Aim To study the relationship between the expressions of Fas,Fas-L and the spontaneous apoptosis of thymocytes in vitro. Methods The expressions of Fas,Fas-L and the apoptotic rate of thymocytes were assayed by flow cytometry. Results The apoptosis and the Fas expression of thymocytes of different culture times in vitro were increased time-dependently in vitro. The Fas-L expression of thymocytes cultured for 24 h was also significant increase. There was significant corelation between the increase of Fas expression and the increase of apoptosis of thymocytes in vitro. Conclusion Fas is an important molecule which mediates spontaneous apoptosis of thymocytes cultured in vitro.

Objective To screen the differential proteins in the brain (neocortex and hippocampus) between the rats with cortical dysplasia (CD) and control ones,and investigate the role of their alteration in the development of epilepsy in CD.Methods Cortical dysplasia was induced in rat pups via in utero delivery of BCNU.A two-dimensional electrophoresis (2-DE)-based approach was used to construct the expression profiles of proteins in both the neocortex and hippocampus at different age groups (postnatal day 7 and 60) and to detect proteome changes between CD rats and control ones.Following gel image analysis,protein spots that differed in abundance between CD and control rats were identified by using Matrx-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) and MS/MS.Results A total of 57 kinds of protein were screened out (P ＜ 0.05),in which 35 were found up-regulated and 22 were down-regulated compared with the control,35 from neonatal stage (postnatal day 7) and others from adult stage.Finally,12 among them were identified,including tubulin,alpha-lB,Beta-actin,tubulin beta-2A,GAP-43,UbCKmit,GAPDH and TMBr-3,etc.Conclusions Changed expression of specific proteins which were found in our study are involved in construction of brain 's cytoskeleton,synaptic function,mitochondrial function and so forth.Thus,they may be related to the pathogenic mechanisms of epileptogenicity of CD.

BACKGROUND:Previous studies have indicated that ulinastatin can inhibit RANKL-induced osteoclastogenesis on RAW264.7 cells and also lower matrix metal oproteinase-9 expression and activity. However, it remains be unclear whether ulinastatin has the intervention effect on polymethyl methacrylate (PMMA)-induced MC3T3-E1 mouse preosteoblast apoptosis or not. ＜br> OBJECTIVE:To explore the intervention role of ulinastatin on the PMMA-induced MC3T3-E1 mouse preosteoblast apoptosis and its effects on type I col agen, osteocalcin, matrix metal oproteinase-2 mRNA expression. ＜br> METHODS:MC3T3-E1 mouse preosteoblasts at passages 6 and 7 were divided into four groups:blank group (only cultured MC3T3-E1 mouse preosteoblast), PMMA-induced group (MC3T3-E1 mouse preosteoblast+1 g/L PMMA bone cement suspension), low dose ulinastatin group (MC3T3-E1 mouse preosteoblast+1 g/L PMMA bone cement suspension+500 U/mL ulinastatin) and high dose ulinastatin group (MC3T3-E1 mouse preosteoblast+1 g/L PMMA bone cement suspension+5 000 U/mL ulinastatin). MTT method was adopted to detect the proliferation activity of proliferative activity of MC3T3-E1 mouse preosteoblast;alizarin red staining method was used to observe mineralization nodules of MC3T3-E1 mouse preosteoblast among different groups;the change of apoptosis rate for MC3T3-E1 cells was detected by flow cytometry analysis;semi-quantitative RT-PCR was taken to analyze type I col agen, osteocalcin, matrix metal oproteinase-2 mRNA expression level in MC3T3-E1 mouse preosteoblasts among different groups. ＜br> RESULTS AND CONCLUSION:Compared with the blank group, PMMA significantly inhibited the proliferation activity of MC3T3-E1 mouse preosteoblast (P＜0.05), and however significantly promoted cells apoptosis (P＜0.05). After addition of different concentrations of ulinastatin (500, 5 000 U/mL), the proliferation activity of MC3T3-E1 mouse preosteoblasts significantly raised (P＜0.05), and cells apoptosis rate significantly decreased (P＜0.05), showing the dose and time-dependent relation. Type I col agen and osteocalcin mRNA expression levels both significantly decreased after co-culture in PMMA group compared with the blank group (P＜0.05), matrix metal oproteinase-2 mRNA expression level, however, significantly increased (P＜0.05). After intervention with 5000 U/mL ulinastatin, type I col agen and osteocalcin mRNA expression levels both significantly increased, while matrix metal oproteinase-2 mRNA expression level significantly decreased (P＜0.05). PMMA group showed no obvious mineralization nodules. Yet, mineralization nodules were formed in the blank group, high and low dose ulinastatin groups. These results indicate that ulinastatin could have the inhibitory effect on the PMMA-induced MC3T3-E1 mouse preosteoblast apoptosis, and it could promote type I col agen and osteocalcin mRNA expression and yet suppress matrix metal oproteinase-2 mRNA expression.

BACKGROUND:It is presumed that urinary trypsin inhibitor could have protective effects on local and systemic tissues and could inhibit osteoclast proliferation and activation under long-term chronic inflammation conditions and in ischemic and anoxic environment which was induced by prosthetic wear. OBJECTIVE:To investigate the inhibitory effect of ulinastatin on receptor activator for nuclear factor-κb ligand-induced differentiation, proliferation and osteoclastogenesis of RAW264.7 cells and its effects on matrix metal oproteinase-2, matrix metal oproteinase-9 expression level and activity. METHODS:Mouse monocyte/macrophage cellline RAW264.7 was treated with different concentrations of urinary trypsin inhibitor (0, 500, 5 000 U/mL) for 24, 48 and 72 hours. Experiments were divided into four groups:the blank group (RAW264.7 cells), receptor activator for nuclear factor-κb ligand-induced group (0 U/mL ulinastatin), 500 U/mL ulinastatin group and 5 000 U/mL ulinastatin group. RESULTS AND CONCLUSION:(1) MTT results indicated that there was no significant difference on the proliferation of RAW264.7 cells treated with urinary trypsin inhibitor at 0-5 000 U/mL (P>0.05) (2) Tartrate-resistant acid phosphatase staining results revealed that compared with receptor activator for nuclear factor-κb ligand-induced group, the number of tartrate-resistant acid phosphatase-positive cells was significantly less in the ulinastatin group (P<0.05), showing a time-dose dependent manner. (3) Immunohistochemisical results found that compared with receptor activator for nuclear factor-κb ligand-induced group, the percentage of matrix metal oproteinase-9-positive cells was apparently lower in the ulinastatin group. (4) Western blot assay results demonstrated that matrix metal oproteinase-9 expression was low in the RAW264.7 cells alone. At 48 hours after addition of receptor activator for nuclear factor-κb ligand, matrix metal oproteinase-9 protein expression was large. At 72 hours after culture in the 5 000 U/mL ulinastatin group, matrix metal oproteinase-9 protein expression was evidently reduced. (5) Gelatin zymography results showed that compared with the receptor activator for nuclear factor-κb ligand-induced group, matrix metal oproteinase-9 expression was significantly lower in the 5 000 U/mL ulinastatin group (P<0.05). Results suggested that urinary trypsin inhibitor inhibited receptor activator for nuclear factor-κb ligand-induced osteoclastogenesis and diminished matrix metal oproteinase-9 expression and activity.

Based on the conserved region nucleotide sequences of five potato viruses/viroid(Alfalfa Mosaic Virus,ALMV;Cucumber mosaic virus,CMV;Cucumber mosaic virus-satellite,CMV-sat;Potato virus Y,PVY;Potato spindle tuber viroid,PSTVd) and one inner control(18S rRNA),the microarray containing specific oligonucleotide probes and PCR probes were designed and fabricated.The effects of probe concentration,hybridization time,hybridization temperature and spotting solutions on microarray hybridiazation were evaluated.Finally the specificity of optimized plant virus detection array was validated.No significant effect on hybridization signal intensity was observed when the concentration of the oligonucleotide probes ranged from 5 to 20 ?mol/L,there was a linear relationship between the concentration of PCR probe and hybridization signal intensity.The greatest signal intensity were obtained when hybridized at 45℃ for 4 h,and the oligonucleotide probes and PCR probes had a similar effect on microarray hybrization.Among the different spotting solutions,DMSO produced a good reproducibility.The plant virus could be detected specifically by oligonucleotide probe microarray and PCR probe microarray after optimization.

OBJECTIVETo find out an effective therapy for obesity induced by antipsychotic agents.

METHODSOne hundred and one cases of obesity who were being treated with antipsychotic agents were randomly divided into an observation group and a control group. The observation group were treated with electroacupuncture at Quchi (LI 11), Zusanli (ST 36), Fenglong (ST 40), Shangjuxu (ST 37) and Xiajuxu (ST 39), once a day for 8 weeks; and no treatment was given to the control group. The Brief Psychiatric Rating Scale (BPRS) was used for assessment of therapeutic effect and the Treatment Emergent Symptom Scale (TESS) for adverse effects.

RESULTSThe clinically effective rate of the observation group (54.9%) was superior to the control group (10.0%) with a significant difference between the two groups (P<0.05). The BPRS score-reducing rate was 24.92% and 28.62% in the both groups respectively, and with no adverse effects.

CONCLUSIONElectroacupuncture has a good therapeutic effect on obesity induced by antipsychotic agents, and it improves the patient's compliance with no adverse effect.

Acupuncture Points ; Antipsychotic Agents ; Electroacupuncture ; Humans ; Obesity

OBJECTIVEThe aim of this survey was to investigate the level of contamination of the most consumed foods in China with 16 rare earth elements (REEs), and to provide the basic data for establishing and revising food safety standards for REEs.

METHODSSixteen REEs in foods were measured by inductively coupled plasma-mass spectrometry (ICP-MS) in the labs of the Centers for Disease Control and Prevention of four provinces and two municipalities, during 2009-2010.

RESULTS1 231 samples were analyzed and 19 121 concentration data of 16 REEs were collected. The REEs levels in the investigated foods varied significantly. The concentrations of cerium (Ce), dysprosium (Dy), yttrium (Y), lanthanum (La), and neodymium (Nd) were relatively high, while the remaining eleven REEs were at low levels. The mean values of total rare earth element oxides (REOs) in cereals, fresh vegetables, fresh aquatic products, fresh meats and eggs varied from 0.052 mg/kg to 0.337 mg/kg.

CONCLUSION16 REEs in the major foods were at very low contamination levels in the investigated regions.

Animals ; China ; Edible Grain ; chemistry ; Eggs ; Fishes ; Food Analysis ; Food Contamination ; Meat ; analysis ; Metals, Rare Earth ; chemistry ; Mollusca ; Swine ; Vegetables ; chemistry

Objective To determine the effect of mitochondria on the protection of heat-shock proteins B1(HSPB1) from oxidative damage in rat cardiac cell.Methods HSPB1 gene-transfected rat cardiomyocytes cell line H9c2 (HSPB1 H9c2) and empty vector transfected H9c2 (control) were established,and treated by 0-1000?mol/L H_2O_2 for 2h.And then the cell morphology, mitochondrial membrane potential and endogenous reactive oxygen species (ROS) were detected. Results (1)HSPB1 inhibited the morphological changes induced by H_2O_2 markedly.(2)HSPB1 inhibited the loss of mitochondrial membrane potential induced by H_2O_2.Following the stimulation of 0,75,150,300,500,1000?mol/L H_2O_2,mitochondrial membrane potential in HSPB1 and control H9c2 cells were (10.0?0.11)vs (7.01?0.26),(9.11?0.17)vs (6.05?0.19),(7.69?0.28)vs (5.14?0.28),(6.95?0.13)vs (4.66?0.11),(6.61?0.20)vs (1.85?0.35),(6.60?0.05)vs (1.19?0.01),respectively (all P0.05).Conclusions HSPB1 protects rat cardiomyocytes cell line(H9c2) from oxidative damage,which suggests that stabilization of mitochondrial membrane potential and the decreased endogenous reactive oxygen species after oxidative stress may be involved in the protection of HSPB1 against oxidative stress in H9c2.

Objective To investigate the effects of different concentrations of sodium fluoride on the morphologic characteristics of primarily cultured thyroid cells of SD rats and in order to obtain important proof for approaehing the mechani8m of thyroid gland damage caused by fluoride.Methods Thyroid cells of SD rat were primarily culture for 96 hours,and cell density was adjusted to 5.0×108/L Cell suspension with 5 ml Wills seeded into 6 weII plates,after 12 hours,0(contr01),10.100,1000 μmol/L of sodium fluoride was added into the well, witll each well representing different level of treatment group.Finally the cultured thyroid cells were collected for morph010gic study.Results Under microscope,the transparency of the control thyroid cells Was good,and cells gathered in cluster and adhered to wall.But a lot of cells treated with fluoride suspended,and lost their transparency-under scaning delectron microscope,the control calls showed integrated membrane and tightness to each other,as well as clear boundary between cells normal proliferation.While the thyroid cells treated with 10,100 μmol/L sodium fluoride 0bviouslv shrinked and deformed,and the cells treated with 1000 μmol/L of sodium fluoride were broken-Conclusions nuoride can affect the growth and development of thyroid cell and damage the structure and morphology.Sodium fluoride affects the morphologie characteristics of thyroid cells in a dose-response manner.

Objective To investigate the relationship between the composition of vaginal microbiota and the course of cervical precancerous lesion.Methods A total of 64 vaginal swabs were collected from 22 healthy women, 18 CINⅠ patients and 24 CINⅡ/Ⅲ patients who visited Obstetrics and Gynecology of the Second Xiangya Hospital of Central South University during July 2014 and July 2015.The Bacterial genomic DNA was extracted and the V3 and V4 hypervariable regions of 16S rRNA were amplified and high-throughput sequenced.The abundance and composition of vaginal microbiota were analyzed by Uparse, Mothur and LefSe statistical software.Results There was no significant difference in Alpha diversity index between CINⅡ/Ⅲ group(Chao:63±32;ACE:72±38;Simpson:0.70±0.27;Shannon:0.70±0.63) and control group ( Chao:48±24;ACE:54±25;Simpson:0.71±0.27;Shannon:0.65±0.58)(W=192,P=0.11;W=189,P=0.10;W=281,P=0.72;W=241,P=0.62).The ACE(85±37) and Chao(66±25) values of CINⅠgroup were significantly different from those of the control group (ACE:54±25;Chao:48±24)(W=99,P=0.006;W=113,P=0.02).At the phylum level, 78.69%(309 020/392 722) of the vaginal microbiota in the control group was Firmicutes, 16%(62 846/392 722) was Actinobacteria.Firmicutes was reduced to 64.86%(208 422/321 318) and Actinobacteria increased to 27.71%(89 040/321 318) in CINⅠgroup.The composition of vaginal microbiotain in CINⅡ/Ⅲ group was similar to those of control group.At the genus level, the composition of vaginal microbiota were similar between CINⅡ/Ⅲ group and control group, with Lactobacillus as predominant genus[71.81%(307 658/418 424)], Gardnerella[12.91%(55 299/428 424)], others such as Prevotella, atopobium were less.In the CINⅠ group, the abundance of Lactobacillus was decreased to 56.26%(180 787/321 318), Gardnerella was increased to 19.62%(63 057/321 318), and Listeria was increased to 7.7%(24 746/321 318).The composition of vaginal microbiota in the most samples was classified as CSTⅢ and CSTⅠ, with Lactobacillus inersand and Lactobacillus crispatus were dominant respectively.There was no significant difference in the composition of vaginal microbiota between the three groups(χ2=2.72, P=0.949).LEfSe analysis showed that the abundance of bacteria in CIN group and control group were varied.At the genus level, there were significant differences in the abundance of Geobacter, Atopobium and Ureaplasma (P<0.05, P<0.05, P<0.01, respectively).At the species level, there was significant difference in the abundance of Ureaplasma urealyticum serotype 9 (P<0.01).Conclusion The diversity and the composition of vaginal microbiota were similar between CIN patients and healthy women, but the abundances of some bacteria were varied, with Ureaplasma increased in patients with CIN.