCα mRNA expression was down-regulated in group I/R, the recovery rates of SP and ± dp/dtmax were significantly decreased, the CK-MB activity was significantly increased, and the expression of HSP70 and MHCα mRNA was up-regulated in groups LS1-4, and the serum thyroid hormone level was significantly increased and the expression of TRa, mRNA was up-regulated in groups LS2-4 ( P ＜ 0.05) . Compared with group I/R, the recovery rates of HR and ± dp/dtmax were significantly increased, the pression of HSP70 and MHCa mRNA was up-regulated, and the MHCJ3 mRNA expression was down-regulated in groups LS1-4 the CK-MB activity was significantly decreased in groups LS1-3, and the serum thyroid hormone level was significantly increased and the expression of TRα1, mRNA was up-regulated in groups LS2-4 ( P ＜ 0.05) . The serum thyroid hormone level increased gradually with the increase in the dose of levothyroxine sodium in groups LS1-4 ( P ＜ 0.05) . The CK-MB activity was significantly higher, while the HSP70 expression lower in groups LS3-4 than in groups LS1-2 (P ＜ 0.05). Conclusion Preconditioning with levothyroxine sodium 10 μg/100 g can alleviate the myocardial I/R injury in immature rats and does not lead to the increase in the level of thyroid hormone, and the up-regulation of HSP70 and MHCa mRNA expression may be involved in the mechanism.

Objective To evaluate the protective effects of fentanyl postconditioning and remote limb ischemic postconditioning (RLIP) against myocardial ischemia-reperfusion (I/R) injury in rats. Methods Thirty-nine male SD rats aged 8 weeks weighing 250-350 g were randomly allocated into 5 groups: Ⅰ group sham operation (group S, n = 5); Ⅱ group I/R ( n = 7); Ⅲ group fentanyl postconditioning (group F, n= 9); Ⅳ group RLIP (group R, n = 9) and group Ⅴ fentanyl postconditioning + RLIP (group F-R, n = 9). The animals were anesthetized with intraperitoneal 3% pentobarbital 50 mg/kg, intubated and mechanically ventilated. Myocardial I/R was induced by occlusion of anterior desending branch of left coronary artery for 30 min followed by 180 min reperfusion. Fentanyl 30 μg/kg was injected iv at 15 min of myocardial ischemia in group F and F-R In group R and F-R the animals underwent 10 min ischemia of bilateral hind limbs starting from 15 min of myocardial ischemia. HR and MAP were recorded at 5,60,120 and 180 min of reperfusion and rate-pressure product( RPP, HR × MAP) were calculated. At the end of 180 min reperfusion, arterial blood samples were obtained for measurement of the activities of plasma lactate dehydrogenase (LDH) and creatine kinase isoenzyme MB (CK-MB), and the concentration of serum cardiac troponin Ⅰ (cTnI). The animals were then sacrificed. The infarct size was evaluated by double staining with Evans blue and triphenyl tetrazolium chloride. Results Myocardial I/R significantly increased plasma LDH and CK-MB activities and serum cTnI concentration and decreased HR,MAP and RPP as compared with group S.Fentanyl postconditioning and RLIP both decreased plasma CK-MB activity, serum cTnI concentration and infarct size and increased HR, MAP and RPP in group F, R and F-R as compared with group I/R. Plasma CK-MB activity,serum cTnI concentration and RPP were significantly lower and infarct size was smaller in group F-R than in group F. The infarct size was significantly smaller and MAP and RPP were higher in group F-R than in group R.Conclusion Fentanyl postconditioning can provide a myocardial protection against I/R injury. Myocardial protection is enhanced by combination of fentanyl postconditioning and RLIP.

Objective To improve the recognition and management of primary breast lymphoma(PBL) by discussing the clinical pathological characteristics and the prognosis of PBL. Methods The clinical and treatment records of PBL pa-tients admitted in Boai hospital of zhongshan from January 2001 to December 2012 were analyzed. Results All cases were B-cell origin(25/25),61.5%(16/25)were diffuse large B-cell lymphoma origin,The 5-year overall survival was 44%. There were no significant difference in the 5-year overall survival rates, 5-year local control rates and 5-year distant control rates in patient treatment with any surgery compared with no-Surgery(P>0.05).There were no significant difference in the 5-year overall survival rates and 5-year distant control rates in patient treatment with any RT com-pared with no-RT(P>0.05),but the 5-year local control rates was significantly different(P<0.05);There were no sig-nificant difference in the 5-year overall survival rates and 5-year local control rates in patient treatment with any chemotherapy compared with no-chemotherapy (P>0.05). but the 5-year distant control rates was significant different(P<0.05). Conclusion The prognosis of PBL is related with the clinical stage and pathological type. Radiotherapy and chemotherapy respectively promote the local control and distant control. The optimal treatment is surgery combined with chemotherapy and/or radiotherapy.

Objective To evaluate the preventive efficacy of oral small-dose thyroid hormone tablet premedication for a short time on euthyroid sick syndrome (ESS) in children undergoing open heart surgery under cardiopulmonary bypass (CPB) .Methods Forty ASA Ⅰ or Ⅱ children aged 3-12 yr, weighing 10-30 kg, scheduled for elective congenital heart disease surgery under CPB, were randomly allocated into 2 groups ( n = 20 each):placebo group (group P) and thyroid hormone tablet group (group T). Group T received oral thyroid hormone tablets 0.4 mg/kg every day for 4 consecutive days before surgery, while group C were given placebo. CPB was routinely established, and mild hypothermia, moderate hemodilution and high flow perfusion were adopted. Blood samples were taken from radial veins before administration (baseline) and on 1st, 2nd and 4th day after surgery to detect the serum concentrations of triiodothyronine (T3), thyroxine (T4) and thyroid stimulating hormone (TSH).SP, DP and HR were recorded before administration, immediately after surgery, and on 1st and 2nd day after surgery. The endotracheal extubation time, length of ICU stay, application of positive inotropic agents and occurrence of ESS were recorded. Results No significant difference was found in hemodynamic parameters, endotracheal extubation time and length of ICU stay between the two groups ( P ＞ 0.05). As compared with the baseline values,the serum T3 levels on 1st, 2nd and 4th day after surgery, and the serum TSH levels on 1 st day after surgery were significantly decreased in the two groups, and the serum T4 levels were significantly decreased on 1 st day after surgery in group P ( P ＜ 0.05). The serum levels of T3 and T4 were significantly higher, the severity of postoperative ESS and the number of positive inotropic agent administration were significantly lower in group T than in group P (P ＜ 0.05 ). Conclusion Although oral small-dose thyroid hormone tablet premedication for 4 days (0.4 mg/kg per day) can reduce the severity of postoperative ESS, but it can not prevent the occurrence of ESS in children undergoing open heart surgery under CPB.

¹⁷⁷Lu- prostate specific membrane antigen (PSMA) radio-ligand therapy has been approved abroad for advanced prostate cancer and has been in several clinical trials in China. Based on domestic clinical practice and experimental data and referred to international experience and viewpoints, the expert group forms a consensus on the clinical application of ¹⁷⁷Lu-PSMA radio-ligand therapy in prostate cancer to guide clinical practice.