Objective To investigate the effect of morphine on the expression of p53 mRNA and E2F-1 mRNA in human gastric carcinoma cell line MGC-803 .Methods The human gastric cancer cell line MGC-803 was purchased from Cell Biology Research Institute, Chinese Academy of Sciences, and cultured in DMEM liquid culture mediun. The cells were seeded in 6-well plates (1 × 103/ml or 2 × 105/ml, 1 ml/well) and divided into 2 groups (n = 18 wells each):group Ⅰ normal control (group C); group Ⅱ was exposed to 10 μmol/L morphine (group M). The proliferation of the cells was determined by colony formation assay at 7 day of incubation with morphine. The expression of p53 mRNA and E2F-1 mRNA was detected and the ulrastructure of the cells examined with transmission electron microscope after being incubated with morphine for 24 h. Results The proliferation of the cells and E2F-1 mRNA expression were significantly lower and p53 mRNA expression was significantly higher in group M than in group C (P ＜ 0.05). The nuclear evelope was intact and the nucleolus and chromosomes were clearly visible in group C, while in group M fragmentation of nuclear envelope and nucleolus and apoptotic bodies were observed. Conclusion Morphine can inhibit the proliferation of the cells and accelerate the cell apoptosis through up-regulating the expression of p53 gene and down-regulating the expression of E2F-1gene in human gastric carcinoma cell line MGC-803.

ObjectiveTo investigate the effects of propofol and etomidate on apoptosis in hippocampal neurons in rats.MethodsOne hundred and forty male 4 weeks old SD rats were randomly divided into 7 groups (n =20 each):control group (group C) ; groups P1,2,3 received intraperitoneal (IP) propofol 50,100 and 200mg/kg and groups E1,2,3 received IP etomidate 10,30 and 60 mg/kg respectively.Arterial blood samples were obtained at 2 h after the animals were fully awake for blood gas analysis.The animals were then sacrificed and their brains removed for microscopic examination of the ultrastructure of neurons in hippocampal CA1 area and detection of Survivin and Caspase-3 mRNA and protein expression in hippocampus by RT-PCR and Western blot analysis.ResultsThere was no significant difference in PaO2,PaCO2,SaO2,HCO3-,BE and pH value among the 7 groups.The neurons in CA1 area were basically normal in groups C,P1 and E1 while condensation of the chromatin of the nucleus and apoptotic bodies were observed in groups P3 and E3.Caspase-3 mRNA and protein expression was significantly up-regulated while Survivin mRNA and protein down-regulated in groups P3 and E3.Conclusion High dose of propofol and etomidate may induce apoptosis in hippocampal neurons in rats by up-regulation of Caspase-3 expression and down-regulation of Survivin expression.

Objective To investigate the effects of different doses of propofol anesthesia on the long-term cognitive function in neonatal rats.Methods One hundred 7-day-old male Sprague-Dawley rats,weighing 9-18 g,were randomly divided into 5 groups (n =20 each) ∶ control group (C group) and propofol 25,50,100 and 200mg/kg groups (groups P1-4,respectively).Groups P1 and P2 received intraperitoneal propofol 25 and 50 mg/kg,respectively.Groups P3 and P4 received intraperitoneal propofol 100 and 200 mg/kg,respectively,and after righting reflex completely recovered,an increment of propofol 50 mg/kg was given until the total amount was finished.Five animals in each were chosen and arterial blood samples were obtained immediately after the animals were fully awake for blood gas analysis.The rats were then continuously fed.When the rats were 9 weeks old,the spatial learning and memory function was tested by Morris water maze.The animals were then sacrificed and their brains were removed for detection of the expression of nerve growth factor (NGF) and Caspase-9 protein and mRNA in hippocampal tissues (by Western blot analysis and RT-PCR) and for microscopic examination of the ultrastructure of hippocampal neurons.Results There was no significant difference in blood gas analysis index between the 5 groups (P ＞ 0.05).Compared with group C,no significant change was found in the escape latency and frequency of crossing the original platform in group P1,the escape latency was prolonged and the frequency of crossing the original platform was decreased in groups P2-4,the expression of NGF protein and mRNA was down-regulated and the expression of Caspase-9 protein and mRNA was up-regulated in groups P1-4 (P ＜ 0.05).The escape latency was gradually prolonged,the expression of NGF protein and mRNA was gradually down-regulated and the expression of Caspase-9 protein and mRNA was gradually up-regulated in groups P1-4.The frequency of crossing the original platform was decreased in groups P2-4 compared with group P1 (P ＜ 0.05).There was no significant difference in the frequency of crossing the original platform between groups P2-4 (P ＞ 0.05).Nucleus condensation,chromatin condensation,nuclear fragmentation and apoptotic bodies were observed in groups P2-4.Conclusion Propofol anesthesia can impair the long-term cognitive function in neonatal rats,the effect is related to the dose,and inhibition of NGF expression and increase in the activity of Caspase-9 may be involved in the mechanism.

Objective To evaluate the effects of propofol anesthesia on hippocampal protein kinase A (PKA)-cAMP response element binding protein (CREB) signaling pathway in neonatal rats.Methods One hundred and seventy-five male Sprague-Dawley rats,aged 7 days,weighing 8-15 g,were randomly divided into 5 groups (n =35 each) using a random number table:control group (C group) and propofol 25,50,100 and 200 mg/kg groups (P~ groups).Groups P1 and P2 received intraperitoneal propofol 25 and 50 mg/kg,respectively.Groups P3 and P4 received intraperitoneal propofol 100 and 200 mg/kg,respectively,and after righting reflex completely recovered,an increment of propofol 50 mg/kg was given until the total amount was finished.Five animals in each group were chosen and arterial blood samples were obtained immediately after the animals were fully awake for blood gas analysis.Five animals in each group were chosen at 2 h after fully awake and the age of 9 weeks,the rats were sacrificed and their brains were removed for microscopic examination of the ultrastructure of hippocampal neurons and for determination of PKA mRNA,CREB mRNA,PKA protein and pCREB protein in hippocampus (using RT-PCR and Western blot analysis).Results There was no significant difference in the indexes of blood gas analysis anong the five groups (P ＞ 0.05).Nuclear swelling and fragmentation,chromatin condensation,apoptotic bodies,decreased number of synapses and widened synaptic space were observed in P2,P3 and P4 groups.Compared with group C,the expression of PKA mRNA,CREB mRNA,PKA protein and pCREB protein was significantly down-regulated at 2 h after fully awake and the age of 9 weeks in P1,P2,P3 and P4 groups (P ＜ 0.05).Conclusion The mechanism by which propofol anesthesia induces neurotoxicity in neonatal rats may be related to inhibition of the activity of PKA-CREB signaling pathway.

Objective To analyze clinical outcomes of the spouse donor kidney transplantation for evaluating the security.Methods Sixty-three cases of living-renal transplantation were divided into two groups, spouse donor group (12 cases) and the other related donor group (53 cases). Twelve cases of spouse-donor(SD)renal transplantation were summarized, that were compared with the nospouse transplantation cases of the other living-related renal transplantation in the same period, which were similar in basic conditions and in immunosuppressant scheme. The observational parameters included average hospitalization time, rate of acute renal necrosis, acute rejection incidence within 1 year, serum creatinine at 7 d, 30 d and 1 year after transplantation.Results The recipient age of the 2 groups was (39±3)years and (37±3)years(P=0.05), dialysis time was (4.7±3.2)months and (4.4±2.9)months(P=0.78), the average hospitalization time was (20.9±8.3)d and (23.0±7.8)d(P=0.41). There was no significant difference between the spouse donor group and the no spouse related donor group. The acute rejection incidence within 1 year was 33.3%(4/12) in spouse group and 3. 9%(2/51) in the other related group, there was significant difference between the 2 groups(P＜0.05). The rate of acute renal necrosis was 16.7%(2/12) in spouse group and 3.9%(2/15)in no spouse related group, there was no significant different between the 2 groups(P＞0.05).SCr was (206.47±47.22)μmol/L and (163. 75±25.91)μmol/L in spouse group at post-operation 7 d and 30 d, and was (142.79±89.42)μmol/L and (119. 99±15.03)μmol/L in no spouse group. There was significant difference between the groups(P=0.02, P=0.00). One year after operation, SCr was (133. 40±6. 11)μmol/L in spouse group and (121. 00±34.12)μmol/L in no spouse group,there was no significant difference between the 2 groups (P=0.25).Conclusions Preoperative comprehensive assessment of the spouse donors and recipients renal transplantation is the guarantee for the success.Short-term outcomes of spouse donor renal transplantation is ideal, but rate of acute rejection within 1 year is higher than that of the other living-relative donor kidney transplantation, which dose not influence the long-time survival of spouse recipients.

Objective To analyze the clinical outcomes and evaluate the safety of living related live donors in kidney transplantation.Methods One hundre and thirty-two cases of living related donors were studied retrospectively for psychological and physiological parameters. The parameters including life quality, urinalysis, serum biochemistry tests, glomerular filtration rate (GFR) and endogenous creatinine clearance rate (CCr) were analyzed.Results There was no significant difference between living related donors and normal people in the aspect of life quality. In living related donors,SCr was (78.33±15.94)μmol/L before operation and was (108.49±19.88)μmol/L at 7 days postoperation, P＜0.05. SCr was (112.47±20. 38)μmol/L at 6 months post-operation. There was no significant difference in SCr levels between 7 days and 6 months post-operation (P=0. 109). CCr was (95.80±20.92)ml/min in pre-operation and was (57.36±14. 92)ml/min at 7 days post-operation,P＜0.05. CCr was (65. 49±8. 25) ml/min at 6 months post-operation. There was no significant difference in CCr between 7 days and 6 months post-operation.The pre-operative total GFR was(74.08±18.51)ml/min. Of which, the right kidney GFR was (38.43±10.33)ml/min. The residual right kidney GFR was (56. 49±13. 01 ) ml/min 6 months after operation, which decreased 17. 59 ml/min (23.8%) compared with pre-operative total GFR (P＜0.05) and increased 18. 06 ml/min (47.0%) compared to the pre-operative right kidney GFR. Surgical complications included 1 case of splenectomy, 1 case of descending colon rupture and 5 cases of wound fat liquefactions.Conclusion Pre-operatively systemic psychological and physiological evaluation on living related donors, detailed consent consultation, standardized operating techniques, careful perioperative management and strict follow-up can improve the safety of living related donors.

Objective To evaluate the influence of peritoneal lavage with different concentrations of povidone-iodine on hemodynamics and acid-base balance in rabbits.Methods Twenty-four clean-grade healthy adult male New Zealand white rabbits,aged 3 months,weighing 2.8-3.2 kg,were divided into 4 groups (n=6 each) using a random number table:control group (group C),povidone-iodine 1/3 of original concentration group (group TI),povidone-iodine l/2 of original concentration group (group HI) and povidone-iodine of original concentration group (group OI).Rabbits were anaesthetized with intraperitoneal 3％ pentobarbital 1.5 ml/kg,the left femoral artery was cannulated for invasive blood pressure monitoring,and the abdominal cavity was opened.Peritoneal lavage was performed with normal saline,povidone-iodine diluted with normal saline (1 ∶ 2),povidone-iodine diluted with normal saline (1 ∶ 1) and original povidone-iodine 20 ml at 10 min after opening abdominal cavity in C,TI,HI and OI groups,respectively.The fluid for peritoneal lavage was sucked out using a sterile gauze 2 min later and then the abdominal cavity was closed.Mean arterial pressure (MAP) and heart rate (HR) were recorded immediately before lavage (T0) and at 5,10 and 20 min and 1 and 2 h after the end of lavage (T1-5).Arterial blood samples were collected at T0,T4 and T5 for blood gas analysis,and the pH value,BE and lactic acid level were recorded.The duration of anesthesia before peritoneal lavage,cumulative dose of anesthetics,fluid volume and urine volume at 2 h after anesthesia,and mortality at 3 h after peritoneal lavage were recorded.Results Compared with group C,MAP at T1-5 and HR at T3-5 were significantly decreased in TI,HI and OI groups,pH value was significantly decreased and BE negative value was increased at T4,5 in HI and OI groups,the lactic acid level was significantly increased at T5 in group OI,and the mortality rate were significantly increased in HI and OI groups (P＜0.05).Compared with group TI,MAP at T4,5 and pH value at T5 were significantly decreased,BE negative value was increased at T5,and the lactic acid level was increased at T4,5 in group Ol (P＜0.05).There was no significant difference in each parameter between group OI and group HI (P＞0.05).Conclusion Peritoneal lavage with povidone-iodine dose-dependently leads to hemodynamic deterioration and acid-base imbalance in rabbits.

Objective To investigate the prognostic factors of transcatheter arterial chemoembolization (TACE) in the treatment of primary hepatocellular carcinoma (HCC).Methods A retrospective analysis was performed on 326 HCC patients treated with TACE.Kaplan-Meier method was used to calculate the 1-year,2-year and 3-year cumulative survival rates.Log-rank test and Cox proportional hazards model were used to analyze univariate and multivariate prognostic factors,respectively.Results The 1-year,2-year and 3-year cumulative survival rate of HCC patient was 73.90％,40.20％and 22.20％,respectively.The median survival time was 21 months.Univariate analysis showed that the alpha-fetoprotein (AFP),gamma-glutamyl transpeptidase (GGT),tumor size,tumor number,Child-Pugh grade,Barcelona clinic liver cancer (BCLC) stage,portal vein thrombosis,arteriovenous fistula and distant metastasis were factors affecting the prognosis of HCC patient (all P＜0.05).Multivariate COX regression analysis showed that AFP,GGT,tumor size,tumor number,BCLC stage,arteriovenous fistula were the independent prognostic factors of HCC patients (all P＜0.05).Conclusion AFP,GGT,tumor size,tumor number,BCLC stage and arteriovenous fistula are independent prognostic factors of HCC patients treated with TACE.