Objective To explore the correlation of the apoptosis of peripheral blood mononuclear cell (PBMC) with plasma 8-iso-prostaglandin F2? (8-iso-PGF2?) level in order to confirm the peroxidation damage of hyperbilirubinemia in neonates with jaundice.Me-thods One hundred and twenty-nine neonates who were 2 to 7 days old were divided into 4 groups:slight jaundice (43 cases),moderate jaundice (38 cases),serious jaundice (18 cases),and healthy control(30 cases)groups.Plasma 8-iso-PGF2? levels were assayed by enzyme immunoassay(EIA),and the apoptosis rates of PBMC were determined by flow cytometry.Results 1.There was no significant difference in the apoptosis rate of PBMC between normal and slight jaundice neonates (P=0.108);apoptosis rate of PBMC was increasing with the aggravation of jaundice.2.Plasma 8-iso-PGF2? level in normal neonates was 14.74?6.71 ng/L,there was no difference between normal neonates and neonates with slight jaundice(P=0.502).Plasma 8-iso-PGF2? levels in neonates with moderate and serious jaundice were much higher(Pa=0).3.Positive correlation existed between plasma 8-iso-PGF2? level and PBMC apoptosis rate (r=0.602 P=0).Conclusions Hyperbilirubinemia can induce peroxidation damage,resulting in increase of PBMC apoptosis.Plasma 8-iso-PGF2? level can accurately eva-luate the peroxidation damage in neonates with jaundice.

Objective To evaluate myocardial apoptosis with 99Tcm-C2A-GST myocardial imaging using the recombined C2A domain of Synaptotagmin Ⅰ by gene engineering. Methods ( 1 ) The C2A gene was inserted into the prokaryotic glutathione S-transferate (GST) fusion protein expression plasmid pGEX-6P-1. The recombinant plasmid was transformed into E. coli BL21. C2A-GST fusion protein was purified after BL21 was induced with isopropyl-β-D-1-thiogalactopyranoside (IPTG). (2)The activity of fusion protein was identified by cell binding test with fluorescein-5-isothiocyanate (FITC)-C2A-GST. (3) The C2A-GST fusion protein was labeled with 99Tcm using 2-iminothiophene hydrocoride method. Radiochemical purity was determined with thin layer chromatography. (4)99Tcm-C2A-GST (7.4 MBq) was injected to ischemia-reperfusion rat models through tail vein. The image was acquired with SPECT at 1 h after injection, and then hearts were removed, rinsed with saline and dyed with triphenyl tetrazolium coride (TTC). The ischemic myocardium was separated from the viable myocardium and was weighted. Its radioactivity was measured by gamma counting. The difference of uptake of radiotracer between ischemic myocardium and normal myocardium was compared using percentage activity of injected dose per gram of tissue ( % ID/g) with standard deviation. SPSS 12.0 and t-test were used for data analysis. Results ( 1 ) C2A-GST fusion protein wassuccessfully expressed and its relative molecular weight was 3.8 × 104. (2) FITC-C2A-GST binding to apoptotic cells could be observed by fluorescent microscopy. (3) The radiochemical purity of 99Tcm-C2A-GST was (98.90 ±0.43)%. (4)The imaging studies showed that there was focal uptake of radioactivity in the ischemic myocardium. In vitro uptake of 99Tcm-C2A-GST was (2.41 ±0.32) % ID/g by the ischemic myocardium, however 99Tcm-C2A-GST-N-hydroxysuccinimide (C2A-GST-NHS) was (0. 82 ± 0. 24) % ID/g. There was statistically significant difference between those two groups (t = 10. 6, P ＜0.01 ). Conclusion The C2A domain of Synaptotagmin Ⅰ expressed by gene engineering can be used as the tracer for noninvasive detection of ischemic myocardium in the ischemia-reperfusion rat model.

Objective:To investigate the impact of self-concealment,self-disclosure,coping style and per- ceived social support on university students'loneliness.Methods:Loneliness and related factors were assessed among 482 university students using scales including UCLA Loneliness Scale,Self-concealment Scale(SCS),Self-disclosure Index(SDI),Simplified Coping Style Questionnaire(SCSQ)and Perceived Social Support Scale(PSSS).Results: The level of university students'loneliness was not high(36.5?7.4);males experienced more loneliness than fe- males(37.4?7.5/35.4?7.3,F=8.25,P0.05). Regression analysis showed that SCS,SCSQ and PSSS predicted UCLA effectively(?=0.207,-0.218,0.157, -0.380).The testing of mediating effect indicated that SCS had direct and indirect impact on UCLA through nega- tive coping style and PSSS;SDI had only indirect impact on UCLA through positive coping style and PSSS.Conclusion:SCS,SDI,SCSQ and PSSS are important factors influencing UCLA,and the intervention of univer- sity students'loneliness should focus on these variables.

Objective To evaluate the effect of dexmedetomidine on autophagy in the hippocampal neurons of rats with traumatic brain injury (TBI).Methods Adult male Sprague-Dawley rats,aged 12-16 weeks,weighing 340-370 g,were randomly divided into 3 groups (n=80 each) using a random number table:sham operation group (group S),traumatic brain injury group (group TBI) and dexmedetomidine group (group Dex).The rats were subjected to a diffuse cortical impact injury caused by a modified weight-drop device to induce TBI.Dexmedetomidine 15 μg/kg was injected intravenously immediately after TBI in Dex group.At 24 and 48 h after TBI,neurological deficit score (NDS) was assessed,Morris water maze test was performed,and brains were removed for detection of brain water content in the brain tissue.At 6,12,24 and 48 h after TBI,the expression of hippocampal LC3]Ⅱ was determined using Western blot analysis.Results Compared with group S,brain water content and NDS were significantly increased at 24 and 48 h after TBI,the escape latency was prolonged,and the expression of hippocampal LC3 Ⅱ was upregulated at 6,12,24 and 48 h after TBI in TBI group.Compared with TBI group,brain water content and NDS were significantly decreased at 24 and 48 h after TBI,the escape latency was shortened,and the expression of hippocampal LC3 Ⅱ was down-regulated at 6,12,24 and 48 h after TBI in Dex group.Conclusion The mechanism by which dexmedetomidine reduces TBI is related to inhibition of autophagy in the hippocampal neurons of rats.

Amifostine ; pharmacology ; Animals ; Benzene ; toxicity ; Blood ; drug effects ; Blood Cell Count ; Male ; Mice ; Random Allocation

The growing status of Anoectochilus roxburghii seedling was observed and the survival rate of seedlings, height, stem diameter and plant fresh weight under the conditions of different transplanting substrate compositions, planting density, shading rate were measured. The results showed that the effects of different transplanting substrates, planting densities, shading rates and nutrient solutions on the growing status of A. roxburghii plantlets varied greatly. A. roxburghii plantlets demonstrated a high survival rate and better growing status under the Following conditions: the ratio of peat and river sand as 2: 1, the planting density as 3 cm x 3 cm, the shading rate as 70%, and the nutrient solution as 1/4MS. The findings of the study provide a solid technical solution for the artificial cultivation of A. roxburghii plantlets.
Breeding ; methods ; Culture Media ; chemistry ; pharmacology ; Orchidaceae ; drug effects ; growth & development ; Seedlings ; drug effects ; growth & development ; Survival Analysis

Objective: To explore sarcoplasmic reticulum ryanodine receptor2 (RyR 2) expression and calcium releasing function in chronic heart failure (CHF) rabbits and to study the impact of long term valsartan treatment in relevant animals. Methods: HF model was established by volume overloading with pressure overloading in experimental rabbits. 27 rabbits were divided into 3 groups: Sham group, HF group and HF+valsartan group. n=9 in each group and the animals were treated for 7 weeks. Left ventricular structure, hemodynamic parameters, expression and functional changes of myocardiocyte sarcoplasmic reticulum RyR 2 were observed and compared among different groups. Results: Compared with Sham group, HF group had increased left ventricular mess index (LVMI), left ventricular end diastolic pressure (LVEDP) and decreased left ventricular shortening fraction, LVEF, all P<0.05. Compared with HF group, HF+valsartan group showed decreased LVMI, LVEDP and increased left ventricular shortening fraction, LVEF, all P<0.05. Sarcoplasmic reticulum RyR 2 expression and calcium releasing function were lower in HF group than Sham group, P<0.05; while they were both higher in HF+valsartan group than HF group, P<0.05. Conclusion: Long term application of valsartan could improve the cardiac function which might be related to increased myocardial sarcoplasmic reticulum RyR 2 expression and calcium releasing function in experimental CHF rabbits.

OBJECTIVETo investigate the influence of fluorescent protein expression on the proliferation of murine NIH3T3 cells, so as to provide a theoretical basis for cell tracing technology.

METHODSNIH3T3 cells were cultured in vitro, and were randomly divided into control, pLEGFP-N1 (with transfection of pLEGFP-N1 retroviral vector), pEGFP-N1 (with transfection of pEGFP-N1 vector) and pDsRed2-C1 (with transfection of pDsRed2-C1 vector) groups. Then the cells were screened by G418 for 3 weeks. The changes in cell adhesive rate were observed and the population doubling times was determined by growth curve.

RESULTSThere was obvious fluorescent protein expression in the transfected NIH3T3 cells after G418 selection, and the highest percentage of labeled NIH3T3 cells was found in pLEGFP-N1 group. The population doubling time in pDsRed2-C1 (40.3+/-0.7 h) , PEGFP-N1 (39.6 +/- 0.6 h) and pLEGFP-N1 (36.5 +/- 0.7 h) groups was evidently longer than that in control (27.9 +/- 0.6 h, P < 0.01), with high adhesive rate in each group.

CONCLUSIONThe expression of fluorescent protein exhibited some inhibitory effect on the proliferation of NIH3T3 cells in vitro. Since the inhibitory effect by retroviral vector was weaker compared with eukaryotic vector, it should be the first choice for fluorescent protein labeling during cell transplantation.

Objective To introduce an improved method for collecting the blood of rat by cardiac puncture .Methods The study se-lected 90 Wistar rats,half male and half female ,and then the rats were anesthetized by intraperitoneally injecting 10％chloral hydrate solution according to 0.3 mL/kg body weight.Then using three-line positioning method to determine the puncture site ,the blood collection needle and blood collection tube were used to collecting the blood ,Finally,the blood volume of each rat was recorded .Results The success rate was cal-culated according to the single blood collection of 5 mL or more,and our 90 rats were collected 90 times,the success rate was about 82.2％. After centrifugation,the total serum was about 260 mL,the average serum separation of each rat was about 2.8 mL.Conclusion The im-proved positioning method of heart blood collection is intuitive and easy to learn ,and the total volume and quality of blood is high ,besides ,the new method has a small surgical trauma and a high efficiency ,which is worth to extend .

Objective To select the optimum process of granule spray for Tangwang Mingmu Granules, To improve the yield and finished product quality. Methods The effects of the relative density, the type of excipients, the ratio of the extract and the excipient, the inlet temperature-the material temperature, the spray rate and the atomization pressure were selected by single factor and orthogonal test respectively to determine the optimum preparation process. Validation for three batch sample, particle flow, hygroscopicity and other related content of the preparation were studied. Results The optimum spray granulation process of Tangwang Mingmu Granules: the relative density of extract (60 ℃ determination) was 1.15–1.20; the excipient was selected as dextrin; the ratio of dry extract and excipient was 58:42; inlet temperature - material temperature was 90-75 ℃; atomization pressure was 0.12 mPa;spray rate was 10 Hz.Conclusion The preparation technology of Tangwang Mingmu Granules optimized in this study is with high preparation rate, good fluidity and hygroscopicity, which can be used for the preparation of Tangwang Mingmu Granules.