Objective To investigate the effects of propofol on mitochondrial membrane permeability during hypoxia/reoxygenation (H/R) in rat hippocampal neurons. Methods Primary cultured hippocampal neurons of fetal rats obtained from Wistar (17-18 days of gestation) were randomly divided into 3 groups: Ⅰ control group (group C), Ⅱ H/R group and Ⅲ propofol + H/R group. Neurons were cultured in the culture medium with combined oxygen glucose deprivation for 2 h followed by reoxygenation in group Ⅱ and Ⅲ. In group Ⅲ propofol was added to the culture medium with the final concentration of 20 μ mol/L before combined oxygen glucose deprivation.Neuronal viability was detected by MTT assay and mitochondrial membrane potential (MMP) with flow cytometry at 0, 4, 8, 12 and 24 h after reoxygenation (T1-5) and the permeability of cell membrane and mitochondrial membrane was monitored at T5 using laser confocal scanning microscope. Results The neuronal viability and MMP were significantly decreased (P ＜ 0.05), while the permeability of cell membrane and mitochondrial membrane was increased at T5 in group Ⅱ as compared to group Ⅰ . The neuronal viability at T1-4 and MMP at T1-5 were significantly increased (P ＜ 0.05), while the permeability of cell membrane and mitochondrial membrane was decreased at T5 in group Ⅲ compared to group Ⅱ . Conclusion Propofol can protect rat hippocampal neurons against H/R injury through increasing MMP, improving the cell and mitochondrial membrane permeability, and increasing the neuronal viability.

Objective The present study was designed to attempt a continuous assessment of cerebral autoregulation by continuously monitoring jugular venous bulb oxygen saturation (SjO 2) in patient undergoing cardiac operation. Methods Twenty-four patients undergoing cardiac surgery under cardiopulmonary bypass (CPB) were included. Among them 16 were male, 8 female. No neuropsychological deficit was found in any of them before or after operation. Patients had neither diabetes mellitus nor immunological diseases. Normocapnia was maintained during operation. SjO 2 and other routine parameters, such as blood pressure, temperature, heart rate, pulmonary atery pressure and cardiac output etc were monitored and recorded continuously via a computerized anesthetic recording system. The numerical data of SjO 2 and mean arterial pressure (MAP) were sampled every 30s. Correlation between SjO 2 and MAP was evaluated before, during and after CPB. According to the slope "a" of the regression line "SjO 2= b + a MAP", cerebral autoregulation was estimated. The cerebral autoregulation was taken as deteriorated if the two parameters were highly correlated with a ≥0.4, showed as dysautoregulation (+). Those with a

Objective To investigate the alteration of nuclear factor kappa B( NF-κB) and tumor necrosis factor-a (TNF-α) mRNA and protein in hippocampus in chronic constrictive injury (CCI) model of rats. Methods Seventy-six male Wistar rats were randomly divided into 2 groups ( n = 38): the CCI group which received the chronic constriction injury and the sham group which received the sham operation as control. The mechanical and thermal nociceptive thresholds were assessed with paw withdrawal latency (PWL) to von Frey filaments and radiant heat at 1d before and ld,4d,7d,14d and 28d after CCI operation. Five animals were sacrificed at each time point for real-time polymerase chain reaction (real-time PCR) and another three animals sacrificed at 7d postoperation for immunofluorescence histochemical staining. Results The thresholds to mechanical and thermal stimuli decreased obviously after operation in CCI group. The expressions of TNF-α and NF-κB mRNA began to increase at ld( (2.079 ±0. 104)times and 4d( ( 1.640 ± 0.064) times) after operation and reached the peak at 7d ((2.748 ±0.147)times, (2.010 ±0.096)times) ,then the expressions of TNF-a mRNA began to decrease,while the expressions of NF-kB mRNA maintained at a high level throughout the experiment. The result of immunofluorescence histochemical staining revealed that NF-kB and TNF-α protein expressions at 7 day increased significantly on the hippocampus,which was consisted with NF-κB and TNF-a mRNA levels. Conclusion The activation NF-κB and TNF-α in hippocampus may be involved in the procession of neuropathic pain.

Background and purpose:It has been reported that gap junctional (GJ) function was signiifcantly decreased in hepatocellular carcinoma (HCC) tissues and cell lines. However, the increased GJ suppress tumorigenesis and the development of liver cancer. This study therefore aimed to examine the effect of simvastatin on GJ function between Hep3b cells. Thus, the exploition of drugs to increase GJ function between liver cancer cells will provide an efifcient approach to ifght against liver tumor as well as increase cytotoxicity of antitumor agents.Methods:SRB was used to assay the toxicity of simvastatin. The effect of simvastatin on GJ function was determined by “Parachute” dye-coupling assay and scrape loading/dye transfer assay.Results:Pretreated Hep3b cells with simvastatin at the concentration of 1, 5 or 10 μmol/L for 24 h did not induce the cytotoxicity. So simvastatin at the concentration of 5 and 10 μmol/L would not reduce the amount of GJ on cell membranes. “Parachute” dye-coupling assay showed that the treatment with 5 and 10 μmol/L simvastatin for 4 h enhanced the dye spread through GJ in Hep3b cells. Similarly, scrape loading/dye transfer assay showed that simvastatin could induce the increasing spread of lucifer yellow (Ly, Sigma) around the scoifng cells with increasing concentrations.Conclusion:Simvastatin could increase the GJ function of Hep3b cells.

BACKGROUND:Osteoporosis caused by diabetes melitus as common secondary osteoporosis has been paid more and more attention recently. Zoledronic acid serves as a novel drug for osteoporosis, and its effect on osteoblasts in vivo remains unclear. OBJECTIVE:To investigate the changes of the expression of bone morphogenetic protein 2 andNoggin in the femur of type 1 diabetes melitus rats and the effect of zoledronic acid on them. METHODS:Models of type 1 diabetes melitus were established by intraperitoneal injection of streptozotocin in 130 Wistar rats. 3 days later, rats with blood sugar > 16.7 mmol/L for three consecutive times were considered as successful models, 120 in total. These models were randomly divided into model, prevention and treatment groups. Rats in the prevention and treatment groups were intravenously administered zoledronic acid (0.1 mg/kg) on the day of modeling and 2 weeks after model establishment. An additional 40 rats were injected with citrate buffer solution as control group. RESULTS AND CONCLUSION: Compared with the control group, femur bone mineral density, serum alkaline phosphatase levels, and femur bone morphogenetic protein 2 mRNA expression levels were significantly lower in the model group (P < 0.05), butNoggin mRNA expression significantly increased (P < 0.05). Compared with the model group, bone mineral density and bone morphogenetic protein 2 mRNA expression levels were significantly higher in the prevention and treatment groups (P < 0.05), butNoggin mRNA expression significantly lower (P < 0.05), and serum alkaline phosphatase levels gradualy restored. These results indicated that the bone metabolic disturbance occurs in early stage in rats with type 1 diabetes melitus. Zoledronic acid can promote bone formation, increase bone density, and improve bone metabolism.

Objective:To evaluate the application of analyzing neutrophil cell surface marker CD64 in diagnosis of premature infants infection.Methods:109 infants inpatient in neonatal department(including NICU)were enrolled in the study.CD64 was measured by FCM,which was compared with C-reactive protein(CRP)and IL-6.Results:There was a statistically significant difference in quantitation of CD64 on neutrophil cells(P

In the item bank construction of medical microbiology examination questions,the accurate difficulty coefficient is an important parameter to guarantee the quality of the bank.Thus,the accurate difficulty estimate plays an important role in the item bank construction of examination questions.Methods of estimating the difficulty of medical microbiology test questions were explored,based on the knowledge points,the type of test questions,the structure of test questions and the degree of students’ familiarity with the questions and so on.Finally,the feasibility of methods was confirmed through the simulated tests.

Objective To investigate the effects of a pulsed electromagnetic field (PEMF) on cell proliferation,differentiation and the genetic expression of osteogenesis specific transcription factor-Osterix (OSX) in the calvaria-derived osteoblasts of SD rats.MethodsPrimary osteoblasts were separated from the calvarias of Sprague-Dawley rats 24 hours after birth by trypsinazation and collagenase digestion.The osteoblasts at passage 4 were randomly divided into a control group and a PEMF group,Cells in the PEMF group were placed onto PEMF therapeutic apparatus and exposed to 11 mT radiation at 12 Hz for 1 h per day for 3 days.The osteoblasts' proliferation ability was then detected via methylthiazdyl tetrazolium assay.Alkaline phosphatase (ALP) activity in the cell lysate and the supernatant fluid was assayed through disodium phenyl phosphate colorimetric determination to determine the cells'differentiation ability.OSX mRNA expression was determined using real time reverse transcription polymerase chain reaction (RT-PCR).ResultsThe proliferation of osteoblasts in the PEMF group was significantly greater than in the control group.PEMF exposure suppressed ALP activity in both the lysate and the supernatant of the osteoblasts.The OSX mRNA expression in the PEMF group was significantly down-regulated compared with the control group.ConclusionPEMF at 12 Hz and 11 mT can promote the proliferation of osteoblasts,but it inhibits cellular differentiation and down-regulates the mRNA expression of OSX.

Objective To investigate the correlation between spot albuminuria to creatinine ratio (ACR) and 24 h urinary protein excretion in women with preeclampsia and determine the optimal cut-off values of spot ACR in mild preeclampsia and severe preeclampsia.Methods Twenty-eight women with mild preeclampsia and 22 with severe preeclampsia at Nanfang Hospital,Southern Medical University between October 2010 and June 2011 were recruited.Maternal serum cystatin,uric acid,mea nitrogen,creatinine and albumin levels were collected and analyzed.Twenty-four hours urinary protein excretion was measured with immunoturbidimetric assay and ACR with automatic analyzer DCA2000.The correlation between ACR and 24 hours urinary protein excretion was explored.And the optimal cut-off values of the spot ACR for mild and severe preeclampsia were determined with receiver operating characteristic curve.Results ( 1 )Maternal serum biochemical parameters:uric acid levels in mild and severe preeclampsia were (359 ± 114)μmol/L and (450 ± 132) μmol/L,while cystatin levels were ( 1.3 ±0.3) mg/L and ( 1.6 ±0.5) mg/L respectively.The differences were statistically significant ( P ＜ 0.05 ).Serum urea nitrogen,creatinine and albumin in mild preeclampsia were(3.6 ± 1.6) mmol/L,(52 ± 38 ) μmol/L and ( 33 ± 3 ) g/L,while in severe preeclampsia were( 6.2 ± 3.1 ) mmol/L,( 78 ± 59 ) μmol/L and ( 29 ± 6 ) g/L respectively.There were no statistical significant differences ( P ＞ 0.05 ).(2) Twenty-four hours urinary protein excretion and ACR:24 hours urinary protein levels in mild and severe preeclampsia was (700 ± 160) mg and (4800 ±2200) mg (P＜0.05).ACR in mild and severe preeclampsia was (72.7 ± 12.4) mg/mmol and (401 ±245) mg/mmol respectively (P ＜ 0.05 ).(3) There was a strong correlation between the spot ACR and 24hours urine protein excretion ( r =0.938 ; P ＜ 0.05 ).( 4 ) The optimal spot ACR cut-off point for the diagnosis of preeclampsia:the optimal spot ACR cut-off point was 22.8 mg/mmol for 300 mg/24 hours of protein excretion in mild preeclampsia,the area under curve was 0.956,with a sensitivity,specificity of 82.4％,99.4％ respectively.And the optimal spot ACR cut-off point was 155.6 mol for 2000 mg/24 hours of protein excretion in severe preeclampsia,the area under curve was 0.956,with a sensitivity,specificity of 88.6％,91.3％ respectively.Conclusions Compared with 24 hours urinary protein excretion,the spot ACR may be a simple,convenient and accurate indicator of early diagnosis of preeclampsia.Spot ACR may be used as a replacement for 24 hours urine protein excretion in assessment of preeclampsia.The optimal spot ACR cut off points were 22.8 mg/mmol for mild preeclampsia and 155.6 mg/mmol for severe preeclampsia.

ObjectiveTo investigate the effects of different time administration of propofol on cytochrome c (Cyt c) in the cytoplasm in rat hippocampal neurons with hypoxia-reoxygenation (H/R) injury.MethodsPrimary cultured hippocampal neurons were randomly divided into 5 groups ( n =5 each):control group (group C),model of H/R injury group (group M),and different time administration of propofol groups (group Ⅰ,Ⅱ,Ⅲ ).In groups M,Ⅰ,Ⅱ and Ⅲ,the neurons were exposed to 95％ N2 + 5％ CO2 for6 h followed by 12 h reoxygenation.In groups Ⅰ, Ⅱ,Ⅲ,propofol was added to the culture medium before hypoxia,immediately after reoxygenation and at 2 h of reoxygenation (T0-2) respectively,with the final concentration of 20 μmol/L.The cell apoptosis was observed at T1,2 and at 24 h of reoxygenation ( T3 ) and the concentration of Cyt c in the cytoplasm was detected at T1-3.ResultsCompared with group C,the concentration of Cyt c in the cytoplasm was significantly increased at T1-3 in group M and at T1,2 in groups Ⅰ,Ⅱ,Ⅲ (P ＜ 0.05).Compared with group M,the concentration of Cyt c in the cytoplasm was significantly decreased at T1-3 in group I and at T1,2 in groups Ⅱ and Ⅲ ( P ＜0.05).The concentration of Cyt c in the cytoplasm was significantly higher at T1,2 in groups Ⅱ and Ⅲ than in group Ⅰ,and at T2 in group Ⅲ than in group Ⅱ ( P ＜ 0.05).The neuronal apoptosis was significantly decreased in groups Ⅰ,Ⅱ and Ⅲ as compared with group M.ConclusionDifferent time administration of propofol can reduce the mitochondrial Cyt c release to the cytoplasm,inhibit apoptosis in hippocampal neurons,and reduce H/R injury in rats,with better effect when given before hypoxia.