Objective To investigate whether the μ- and δ-opioid receptors were involved in mediating the a ntinociceptive effect of opioid injection into the nucleus submedius (Sm). Methods Nociceptive behavior produced by subcutaneous injection of formalin (65 mmol/L, 50 μL) into the hind paw of the rat was assessed quantitatively using an automated movement detection system. The effects of morphine and selectiveμ- and δ-opioid receptor antagonists microinjected unilaterally into the Sm were determined in the awake rats. Results Morphine (31 mmol/L, 0. 5 μL) depressed the nociceptive behavior elicited by formalin, and this effect was antagonized completely by the selective μ-receptor antagonist β-funaltrexamine (β-FNA, 0. 4 mmol/L, 0. 5 μL) and naloxonazine (0.8 mmol/L, 0.5 μL), and partly by the δ-receptor antagonist naltrindole (0.4 mmol/L, 0.5μL).Administration of morphine into thalamic regions more than 0. 5 mm dorsal to the Sm had no effect on the nociceptive behavior. Conclusion Antinociceptive effects produced by opioid acting on Sm neurons are mediated mainly by μ-opioid receptors, and partly by δ-receptors.

Objective To observe the changes of hippocampal NogoA-NgR1 signaling on postoperative cognitive function (POCD) in aged mice, and explore the potential underling mechanism.Methods Isoflurane anesthesia and laparotomy were applied to establish the POCD model.Forty aged male C57BL/6 mice were randomly divided into the following four groups (n=10): group O2+saline (group OS), group O2+NEP1-40 (group ON), group isoflurane anesthesia+laparotomy surgery+saline (group SS), and group isoflurane anesthesia+laparotomy surgery+NEP1-40 (group SN).Cannula placement was performed into lateral ventricle 7 days before the surgery.Animals were subjected to an administration of NEP1-40 (20 μg/2 μl) or isochoric saline via intracerebroventricular injection once daily for 8 consecutive days, injection was given from 2 h before isoflurane anesthesia to the last behavioral test.Open field test was performed at 5th d after operation.Contextual and cued fear conditioning training and testing were exhibited at 6th and 7th d after operation, respectively.The hippocampus was harvested 2 h after the behavioral test.Western blot was used to detect the expressions of NogoA, NgR1, RhoA, ROCK2 and GAP43.Golgi staining was applied to measure the changes of dendritic spines in hippocampal CA1 area.Results Compared with the groups OS and ON, the freezing time in the contextual fear conditioning test was significantly decreased, the contents of NogoA, NgR1, RhoA and ROCK2 were significantly increased, the content of GAP43 and the number of dendritic spine were significantly decreased in group SS (P<0.05).Compared with the group SS, the freezing time in the contextual fear conditioning test was significantly increased, the contents of RhoA and ROCK2 were significantly decreased, the content of GAP43 and the number of dendritic spine were significantly increased in group SN (P<0.05).Conclusion Over-activated of hippocampal NogoA-NgR1 signaling participated in the pathogenesis of POCD in aged mice.

OBJECTIVETo analyze causes of missed diagnosis of hiding post-malleolar fractures in treating ankle joint fractures of pronation-external rotation type according to Lauge-Hansen classification and assess its medium-term outcomes.

METHODSAmong 103 patients with ankle joint fracture of pronation-external rotation type treated from March 2002 to June 2010,9 patients were missed diagnosis,including 6 males and 3 females,with a mean age of 35.2 years old (ranged, 18 to 55 years old) . Four patients were diagnosed during operation, 2 patients were diagnosed 2 or 3 days after first surgery and 3 patients came from other hospital. All the patients were treated remedially with lag screws and lock plates internal fixation. After operation,ankle joint function was evaluated according to American Orthopaedic Foot and Ankle Society (AOFAS).

RESULTSAll the 9 patients were followed up, and the duration ranged from 14 to 30 months (averaged, 17 months). No incision infection was found, and all incision healed at the first stage. At the latest follow-up, AOFAS was 83.0 +/- 4.4, the score of 4 patients diagnosed during operation was 85.0 +/- 2.9, and the score of 5 patients treated by secondary operation was 81.0 +/- 5.3. All the patients got fracture union observed by X-ray at a mean time of 2.2 months after operation. There were no complications such as internal fixation loosing, broken and vascular or nerve injuries.

CONCLUSIONAnkle joint fracture of pronation-external rotation type may be combined with hiding post-malleolar fractures. So to patients with ankle joint fracture of pronation-external rotation type, lateral X-ray should be read carefully, and if necessary, CT or MRI examination should be performed. If adding lateral X-ray examination after reduction of exterior and interior ankle joint fixation, the missed diagnosis may be avoided.

Adolescent ; Adult ; Ankle Fractures ; False Negative Reactions ; Female ; Fractures, Bone ; diagnosis ; diagnostic imaging ; physiopathology ; surgery ; Humans ; Male ; Middle Aged ; Pronation ; Rotation ; Tomography, X-Ray Computed ; Treatment Outcome ; Young Adult

Aim To determine the protective effects of IMD on ischemia/reperfusion(I/R) injury and its possible mechanism.Method Isolated rat hearts were perfused by Langendorff mode,and after 45 min global ischemia and 30 min reperfusion ventricular function was measured on a Power Lab,and adequate amount of ventricular tissues and perfusate were collected for biochemical measurement.Results Treatment with IMD during the reperfusion period significantly attenuated the effects of I/R on cardiac function inhibition and tissue injury.Compared with I/R group,IMD induced increase in △LVP,LV(dp/dtmax,HR and CF,whereas induced decrease in LVDP.Reperfusion with IMD exerted decrease in LDH,total protein,Mb and MDA content compared with I/R group,but increased myocardial cAMP content.All these values were similar to the effects of ADM.Furthermore,I/R induced significant increase in Bmax and Kd value.Conclusion IMD exerted beneficial effects on cardiac injury induced by I/R which might be mediated by cAMP pathway.And the cardioprotective effects of IMD were equal to ADM,a potent cytoprotective factor.

OBJECTIVETo investigate the effect of Smad7 on the expressions of collagen I and alpha-smooth muscle actin (alpha-SMA) in HSC-T6 cell line activated by transforming growth factor-beta1 (TGF-beta1).

METHODSHSC-T6 cells stably expressing M2-flag protein were selected after co-infection of the cells with pTRE-Smad7-M2-flag and pTet-on. The optimal dose of doxycycline for inducing Smad7 was determined, and the effects of Smad7 over-expression on the expressions of collagen I and alpha-SMA in the cells activated by TGF-beta1 and on Smad2/3 phosphorylation were evaluated using Western blotting.

RESULTSThe optimal dose of doxycycline for inducing Smad7 expression was 2 mg/L. Smad7 over-expression induced by doxycycline decreased the expressions of collagen I and alpha-SMA in HSC-T6 cells activated by TGF-beta1, and down-regulated the level of Smad2/3 phosphorylation.

CONCLUSIONSmad7 over-expression inhibits Smad2/3 phosphorylation, and decreases the expression of collagen I and alpha-SMA in HSC-T6 cells induced by TGF-beta1 to inhibit the progression of liver fibrosis.

Actins ; genetics ; metabolism ; Cells, Cultured ; Collagen Type I ; genetics ; metabolism ; Genetic Therapy ; Hepatocytes ; cytology ; metabolism ; Humans ; Liver Cirrhosis ; metabolism ; therapy ; Smad7 Protein ; pharmacology ; Transforming Growth Factor beta1 ; pharmacology

Animals ; Cell Proliferation ; DNA ; genetics ; immunology ; Female ; Hepatitis B Core Antigens ; genetics ; Hepatitis B virus ; genetics ; Interferon-gamma ; blood ; genetics ; Killer Cells, Natural ; immunology ; Lymphocyte Activation ; Mice ; Mice, Inbred BALB C ; Plasmids ; genetics ; immunology ; Random Allocation ; Recombination, Genetic ; Spleen ; cytology ; immunology ; metabolism ; T-Lymphocytes ; immunology

Objective To understand the epidemiological distribution and epidemic situation of brucellosis cases in Zhejiang Province in 2003 - 2012. Methods Questionnaires of confirmed brucellosis cases, annual reports of prevention and control work of brucellosis in the cities and monitoring points were collected and analyzed in 2003 - 2012. Population distribution, regional distribution, infection sources and routes of infection, aetiology and clinical symptoms and signs of the brucellosis cases were analyzed descriptively. Results A total of 323 brucellosis cases were reported in 2003 - 2012, the average annual incidence rate was 0.070/one hundred thousand, the incidence rate in 2012(0.190/one hundred thousand) was high. There were 272 people of the 323 brucellosis cases were from occupational populations, accounting for 84.21%(272/323); people worked in buying, slaughtering and trafficking of livestock products were majority of the cases, accounting for 65.02%(210/323); and unoccupational population was accounting for 15.79%(51/323). Regional distribution of brucellosis was gradually spreading; the average incidence rate of Shaoxing City was the highest ( 0 . 226/one hundred thousand ) . Three hundred patients were infected by sheep, accounting for 92.88%(300/323), and only 7.12%(23/323) of the patients were infected by cattle. Two hundred and eighty-seven acute phase patients of brucellosis were checked by blood culture, and thirty-nine Brucella melitensis strains and two Brucella abortus strains were detected. The chronicity survey of 120 confirmed cases of brucellosis were conducted, chronicity rate was 3.33%(4/120). Conclusions Trends in the epidemic situation of brucellosis in Zhejiang Province has continued to spread. The main source of infection is sheep from the North. Occupational populations are major populations at risk. The key of prevention and control is to strengthen the active monitoring of occupation personnel , health education and behavior intervention.

Objective To explore the effect and the possible pathway of different concentrations of QLT0267,which was the inhibitor of the integrin-linked kinase (ILK),on the process of high glucose-induced tubularepithelial-myofibroblast transdifferentiation (TEMT) in human renal tubular epithelial cells (HK-2).Methods HK-2 cells were exposed to 30 mmol/L GS,and TEMT model was established.After excluding the effect of high osmotic in TEMT,HK-2 cells were divided into 6 groups by different concentrations of GS and QLT0267 for 48 hours.The rate of the cell proliferation was calculated by MTT.The expression of ILK and α-smooth muscle actin (α-SMA) were determined by immunofluorescence and Western blot,and the expression of protein kinase B (AKT),phosphorylated protein kinase B (p-AKT),and E-cadherin were determined by Western blot.Results (1) The expression of ILK,p-AKT,and α-SMA in HK-2 cells were unregulated and the expression of E-cadherin was downregulated for 48 hours with glucose treating vs control (P ＜ 0.05);(2) The proliferation rate in high glucose group was higher than the group which concentration of QLT0267 was greater than 5 μmol/L (P ＜ 0.05);(3) With the concentrations of QLT0267 increased,the expression of p-AKT,α-SMA was gradually decreased (all P ＜ 0.05),and the expression of E-cadherin was gradually increased (all P ＜ 0.05).Conclusions 30 μmol/L of GS can lead to TEMT in HK-2 cell.The QLT0267 with concentration greater than 5 μmol/L may prevent the activation of ILK downstream proteins,then partially inhibits cell proliferation and TEMT in HK-2 cell.

Objective To observe the effects of different concentrations of SB203580, the inhibitor of P38MAPK, in process of high glucose (GS)-induced renal tubular epithelial-myofibroblast transdifferentiation (TEMT). Methods The cultured human renal tubular epithelial cells (HK-2) were divided into control group (5.5 mmol/L GS), GS (30 mmol/L GS) group and different concentrations of SB203580 (30 mmol/L GS +5, 10, 20 and 30 μmol/L SB203580) groups. The treat?ments were for 48 hours. MTT assay was used to observe cell proliferation. The median inhibiting concentration (IC50) was cal?culated. Western blot assay was used to detect the expressions of P38MAPK, P-P38MAPK andα-smooth muscle actin (α-SMA) in control group, high-glucose group and S30 group. The expression ofα-SMA was also detected by the method of im?munofluorescence. Results 1.Compared with control group, there was no significant inhitory effect on proliferation rate in DMSO group (P>0.05). There were increased HK-2 cells in high glucose group and S5group (P<0.05). Proliferation rates were significantly decreased in S20 and S30 groups (P<0.05). Compared with high glucose group, the proliferation rates of HK-2 cells were inhibited in S5, S10, S20 and S30 groups (P<0.05). 2. The expression of P-P38MAPK was significantly higher in high glucose group and S30 group than that of control group (P<0.05). Compared with high glucose group, the ex?pression of P-P38MAPK was significantly decreased in S30 group (P<0.05), whereas no significant difference in the expres?sion of P38MAPK between the two groups (P>0.05). 3. Compared with control group, the expression ofα-SMA was signifi?cantly increased in high glucose group and S30 group (P<0.05). Compared with high glucose group, the expression of α-SMA was significantly decreased in S30 group (P < 0.05). Conclusion The 30 mmol/L GS can lead to TEMT in HK-2 cells. The more suitable inhibitory concentration of SB203580 in the process of TEMT is 30μmol/L. SB203580 can slow down the process of TEMT by inhibiting P38MAPK activation and inhibiting proliferation and the expression ofα-SAM s of HK-2 cells.

Objective To investigate the effects of 1,25-dihydroxyvitamin D3[1,25 (OH)2D3] on cell proliferation in hu?man glomerular mesangial cells and it′s effects on the regulation of mTOR/p70s6K signaling pathway in this cell line. Meth?ods The cultured human mesangial cells at passage 3-7 were divided into four groups:control group,VD group (addition of 10-8 mol/L of 1,25-dihydroxyvitamin D3 ),R group (addition of 5 mg/L of rapamycin) and R+VD group(addition of 5 mg/L ra?pamycin combined with 10-8 mol/L of 1,25-dihydroxyvitamin D3). Drug incubation last 48 h. The effect of mesangial cell pro?liferation was measured by CCK-8 colorimetric assay. The cell cycles were measured by flow cytometry. The expression of mTOR and p70s6K were detected by immunofluorescence. Results (1) The absorbance of A450 was higher in control group than that in VD group than that in R group than that in R+VD group. But the inhibition rate (IR) was lower in control group than that in VD group than that in R group than that in R+VD group. All comparisons were of statistic significance. ( 2) Cells in G1 phase were higher while cells in G2/M and S phases as well as proliferation rate (PI) were lower in control group than those in VD group than those in R group than those in R+VD group. All comparisons were of statistic significance except in?dexes between group R and group VD. (3) mTOR and p30s6K expressions in mesangial cells were higher in control group than those in VD group than those in R group than those in R+VD group. All comparisons were of statistic significance ex?cept indexes between group R and group VD. Conclusion 1,25-dihydroxyvitamin D3 might inhibit mesangial cell prolifera?tion significantly through mTOR/p70s6K signaling pathways.