Objective To investigate the distribution relationship of microglia and neuron in rat medullary visceral zone(MVZ) after stomatic nociception. Methods After 2ml formalin(2.5%)injection in rat stomach by tiny plastic tube,Anti-Fos protein(labeling neuron)and anti-OX 42(labeling microglia)single and double immunohistochemical ABC method staining on model medulla oblongata sections in different time point(0.5h,1h,2h and 3h) were used. Results 1.After wakefulness,the animal models appear restlessness and agitation in 1-2h.2.We found hemorrhage and exfoliation in model's gastric mucosa.3.Fos positive neurons and OX 42 positive microglias were observed in MVZ,whose distribution was congruous and appeared clearly character of locations.4.The results of immunochemical diplex staining indicate that many Fos positive neurons were surrounded by intensive OX 42positive microglias.5.OX 42 positive microglia showed a tendency,which increased in 0.5-2h and decreased in 3h.Conclusion Neurons and microglia in MVZ may be involved in visceral nociception transfer and adjustment function.

Objective To investigate the expression of GFAP in rat hypothalamus after acute heat stress. Methods The rats were caged in a experimental incubator for 60 minutes,the temperature within the incubator was adjusted to 24℃,34℃,38.5℃ or 42℃,the humidity was 60%.Single anti-GFAP immunohistochemical(ABC) method and anti-Fos and GFAP double immunohistochemical method were used to observe the expression of GFAP in hypothalamus in different ambient temperatures after heat stress. Results The GFAP-positive cells were rare in hypothalamus at 24℃,however it was increased in many nuclei(anterior hypothalamic area,paraventricular nucleus,arcuate nucleus,suprachiasmatic nucleus and supraoptic nucleus)at 34℃ and peaked when ambient temperature was 38.5℃,and then decreased.However,Fos/GFAP-IR double labelled astrocytes were observed at 42℃.Conclusion Astrocytes participate in the pathophysiological process of heat stress.

Objective To investigate the mechanisms of astrocytes modulating neurons in rat supraoptic nucleus induced by hypotonic stimulation and the effect of 6-aminomethyl-3-methyl-4H-1,2,4-benzothiadiazine-1,1dioxide HCl(TAG,a antagonist for taurine) or carbenoxolone(CBX,a gap junction blocker)on the responses of astrocytes and neurons in SON.Methods Adult male Sprague-Dawley rats were divided into four groups: the control group was injected with 5.5ml/kg 0.9% NaCl solution into the caudal vein;the hypotonic group was injected with 5.5 ml/kg hypo-saline(0.83% glucose plus 0.3% NaCl);TAG + hypotonic and CBX + hypotonic groups were injected with TAG(100?mol/L) or CBX(10g/L) into the lateral ventricle respectively,and were injected 2 hours later with hypotonic saline into the caudal vein.With anti-Fos,anti-vasopressin(VP),anti-glycine receptor(GlyR),anti-glial fibrillary acidic protein(GFAP) and anti-connexin43(Cx43) immunofluorescent staining methods,the responses of neurons and astrocytes in SON were studied.Results In control rats,Fos-,VP-,and GlyR-expression in the neurons and GFAP-or Cx43-expression in the astrocytes were lower.In hypotonic rats,GFAP-,Cx43-and GlyR signals were more than those in control rats,while Fos-and VP-signals were less.Compared with those in hypotonic rats in TAG + hypotonic or CBX + hypotonic rats,GFAP-and Cx43-signals in the astrocytes were the same,GlyR-signals in the neurons decreased,and Fos-and VP-signals increased.Conclusion Hypotonic stimulation activates SON astrocytes,which then release taurine through gap junction signaling to the neurons and inhibit the release of VP from the neurons.

AIM: To investigate the cause and the description of pathogenic characteristics of chronic infective keratitis in different age groups.

METHODS: Totally 89 patients(89 eyes)with chronic infective keratitis were diagnosed in our hospital from January 2015 to December 2017 were selected as the research objects and the cause of the disease and pathogenic characteristics of different age groups, including 13-32 years group(35 eyes in 35 patiens), 33-52 years group(28 eyes in 28 patiens)and 53-73 years group(26 eyes in 26 patiens), were retrospectively studied.

RESULTS: There were no significant differences in the etiological distribution of patients in the three groups(P=0.290). There was no significant difference in the distribution of microbial composition among the three groups(χ²=0.258, P=1.000). The incidence of chronic infectious keratitis caused by fungal infection in the three groups was higher than that of other pathogenic microbes. In this study, 26 cases were positive in bacterial culture, including Staphylococcus epidermidis(50%), Pseudomonas aeruginosa(15%), Streptococcus salivarius(12%), Streptococcus pneumoniae(8%), Escherichia coli(8%)and Neisseria gonorrhoeae(8%). Pseudomonas aeruginosa and Staphylococcus epidermidis were the common bacterial strains in the positive culture results. In the cultivation of fungal strains 24 were positive cultures including Fusarium(46%), Aspergillus(17%), Alternaria(21%), Mycelium(no spore,4%), Penicillium(4%)and Curvularia(8%). Two strains of Fusarium spp. and Alternaria spp. were high of all patients.

CONCLUSION: There were no significant differences in the etiology of infectious keratitis among different age groups. Fungal infection is the main pathogen in the distribution of pathogens, the bacteria were mainly Streptococcus epidermidis in etiological features but in fungi, Fusarium dominates. So early prevention and prevention of infective keratitis should be strengthened and early treatment should be given.

OBJECTIVETo investigate whether hypertonic saline (HS) can induce the synthesis and release of glutamate in cultured hypothalamic astrocytes or C6 cell line.

METHODSAstrocytes were isolated, cultured, purified and identified from the hypothalamus of newborn rat (1 day). The astrocytes were randomly divided into five groups: isotonic (IS) and HS groups, astrocytes were incubated by IS and HS (320 mosM NaCl) medium, respectively, for 1, 3, 5, 10 or 15 min; carbenoxolone (CBX)+IS and CBX+HS groups, astrocytes were pre-treated with CBX (100 mmol/L) for 1 h at 37 degrees C in a 5% CO(2) / 95% atmosphere, then removed to IS and HS medium, respectively, for 1, 3, 5, 10 or 15 min; Ca(2+)+HS group, astrocytes were pre-incubated with Ca Ca(2+) (1,000 micromol/L) for 1 h at 37 degrees C in a 5% CO(2) / 95% atmosphere, followed by a wash with isotonic FBS/DMEM, and then removed to hypertonic saline for 1, 3, 5, 10 or 15 min. The media of five groups were collected to analyze the medium glutamate concentration with high performance liquid chromatography. The astrocytes were fixed and double immunofluorescent stained with anti-glial fibrillary acidic protein (GFAP) and anti-glutamate. The C6 cells were divided into four groups: IS, HS, CBX+IS and CBX+HS groups, and used for quantitative measurement of glutamate in cells by flow cytometry (FCM).

RESULTS(1) Anti-GFAP immunofluorescent signal revealed no significant difference among various time points in each group, or among the five groups. (2) The anti-glutamate immunofluorescent signal was increased in HS group and peaked at 5 min, and decreased and returned to the level of IS group at 15 min (P < 0.01 vs the 5 min of HS group). In CBX+HS group, the glutamate intensity was higher than that in CBX+IS and HS groups. (3) The medium glutamate concentration had no change after treatment with HS for 1 and 3 min, while increased markedly after treatment for 5 min to 15 min (P< 0.01 vs 1 min and 3 min). On the contrary, the medium glutamate concentrations in the CBX+HS or Ca(2+)+HS group were significant lower than that in the HS group (P < 0.01). (4) FCM showed HS and CBX+HS induced glutamate increase in C6 cells.

CONCLUSIONHS induced cultured rat hypothalamic astrocytes or C6 cells to synthesize and release glutamate; CBX could block glutamate release, but could not disrupt glutamate synthesis.

Analysis of Variance ; Animals ; Animals, Newborn ; Astrocytes ; drug effects ; metabolism ; Calcium ; pharmacology ; Carbenoxolone ; pharmacology ; Cells, Cultured ; Chromatography, High Pressure Liquid ; methods ; Flow Cytometry ; Gene Expression Regulation ; drug effects ; Glial Fibrillary Acidic Protein ; metabolism ; Glutamic Acid ; metabolism ; Hypothalamus ; cytology ; Rats ; Saline Solution, Hypertonic ; pharmacology ; Time Factors

OBJECTIVEGap junctions, the clusters of intercellular channels, play an important role in synchronizing electrical activity. This study investigated the effect of gap junction blocker carbenoxolone (CBX) on epileptic activity in pentylenetetrazo (PTZ)-kindled rats.

METHODSThirty adult male SD rats were randomly divided into three groups: control, PTZ-kindled and CBX-treated groups (n=10 each). The rats from the PTZ-kindled and the CBX-treated groups were intraperitoneally injected with PTZ (35 mg/kg x d) to induce epilepsy. After epilepsy kindling, they were intraperitoneally injected for 3 days with CBX (10 mg/kg) (CBX-treated group) or with normal saline (PTZ-kindled group). The control group received intraperitoneal injections of normal saline. Anti-GFAP, anti-Fos, and anti-NMDARZ immunohistochemical ABC methods were used to detect the expression of GFAP-Li, Fos-Li and NMDAR2-Li in the hippocampus respectively.

RESULTSSpontaneous seizures occurred in PTZ-kindled epileptic rats. CBX administration reduced spontaneous seizures. The NMDAR2-Li and Fos-Li neurons as well as GFAP-Li astrocytes in hippocampi increased in PTZ-kindled epileptic rats compared with controls. The numbers of Fos-Li (93.75 +/-7.94 vs 165.25 +/-15.87, P < 0.05) and NMDAR2-Li neurons (61.47 +/-3.62 vs 148.72 +/-14.53, P < 0.01) in the CBX-treated group were significantly less than in the PTZ-kindled group. There were no significant differences in the GFAP-Li expression between the CBX-treated and the PTZ-kindled groups.

CONCLUSIONSCBX may inhibit spontaneous seizures and decrease the numbers of Fos-Li and NMDARZ-Li neurons, thus providing anti-epileptic effects.

Animals ; Carbenoxolone ; pharmacology ; Epilepsy ; drug therapy ; metabolism ; Gap Junctions ; drug effects ; Glial Fibrillary Acidic Protein ; analysis ; Hippocampus ; drug effects ; metabolism ; Immunohistochemistry ; Kindling, Neurologic ; drug effects ; metabolism ; Male ; Pentylenetetrazole ; Proto-Oncogene Proteins c-fos ; analysis ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; analysis

OBJECTIVETo assess the relationship between overweight, obesity and blood lipid profiles of children and adolescents and to validate body mass index (BMI) cutoff points for overweight and obesity screening to Chinese children and adolescents, recommended by Working Group of Obesity, China (WGOC), International Life Science Association.

METHODS2293 children and adolescents (1124 males and 1169 females), aged between 10 and 18 years, were randomly selected as samples from 6 schools in Beijing area. Fasting serum lipids including total cholesterol (TC), total triglycerides (TG), high density lipoprotein cholesterol (HDLC), thropometrical index as weight and height were measured. BMI equals to weight in kilograms were then divided by the square of height in meters.

RESULTSAccording to BMI cutoff points recommended by WGOC, samples fell into 3 groups including normal group (BMI < 85 percentiles), overweight group (BMI >or= 85 and < 95 percentiles) and obesity group (BMI >or= 95 percentiles). Results clearly showed an increase of both serum TC and TG and a decrease of HDLC when BMI was increasing, among most age groups regardless of sex difference and the difference among BMI groups was statistically significant (P < 0.05 or P < 0.01).

CONCLUSIONSResults of this study indicated that there was an obvious dose-effect relationship between BMI and lipid profiles which accounted for some rationality of the BMI cutoff points recommended by WGOC. The authors reckoned the findings important to managing relevant adult diseases during childhood, in China.

Adolescent ; Biomarkers ; blood ; Body Mass Index ; Child ; China ; Cholesterol ; blood ; Cholesterol, HDL ; blood ; Female ; Guidelines as Topic ; standards ; Humans ; Lipids ; blood ; Male ; Obesity ; diagnosis ; Reproducibility of Results ; Triglycerides ; blood

OBJECTIVETo study the changes in the plasticity of the neurons and astrocytes in the paraventricular nucleus (PVN) and supraoptic nucleus (SON) of the hypothalamus of rats exposed to a humid and hot environment.

METHODSThe rats were subjected to stimulation with a humid and hot environment for 120 min in a climate chamber (dry bulb temperature of 40.0-/+0.5 degrees C with relative humidity of 60-/+5%). During the exposure, the behavioral responses of the rats were observed, and the changes in the expressions of Fos and GFAP in the PVN and SON in response to the exposure evaluated using immunohistochemical ABC methods.

RESULTSExposure to a humid and hot environment caused restlessness and agitation in the rats, which showed increased respiratory frequency and scratching of the face with the forelimbs. Two rats died after the 120-min exposure. Significantly increased expressions of Fos and GFAP were detected in the PVN and SON following the exposure as compared with the control group.

CONCLUSIONThe neurons and astrocytes in the PVN and SON both participate in the regulation of responses to exposure to a humid and hot environment.

Animals ; Astrocytes ; cytology ; physiology ; Glial Fibrillary Acidic Protein ; analysis ; Hot Temperature ; Humidity ; Hypothalamus ; cytology ; metabolism ; Immunohistochemistry ; Male ; Neuronal Plasticity ; physiology ; Neurons ; cytology ; physiology ; Oncogene Proteins v-fos ; analysis ; Paraventricular Hypothalamic Nucleus ; cytology ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Supraoptic Nucleus ; cytology ; metabolism

OBJECTIVETo investigate the effects of liver X receptor (LXR) agonist on adipose-derived mesenchymal stem cells (AD-MSCs) implantation into infarcted hearts of mice.

METHODAD-MSC(Fluc+) which stably expressed firefly luciferase (Fluc) were isolated from β-actin-Fluc transgenic mice and characterized by flow cytometry. Male FVB mice were randomly allocated into the following four groups (n = 10 each): (1) sham group; (2) MI + PBS group; (3) MI + AD-MSC(Fluc+) group; (4) MI + AD-MSC(Fluc+) + LXR agonist (T0901317) group. AD-MSC(Fluc+) or PBS were injected intramyocardial into peri-infarcted region of mice heart after permanent left anterior descending (LAD) artery ligation. Bioluminescence imaging (BLI) was performed for quantification of injected cells retention and survival. Cardiac function was evaluated by echocardiography.

RESULTSThe AD-MSC(Fluc+) were positive for CD44 and CD90 by flow cytometry. BLI evidenced the firefly luciferase expression of AD-MSC(Fluc+) which was positively correlated with cell numbers (r(2) = 0.98). The results of BLI in vivo revealed that LXR agonist could improve the survival of AD-MSC(Fluc+) at day 7, 14 and 21 after transplantation compared with AD-MSC(Fluc+) alone group. Cardiac function was further improved in combination therapy group compared with AD-MSC(Fluc+) alone group (P < 0.05).

CONCLUSIONSLXR agonist T0901317 can improve the retention and survival of intramyocardial injected AD-MSC(Fluc+) post-MI, and the combination therapy of T0901317 and AD-MSC(Fluc+) has a synergetic effect on improving cardiac function in this model.

Animals ; Hydrocarbons, Fluorinated ; therapeutic use ; Liver X Receptors ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Mice ; Mice, Transgenic ; Myocardial Infarction ; mortality ; surgery ; Orphan Nuclear Receptors ; agonists ; Sulfonamides ; therapeutic use ; Treatment Outcome

OBJECTIVETo investigate prevalence of Saffold virus (SAFV) in Changsha area of hospitalized children with respiratory tract infection, and to discuss whether this virus is related to respiratory tract infection of children.

METHODS643 nasopharyngeal aspirates samples were collected from hospitalized children with respiratory tract infection of the first affiliated hospital of Hunan nomal university during Nov. 2007 to Oct. 2008. Real-time fluorescent quanti-tative PCR(FQ-PCR) performed to screen the 5'UTR gene. And then analyze clinical data.

RESULTSSAFV were detected in 67 patients (10.42%) out of the 643 children, it was not detected over 5 years of age. The virus were detected in 8 patients (25.81%) out of the 31 children with persistent pneumonia and chronic pneumonia, there was statistically significant.

CONCLUSIONThere existed SAFV infection in hospitalized children with lower respiratory infection in Changsha area; SAFV maybe related to disease onset with lower respiratory tract infection of children.

Adolescent ; Cardiovirus ; genetics ; isolation & purification ; Child ; Child, Preschool ; China ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Polymerase Chain Reaction ; Respiratory Tract Infections ; virology