Objective To investigate the contents of cAMP and cGMP in the central nervous system in morphine dependent and withdrawal rats.Methods A physical morphine dependent model in rats was established by subcutaneous injection of morphine in gradually increasing doses.The cAMP and cGMP contents in the brain regions were determined by radioimmunoassay.Results Compared with control group,morphine dependence could significantly decrease the cGMP content or profoundly increase the cAMP content and the cAMP/cGMP ratio in rat striatum,diencephalons,midbrain,pons and hippocampus,but these changes described above were not detected in cerebellum.Compared with morphine dependent group,naloxone induced withdrawal could significantly decrease the cGMP contents or increase the cAMP contents and the cAMP/cGMP ratio in striatum and hippocampus,but these changes described above were not observed in the other regions.Conclusion The changes of the cAMP and cGMP contents may be one of the important molecular mechanisms leading to morphine dependence and abstinence.

Objective To investigate the role of Stat1 in pathological process of nerve cells apoptosis induced by diffuse axonal injury (DAI) on rats. MethodsThe DAI model was established by using an injury model adapted from Marmarou et al. in 1994. All animals were divided into three groups, including control group, mock group and test group sacrificed on 6, 12, 24, 48, 72, 120 and 240 hours post injury (hpi). The paraffin-embedded sections of brain tissue were processed for HE staining and Bielschowsky’s silver method. Cell apoptosis was examined by flow cytometry and the expression of bax and bcl-2 were analyzed by RT-PCR. And Phospho-Ser727 Stat1 expression was examined by immunohistochemistry in different brain regions. ResultsThere was no brain contusion within HE staining, however, waving and enlargement of axons were observed within Bielschowsky’s silver method. The apoptotic rate of brain cells as well as PCR products ratio of bax to bcl-2 was highest at 24 hpi and decreased with time. An up-regulation of Phospho-Ser727 Stat1 at 6 hpi was discernible, and then reached the top at 24 hpi in cortex, cerebellum, brain stem and corpus callosum, and at 12 hpi in hippocampus. This increase was associated with the nerve cells apoptosis, r=0.921. In addition, the Phospho-Ser727 Stat1 positive cells were neurons and glial cells assessed from morphous. ConclusionsOur data indicate that Stat1 may contribute to the apoptosis of DAI on rats. In addition, the expression of Phospho-Ser727 Stat1 in glial cells suggested that glial cells may play an important role in the pathogenic mechanism of DAI.

Objective To investigate the changes of the white matter in adolescent depression by using the method of tract-based spatial statistics (TBSS). Methods We employed TBSS to examine WM microstructure in 35 treatment-naive adolescents with clinical depression and in 40 matched controls. By using the TBSS, we compared the difference of fractional anisotropy (FA), axial diffusivity (AD), radial diffusivity (RD) and mean diffusivity (MD) between theadolescent patients with depression and the controls. Results Our analysis revealed the abnormal WM microstructures in the clinically depressed adolescents. The whole-brain analysis revealed that patients, with lower FA values in the body of the corpus callosum (CC) (P < 0.01), had elevated RD and MD (P < 0.01), and preserved AD (P > 0.05). The FA values in the body of the corpus callosum was negatively correlated with the severity of depression (P < 0.01). Conclusions Our findings suggest that WM abnormalities are involved in the path-physiology of depression. Importantly , our findings show that these WM abnormalities present early in the course of the disorder.

Aim To investigate the hippocampal neuronal injury induced by ?-amyloid peptide ( A?) in rats and the candidate contribution of JNK signal transduction pathway to A? toxicity. Methods Rats were bilaterally injected with A?1 -40 into hippocampi CA1 area. The pathological changes,survival and apoptosis,and ultrastructure of hippocampal neurons,as well as p-JNK positive cells were observed by HE staining,Nissl staining,TUNEL staining,immunohistochemistry and transmission electron microscopy,respectively. Results Thepyramidal cells in hippocampus arranged loosely with decreased cell number. The gliacytes significantly proliferated. Significant apoptotic features were observed in CA1 ultrastructure. TUNEL-positive cells and p-JNK-positive cells significantly increased ( P

Objective To evaluate the effects of dexmedetomidine on the expression of Toll-like receptor 2 (TLR2) and TLR4 in monocytes of blood during the perioperative period in the patients undergoing pulmonary lobectomy.Methods Fifty patients of both sexes, aged 40-64 yr, with body mass index of 20-24 kg/m2 , of ASA physical status Ⅰ or Ⅱ , scheduled for elective pulmonary lobectomy under general anesthesia, were randomized into 2 groups (n =25 each) using a random number table: control group (group C) and dexmedetomidine group (group D).In group D, dexmedetomidine was infused intravenously at dose of 1.0 μg/kg over 10 min before induction of anesthesia, followed by continuous infusion at a rate of 0.3 μg · kg-1 · h-1 until 30 min before the end of operation, while the equal volume of normal saline was given in group C.After admission to the operating room, at 1.5 h after beginning of operation, at the end of operation, and at 12 and 24 h after operation, blood samples were taken for blood gas analysis, and for determination of the expression of TLR2 and TLR4 in monocytes.Oxygenation index was calculated.The consumption of propofol and remifentanil and pulmonary complications within 48 h after operation were recorded.Results Compared with group C, the expression of TLR2 and TLR4 in monocytes of venous blood was significantly down-regulated, oxygenation index was increased, and the incidence of pulmonary complications after operation was decreased in group D (P＜0.05).Conclusion The mechanism by which dexmedetomidine attenuates acute lung injury is associated with down-regulation of TLR2 and TLR4 expression in monocytes of blood in the patients undergoing pulmonary lobectomy.

AIM: To investigate in vitro effects of cholecystokinin octapeptide(CCK-8) on the expressions of B7.1 and B7.2 and the costimulatory activity of T lymphocytes in unstimulated macrophages.METHODS: Mouse peritoneal macrophages were isolated and incubated with CCK-8(10-12-10-6 mol/L) for indicated time.The B7.1 and B7.2 expressions of murine peritoneal macrophages were analyzed by flow cytometry.CD4+T cells were isolated from mouse spleen using immunomagnetic beads,and cultured with 1/4 numbers of macrophages which were pretreated with CCK-8 and/or anti-B7.1 antibody,anti-B7.2 antibody,CCK1R antagonist CR1409,CCK2R antagonist CR2945 for 24 h.ConA was added into the culture medium to stimulate CD4+T cell proliferation.The proliferation was determined by measuring -TdR incorporation in a ?-scintillation counter.RESULTS: B7.1 and B7.2 expressions and costimulatory activity of peritoneal macrophages were enhanced by CCK-8 in a dose-dependent manner,and the maximal effects occurred at the concentrations of 10-9 mol/L to 10-7 mol/L.Anti-B7.2 antibody,but not anti-B7.1 antibody,reduced the modulatory role of CCK-8 on costimulatory activity.Both CR1409 and CR2945 reversed the effect of CCK-8 on costimulation,and the role of CR1409 was more significant.CONCLUSION: CCK-8 enhances macrophage costimulatory activity by upregulating B7.2 expression,which is mediated by CCK1R and CCK2R.CCK1R might be the major receptor responsible for the modulation of CCK-8 on costimulation.

Objective To detect Chlamydia trachomatis phage Vp1 gene in clinical swab specimens and anti-Vp1 antibodies in serum specimens.MethodsCervical and urethral swab as well as serum specimens were collected from attendees to the sexually transmitted disease(STD) clinic in the Tianjin Institute of STD,during March 2008 to March 2011.PCR was conducted to detect chlamydial phage Vp1 gene in swab samples,enzyme linked immunosorbent assay(ELISA) and Western blot to detect anti-Vp1 antibody in sera.The swab specimens positive for Vp1 gene were subjected to cell culture followed by the detection of Vp1 protein with an immunofluorescence-based method.ResultsTotally,36 out of 1542 swab specimens turned out to be positive for Vp1 gene,and 23 out of 453 serum specimens for anti-Vp1 antibody.No positive results were obtained in the Vp1 gene-positive swab specimens by cell culture and immunofluorescence-based assay.ConclusionThe Vp1 gene of Chlamydial trachomatis phage and anti-Vp1 antibody are successfully detected from clinical swab and serum specimens respectively.

Objective To observe the effect of Zusanli acupoint injection of Astragalus Membranaceus Injection on patients with tumor leukopenia after radiotherapy and chemotherapy and its influence on cellular immune function.Methods According to clinical case of random table,60 cases of malignant tumor patients in Hubei province Xiangyang city hospital from March 2010 to August 2011 were randomly divided into two groups of 30 patients in each group.The treatment group was treated with Zusanli acupoint injection of Astragalus Injection therapy,while the control group was taken Leucogen,vitaminB4,batyl alcohol orally.Venous blood and peripheral white blood cell were taken in both groups before and after treatment.Direct immunofluorescence staining and flow cytometry were used for the detection of peripheral blood T lymphocyte total number and its subsets.Results ① clinical efficacy:Total effectiveness was 77.5％in the treatment group,and 52.5％ the control group,the difference was statistically significant between the two groups (x2=8.027,P＜0.05).Comparison of the white blood cell count and T lymphocyte subsets of peripheral blood:the white blood cell count was (3.56～4.21) × 109/L in the treatment group,and (2.89～3.39) × 109/in the control group,there was significant difference between two groups (t=2.435,P＜0.05); there was significant elevation of CD3 +,CD4+,CD4+/CD8 + [(62± 1.63) ％,(43 ± 0.80) ％,(1.48 ± 0.23) in the treatment group after the treatment comparing with (48± 1.25)％,(31 ± 0.74)％,(0.93± 0.12) before the treatment (P＜0.05 or 0.01),while there was no such changes in the control group.Conclusion Zusanli acupoint injection of Astragalus Injection may modulate the cellular immune function of patients with cancer.It is effective in the treatment of leukopenia after radiotherapy and chemotherapy.

Objective To study the role of hydrogen sulfide(H2S)in the pathogenesis of acute lung injury (ALI)-induced oleic acid(OA)and its regulatory effects on the inflammatory msponse.Method The ALI models in SD rats were produced by intravenous injection of OA(0.1 ml/kg).Forty-nine male SD rats were divided into three groups randomly:control group,OA group and OA+sodium hydrosulfide(NaHS)group.Rats of OA group and OA+NaHS group were sacrificed at 2,4 and 6 hours after the administration of OA,respectively.Rats of control group were sacrificed at 6 hours after intravenous normal saline(0.1 ml/kg).PaO2,lung tissue wet/dry(W/D)ratio and IQA score were determined.The concentrations of H2S in lung tissue and plasma were tested by sensitive sulphur electrode.The IL-6.IL-8 and IL-10 levels were measured by doubleantibody sandwich ELSIA.Results Compared with control group,of lung tissues was observed in rats treated with OA induced ALI,which led PaO2 dropped,and lung tissue W/D ratio,PMN%in alvedolar lavage and IQA raised score in the lung at 2,4 or 6 hours after OA injection(P＜0.01).In addition,IL-6,IL-8,and IL-10significantly increased,IL-6[plasma,control:(73.95±14.68)pg/ml,2 h:(186.70±23.85)pg/ml,4 h:(238.50±26.46)pg/ml,6 h:(215.95±25.86)pg/ml,P＜0.01;lung tissue,control:(60.58±12.91)pg/ml,2 h:(160.32±24.57)pg/ml,4 h:(195.27±46.28)pg/ml,6 h:(185.66±17.42)pg/ml,P all =0.000)],IL-8[plasma,control:(80.69±20.42)pg/ml,2 h:(184.11±19.51)pg/ml,4 h:(286.20±53.34)pg/ml,6 h:(241.30±45.85)pg/ml,P＜0.01;lung tissue,control:(69.14±15.96)pg/ml,2 h:(174.10±20.36)pg/ml,4 h:(249.02±31.17)pg/ml,6 h:(237.74±34.18)pg/ml,P＜0.01] and IL-10[plasma,control:(39.78±8.97)pg/ml,2 h:(111.18±11.46)pg/ml,4 h:(115.60±13.91)pg/ml,6h:(102.41±9.93)pg/ml,P＜0.01;lung tissue,control:(71.86±14.19)pg/ml,2 h:(126.96±18.72)pg/ml,4 h:(151.88±27.61)pg/ml,6 h:(137.28±14.22)pg/ml,P all=0.000] levels.in association with a decreased H2S content of plasma lung tissue decreased in[plasma,control:(36.58±6.80)μmol/L,2 h:(21.30±2.75)μmol/L,4 h:(20.63±1.26)μmol/L,6 h:(20.00±1.60)μmol/L,P＜0.01;lung tissue,control:(27.61±2.20)μmol/L,2 h:(20.67±1.37)μmol/L,4 h:(20.79±1.10)μmol/L,6 h:(18.92±0.75)μmol/L,P＜0.01]were observed in the plasma and lung tissue of OA-treated rats compared to controls.Administration of NaHS prior to OA treatment could lessen the lung pathologic changes induced by OA and elevate the concentrations of H2S in plasma[4 h:(26.67±3.44)μmol/L vs(20.63±1.26)μmol/L,P=0.042;6 h:(26.98±4.93)μmol/L vs(20.00±1.60)μmol/L,P＜0.05]and lung tissue[4 h:(23.20±1.48)μmol/L vs(20.79±1.10)μmol/L,P=0.011;6 h:(21,43±1.79)μmol/L vs(18.92±0.75)μmol/L,P=0.016].At the same time,the levels of IL-6 in plasma(185.37±21.98)pg/ml vs(238.50±26.46)pg/ml,4 h,P=0.000;(124.22±21.84)pg/ml vs(215.95±25.86)pg/ml,6 h,P＜0.01]and IL-8(199.40±34.56)pg/ml vs(286.20±53.34)pg/ml,4 h,P＜0.01;(146.58±20.23)pg/ml vs(241.30±45.85)pg/ml,6 h,P＜0.01]decreased significantly,but the level of IL-10(154.48±18.08)pg/ml vs(115.60±13.91)pg/ml,4 h,P=0.000;(138.06±20.01)pg/ml vs(102.41±9.93)pg/ml,6 h,P＜0.01]increased significantly.Conclusions The levels of endogenous H2S drop during the course of ALI oleic acid induced in rats.Exogenous H2S could decrease the levels of inflammatory factors(IL-6and IL-8),but increase the level of anti-inflammatory factors(IL-10).It could change the ratio of inflammatory factors/anti-inflammatory factors and therefore play a protective role against oleic acid induced ALI in rats.

Aim To investigate the inhibitory effect of hydrogen sulfide on expression of elastin in hypoxic pulmonary hypertensive (HPH) rats. Methods Twenty-four rats were randomly divided into three groups:control group (n=8), hypoxic group (n=8) and hypoxia+NaHS group (n=8). Pulmonary artery mean pressure (mPAP) of three group rats was measured via right ventricular catheterization. Right ventricle and left ventricle+septum of each rats were isolated from intact rats and weight of right ventricle/(weight of left ventricle+septum) ratio was calculated. The percentage of muscularized arteries of small pulmonary arteries was measured using a light microscope. The expression of elastin and TGF-? in pulmonary artery smooth muscle cells was observed with immunohistochemistry. Results mPAP and weight of right ventricle/(weight of left ventricle+septum) ratio were significantly higher in rats of hypoxic group than those of control group (P