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 clinical effect of magnesium sulfate combined with nifedipine in the treatment of patients with pregnancy induced hypertension( HDCP) .Methods Retrospective study was used in this study and 116 patients with HDCP from January 2013 to July 2015 in department of obstetrics and gynecology from our hospital were divided into two groups, including routine group of 62 patients who received routine treatment +magnesium sulfate) and combination group of 54 patients who received routine treatment +magnesium sulphate +nifedipine.The clinical effect was analyzed after five days’ continuous treatment.Results The systolic blood pressure, diastolic blood pressure,24h urinary protein, random urine protein /creatinine,serum homocysteine (Hcy) and CRP values in combination group were lower than routine group (P<0.05).There were no statistical difference in maternal uterine inertia, neonatal asphyxia, fetal distress, postpartum hemorrhage rate between the two groups after the treatment.But the rate of cesarean section in the combination group(50.00%)was significantly lower than that in the routine group(68.25%)(P <0.05).Conclusion Magnesium sulfate combined with nifedipine in the treatment of patients with HDCP had better antihypertensive effect, and would not increase fetal adverse birth outcomes incidence and significantly reduce the rate of cesarean section.

To evaluate the efficacy and safety of Tanreqing Injection, a compound traditional Chinese herbal medicine, for community-acquired pneumonia.

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.

BACKGROUND: Several studies have demonstrated that many American women who are at high risk of developing osteoporosis have higher levels of serum phosphorus. This indicates that some substances which can lower the serum level of phosphorus will supply a new and effective method to prevent and treat osteoporosis.OBJECTIVE: To observe the influences of porcine bone protein on bone mineral density (BMD) and serum levels of calcium and phosphorus in a rat model of osteoporosis.METHODS: Wistar rat models of osteoporosis were established by intramuscular injection of dexamethasone. Rat models were randomly divided into physiological saline, Jiegu Qili tablet, 50, 100, 200 mg/kg porcine bone protein groups. Rats that did not receive any treatments served as normal controls. After 12 weeks of treatment, serum was collected and serum levels of phosphorus and calcium were determined by biochemistry method. At the same time, tibia sections were made to determine tibial DMD by QDR-400 dual energy X-ray absorptiometry and to observe tibia marrow cavity by hematoxylin-eosin staining.RESULTS AND CONCLUSION: There was no significant difference in serum level of calcium among groups (P＞0.05).Compared with the physiological saline group, serum level of phosphorus in the 50, 100, 200 mg/kg porcine bone protein groups was significantly decreased (P ＜ 0.05). BMD was significantly higher in the 50, 100, 200 mg/kg porcine bone protein, Jiegu Qili tablet groups than in the physiological saline group (P ＜ 0.05). The tibia marrow cavity was smallest in the normal control group and largest in the physiological saline group. The tibia marrow cavity was larger in the 50, 100, 200 mg/kg porcine bone protein,Jiegu Qili tablet groups than in the physiological saline group. These results indicate that porcine bone protein cannot change the serum level of calcium, but it lowers serum level of phosphorus, and increases BMD, in a rat model of osteoporosis. However, the dose-dependent effect of porcine bone protein was not observed within the present experimental dosage. In addition, porcine bone protein can also reduce the marrow cavity of the tibia of rats with osteoporosis.

To evaluate the efficacy and safety of new drugs of traditional Chinese medicine (TCM) for acute upper respiratory tract infection (common cold).

OBJECTIVETo identify the relationship between coagulation factor V (FV) gene single nucleotide polymorphisms (SNPs) and venous thromboembolism (VTE).

METHODSThe FV clotting activity (FV: C) and FV antigen (FV: Ag) in plasma of VTE group (111 patients) and normal control (110 patients) were detected using one-stage clotting assay and ELISA, respectively. Five pairs of primers of the F V polymorphisms including Asp79His, Arg306The, Arg306Gly, Arg506Gln and Ile359The/His1299 Arg were synthesized and amplified by PCR. The PCR products were digested by restriction enzyme using PCR-RFLP. The detected polymorphisms were confirmed by direct sequencing. The samples containing the polymorphisms were screened for coding regions of all F V exons with direct sequencing.

RESULTSThe plasma levels of F V: C and F V: Ag of VTE group and normal control were (106.9 +/- 28.0)%, (110.4 +/- 33.3)% and (102.4 +/- 30.9)%, (102.1 +/- 24.1)%, respectively. The plasma level of FV: Ag was significantly different between VTE group and normal control. However, there was no difference in F V: C levels. Polymorphisms for the fore mentioned 5 primer pairs were not found in either patients or normal controls. Polymorphism of His1299Arg was identified in 5 patients with VTE and 3 normal controls. And these 5 cases also combined Met1736Val polymorphism, 3 of them combined another Asp2194Gly polymorphism.

CONCLUSIONThe higher plasma level of F V: Ag contribute to venous thromboembolism. There is no relationship between polymorphisms of Asp79His, Arg306The, Arg306Gly, Arg506Gln, Ile359The and venous thromboembolism in Chinese studied. Polymorphism His1299Arg is higher in VTE group than in normal control, but has no statistical difference. Polymorphisms of His1299Arg, Met1736Val and Asp2194Gly are linked disequilibrium in Chinese Han population.

Factor V ; genetics ; Female ; Gene Frequency ; Humans ; Linkage Disequilibrium ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Venous Thromboembolism ; genetics

Objective:To investigate the expression, prognostic value of acylphosphatase 1 (ACYP1) and its relationship with hepatocellular carcinoma (HCC) immune cell infiltration.Methods:The expression of ACYP1 in 374 cases of HCC was analyzed by using The Cancer Genome Atlas (TCGA) database. According to the median expression level of ACYP1 in HCC, the patients were divided into high expression (187 cases) and low expression group (187 cases). Fifty normal liver tissue were used as negative control. The differential expression, Kaplan-Meier survival analysis, univariate and multivariate Cox analysis were performed to evaluate the role of ACYP1 in the diagnosis and prognosis of HCC. The relationship between the expression level of ACYP1 and immune cell infiltration and immune checkpoint were analyzed through the TIMER database.Results:The expression level of ACYP1 in HCC (2.18±0.69) was higher than that in normal liver tissue (1.02±0.31), and the difference was statistically significant ( t=11.76, P<0.001). The survival of HCC patients with high ACYP1 was shorter than those HCC patients with low ACYP1 expression, and high expression of ACYP1 was an independent risk factor for poor prognosis of HCC patients ( HR=2.402, 95% CI: 1.483-3.891, P<0.05). The area under the curve of ACYP1 expression level in diagnosis of HCC was 0.965. The expression level of ACYP1 was significantly positively correlated with immune cell infiltration and immune checkpoints programmed cell death protein 1( r=0.288, P<0.001) and cytotoxic T lymphocyte-associated antigen 4 ( r=0.311, P<0.001). Conclusion:ACYP1 is a potential biomarker for HCC diagnosis and prognosis , as well as a potential therapeutic target.

Objective To establish an HPLC method for the determination of alkaloids(epiberberine,coptisine,palma-tine,berberine)and catalpol in different ratios(1∶1,1∶10)of ancient and modern Qianjin Huanglian Pills,and to compare the differences in their contents.The content differences were compared to preliminarily evaluate the differences in the efficacy of Qianjin Huanglian Pills in the treatment of diabetes under different preparation processes and different ratios.Methods The alkaloid solvent was methanol∶ hydrochloric acid(100∶1).The detection conditions were as follows:C18 column,acetonitrile-0.05 mol·L-1 potassium dihydrogen phosphate solution(50∶50),detection wavelength 345 nm,column temperature 30℃,flow rate 1 mL·min-1,injection volume 10 μL.The catalpol solution was methanol∶ water(20∶80).The detection conditions were as follows:chromatographic column C18 column,methanol-0.1%phosphoric acid solution(1∶ 99),detection wavelength 210 nm,column temperature 30℃,flow rate 1 mL·min-1,injection volume 10 μL.Results The established method was spe-cific,and the separation effect of the five components was good.It exhibited a good linear relationship(R2＞0.999)in their respec-tive linear ranges.The repeatability,precision,stability,and sample recovery rate all met the requirements.The content of four alka-loids in the ancient method 1∶1 was the highest,and the content of catalpol was the lowest.The content of four alkaloids in the ancient method 1∶10 was the lowest;the content of 1∶1 in the present method was higher than that in the ancient method 1∶10,and the content of berberine in the present method 1∶10 was slightly lower than that in the present method 1∶1,and the rest were higher than that in the present method 1∶1.The PCA results showed that the chemical composition contents of the four kinds of Qianjin Huanglian pills were very different.Conclusion The method is simple,accurate,and reproducible,making it suitable for the quality control of Qianjin Huanglian Pills.It provides a theoretical basis for exploring the difference in efficacy of Qianjin Huanglian Pills.