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 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.

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.

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

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

OBJECTIVETo investigate the value of Diffusion-Weighted Imaging (DWI) in the diagnosis of early stage liver diffuse lesions.

METHODSDiethylnitrosamine (DEN) was used to induce liver lesions in rats. Sequential DWI studies were performed on the livers from 1 to 14 weeks after DEN was administered through drinking water. Comparing studies with a blank control group was set and pathohistological examinations of the livers were performed.

RESULTSNo obvious routine MRI morphological change was found in either group during this period, but DWI demonstrated heterogeneous changes in the test group at the cirrhosis stage. There was no significant alteration of the apparent diffusion coefficient (ADC) value in the control group during this period (P > 0.05). The ADC values of the test group began to decline from the fifth week. Until the tenth week, the ADC value of the test group decreased drastically and when b = 300 s/mm2 statistic, the results showed an obvious difference between the two groups. There were also differences between the ADC values at the 10th, the 9th and the 1st weeks of the test group (P < 0.05). When b = 600 s/mm2 and 1000 s/mm2, significant differences were found after the sixth week between the two groups (P < 0.01). The main pathohistological liver change in the test group during the 1 to 4 week period after DEN was administered was swelling of hepatocytes; during the 5 to 8 week period it was fibrous tissues hyperplasia, and in the 9 to 14 week period it was cirrhotic nodule formation.

CONCLUSIONMR functional DWI could detect liver diffuse lesions earlier than conventional MR imaging. Measurement of ADC value may be of use in early diagnosis of liver diffuse diseases and for monitoring the changes of the lesions.

Animals ; Chemical and Drug Induced Liver Injury ; Diethylnitrosamine ; Diffusion Magnetic Resonance Imaging ; methods ; Liver Diseases ; diagnosis ; pathology ; Male ; Rats ; Rats, Wistar

OBJECTIVETo study the inhibition of the expression of bcl-xL gene induced by RNA interference in CNE-2Z cell line in addition to the inhibition of its proliferation and apoptotic induction.

METHODSSmall interfering RNAs targeting bcl-xL gene were synthesized by using web design software provided by Amnion and the silencer short interfering RNA (siRNA) construction kit; fluorescein-labeled siRNAs were done by FAM-silencer siRNA labeling kit; siRNAs were transfected into CNE-2Z cells by using lipofectamine 2000 reagent; siRNA transfection efficiencies were analyzed by fluorescent microscopy; down-regulation of bcl-xL was detected by RT-PCR; thiazolyl blue (MTT) assay was used to assess the cell growth; apoptosis of CNE-2Z cells was analyzed by flow cytometry.

RESULTSGreen fluorescence in the cells was seen clearly in FAM-labeled siRNA transfected group under the fluorescent microscope while none in the untransfected group. Different down-regulations of bcl-xL mRNA expression were found in the transfected groups. The expression of bcl-xL mRNA decreased by 10% - 70% in the siRNAs transfected CNE-2Z by RT-PCR scan analysis. The inhibitory rate of cell proliferation depended on time and concentrations to some extent. Different cell apoptosis could be induced by different concentrations of siRNA4.

CONCLUSIONSThe synthesized siRNAs in vitro were able to down-regulate the expression of bcl-xL There were different capabilities of the specific siRNAs down-regulation. The transient transfected bcl-xL siRNA4 could effectively inhibit the growth of the cancer cells and induce theirs apoptosis. It was suggested that the siRNA technique provide not only an extremely powerful tool for the functional analysis of genome but also a new method for anti-nasopharyngeal carcinoma gene therapy.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Humans ; Nasopharyngeal Neoplasms ; genetics ; pathology ; RNA Interference ; RNA, Messenger ; genetics ; Transfection ; bcl-X Protein ; genetics

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.

Animals ; Cell Culture Techniques ; Cell Proliferation ; Green Fluorescent Proteins ; biosynthesis ; Mice ; NIH 3T3 Cells ; Transfection

OBJECTIVETo investigate the efficacy of transcutaneous electrical nerve stimulation (TENS) on four specific acupuncture points Hegu (LI4), Neiguan (PC6), Danshu (BL19) and Weishu (BL21) for reducing pain in labor.

METHODSA total of 160 voluntary nulliparous women who were willing to receive TENS for analgesia were assigned to the treatment group after cervical dilation of more than 2 cm. Another 145 matched nullipara were recruited as the control group. Visual analogue scale (VAS) was used to assess the pain before and 0.5 h after the application of TENS. Then, VAS was assessed every one hour until delivery. Percentage of VAS score decreased by > 25% was the primary outcome, the delivery mode and neonatal outcome were measured as secondary outcomes. Adverse reactions were also recorded during TENS.

RESULTSThe percentage of VAS score decreased by > 25% was 68.6% in the TENS treatment group. Maternal delivery mode and neonatal outcomes were not significantly different between the two groups. In addition, the incidence of postpartum hemorrhage in the TENS treatment group was less than the control group (P<0.05). There was no adverse reaction recorded with TENS on acupoints.

CONCLUSIONAs a novel and non-invasive approach, TENS on specific acupoints including Hegu (LI4), Neiguan (PC6), Danshu (BL19) and Weishu (BL21) was an effective method for analgesia in labor.

Acupuncture Points ; Case-Control Studies ; Delivery, Obstetric ; Demography ; Female ; Humans ; Infant, Newborn ; Labor, Obstetric ; blood ; Pain Management ; Pain Measurement ; Postpartum Period ; blood ; Pregnancy ; Time Factors ; Transcutaneous Electric Nerve Stimulation ; adverse effects ; Treatment Outcome