Objective To investigate the expression of regeneration gene protein-2(Reg-2) after the transection injury in rat spinal cord. Methods Forty five SD rats were randomly divided into two groups: in the injury group,the SCI was produced by complete transection of the cord at the 9(th) thoracic level,and the sham operation rats were taken as the normal control.The animals were perfused at 1h,1d,3d,7d,14d after the operation,and the spinal cord was taken out at different time points.The expressions of Reg-2 were tested by immunohistochemical analysis and Western blotting,the double staining of Reg-2 and different neural cells special markers(GFAP,Olig2,NPY,CGRP,GAP-43,et al)was done with immunofluorescence method. Results The expression of Reg-2 was found at 1h after the spinal cord injury,reached the peak on day 3 after the injury in the neurons of dorsal horn and day 7 in the neurons of ventral horn.The high expression persisted for 1 week,then decreased gradually.Conclusion Reg-2 may work as an important growth factor and participate in the regeneration and rehabilitation process after the spinal cord injury.

Objective To study the expression of substance P(SP) and substance P receptor(SPR) during the development of mice brains. Methods The expression of SP and SPR during the development of mice brains from embryonic day(E) 11 to postnatal day(P) 0 days was analyzed by immunohistochemical method. Results The expression of SP began at E11 and gradually increased until birth. The expression of SPR began at E11 and maintained stable expression until birth. SP mostly expressed at striatum and SPR mostly expressed at medullary raphe.Conclusion The expression of SP and SPR during the embryo brain stage may indicate that SP could be an important factor involved in the early organization and maturation of neuron.

Acupuncture Therapy ; Adult ; Aged ; Female ; Humans ; Irritable Bowel Syndrome ; therapy ; Male ; Middle Aged ; Scalp ; Young Adult

OBJECTIVE This study was to investigate the effects of CP- 25 on the functions of activated human B cells through regulating BAFF and TNF-alpha signaling. METHODS B cells from peripheral blood mononuclear cells (PBMCs) of normal human were isolated using magnetic cell separation (MACS) by a positive selection. B cells (107 cells·mL-1) were stimulated by BAFF (100 ng·mL-1) or TNF-alpha (100 ng·mL-1) for two hours, and then were treated with CP-25 (10-5 mol·L-1) or Rituximab (5 μg·mL-1) or Etanercept (10 μg·mL-1). B cell proliferation was detected by CCK-8. B cell subsets and BAFF receptors (BAFFR, BCMA and TACI) were analyzed by flow cytometry. The expression of TNFR1 and TNFR2 on B cells was analyzed by flow cytometry. The expression of MKK3, MKK6, P-p38, P-p65, TRAF2 and p100/52 was analyzed by Western blotting. RESULTS CP-25 inhibited B cells proliferation stimulated by BAFF or TNF- alpha. CP- 25, Rituximab and Etanercept reduced the percentage and numbers of CD19+ B cells, CD19+CD20+ B cells, CD19+CD27+ B cells and CD19+CD20+CD27+ B cells induced by BAFF or TNF-alpha. CP-25 down-regulated the high expression of BAFFR, BCMA and TACI stimulated by BAFF or TNF-alpha. CP-25, Rituximab and Etanercept down-regulated significantly the expression of TNFR1 and TNFR2 on B cell stimulated by BAFF or TNF-alpha. CP-25, Rituximab and Etanercept down-regulated the expression of MKK3, P-p38, P-p65, TRAF2 and p52 in B cells stimulated by BAFF and the expression of TRAF2 and P- p65 in B cells stimulated by TNF-alpha. CONCLUSION CP- 25 regulated moderately activated B cells function by by regulating the classical and alternative NF-κB signaling pathway mediated by BAFF and TNF-alpha-TRAF2-NF-κB signaling pathway. This study suggests that CP-25 may be a promising anti-inflammatory immune and soft regulation drug.

OBJECTIVETo investigate the expression of recombination activating gene-1 (RAG-1) and its localization in the mouse brain during the embryonic development.

METHODSThe brain tissues of E (embryonic day) 11, E13, E15, E17, E19, P0 (the birth day) and adult mice were taken, the total RNA of brains were extracted and the changes of RAG-1 expression were detected with the method of RT-PCR. The freeze sections of brain tissues from each group were stained with immunohistochemistry method.

RESULTThe expression of RAG-1 persisted from E11 to P0 brain and was steadily increased from E11 to E19; the results of RT-PCR were similar to that of immunohistochemistry. The positive-cells mainly appeared in the nucleus amygdalae, hypothalamus, thalamus and hippocampus at developmental stage. The expression began to appear in ventricular zone (VZ) and intermediate zone (IZ) of telecephalic vesicle, then gradually increased in subventricular zone (SVZ), corticle plate (CP) and subcorticle plate (SP).

CONCLUSIONThe expression of RAG-1 in mouse embryonic brain tissue is higher than that in the adult mouse, which may be related to the process of neuron development.

Animals ; Brain ; cytology ; embryology ; metabolism ; Female ; Gene Expression Regulation, Developmental ; Homeodomain Proteins ; genetics ; metabolism ; Immunohistochemistry ; Mice ; Mice, Inbred ICR ; Neurons ; cytology ; metabolism ; Pregnancy ; Reverse Transcriptase Polymerase Chain Reaction ; Time Factors

Objective To investigate the effects of matrix metalloproteinase-9 (MMP-9) inhibitor minocyclin hydrochloride in Lewis rats with experimental autoimmune myocarditis (EAM). Methods EAM was induced by injection of cardiac C protein emulsified in completed Freund adjuvant in double footpad and intra peritoneal injection of pertussis toxin on 6- to 8-week old Lewis rats. Sixty EAM Lewis rats were dividedinto 3 groups (early, middle and late intervention groups, n =20 each: 10 minocyclin treated and 10 control rats). In early intervention group, rats in treatment group received intraperitoneal injection of minocyclin hydrochloride from 1st to 21st day after immunization; in middle intervention group, rats were treated from 8th to 28th day after immunization and in late intervention group, rats were treated from 15th to 35th day after immunization (50 mg/kg body weight, once daily). Control rats received intraperitoneal injection of same volumetric physiological saline at corresponding time periods. At the end of intervention, rats were euthanatized and hearts were harvested. Paraffin sections were used for hematoxylin and eosin stain to determine the inflammatory score, for picrosirius stain to determine fibrosis score and collagen content, and for immunohistological stain to determine macrophages and T lymphocytes. Real time PCR was used to detect mRNA expression of myocardial MMP-2 and MMP-9. Cryostat sections were used for in situ zymography to detect protein activity of gelatinase. Results Inflammatory score in cardiac paraffin slides, number of cardiac macrophages and T lymphocytes, cardiac interstitial fibrosis score and content, expression of MMP-2, 9 mRNA and activity of gelatinase in treatment group were all significantly lower than in control group for early and middle intervention groups ( inflammatory score: early control group vs. treatment group: 3.03 ± 1.35 vs. 1.51 ±0. 36,P ＜0. 05, middle control group vs. treatment group: 3.75 ±0. 29 vs. 2. 11 ±0. 82,P ＜0. 01; cardiac interstitial fibrosis score, early control group vs. treatment group: 2. 75 ±0. 29 vs. 1.51 ± 0.35, P＜0.01, middle control group vs. treatment group: 2.50 ±0.41 vs. 1.61 ±0.42, P＜0.05;gelatinase, early control group vs. treatment group: 162 367 ±5095 vs. 62 366 ±2131, P ＜0. 01, middle control group vs. treatment group: 184 256 ±5427 vs. 113 197 ±4809, P ＜0. 01 ) while these parameters were similar between minocyclin-treated and control rats in late intervention group ( all P ＞ 0. 05 ).Conclusions MMP-9 plays an important role in the pathogenesis of autoimmune myocarditis. Inhibition of MMP-9 in early and middle stage could significantly attenuate inflammatory responses and myocardial fibrosis in this experimental EAM model.

Objective To study changes of glial fibrillary acidic protein (GFAP) expression and neural apoptosis in rat hippocampus and cortex of cesarean delivered offspring.Methods Thirty-eight pregnant SD rats were randomly allocated into 2 groups: 19 rats in vaginal delivery (VD) and 19 rats in cesarean section (CS).Forty-eight fetuses born by VD were kept intact, 40 fetuses were delivered by CS on day 21 of gestation.The fetal brain tissues were taken out on postnatal day 30 and 115, the expression profiles of GFAP in hippocampus and cortex were measured by immunohistochemical staining and western blot Apoptotic cells were detected using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining.Results (1) The expression profiles of GFAP: on postnatal day 115, the mean number of GFAP-immunoreactive astrocytes of hippocampus 29.7 ± 10.9 in VD group was significantly lower than 36.2 ± 2.8 in CS group ( P ＜ 0.05 ).The average GFAP-positive cells in the cortex of frontal lobe of 23.2 ±4.6 in VD group was significantly lower than 36.8 ± 5.9 in CS group (P ＜0.01 ).Likewise, on postnatal day 30, the mean number of GFAP-immunoreactive astrocytes of frontal cortex of 27.8 ± 6.0 in VD group was remarkably lower than 39.4 ± 4.5 in CS group ( P ＜ 0.01 ).The average GFAP-positive cells in the hippocampus of 31.5 ±3.5 in VD group were not significantly lower than 37.2 ±7.0 in CS group ( P ＞0.05 ).The expression of GFAP was detected in hippocampus and frontal cortex by western blot, however,there was no significant different expression of GFAP between VD group and CS group.(2) Neuronal apoptosis: TUNEL staining results indicated that, on postnatal day 115, fewer apoptotic cells scattered in offspring hippocampas subregion were only shown in CS group, never in VD group.No TUNEL positive staining cells were labeled in hippocampal subregion in VD group, therefore significantly lower than that of CS group ( P ＜ 0.05 ).Conclusions There were different influences of cesarean section on GFAP expression in hippocampus or cortex in different developmental stage of offspring Cesarean section might increase GFAP expression in the hippocampus and frontal cortex, even trigger neuronal apoptosis of hippocampus region.

OBJECTIVETo detect the expression of CD95 and CD95L mRNAs after liver transplantation and investigate the role of CD95 and CD95L in acute liver allograft rejection.

METHODSThe expressions of CD95 and CD95L mRNAs of peripheral blood lymphocyte from 56 liver allograft recipients were examined using SYBR real-time PCR.

RESULTSCD95 and CD95L mRNA levels in the recipients with acute rejection were significantly higher than those without rejection (P<0.01), and the elevation occurred about 2 days earlier than that of alanine aminotransferase and aspartate aminotransferase.

CONCLUSIONCD95 and CD95L are related to acute liver allograft rejection and their mRNA expression level may serve as an indicator for prediction and diagnosis of acute rejection episodes.

Acute Disease ; Adult ; Aged ; Fas Ligand Protein ; genetics ; Female ; Gene Expression ; Graft Rejection ; diagnosis ; genetics ; Humans ; Liver Cirrhosis ; surgery ; Liver Neoplasms ; surgery ; Liver Transplantation ; Lymphocytes ; metabolism ; Male ; Middle Aged ; Organic Chemicals ; Polymerase Chain Reaction ; methods ; RNA, Messenger ; biosynthesis ; genetics ; fas Receptor ; genetics

Objective: To explore the determinants for health-seeking behavior of the residents after cough for more than 3 weeks in Yiwu, Zhejiang Province, in order to provide reference for prevention and control of respiratory diseases. Methods: A multi-stage cluster random sampling method was used to recruit the community residents aged 5 years and above in Yiwu. A face-to-face questionnaire survey was conducted to collect demographic information, features of cough and health-seeking behaviors in the past month. The multivariate logistic regression model was employed to analyze the associated factors for health-seeking behavior after a cough for more than 3 weeks. Results: Among 6 374 residents investigated, 152 cases had a cough for more than 3 weeks in the past month, accounting for 2.48%. They were( 45.00±21.15 ) years old, including 70 ( 46.05% ) males and 82 ( 53.95% ) females. About 58.55% ( 89 ) of them sought medical treatment. The results of multivariate logistic regression analysis showed that females ( OR=2.100, 95%CI: 1.005-4.391 ), middle school education level ( OR=0.406, 95%CI: 0.168-0.983 ), family annual income of 100 000 to 199 999 yuan ( OR=2.993, 95%CI: 1.215-7.373 ) were associated factors for health-seeking behavior after a cough for more than 3 weeks. Conclusion The rate of health-seeking behavior after a cough for more than 3 weeks among the residents in Yiwu is 58.55%, which is associated with gender, education level and income.

BACKGROUND: Carotid intima media thickness (CIMT) and stiffness are taken as useful surrogate markers of atherosclerosis. In China, the number of elderly patients undergoing hemodialysis has increased year by year, with the increase of dialysis-related cardiovascular events. This study was undertaken to examine carotid stiffness in elderly hemodialysis patients by the ultrasound techniques in order to find out the possible risk factors. METHODS: From January 2006 to February 2010, a total of 87 patients (41 males and 46 females) treated with routine hemodialysis at the 97th Hospital of People's Liberation Army were enrolled in this study. The distensibility coefficient (DC) of the carotid artery was detected by Doppler ultrasonic diagnosis apparatus (Philips HBI5000, frequency 12 MHz) for evaluation of arterial stiffness. Serum albumin, total cholesterol (TC), high density lipoprotein (HDL), low density lipoprotein (LDL), triglyceride (TG), glucose, creatinine, calcium, phosphorus, and intact parathyroid hormone (iPTH) were examined with standard methods. The liner correlation and multiple stepwise regression analysis were used to find correlations between them. RESULTS: In this study, the systolic blood pressure was 153.33±25.98 mmHg, DBP 84.22± 10.39 mmHg, TC 4.39±1.05 mmol/L, TG 1.36±0.72 mmol/L, LDL 2.47±0.77 mmol/L, Cr 889.82± 207.38 mol/L, Glu 5.36±1.87 mmol/L, Ca I 2.00±2.19±0.21 mmol/L, and DC 13.39±5.32×10-3/kPa. DC was associated with age (r =-0.459, P<0.001), SBP (r =-0.527, P<0.001), and serum calcium (r =-0.273, P=0.011). The multiple stepwise regression analysis showed that SBP, age, increased serum calcium level, and diabetes were independent risk factors for decreasing DC. CONCLUSION: Systolic blood pressure, age, increased serum calcium level and diabetes in elderly hemodialysis patients are independent risk factors for increased carotid arterial stiffness.