BACKGROUND: Excessive nitric oxide (NO) release can cause the occurrence and development of brain injury and senile dementia due to the apoptosis induction role of NO at high concentration to nerve cells. Therefore one strategy to prevent and treat senile dementia is inhibiting the apoptosis induced by NO.OBJECTIVE: To observe whether acidic peptide will inhibit the neuron apoptosis caused by NO. DESIGN: An cell and molecule observation experiment by comparisons. SETTING: Department of Biochemistry and Molecular Biology of Basic Medical College in Zhengzhou University and the Second Laboratory of Biological Active Peptide Institute in Zhengzhou University. MATERTALS: The experiment was performed between May 2003 and May 2005, in the Second Laboratory of Biological Active Peptide Institute in Zhengzhou University and the cell culture room of Department of Biochemistry and Molecular Biology of Basic Medical College in Zhengzhou University. The newborn SD male rats within 24 hours after birth were provided by the Animal Center of Henan Province (410117).METHODS: On day 11 of primary cultures, hippocampus neurons of the newborn SD rats were pretreated with different dosages of acidic peptide for six hours. Sodium nitroprusside (SNP) of 50 μmol/L final concentration was added to the cells which were incubated for another 24 hours. Cells were collected and adopted in this experiment of five different groups, namely normal control group, group treated with SNP, group of SNP plus 0.037 5 mg/mL acidic peptide, group of SNP plus 0.075 mg/mL acidic peptide, group of SNP plus 0.15 mg/mL acidic peptide. The cell's survival rate wasmeasure by methyl thiazolyl (MTT) method; The neurofilament protein was stained with the method of immunohisto chemistry. The shape of apoptosis was display with acridine orange fluorescent stain. Then DNA ladder zone of apoptosis cells was analyzed with the method of agarose gel electrophoresis. Western Blot and absorbance scan were used to determine the expression level of Bcl-2 protein and Bax protein.MAIN OUTCOME MEASURES: ①Experimental result of cell survival rate with MTT method;②Observation results of nuclear type of apoptosis; ③DNA electrophoresis analysis of apoptosis; ④Western Blot analysis results of Bcl-2 protein and Bax protein.RESULTS: ①Neuron survival rate was 58.9％ for group treated with SNP, 70.0％ for group of SNP plus 0.037 mg/mL acidic peptide, 72.8％ for group of SNP plus 0.075 mg/mL acidic peptide, and 75.3％ for group of SNP plus 0.15 mg/mL acidic peptide. ②Observation results of nuclear type of apoptosis: Significant characteristics of apoptosis were seen in group treated with SNP. The nucleus of hippocampus neuron treated with different concentrations of acidic peptide plus SNP was similar to that of normal control group in morphology. ③The results of DNA electrophoresis analysis of apoptosis: Only the neuron DNA of group treated with SNP showed clear characteristic DNA ladder zone of apoptosis on agarose gel electrophoresis. ④Analysis results of Bcl-2 protein and Bax protein with Western Blot and absorbance scan: The expression level of Bcl-2 protein in SNP treated group was decreased while that of Bcl-2 protein was increased. Bcl2 protein levels in acidic peptide plus SNP group were increased and Bax protein levels were decreased gradually with the increasing concentrations of acidic peptide compared with SNP treated group. CONCLUSION: Acidic peptide can inhibit neuron apoptosis, increase expression level of neuron Bcl-2 protein and decrease expression level of neuron Bax protein.

Objective To investigate the effects of valsartan on the expression of acyl-coenzyme A: cholesterol acyltransferase-1(ACAT-1) and peroxisome proliferator activated receptor-gamma(PPAR-?) in atherosclerotic plaques on rabbit aortic wall.Methods Twenty-four male Japanese white rabbits were randomly assigned into three groups(8 each): control group,valsartan group and high cholesterol feeding group.All rabbits were fed according to the experimental protocol for 12 weeks.Blood samples were taken from vein for measurement of serum lipids.The ratio of intima/media thickness of the aorta was measured.ACAT-1 mRNA/protein and PPAR-? mRNA/protein were determined by reverse transcription polymerase chain reaction(RT-PCR) and Western blotting,respectively.Results After 12 weeks,the levels of serum total cholesterol(TC),triglyeride(TG) and low density lipoprotein-cholesterol(LDL-C) in valsartan group and cholesterol group were significantly higher than those in control group(P0.05).The intima thickness and the ratio of intima/media in carotid arteries in cholesterol group were significantly higher than those in control group and valsartan group(P

Medical moral education is a most important component of clinical practice teaching.In the department of anesthesiology,the success of medical moral education depends on the participation and concerne of the head of this department,At the same time,we should ask the clinical practice teacher to train the medical students to take the patients as the center and dedicate themselves to the duty of their profession.

Objective To evaluate the therapeutic responses to transsphenoidal surgery and medical therapy in terms of normalization of prolactin(PRL),mortality,morbidity and the cost-effectiveness of PRL normalization in order to establish an individualized therapeutic protocol for the patients with prolactinoma.Methods A retrospective study was undertaken of a consecutive series of patients with prolactinoma who were followed for at least 1 year after transsphenoidal surgery or medical treatment.The clinical characteristics and the long-term outcomes(normalization of PRL,morbidity or mortality)were assessed.Utilizing the principle of medical economics and data from the two types of treatment,we worked out a Markov chain and calculated the lowest cost of two kinds of therapeutic protocols.Results(1)The success rate of normalizing serum PRL through surgical treatment in microadenoma was 85%(22/26),and that of medical treatment was 95%(19/20).There was no statistical difference between the two therapies(P＞0.05).The success rate of normalizing serum PRL through surgical treatment in macroadenoma was45%(19/42),and that of medical treatment was 5/5.There was a statistical difierence between the two therapies(P＜0.05).(2)According to the Markov model,it would cost a microprolactinoma patient 25 129.25 yuan to normalize serum PRL by surgical treatment.This is comparable to the cost of medical treatment which would be 24 943.99 yuan.Whereas for a macroprolactinoma patient surgery would cost 35 208.20 yuan and medical treatment would cost 25 344.38 yuan.Conclusions Medical therapy is superior to surgical treatment in regard to complication rate and cost-effectiveness for macro-and extra big prolactinomas.Transsphenoidal surgery remains an option for patients with microadenomas.Markov model is an effective way to predict the treatment cost for patients with hyperprolactinoma at different ages and with different canses

Objective To investigate the changes of serum protein fingerprint in patients with lung cancer with deep venous thrombosis.Methods Eighteen case patients with lung cancer were selected,including 8 case of lung cancers with thrombosis and 10 cases of lung cancers with no thrombosis.Surface enhanced laser desorption ionization protein-time of flight mass spectrometry (SELDI-TOF-MS) was used to analyze serum protein content of two groups in the same mass to charge ratios(M/Z),then drew the protein peaks that content difference was statistically significant.Results The M/Z of lung cancer with thrombus group and control group were 5911,1216,4187,1019,4293,the protein peaks had significant differences between the two group (43.81±7.74,6.37±5.02,2.97±0.35,35.96± 12.10,9.65±4.37;15.35± 12.69,2.06±0.37,4.67± 1.35,15.94±6.47,14.65±8.80;t =5.334,4.800,2.981,4.639,4.596;P＜ 0.05).Compared with the control group,the decrease of protein peak M/Z were 5911,1019,1216,and the increase of protein peak M/Z were 4293,4187 in the deep venous thrombosis group.Conclusion In the serum of patients with tumors SELDI profiles M/Z are 5911,4293,4187,1019,1216 of SELDI protein fingerprinting can be considered in patients with thrombosis of tumor specific markers.

Objective:To investigate the effects of Daidzein on behavior of chronic stress depression rats and the expression of hippocampal brain-derived neurotrophic factor ( BDNF ) , neuropeptide Y ( NPY ) and non-specific immune regulation.Methods: 40 healthy adult male SD rats with body weight(210±19)g,clean grade,were chosen and fed with 1%sucrose solution for 4 d to change drinking habits.On the fifth day rats were subjected to water deprivation for 24 h without fasting.On the sixth day rats were fed with 1%surcrose solution.4 h later, preference of 1% surcrose solution was examined.According to the 1% sucrose solution preference and weight rats were randomly divided into 4 groups,normal control group(CG),model control group,(MG),fluoxetine group(FG,10.0 mg/kg),daidzein group(DG,80.0 mg/kg).At the same time of establishing model,rats were administered orally once a day for 32 d.The depression model was established by chronic unpredictable mild stress model and separation.The behavioral changes of the rats were observed, and expression of BNDF in hippocampus and NPY was measured by Western blot technology and immunohistochemistry.It was observed the proliferation function of lymphocytes,spleen index,the number of peripheral blood leukocytes and antibody-secreting cell function.Results: Compared with the normal control group(CG),the weight of rats with chronic stress protocol was lower, 1%sucrose consumption decreased,scores of rats in the open field test dropped significantly,the immobility time in the forced swimming test prolonged,the level of expression of BNDF and NPY decreased,all the differences above were statistically sig-nificant(P<0.01).Compared with model group,weight of rats in fluoxetine treatment group(FG) and daidzein treatment group(DG)in-creased,sugar consumption,scores in the open field test and the levels of expression of BNDF and NPY significantly increased,the differences were statistically significant( P<0.05 or P<0.01 ).The number of peripheral blood leukocytes and antibody-secreting cell function and proliferation of lymphocytes force in daidzein treatment group was significantly higher than the model group,daidzein dose spleen index was significantly higher than the model group(P<0.01).Conclusion: The daidzein can antagonize depressive symptoms in chronic stress mice,daidzein may increased content of BDNF in hippocampus and NPY protein, and enhanced the role of humoral immune response and lymphocyte proliferation in rats with chronic stress model.The mechanisms of antidepressant effects of daidzein might be related to the increase of content of BDNF in hippocampus and NPY protein and non -specific immune regulation.

BACKGROUND: Nerve growth factor (NGF) and brain-derived neurotrophic factor(BDNF) are very important to the survival and proliferation of nerve cells. In the patients with Alzheimer disease (AD), the levels of NGF and BDNF are low.OBJECTIVE: To investigate whether acidic peptide can stimulate rat astrocytes to secrete NGF and BDNF.DESIGN: A randomized control animal experiment.MATERLALS: The experiment was finished in the First Laboratory of Institute of Biopeptide, Zhengzhou University; Cellular Culture Center,School of Basic Medical Sciences of Zhe ngzhou University from September 2003 to May 2005. Fifteen neonatal SD rats within 2 days after birth were selected.METHODS: ① The cerebral cortex of the neonatal SD rats was removed under sterile condition, the astrocytes were isolated and cultured, and then identified with the glial fibriliary acidic protein immunohistochemical staining. ② The cultured astrocytes were randomly divided into six group:blank control group, serum control group, positive control group and acidic peptide treated groups. No treatment was given in the blank control group,serum of 0.2 in volume fraction and 1 000 U/mL interferon were added in the serum control group and positive control.group, 37.5, 75 and 150 mg/L acidic peptides were added in the acidic peptide treated groups respectively. ③ The astrocytes of the 2nd generation, which covered the whole bottom of bottle, were digested to single cell suspension, and then inoculated to three 12-well plates equally at 5×105 /mL. The survival rate and the contents of NGF and BDNF in the supernatant of each group were determined at 24, 48 and 72 hours respectively.MAIN OUTCOME MEASURES: ① Cell numbers and survival rates at different culture time-points; ② Effect of acidic peptide on the proliferation of astrocytes in rats; ③ Changes of NGF and BDNF in the supernatant of astrocytes at different culture time-points.RESULTS: ① As compared with the blank control group, the cell numbers and survival rates at 24, 48 and 72 hours were obviously increased in the acidic peptide groups treated with 75 and 150 mg/L (P＜0.05, 0.01,0.001), but not obviously increased in the acidic peptide group treated with 37.5 mg/L. ② As compared with the blank control group, the rates of proliferation in the acidic peptide groups treated with 37.5, 75 and 150 mg/L were all significantly increased (17.5%, 45.5%, 72.5%, P＜0.001). ③ As compared with the blank control group, the absorbance (A) values of NGF in the supernatant at 24, 48 and 72 hours were all markedly increased in the acidic peptide groups treated with 37.5, 75 and 150 mg/L (P＜0.001),and the A values of BDGF in the supernatant at 48 and 72 hours were significantly increased (P＜0.05, 0.01).CONCLUSION: Acidic peptide can increase the secretions of NGF and BDNF of rat astrocytes to different extent.

Objective To discuss the influence of microRNA(miR)-155/miR-21 on toll-like receptor 4 (TLR4) in children with sepsis.Methods Fifty children with sepsis who were hospita-lized in Pediatric Intensive Care Unit,Shenzhen Children's Hospital,were enrolled in the study,and 15 healthy children at the same age were selected as healthy control group.Expression levels of TLR4 protein and human leukocyte antigen(HLA)-DR in CD14 + monocytes (MC) were detected by using flow cytometry,and sepsis patients were divided into 2 groups according to whether they exceeded the value of HLA-DR by 30％ or not.Expression level of programmed cell death factor 4 (PDCDM) and inositol phosphatases 1 containing SH2 (SHIP1) were detected at the same time.MC were separated by CD14 + immune magnetic bead,and expression level of miR-155,miR-21 and tumor necrosis factor-α (TNF-α),interleukin-10 (IL-10) mRNA in CD14 + MC were detected by using real-time fluorescent quantitative PCR.Results Sepsis group consisted of 27 male and 23 female,and their ages were (2.34 ± 0.79) years old,among whom 9 patients died.There were 36 patients in the HLA-DR increase group and 14 patients in the HLA-DR decrease group.Expressions ofTLR4(2.33±0.90),miR-155[(7.19±3.75) ×10 3] and TNF-α[(21.98±14.15) ×10-2 pg/L] in CD14 + MC were higher in the HLA-DR increase group than those in the HLA-DR decrease group [1.24±0.60,(4.83 ±1.17) × 10-3,(14.18±5.45) ×10-2 μg/L] and healthy control group[1.57±0.55,(3.99 ± 1.29) × 10-3,(1.61 ± 0.84) × 10 2 pg/L],and the differences were statistically significant(F =11.943,7.583,18.538,all P ＜0.05),while the expressions of miR-21 (12.10 ±5.66),IL-10[(29.74 ± 12.55) × 10-4 μg/L] in CD14 + MC were lower in the HLA-DR increase group than those in the HLA-DR decrease group[4.68 ± 2.07,(12.50 ± 5.73) × 10-4 μg/L] and healthy control group [2.39 ± 0.86,(2.04 ± 0.92) × 10-4 μg/L],and the differences were statistically significant(F =41.673,54.991,all P ＜ 0.05).The levels of SH1P1 and PDCD4 decreased in sepsis compared with healthy control group[0.70 ±0.36)vs.(1.59 ±0.48);(1.55 ±0.56) vs.(3.01 ±0.70)],and the differences were statistically significant (t =7.682,8.339,all P ＜ 0.05),but SHIP1 decreased more significantly in the HLA-DR increase group than that in the HLA-DR decrease group [(0.60 ± 0.34) vs.(0.97 ± 0.26)],and the difference was statistically significant (F =39.214,P ＜ 0.05).PDCD4 decreased more significantly in the HLA-DR decrease group than that in the HLA-DR increase group (0.94 ±0.19 vs.1.79 ±0.47),the difference was statistically significant(F =65.367,P ＜ 0.05).Conclusions Regulation imbalance of miR-155/miR-21 may be one of the reasons for abnormal expression of TLR4 in children with sepsis,and it plays a role in enlarged or inhibited expression of TLR4 in the sepsis process which results in different immune status in sepsis patients.

Objective To evaluate the potential value of IgG antibodies against recombinant PPE65 protein (rPPE65) of Mycobacterium tuberculosis in serodiagnosis of tuberculosis.Methods The gene encoding PPE65 protein of M.tuberculosis was cloned into the PET-28a vector and then expressed in Escherichia coli.The rPPE65 was purified with Ni-NTA affinity and ion exchange chromatography.After dialysis renaturation, the concentration of rPPE65 was determined using Lowry assay.ELISA was used to detect the levels of specific IgG against rPPE65 and recombinant PstS1 protein (rPstS1) in sera from 144 patients with pulmonary tuberculosis (PTB patients), 144 health controls, and 56 patients with non-tuberculosis pulmonary diseases.ROC curves were used to determine cut-off values with the results of IgG antibodies against rPPE65 and rPstS1 for 144 PTB patients and 97 controls with negative PPD skin test.The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy of rPPE65 and the combination of rPPE65 and rPstS1 were counted.Results The PPE65 protein of M.tuberculosis was successfully expressed in E.coli. The purity and concentration of rPPE65 were 95% and 0.5 mg/ml, respectively.ROC analysis showed that the cut-off of ELISA using rPPE65 was 0.64.The sensitivity, specificity, PPV, NPV, and accuracy of rPPE65 were 34.7%(50/144), 93.5%(187/200), 79.4%(50/63), 66.5%(187/287), and 68.9%(237/344), respectively.The sensitivity, specificity, PPV, NPV, and accuracy of the combination of rPPE65 and rPstS1 were 59.0%, 91.0%, 82.5%, 75.5%, 77.6%, respectively.Conclusions The rPPE65 of M.tuberculosis appears to be a candidate antigen for serodiagnosis of tuberculosis.Detection of IgG antibodies against the combination of rPPE65 and rPstS1 can increase the sensitivity of serological test for tuberculosis.

OBJECTIVE To investigate the infection episode and related risk factors in continuous hemodialysis patients. METHODS The relationship among infection and etiologies of infection,nutritional status,pathogens and causes of chronic renal failure(CRF) were retrospectively analyzed in 180 continuous hemodialysis patients. RESULTS Totally 113 times infections were observed among the 86 inpatients under continuous hemodialysis.The main infectious site in hemodialysis patients was lungs.Thirty eight times were positive in 50 times of etiologic detection,Gram-negative germ was the most common(60.3%).Hemoglobin and serum albumin decreased obviously in infectious patients.Diabetes and systemic lupus erythematosus patients were more susceptible to infection.The hepatitis virus infections rate in hemodialysis patients was relatively high. CONCLUSIONS There is higher infections rate in continuous hemodialysis patients.Diabetes and systemic lupus erythematosus patients are more susceptible to infection.Anemia,lower serum albumin,old age and bad compliance are the susceptible factors.