Objective:To investigate the potential value and feasibility of MG132 as a new therapy method for the ovarian cancer and a cisplatin chemotherapy-synergistic agent.Methods:The mode of the nude mice transplanted tumor tissues of ovarian epithelial carcinoma were established,20 nude mice were randomly divided into four groups and were all intrapedtoneal injected,once a day,a total of seven days:①Control group(0.2 ml saline),②MG132 group(2 mg/kg),③Cisplatin group(1 mg/kg),④Combination group[MG132(2 mg/kg) + cisplatin (1 mg/kg)].The weight inhibitory rates of tumors in each group were compared after four weeks.The expressions of Caspase3,Beclin1 in each group were detected by IHC,FIA,western blot and RT-PCR.Results:①The inhibitory rate of tumors in cisplatin group,MG132 group,and combination group was 53.85％,15.38％,88.46％,respectively,the additive effect of cisplatin and MG132 combination therapy was 60.95％.②IHC,FIA,western blot detected that compared to control group,the positive expressions of Beclin1,Caspase3 were increased in cisplatin group,MG132 group,and combination group,among which combination group increased more.③RT-PCR detected that the mRNA relative quantity of Beclin1 in cisplatin group,MG132 group,combination group respectively were higher than that of control group(P＜0.05);and it was higher in combination group than that of cisplatin group and MG132 group(P＜0.05).Conclusions:The growth of nude mice transplanted tumor tissues of ovarian epithelial carcinoma can be inhibited by MG132,and has a synergistic effect for treating ovarian cancer by combination with cisplatin,it is expected to be an effective anti-tumor drug for platinum resistant refractory ovarian cancer.

Objective To study the effects of ulinastatin on oxidative stress factors and renal function in rats with severe heatstroke. Methods-Forty-two male SPF Wistar rats were randomly divided into normal control group (group C, n=6), heat stroke group (HS group, n=18) and ulinastatin treatment group (UTI group, n=18). Rats in the HS group and UT group were placed in an artificial climate chamber to induce HS, while rats of group C were kept at room temperature (23±0.2°C). UT rats were given by intraperitoneal injection of ulinastatin before model reproduction in a dose of 100 thousand units/kg, and it was repeated every 12h. The other in the rest two groups were injected with equal amount of normal saline. All the rats were sacrificed in batches at 0, 6 and 24h after successful modeling. Before death, blood samples were collected to determine serum creatinine and urea nitrogen with automatic biochemical analyzer. Meanwhile the concentrations of renal superoxide dismutase (SOD), glutathione peroxidase (GPx) and malondialdehyde (MDA) were determined with colorimetric method. Pathological changes in renal tissue of all animals were observed at 24h after model replication with light and electron microscopy. Results-Compared with rats in group C, the MDA levels at 0h after onset of heatstroke increased significantly (P<0.05) in the renal tissue of HS and UTI group, and they were also significantly higher at 6 and 24h although they began to fall afterward (P<0.05). The MDA level was significantly lower in the UTI group than that in the HS group at each time point (P<0.05). The levels of SOD and GPx in the rat kidney tissue of HS group were lower than that of group C at 6h after successful model replication (P<0.05), while there was no significant decline in UTI group compared with group C (P>0.05). At the same time, compared with HS group, the elevated levels of serum Cr and BUN in the UTI group were lower at 6 and 24h after heatstroke onset (all P<0.05). The pathological damage of the kidney at 24h after heatstroke was significantly ameliorated in the UTI group. Conclusions-Oxidative stress is involved in the pathogenesis of the renal injury during heatstroke, UTI may protect rats with heatstroke from renal injury by inhibiting oxidative stress in the kidney.

Objective:To develop an economical,simple and portable device for imitation of ship shock motion produced by non-contacting under water explosion.Methods:The device was developed based on aerodynamical principles and sensor detection technology,and was tested with animal experiments to verify its performance.The injuries caused by 4 grades of shock motions with different accelerations,duration and displacement were observed in 40 black rabbits(10 for each grade).Results:The device was safe and functionally stable,with a peak acceleration of 1 000 m/s~2,a shock duration of about 2 ms and a device displacement of 100 mm.The accelerations of 4 shock motion grades were significantly different and with good reproducibility.The damage to the rabbits were mainly haemorrhage of different extents in organs of pleuroperitoneal cavity,especially in the lungs,but no rupture of organs was found.The degree and involvement of damage had an increasing tendency with the increase of acceleration of shock motion.Conclusion:The device we developed can imitate the effects of ship shock motion.

Objective To study the expression of interferon regulatory factor 1 (IRF-1) in patients with systemic lupus erythematosus (SLE) and the influence of prolactin (PRL) upon it. Methods The level of serum PRL in quiescent condition was examined by electrochemiluminescence-meter in 30 patients with SLE and 20 healthy controls. Peripheral blood mononuclear cells (PBMC) were separated from all the subjects by gradient centrifugation density, and cultured with or without the presence of recombinant human PRL (rhPRL) for 24 hours. The expression of IRF-1 gene in cultured PBMC was detected by reverse transcriptase-PCR (RT-PCR) with gel image scanning. Results The relative value of IRF-1 gene expression was significantly higher in SLE patients than in normal controls (0.89±0.21 vs 0.78±0.18, P=0.026), and in SLE patients with high PRL than in those with normal PRL (1.06±0.26 vs 0.82±0.21, P=0.005). However, there was no significant difference between SLE patients with normal PRL and healthy controls in regard to the expression of IRF-1 gene (P=0.514). The stimulation with rhRPL significantly elevated the relative expression of IRF-1 gene in SLE patients with normal PRL (0.99±0.22 vs 0.82±0.21, P=0.036), but had no obvious effect on that in the normal controls. Conclusion The study reveals a high expression of IRF-1 gene in SLE patients, which may be related to the high level of PRL.

Animals ; Carnitine ; pharmacology ; H(+)-K(+)-Exchanging ATPase ; metabolism ; Mitochondria, Muscle ; enzymology ; Muscle, Skeletal ; drug effects ; Physical Conditioning, Animal ; Rats

Adult ; Female ; Humans ; Paraquat ; poisoning

BACKGROUND:The levels of interleukin-1 (IL-1) and tumor necrosis factor-α (TNF-α) are increased during infectious brain edema, and are positively relevant to the degree of brain damage. However, whether TNF-α can enhance blood brain barrier (BBB) permeability remains unclear, especially in vitro.OBJECTIVE: To understand the changes and possible mechanism of the BBB permeability induced by TNF-α in vitro.DESIGN: Randomized controlled cell model study in vitro.SETTING:Department of Pediatrics, Xiangya Hospital, Central South University; Department of Biochemistry, Xiangya Medical College, Central South University.MATERIALS: Twenty 7-day-old healthy Sprague-Dawley rats, of clean grade and either gender, were provided by the Animal Center, Xiangya Hospital, Central South University. TNF-α was purchased from sigma Company; DMEM fluid medium and fetal bovine serum were purchased from Hyclone Company; Y-27632 was purchased from Alexis Company,and rabbit anti-human factor Ⅷ -related antigen was purchased from Zymed Company; Mouse anti-rat glial fibrillary acidic protein (GFAP) was purchased from Neomarkers. Other biochemical reagents were imported (Sigma Company).METHODS: This experiment was carried out in the Xiangya Hospital, Central South University between March 2004 and April 2005. Brain microvascular endothelial cells and astrocytes were co-cultured 10 days to set up rat models of BBB in vitro. Then, the cells were divided into 4 groups: model group(BBB models were prepared), TNF-α group ( BBB model incubated with 0.01 g/L TNF-α for 5 hours), Y-27632 pretreated group ( BBB model incubated with 30 μmol/L Y-27632 for 1 hour before 0.01g/L TNF-α challenge ) and Y-27632 control group (BBB models only incubated with Y-27632 as those in the Y-27632 pretreated group). The effect of TNF-α on BBB permeability was observed by detecting the 125 I -BSA, which passed through the inserts at each time point (30,60,120 and 240 minutes) using .γradioimmunoassay counter.MAIN OUTCOME MEASURES: The 125 I -BSA, which passed through the inserts in rat models of BBB at different time points after intervention.RESULTS: The 125 I -BSA, which passed through the inserts in rat models of BBB, was all significantly higher in the TNF-α group than in the other groups at 30, 60, 120 and 240 minutes after intervention, respectively (P ＜ 0.01), and reached the peak at 240 minutes; The 125 I -BSA, which passed through the inserts, was lower in the Y-27632 pre-treated group than in the TNF-α group at 30 and 60 minutes after intervention (P＜ 0.01). There was also significant difference in 125 I -BSA permeation between Y-27632 pretreated group and Y-27632 control group after 120 minutes (P ＜ 0.05).CONCLUSION: TNF-α can increase BBB permeability, and Y-27632 pretreatment can early reverse the effect of TNF-α on BBB permeability.

Objective To study the variation of serum thyroid hormones in each trimester of pregnancy and their relationship with thyroid peroxidase antibody(TPOAb).Methods Three-hundred and seventy-seven pregnant women in different trimesters were enrolled,the serum TSH,FT3,FT4,TT3,TT4,and TPOAb were determined by ICMA assay.Results Serun TSH in normal pregnant group was gradually increased as the pregnant trimester grew up ( all P＜0.05 ).The median of TSH in positive TPOAb group was higher than that of negative TPOAb group( P＜0.05 ).The prevalences of both clinical and subclinical hypothyroidism in positive TPOAb group were also higher than those in negative TPOAb group( all P＜0.05 ).The prevalences of subclinical hypothyroidism were 23.61％ and 18.04％,while those of clinical hypothyroidism were 0.27％ and 5.84％ by using TT4 and FT4 determination,respectively( P＜0.05 ).Conclusions More attention should be paid to the difference between FT4 and TT4 in hypothyroidism diagnosis during both second and third trimesters of pregnancy.Establishment of normal thyroid hormoues cut-off points in each trimester of pregnancy and screening for TPOAb is necessary.

Objective To study the effects of glycogen synthase kinase-3?(GSK-3?)and free radical on neuron apoptosis of hypoxic-ischemic brain damage(HIBD)in newborn rats.Methods Eighty 7-day-old neonatal rats were randomly divided into 2 groups:normal group and hypoxic-ischemic(HI)group.Rats in HI group were subjected to left common carotid artery ligation,exposed to 80 mL/L oxygen and 920 mL/L nitrogen gas in 37 ℃ closed container for 2.5 h.Rats in 2 groups were killed at 6 hours,24 hours,48 hours,72 hours,5 days after hypoxia respectively.The neuron apoptosis was detected by flow cytometry.The activity of GOD-PX and the contents of SOD and MDA were detected by spectrophotometry,the level of GSK-3? was mensurated by Enzyme-linked immumosorbent assay(ELISA).Results The rates of neuronal apoptosis in HI group were significantly higher than those in normal group at 6,24,48 and 72 hours after hypoxia,respectively(Pa

Objective Overexpression of multidrug resistance(MDR)is one of the mechanisms that will lead to chemotherapy failure.Of the MDR pathways in tumor cells,P-glycoprotein (P-gp) encoded by MDR genes is one of the key points.99Tcm-methoxyisobutylisonitrile(MIBI) is an imaging agent that can detect overexpression of MDR in tumor cells.The aim of this study was to observe the relationship between 99Tcm.MIBI uptake kinetics and P-gp levels in tumor cells,with or without MDR reversing agents.Methods A totle of 2×106 cells of human myelogeneous leukemia cell line K562 and its resistant subline(K562/D) were incubated with 8 MBq 99Tcm-MIBI respectively.99Tcm-MIBl accumulation and emux at various time inter-vats and the uptake difference with the presence of different cyclosporin A(O.1-O.4 ug/ml)were also ob-served.Comparison of different cell lines or without and with cyclosporin A were performed with the t-test, and the daa of different groups were compared by q-test.Results 99Tcm-MIBI uptake in K562 and K562/D cell line were 1.559±0.529 and 0.107±0.036,99Tcm-MIBI uptake in k562 was flitleon times higher than k562/D.As compared with K562 cell line with no expression of P-gp,significantly increased the 99Tcm-MIBI uptake of K562/D to 106%,148% and 163%after treated with cyclosporin A(0.1,0.2.0.4ug/ml)was ob-served(t=4.35,4.83,5.88,P<0.05).Conclusiom 99Tcm-MIBI uptake can reflect the P-gP level and multidrug-resistance inhibitor may impact 99Tcm-MIBI uptake in P-gP overexpressing cells.