Acupuncture Therapy ; Animals ; Female ; Humans ; Polycystic Ovary Syndrome ; therapy

Aim To ascertain whether defibrase has the protective effect against reperfusion injury after cerebral ischemia.Methods 70 renovascular hypertensive rats(RHR) were randomly divided into defibrase group, control group and sham-operated group.Reversible middle cerebral artery occlusion(MCAO) models were produced by the modified. Longa's method,and reperfusion was begun 2 hours after occlusion.Rats in the defibrase group were given defibrase 10 U?kg-1 body weight via femonal intraveneous injection, and in the control group with the same amount of saline. The brain pieces were processed by TTC and HE staining and the infarct size,brain microvessels damage and secondary bleeding were compared between the two groups. Results The volume of infarction in the defibrase group was obviously smaller than in the control group, the damage of brain microvessels was less severe, and the bleeding lesions under optical microscope were less than in the control group. Conclusion Defibrase has protective effect against reperfusion injury post cerebral ischemia.

OBJECTIVE To observe the effect of natrin from Naja naja atra(Chinese cobra)on intracellular free calcium overload,and to discuss the protective effect and the possible mechanism of natrin on myocardium calcium(Ca2+)and potassium(K+)ion channels in the primary cardiomyocytes of SD neonatal rats. METHODS The primary cardiomyocytes of SD neonatal rats were used,which were respectively pretreated with natrin 5,25 and 125 mg · L-1 for 24 h before injury was induced by H2O2 0.3 mmol · L- 1. The dynamic variation of intracellular calcium was monitored by laser confocal microscopy using Fluo-3 as Ca2+fluorescence probe. Additionally,the cardio myocytes of neonatal rats were pretreated for 24 h using different concentrations of natrin 5,25,125 mg · L-1 and verapamil 5 nmol · L-1,followed by exposure to H2O2 0.3 mmol · L-1 for 15 min. Then,the mRNA expressions of calcium channels subunits Cav1.2,Calm,RyR2 and potassium channel Kir6.2 were analyzed by FQ-PCR method. RESULTS Laser confocal microscopy revealed that H2O2 obviously caused calcium overload in cardiomyocytes, giving rise to 49.37% fluorescence increase in intracellular calcium compared with the control group(P<0.01). However,natrin 5,25 and 125 mg·L-1 resulted in 27.52%, 12.71% and 5.15% fluorescence increase in intracellular calcium,respectively,compared with the control group(P<0.01). Moreover, the PCR results showed that the mRNA expressions of Cav1.2, Calm and RyR2 in the myocardial cells treated with H2O2 were increased 2.78,2.26,and 5.34 times as compared with the control group,while Kir6.2 displayed a 1.79-fold expression level(P<0.01). By contrast, the combination of natrin and verapamil significantly decreased the mRNA expression of Cav1.2,Calm and RyR2,compared to the H2O2-treated group(P<0.01). Meanwhile,the expression of Kir6.2 was considerably higher than that of the H2O2-treated group(P<0.05). CONCLUSION Natrin can reduce the intracellular calcium overload of cardiomyocytes induced by H2O2 and shows a protective effect against oxidative damage for cardiomyocytes. The possible mechanism is that natrin can decrease the mRNA expression of Cav1.2,Calm,RyR2 and increase the expression of Kir6.2 of the H2O2-induced cardiomyocytes.

This paper is to report the study of the pharmacokinetics of a fusion protein TAT-haFGF(14-154) for human acidic fibroblast growth factor and transcriptional activator protein in rat plasma, and the investigation of their penetration across blood-brain barrier in mice and rats, in order to provide a basis for clinical development and treatment of Alzheimer's disease. Enzyme-linked immunosorbent assay (ELISA) was used to determine concentration of TAT-haFGF(14-154) in rat plasma and in mouse brain homogenate; and immunohistochemistry was used to analyze the distribution in brain. The concentration-time curve fitted two-compartment open model which was linear kinetics elimination after a single intravenous injection of TAT-haFGF(14-154) in rat at the dose of 300 microg x kg(-1). The half life time was 0.049 +/- 0.03 h for distribution phase and 0.55 +/- 0.05 h for elimination phase, and the weight was 1/C2. The result showed that TAT-haFGF(14-154) could be detected in the brain by ELISA and immunohistochemistry, the elimination of TAT-haFGF(14-154) in rat was swift, and TAT-haFGF(14-154) could penetrate across the blood-brain barrier, distribute in pallium and hippocampus and locate in the nucleus.

Species identification of biological samples is widely used in such fields as forensic science and food industry. A variety of accurate and reliable methods have been developed in recent years. The current review shows common target genes and screening criteria suitable for species identification, and described various DNA-based molecular biology methods about species identification. Additionally, it discusses the future development of species identification combined with real-time PCR and sequencing technologies.
Animals ; DNA/genetics* ; Genetic Techniques/trends* ; Humans ; Oligonucleotide Array Sequence Analysis ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction ; Sequence Analysis, DNA/trends*

Objective To investigate the curriculum construction and evaluation of teaching effectiveness of simulation and comprehensive experiment for clinical nursing. Methods Based on the investigation of hospital, taking working progress and working task of nurses as orientation, we constructed the curriculum of simulation and comprehensive experiment for clinical nursing, and unfold in junior class (before clinical practice) for student nurses. The teaching effect was evaluated. Results Unfolding simulation and comprehensive experiment could effectively enhance the nursing students' ability to transform theoretical knowledge into clinical nursing practice, and was favorable to change the role of nurses and improve the satisfaction degree of clinical practice.Conclusions Unfolding simulation and comprehensive experiment for clinical nursing for student nurses before clinical practice is practicable and essential.

In order to decrease the potential side-effects of human basic fibroblast growth factor (hbFGF) caused by its broadspectrum mitogenic activity, a single residue of hbFGF, the residue serine 108, was replaced with neutral alanine residue to construct a mutant of hbFGF (mhbFGF) with reduced mitogenic activity. The mutant was overexpressed in Escherichia coli BL21(DE3) by IPTG induction. The expression level of mhbFGF was about 30% of the total cellular protein. The expressed mhbFGF was purified by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate. Measured by MTT method, the effect of mhbFGF on Balb/c 3T3 cell proliferation was much lower than that of wild-type hbFGF. The purified recombinant mhbFGF was prepared and sufficient for the following pharmacological and safety studies.

AIM:To study the mechanism of the unique export of one of human basic fibroblast growth factor (hbFGF) forms lacking the N-terminal nuclear localization signal (NLS),we high expressed and purified this hbFGF form in E.coli strain BL21(DE3).METHODS:The cDNA fragment of the hbFGF amplified by polymerase chain reaction (PCR) was cloned into the expression vector pET3c and expressed in BL21(DE3) by IPTG induction.The expressed hbFGF was purified by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate.The mitogenic activity was measured by MTT.RESULTS:The expression level of hbFGF in E.coli was about 20% of the total cellular protein.The appreciable mitogenic activity of the purified hbFGF was comparable to that of hbFGF standard.CONCLUSION:The BL21(DE3)/ pET3c expression system could be used to high express hbFGF lacking NLS.The purified recombinant hbFGF was prepared and sufficient for further study.

AIM:To synthesize the human platelet factor-4(hPF4) gene with a convenient and effective approach, and high express the hPF4 gene in E. coli BL21 (DE3). METHODS: According to the primary structure of hPF4, the nucleotide sequence was synthesized using touch-down PCR method. The resultant gene fragment containing EcoR Ⅰ and Xho Ⅰ overhangs at 5' and 3' ends was cloned into the expression vector pGEX-4T-1 to construct the recombinant plasmid pGEX-4T-1-hPF4,which was then transformed into the E. coli strain BL21 (DE3). RESULTS: hPF4 gene was successfully synthesized by touchdown PCR method. A fusion protein composed of glutathione S-transferase (GST) and the recombinant hPF4 was expressed in BL21(DE3) by IPTG induction. The expression level of the fusion protein in E. coli was about 30% of the total cellular protein. CONCLUSION: Touch-down PCR may provide a convenient and effective approach to obtain other target genes. The expressed fusion protein forms the inclusion bodies, providing sufficient material for further purification and biological activities process.

Objective To investigate the syndrome type distribution of hypertensive patients; To analyze the correlation of characteristics of HRV time domain parameters and its influence factors. Methods Totally 515 cases of hypertensive patients were included and were put under syndrome type distribution. Demographic information, laboratory test parameters, risk factors and clinical symptoms were collected for correlation analysis. HRV time domain parameters were recorded by using 24 h ambulatory electrocardiogram. The differences in SDNN, SDNN Index, HRV Index, PNN50, and RMSSD of different TCM syndrome types were compared. Results Among 515 patients: 160 cases with hyperactivity of yang due to yin deficiency syndrome, 136 cases with turbid phlegm and blood stasis syndrome, 83 cases with overabundant liver-fire syndrome, 69 cases with deficiency of kidney qi, and 67 cases with abundant phlegm-dampness syndrome. By comparing different TCM syndromes, the level of SDNN was significantly reduced in the hyperactivity of yang due to yin deficiency syndrome, overabundant liver-fire syndrome,deficiency of kidney qi syndrome compared with turbid phlegm and blood stasis syndrome, abundant phlegm-dampness syndrome (P<0.05); SDNN Index and HRV Index decreased significantly in the hyperactivity of yang due to yin deficiency and overabundant liver-fire syndrome compared with abundant phlegm-dampness syndrome (P<0.05). SDNN Index decreased significantly in the deficiency of kidney qi compared with abundant phlegm-dampness syndrome (P<0.05). The level of PNN50 was significantly reduced in the deficiency of kidney qi compared with hyperactivity of yang due to yin deficiency syndrome (P<0.05). RMSSD decreased significantly in the hyperactivity of yang due to yin deficiency syndrome, deficiency of kindney qi syndrome, overabundant liver-fire syndrome compared with turbid phlegm and blood stasis syndrome (P<0.05). Discriminant analysis showed that SBP, DBP, MBPS, SDNN, SDNN Index, HRV Index, PNN50, RMSSD were correlated with the diagnosis of five syndrome types. Logistic regression analysis showed that the factors including gender (female), insomnia, elevated systolic blood pressure, MBPS, decreased SDNN Index and PNN50 were positively correlated to hyperactivity of yang due to yin deficiency; other factors including gender (female), advanced age, elevated blood pressure, decreased SDNN, HRV Index and RMSSD were positively correlated with turbid phlegm and blood stasis syndrome. And the study also showed that advanced age, family history of hypertension, elevated blood pressure, decreased SDNN Index, HRV Index and PNN50 were positively correlated to abundant phlegm-dampness syndrome. Conclusion HRV time domain parameters can be significantly reduced in the hyperactivity of yang due to yin deficiency, overabundant liver-fire syndrome, and deficiency of kidney qi syndrome. The autonomic nerve function is damaged seriously. Hyperactivity of yang due to yin deficiency syndrome, abundant phlegm-dampness syndrome turbid phlegm and blood stasis syndrome are closely related to the influencing factors that lead to cardiovascular and cerebrovascular events.