Objective To observe the effects of low dose irradiation (LDI) on autoiogous CIK cell proliferation, phenotype and killing activity in tumor patients, and to provide the evidence for clinical application of adoptive immunotherapy with CIK cells. Methods Peripheral blood mononuclear cells (PBMC) were separated from 10 patients with malignant tumor, and CIK cells were cultured with different cytokines. (1) After 10 d culture, C1K cells were irradiated with different doses as 30, 50, 80, 100 and 200 Gy of X-rays was also detected. The CIK cell proliferation and killing activity were measured with 3H-TdR incorporation assay and 3H-TdR release assay, respectively and the percentage variation of CD3 +CD56 + were measured with flow cytometry after 24 h. ( 1 ) Autologous CIK cells were irradiated with 80 mGy X-rays. At different culture time ( 12, 24, 48, 72 h) after irradiation, the killing activity was measured with 3H-TdR release assay. (3) The effect of 3d low dose irradiation of 80 mGy X-rays on thekilling activity of CIK cells was also detected. Results After the CIK cells were irradiated with different doses as 50, 80, 100, 200 mGy of X-rays, the CPM values were 20 048.6 ± 2332. 2 ( t = 2.2, P ＜0.05), 21 832.2 ±2975.9 (t=3.5, P＜0.01), 21 000.3 ±2451.1 (t=3.3, P＜0.01), 19908.1 ±2051.0 ( t = 2.2, P ＜ 0.05 ), respectively and the proliferation of CIK cells were significantly higher than that of control group. The CD3 + CD56 + cell percentage of 50, 80, 100 mGy groups were ( 30.3 ±3.8)% (t=2.3, P＜0.05), (32.3±3.4)% (t=4.2, P＜0.01), (29.742.9)% (t = 2.4, P＜0.05 ), respectively. The killing activity of CIK cells of 80, 100 mGy groups were 55.2 ± 5.0 ( t = 3.3, P ＜ 0.01 ), 52.8 ± 4.1 ( t = 2.3, P ＜ 0.05 ), respectively. The killing activity of CIK cells up-regulated significantly at 24 h, dropped to normal levels at 48 h and 72 h. After 80 mGy X-ray irradiation for 3 consecutive days, the killing activity of CIK cells at different time points were 55.2 ± 5.3 (t = 2.6, P ＜0.05),61.9 ± 4.4 (t = 4.7, P ＜0.01), 67.2 ±5.7 (t = 5.7, P ＜0.01) for 24, 48, 72 h,respectively. Conclusions LDI might have the hormesis effect on CIK cells.

Objective To investigate the effect of siRNA targeted against Bcl-xl on cell proliferation of rheumatoid arthritis (RA) and the effect on expres-ion of apoptosis relevant factors Bcl-xl,Bax,Caspase-3.Methods Human RA synovial cells were cultured and passed by tissue block collagenase digestion method.The siRNA plasmid targeting Bcl-xl gene was constructed by Bcl-xl cDNA sequence provided by gene banks,while single missense sequences were used as negative controls.LipofectamineTM 2000 was used to transfect synovial cells.The effect of Bcl-xl silencing on proliferation of synovial cells was evaluated by MTT at 24,48,72 hours after transfection.The expressions of Bcl-xl,Bax,Caspase-3 protein,synovial apoptosisrelated factors were determined by Western blotting after transfected at different time points.The average of multiple-sample average was analyzed by single-factor x2 test or LSD-t and Tamhane's T2 test was used for twotwo comparison.Results MTT result s showed that RNA interference that specifically silent Bcl-xl could obviously suppress the proliferation of sliding film cells,which was most eveident at 48 hours.This inhibition was gradually weakened with prolonged time,but when compared was the control group,differences was significant (P＜0.05).Western blotting results displayed that when compared to the control group,Bcl-xl protein expression obviously declined after transfection P＜0.01,which was the least at 48 h time point.Bax,Caspase3 protein expression were obviously increased when compared to the coutrol group.Conclusion The expression of Bcl-xl,a RA synovial cell anti-apoptotic factor,is significantly reduced by specific RNA interference silencing Bcl-xl,which may play an important role in inhibiting the excessive proliferation of synovial cells.

BACKGROUND: Sleep deprivation cannot only cause learning and memory impairment of animal and human, but also lead to increased content of nitric oxide in brain tissue of rats. Melatonin has the effects of antifreeradical and antioxidation. It has been reported that melatonin can improve aluminum chloride and morphine abstinence induced learning and memory impairment of animal, however, whether it has influence on sleep deprivation induced learning and memory impairment is not very clear. OBJECTIVE: To observe the effect of melatonin on memory of rats after sleep deprivation and analyze its possible mechanism. DESIGN: Randomized controlled trial.SETTING: Teaching and Experiment Center of Basic Medicine and Department of Nursing, the Fourth Military Medical University.MATERIALS: The experiment was performed in Teaching and Experiment Center of Basic Medicine of the Fourth Military Medical University in January 2005. A total of 24 adult male Sprague Dawley rats were selected and randomly divided into 3 groups, namely, control group, small dosage of melatonin group and large dosage of melatonin group, with 8 in each group on the basis of random digits table.METHODS: To rats in small dosage of melatonin group and large dosage of melatonin group, the dosage of melatonin was 5 mg/kg and 15 mg/kg respectively, which was made into 2 mL solution and intraperitoneally injected into the rats at 17:00 o'clock every day, while rats in control group were injected with 2 mL physiological sodium at the same time, once a day for continuous 7 days. Then a 3-day sleep deprivation was given to the rats; melatonin or physiological sodium were also given according to different groups during these days. Rat model of sleep deprivation was established by "Flower Ppot" technique; water maze was used for detecting the memory of rats after 48-hour and 72-hour sleep deprivation; took escape latency (s) as indicator of changes of learning and memory of rats; the shorter the escape latency, the better the spacial memory of rats. When sleep deprivation was finished, all the rats were put to death and hippocampus and cerebral cortex were taken out in ice bath. The content of nitric oxide in cerebral cortex and hippocampus was detected with the method of nitrate reduction, and malondialdehyde (MDA) with the method of thiobarbital acid.MAIN OUTCOME MEASURES: Results of escape latency after 48hour and 72-hour sleep deprivation. Contents of nitric oxide and MDA in cerebral cortex and hippocampus of rats.There was significant difference in escape latency in water maze after 48-hour and 72-hour sleep deprivation among each group (F=11.886, P=0.000)and (F=5.440, P=0.012); the escape latency after 48-hour and 72-hour sleep deprivation remarkably decreased both in small and large dosage of melatonin groups as compared with control group, and the latency after 48-hour sleep deprivation was shorter in large dosage group than that in small ide and MDA in brain of rats among each group, namely, nitric oxide in cerebral cortex (F=14.038, P=0.000), MDA in cerebral cortex (F=27.414,P=0.000), nitric oxide in hippocampus (F=22.692, P=0.000), MDA in hippocampus (F=14.316, P=0.000). Compared with control group, the contents of nitric oxide and MDA in cerebral cortex and hippocampus in the two experimental groups decreased significantly, and there was obvious difference in the content of nitric oxide in hippocampus between large and small dosage groups, which showed a dose-effect relationship.CONCLUSION: Melatonin can improve memory impairment of rats after sleep deprivation, which may be closely related to the effect of inhibiting the increase of nitric oxide and MDA in their cerebral cortex and hippocampus.

10.3969/j.issn.1007-3969.2013.05.009

Objective The aim of this study is to evaluate the safety and efficacy of partial pancreatectomy as part of primary cytoreductive surgery in advanced epithelial ovarian cancer (EOC). Methods A total of 8 patients were recruited in this study who underwent partial pancreatectomy during the primary cytoreductive surgeries for advanced EOC in Fudan University Shanghai Cancer Center from April 2009 to July 2015. Their clinicopathological characteristics, diameter of metastatic tumors, the scope of cytoreductive surgeries, residual diseases after cytoreductive surgeries, postoperative complications and survival situation were retrospective analyzed. Results (1) Clinicopathological characteristics:the median age of these patients was 58 years old(range: 39-63 years old). The median value of preoperative serum CA125 was 1 688 kU/L(range: 119-5 000 kU/L). The median diameter of metastatic tumors involved in pancreatic body or tail was 4.5 cm (range:3-10 cm). All the tumors from the 8 patients were confirmed to be high-grade serous carcinoma. Four patients were staged as International Federation of Gynecology and Obstetrics (FIGO)Ⅳ, and the other 4 patients were staged as FIGOⅢc. (2) Tumor metastases and the scope of cytoreductive surgeries:all of these 8 patients had widely disseminated ovarian cancer, with involvement of upper abdominal, middle abdominal and pelvic cavity. Each patient underwent extensive intra-abdominal cytoreductive surgeries, including hysterectomy, bilateral salpingo-oophorectomy, omentectomy, pelvic periton-ectomy, splenectomy, partial pancreatectomy. Each patient had cytoreductive surgeries of 9.6 different sites on average. Of all 8 patients who underwent partial pancreatectomy, 7 patients had pancreatic tails removed;the other 1 patient had pancreatic body and tail removed. The median volume of blood loss during surgery was 1 350 ml(range:300-3 500 ml), blood transfusion was performed in 7 patients with the median volume of 1 150 ml (range: 500-1 800 ml). (3) Residual diseases after cytoreductive surgeries: optimal cytoreduction was achieved in all patients, with microscopic residual disease in 3 patients, residual tumors diameter < 0.5 cm in 3 patients, and residual tumors diameter between 0.5 and 1 cm in 2 patients. (4) Postoperative complications: 4 patients suffered from complications including pancreatic leakage (2/8), intraperitoneal hemorrhage (1/8) and pancreatic pseudocyst accompanied by infection (1/8). These complications were treated successfully by conservative managements. (5) Survival situation: during the median follow-up duration of 17 months (ranged from 2 to 46 months), 5 patients were still alive until the end of follow-up, including 4 cases under treatment and 1 case survived 29 months without relapse after treatment. Three patients were respectively died in 5, 20 and 46 months after surgery. Conclusion There is a higher risk of postoperative complications of pancreas resection as part of primary cytoreductive surgery in advanced epithelial ovarian cancer, but the resection of pancreatic metastases and part of the pancreas is feasible and necessary.

Objective To evaluate the difference of clinical short-term effect and adverse reaction between paclitaxel liposome and paclitaxel in non-small cell lung cancer patients with lymph node metastasis after pulmonary resection.Methods Sixty-eight patients after pulmonary resection were divided into two groups by random digits table method,37 patients in experimental group with paclitaxel liposome (135mg/m2) combined with carboplatin (CBP) at 300 mg/m2 in chemotherapy,and 31 patients in control group with paclitaxel (135 mg/m2) combined with CBP at 300 mg/m2 in chemotherapy.Results All patients were evaluable.In experimental group,5 patients had complete remission,10 patients had partial remission,17patients were stable,5 patients' condition aggravated,the total effective rate was 40.5％(15/37),clinical control rate was 86.5％ (32/37).In control group,2 patients had complete remission,8 patients had partial remission,15 patients were stable,6 patients' condition aggravated,the total effective rote was 32.3％(10/31),clinical control rate was 80.6％(25/31).The treatment effectiveness in experimental group was significantly higher than that in control group (P ＜ 0.05).The main adverse reaction included marrow suppression,hair loss,muscle and joint pain and gastrointestinal symptom,there was no serious hypersensitivity.The rate of hypotension,face flushing,paresthesia,muscle and joint pain,erythra in experimental group was lower than that in control group [0 vs.9.7％ (3/31),5.4％ (2/37) vs.19.4％ (6/31),10.8％ (4/37) vs.22.6％ (7/31),13.5％ (5/37) vs.38.7％ (12/31),5.4％ (2/37) vs.25.8％ (8/31)] (P ＜0.0 1 or ＜0.05).Conclusion The curative effect rate of paclitaxel liposome is better than paclitaxel in patients with lymph node metastasis after pulmonary resection and with lower incidence of side effects.

Objective To investigate the changes of compliance in large arteries and carotid artery intima-media thickness(IMT)in patients with metabolic syndrome.Methods There were 64 patients with metabolic syndrome and 56 age-matched control subjects.Their carotid-femoral pulse wave velocities(C-FPWV)were measured by the Complior SP and their carotid artery IMT were detected by B-mode ultrasound.At the same time their height,weight,waist circumstance,hip girth,blood pressure,blood glucose,blood lipid,BMI and waist to hip ratio(WHR)were measured.Results Compared with the control,the patients with metabolic syndrome had higher C-FPWV(P

This study examined the effect of ischemia-reperfusion injury on the expression of Pim-3 gene in myocardial tissues and their underlying mechanism. Rat models of myocardial ischemia-reperfusion injury were established by ligating the left anterior descending coronary artery of the rats. A total of 30 SD male adult rats were randomly divided into 5 groups: group A (sham operation, n=6); group B (in which the rats were subjected to 15 min of ischemia by ligation of the left anterior descending coronary artery, n=6); group C (in which the rats received 30 min of ischemia, n=6), group D and group E (in which the left anterior descending coronary artery of the rats were ligated for 30 min and then reperfused for 30 min or 120 min, n=6 in each). The left ventricular tissues were removed immediately after the ischemia-reperfusion injury. Neonatal cardiomyocytes were cultured and treated with different concentrations of H(2)O(2) (0, 5, 10, 20 μmol/L) or tumor necrosis factor-α (TNF-α, 0, 1, 5, 10 ng/mL). The mRNA and protein expression of Pim-3 gene was determined by using RT-PCR, western blotting and immunohistochemistry. Additionally, neonatal cardiomyocytes were transfected with Pim-3 siRNA, and induced to develop apoptosis by using H(2)O(2). The results showed that normal myocardial tissues expressed a quantity of Pim-3 gene mRNA and protein. Ischemia-reperfusion injury could up-regulate the mRNA and protein expression of Pim-3 gene in myocardial tissues. Furthermore, H(2)O(2) but not TNF-α up-regulated the Pim-3 gene expression in cultured cardiomyocytes. And Pim-3 silencing failed to strengthen the H(2)O(2)-inducing apoptosis in cardiomyocytes. It was concluded that ischemia-reperfusion injury up-regulated the Pim-3 gene expression through oxidative stress signaling pathway in myocardial tissues.

Pim kinases contribute to tumor formation and development of lymphoma, which shows enhanced DNA replication, DNA recombination and repair. Endothelial cells^(ECs) express all the three members of Pim kinase gene family. We hypothesized that DNA repair gene would regulate Pim expression in ECs. Human umbilical vein endothelial cells (HUVECs) were isolated and maintained in M199 culture medium. The cellular distribution of Pim-3 in ECs was determined by immunofluorescent staining. The siRNA fragments were synthesized and transfected by using Lipofectamine LTX. The total cellular RNA was extracted from the cells by using Trizol reagent. cDNAs were quantified by semi-quantity PCR. The effects of LY294002 and wortmannin on RNA stability in ECs were also examined. Our data showed that LY294002 and wortmannin, phosphatidylinositol 3-kinase (PI3K) and PI3K-like kinase inhibitors, increased Pim mRNA expression in ECs without altering the mRNA stability. RNA interference (RNAi) targeting DNA-dependent protein kinase catalytic subunit (DNA-PKcs) and ataxia telangiectasia mutated (ATM) increased mRNA expression of Pim-3 and Pim-1, respectively. Silencing of Akt decreased Pim-1 instead of Pm-2 and Pim-3 gene expression in ECs. But etoposide, a nucleoside analogue, which could activate DNA-PKcs and ATM, increased Pim expression in ECs. Our study indicates that the expression of Pim kinases is physiologically related to DNA-PKcs and ATM in ECs.

BACKGROUND: Simple polyvinyl alcohol (PVA) has limited ability to cell adhesion. There are not generally accepted studies on improved effects of collagen protein modified polyvinyl alcohol on cell adhesion and proliferation.OBJECTIVE: To investigate the feasibility of PVA/type Ⅰ college (COL-Ⅰ) as anterior cruciate ligament (ACL) scaffolds in tissue engineering.DESIGN, TIME AND SETTING: The controlled observation experiment was performed at the Fourth Affiliated Hospital, Medical College. Ji'nan University, Guangzhou Red Cross Hospital, Guangzhou Institute of Trauma Surgery from August 2006 to October 2007.MATERIALS: COL-Ⅰ gel was produced by Guangzhou Institute of Trauma Surgery.METHODS: PVA filature was used to weave fascicular scaffolds. NIH-3T3 cell line and human ACL cells were in vitro incubated, amplified, and then implanted on the PVA/COL scaffolds.MAIN OUTCOME MEASURES: The growth of NIH-3T3 cell line and human ACL cells on the PVA/COL scaffolds and the secretion of extracellular matrix were observed using scanning electron microscope. Cell compatibility of PVA/COL scaffolds was assessed. Mechanics characteristic of PVA/COL scaffolds was measured by using the electric. tensile force apparatus. Mechanical property of PVA/COL scaffolds was analyzed using the SPSS 11.5 software package.RESULTS: NIH-3T3 cell line and human ACL cells on the PVA/COL scaffolds adhered, proliferated, and secreted extracellular matrix. NIH-3T3 cell line highly grew compared with human ACL cells on the PVA/COL scaffolds. The adhered number of NIH-3T3 cell line and human ACL cells was significantly increased on the PVA/COL scaffolds. NIH-3T3 cell line and human ACL cells presented well morphology on the PVA/COL scaffolds. COL-Ⅰ could promote the secretion of extracellular matrix from NIH-3T3 cells, but its effects on human ACL cells were not significant. Tensile force test showed that load-extension curve of the materials was identical to ACL of human and rabbits, and the scaffolds possessed strong flexibility. The maximal load, ultimate stress and elastic modulus were respectively 52.61 N, 14.96 MPa and 202.08 MPa.CONCLUSION: COL-Ⅰ accelerates the adhesion and proliferation of NIH-3T3 cell line and human ACL cells on the surface and in the pore of the PVA/COL scaffolds, promotes the secretion of extracellular matrix from NIH-3T3, and PVA filature material has mechanical property and good cell compatibility.