Two types of cannabinoid receptors,named cannabinoid receptor 1(CB1) and cannabinoid receptor 2(CB2),have been cloned.CB2 receptors are expressed predominantly in the peripheral immune tissues,but accumulating evidence has revealed that CB2 receptors are also expressed in CNS.Previous studies showed that CB2 agonists can cure and suppress formation of inflammatory and neurophathic pain without central effects after chronic administration.Therefore,they will have good clinical applications in the treatment of pain and neurodegenerative diseases.In this paper,we will review the tissue distribution of CB2 in CNS and pharmacological characteristics of the CB2 agonists have been reviewed.

Objective To evaluate glaucoma and anterior chamber angle changes after intravitreal silicone oil injection(SOI). Methods The intraocular pressure(IOP) and anterior chamber angles of 34 eyes(33 patients)undergone pars plana vitrectomy and SOI were observed. Results Glaucoma occurred in 9 eyes(26%),in which silicone oil glaucoma(SOG)accounted for 77%(7/9).The changes of the superior part of anterior chamber angle were peripheral anterior synechiae in 11 eye(32%) and SO emulsification droplets in 22 eyes(64%) in 1～4.5 months after surgeries.Glaucoma occurred in 6 eyes of 7 eyes undergone peripheral iridectomy at the inferior part of the iris after the closure of iridectomy holes(P

Objective: To explore the relationships between blood levels of osteopontin (OPN), brain natriuretic peptide (BNP) and cardiac function in patient with degenerative heart valve disease (DHVD). Methods: Our research included in 2 groups: DHVD group,n=120 relevant patients treated in our hospital from 2013-12 to 2015-02 and Control group,n=30 healthy subjects from physical examination in the same period of time. Based on blood levels of OPN, DHVD patients were further divided into 2 sub-groups as Normal OPN sub-group, the patients with 18.8 ng/ml ≤OPN≤ 30.0 ng/ml,n=35 and High OPN sub-group, the patients with OPN>30.0 ng/ml,n=85. OPN levels at prior treatment (T0) and 3 days (T1), 1 week (T2), 2 weeks (T3) after treatment were compared between DHVD group and Control group; BNP levels, cardiac outcome (CO), cardiac index (CI) and left ventricular ejection fraction (LVEF) were also compared. The relationships between blood levels of OPN, BNP and cardiac function in DHVD patients were studied by Pearson correlation analysis. Results: Compared with Control group at T0 time point, High OPN sub-group showed increased blood levels of OPN and BNP, while decreased CO, CI and LVEF. Compared with Normal OPN sub-group, High OPN sub-group had the higher levels of OPN and BNP at all 4 time points, while lower levels of CO, CI and LVEF. In DHVD group, compared with T0 time point, OPN and BNP levels were decreased at T2 and T3 time points, while CO, CI and LVEF were increased, allP<0.05. Pearson correlation analysis presented that in DHVD patients, blood levels of OPN were positively related to BNP (r=0.936,P=0.00) and negatively related to CO, CI and LVEF (r=-0.869,r=-0.884 andr=-0.858 respectively, allP=0.00). Conclusion: DHVD patients had increased blood level of OPN which is related to BNP level and cardiac function; this might be because of OPN promoting heart valve calciifcation, inlfammatory reaction and myocardial injury. OPN could be used as a reference index for evaluating the cardiac function in DHVD patients.

BACKGROUND: Adipose tissue-derived mesenchymal stem cells (ADMSCs) have the multilineage differentiation potential, and are relatively easier to be obtained, thus they have attracted more and more attention as a new seed cell for cell engineering.OBJECTIVE: To observe the in vitro culture conditions of ADMSCs isolated from rat's subcutaneous adipose tissue, and identify them using immunohistochemical staining.DESIGN: An animal experiment.SETTING; Department of Cardiology, the General Hospital of Chinese PLA.MATERIALS: One healthy male Wistar rat of clean degree, 4 months old, weighing 200 g, was used. DMEM, fetal bovine serum were from GIBCO; Monoclone antibodies of rabbit-anti-rat CD13, CD34, CD44, CD45, CD105, D-related human leucocyte antigen (HLA-DR), factor-Ⅷ, vov Willebrand factor (VWF), Myosin, SABC kits and DAB staining kit from Wuhan Booster Biological Engineering, Co.,Ltd; Adeno-associated virus encoding green fluorescent protein from Vector Gene Technology Company Limited (Beijing).METHODS: The experiments were carried out in the Department of Internal Medicine, the General Hospital of Chinese PLA in October 2006. ① Cell isolation and culture: 0.3 g adipose tissue was cut from subcutaneous adipose tissue of Wistar rat's groin under aseptic condition, then minced and digested before culture, DMEM was changed at 2-3 days after plenty of fusiform-shap ed attached cells were observed under microscope, and the cell growth was observed. The cell concentration was adjusted to 2×107 L-1, then seeded into 96-well plate, and 100 μL for each well. From the second day, 3 wells were randomly selected every day, the cells were released with tripsin, and counted with blood cell counting chamber under inverted microscope. ② Cell viability assay: ADMSCs of passages 3 to 8 were added to DMSO freeze medium, and thawed after 2-4 weeks, and the cell viability was assessed by trypan blue dye exclusion. ③Immunohistochemical staining and identification: 2 ×107 L -1 cells were seeded to culture plate, then the immunohistochemical (SABC method) identification and Oil red O staining were performed to determine the cell surface antibodies of CD13, CD34, CD44, CD45, CD105, HLA-DR, factor-Ⅷ, HLA-DR and VWF. ④Lineage-specific differentiation and identification: The ADMSCs were plated on multi-well chamber and induced with lineage-specific media supplementation at least two weeks and identified by histologic/immunohistochemical assay of Oil red O for adipogenisis, alkaline phosphatase (ALP) stain for osteogenisis and Myosin monoclonal antibody for myogenisis. ⑤Transfected adenovirus carried green fluorescence protein (AD-GFP) medium: The fourth generation of ADMSCs were seeded on 96-well plate, 3 000 cells for each well, serum-free DMEM was changed after 24 hours, and added by AD-GFP at the same time, then transfected with different multiplicity of infection (MOI) of 1∶50, 1∶100, 1∶150 and 1∶200respectively, and then the transfection was observed.MAIN OUTCOME MEASURES: ① Results of cell isolation and culture; ② Cell viability after freezing and thawing; ③Results of immunohistochemical staining and identification; ④ Results of lineage-specific differentiation and identification;⑤ Results of transfected adenovirus carried AD-GFP.RESULTS: ① About 3.6×105 attached cells were obtained from 0.3 g subcutaneous adipose tissue, and these cells could be subcultured for passages in vitro with stable population doubling time. ② The cells were thawed after freezing for 2-3 weeks, and the trypan blue staining showed that the cell viability was above 90%. ③ The immunocytochemical staining showed that CD13, CD44, CD105 were positive and CD45, factor-Ⅷ, HLA-DR and VWF negative in different generations. ④ From the second generation, a few Oil red O positively stained cells were observed, which were obviously increased after prolonging the refreshing. After lineage-specific differentiation, the cells were all positive by Oil red O staining, ALP staining and Myosin immunohistochemical staining. ⑤ 72 hours after transfection, it was observed under fluorescence microscope that most cells were green fluorescence when the MOI value was 1∶200, the transfection was successful, and it was generally determined that the transfection rate was above 90%.CONCLUSION: A large number of ADMSCs with multilineage differentiation potential can be easily obtained from rat adipose tissue, osteoblast, myoblasts, they can be expanded in large quantity and stored in vitro for long time, AD-GFP were also successfully transfected.

Objective To investigate the effect of endoplasmic reticulum stress (ERS)-associated apoptosis on renal interstitial fibrosis induced by unilateral ureteral obstruction (UUO) in rats. Methods Eighteen healthy male Wistar rats undergoing UUO were sacrificed at 3,7,14 days after operation. Additional seven rats underwent sham operation. Histological changes were observed by HE and Masson staining. Immunohistochemistry was performed on renal tissue for α-smooth muscle actin (α-SMA). Chromatometry was used to detect the content of hydroxyproline. Apoptosis cells were determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and the electrophoresis analysis of genome DNA. The mRNA expression of glucose-regulated protein 78 (GRP78), which are important markers of ERS, were detected by RT-PCR. Western blotting was used to assess the protein level of GRP78 and cysteinyl aspartate specific proteinase-3 (caspase-3). Results Compared with sham operation group, the degree of renal interstitial and the level of hydroxyproline content of UUO group increased significantly (P<0.05). Immunohistochemistry staining indicated that a-SMA extensively expressed in renal tubular and interstitial cells. The apoptotic cells in the renal tubular and interstitium were continuously increased from day 3 to the end of experiment of UUO group. As early as 3 days after surgery, the mRNA level of GRP78 in UUO group increased compared with sham operation group (P<0.01), while the protein expression increased on day 7 after surgery (P<0.01). Prolonged ERS triggered apoptosis, the protein expression of caspase-3 increased significantly on day 3 after surgery (P< 0.05), and the expression sustained high level during the experiment afterwards. There was a positive correlation between GRP78 protein expression and hydroxyproline content (r =0.657, P< 0.01) as well as caspase-3 protein expression (r=0.714, P<0.01). Conclusions UUO induces a significant up-regulation in endoplasmic reticulum molecular chaperones at early stage, indicating that ERS response is activated in the rat kidney. Prolonged ERS can lead to renal tubular and interstitial cell apoptosis, and caspase-3-mediated ERS associated apoptosis may contribute to the fibrosis.

Objectives To investigate the effect of different dosages of low molecular weight heparin on acute pulmonary embolism and inhibition of pulmonary intimal hyperplasia in immature rats. Methods 90 male immature SD rats were randomly divided into ifve groups: sham group, pulmonary embolism group, low-low molecular heparin group (L-LMH), medium-low molecular heparin group (M-LMH) and high-low molecular heparin group (H-LMH). The model of acute pulmonary embolism was established through jugular vein injection with gel-foam solution. The rates in the L-LMH, M-LMH, H-LMH groups were treated with low molecular weight heparin by subcutaneous injection after surgery with a dosage of 0 . 005 ml/kg, 0 . 01 ml/kg, 0 . 02 ml/kg, twice a day. Animals in the control group were given saline injection. Arterial blood gas, pulmonary artery pressure (mPAP), right ventricular pressure (RVP), wall area/tube area, wall thickness/tube diameter, and the expression of PDGF-B and MCP-1 at gene and protein levels in lung tissue were detected on the 7 th ( 7 d), 14 th ( 14 d) and 28 th ( 28 d) after opration. Results There were signiifcant differences of PaO 2 among 5 groups on 7 d, 14 d and 28 d. PaO 2 in group M-LMH ( 105 . 1 ± 4 . 6 mm Hg) were signiifcantly higher than that of embolization group, L-LMH, but not H-LMH group at 28 d. mPAP of M-LMH group was lower than that in the other three intervention groups, but showed no signiifcant difference compared with sham group (P?>0 . 05 ). There were signiifcant differences of RVP on 7 d and 14 d. PDGF-B, MCP-1 of M-LMH group were signiifcantly lower compared with the other three intervention groups (P?0 . 05 ). Wall area/tube area, wall thickness/tube diameter scores of M-LMH group had no signiifcance differences compared with sham group on 28 d (P?>?0 . 05 ). Conclusion Medium dose of low molecular weight heparin could ameliorate the acute pulmonary embolism and inhibit the proliferation of pulmonary arteries in rats.

Objective To investigate the effects of oxidative stress on the expressions of transforming growth factor-?1(TGF-?1) and transforming growth factor-? receptor Ⅰ(TGF-?RⅠ) in rat unilateral ureteral obstruction(UUO) models.Methods Thirfty Wistar rats were randomly assigned into three groups:SOR group(sham-operated group,n=7);UUO group(operation group,n=11);US group(spironolactone 50 mg?kg-1?d-1 by daily gastric gavage after UUO,n=12).All the rats were killed 14 d after surgery.Renal fibrosis was assessed by the determination of tissue hydroxyproline(HYP) content.Malondialdehyde(MDA),superoxide dismutase(SOD) as well as aldosterone(ALD) content were measured.Histological changes were observed by HE and Masson staining.Immunohistochemistry was performed to detect the expressions of TGF-?1 and TGF-?RⅠ.Western blotting was used the determine the expression of TGF?RⅠ protein.Results Compared with sham group,the ALD contents in plasma and kidney tissues in UUO group significantly increased(P

AIM: To investigate effect of sodium ferulate on A?25-35-mediated signaling pathway. METHODS: The isolated peritoneal macrophages from mice were cultured. p38 MAPK protein kinase in nuclear extracts was analyzed by Western blotting. The concentration of TNF-? and NO in supernatant were measured by ELISA and Griess reaction technique. The expression of iNOS protein was detected by immunochemical technique. RESULTS: A?25-35 significantly increased the concentrations of TNF-? and NO in supernatant, expression of iNOS in macrophages and p38 MAPK protein kinase in nuclear extracts, which were blocked by sodium ferulate. CONCLUSION: Sodium ferulate inhibits p38 MAPK activation triggered by A?25-35.

Objective To evaluate the outcome of Stage Ⅱ nasopharyngeal carcinoma(NPC) trea-ted by conventional fractionation radiotherapy. Methods From January 1990 to May 1999,216 patients with untreated stage Ⅱ NPC were irradiated by 6 MV photons and 9-12 MeV electrons with linear accelera-tor. Of 26 patients with residual lesions after 70 Gy irradiation, 18 received intracavitary brachytherapy and 8 received stereotactic irradiation boost. Results The 10-year overall survival (OS) of patients with T2 N0 M0, T1N1M0 and T2N1M0 disease were 90%, 80% and 75.2% (χ2 = 3.26, P =0. 200), respectively. The corresponding disease-free survival(DFS) and distant metastasis-free survival(DMFS) were 79% ,60% and 62.6% (χ2 = 5.87, P = 0.053) ;90% ,74% and 68.0% (χ2 = 7.09, P = 0. 030), respectively. In uni-variate analysis, T stage had no impact on OS, DFS or DMFS (χ2 = 0.44, P = 0. 500 ; χ2 = 0.44, P = 0.500 ; χ2 = 0.25, P = 0. 610), while N stage significantly influenced DFS and DM FS (χ2 = 5.86, P = 0. 015 ; χ2 = 5.31 ,P = 0. 021). Multivariate analysis showed that N stage was an independent prognostic factor for DFS and DMFS(χ2=5.03,P=0.025;χ2=6.47,P=0.01). Conclusions Stage Ⅱ NPCwithposifivelymph node treated by radiotherapy alone is inadequate. In order to improve DFS and DMS, combined chemo-radio-therapy should be considered.

Objective To investigate the clinical features,treatment outcome and prognosis of pa tients with nasopharyngeal adenoid cystic carcinoma(ACC).Methods From March 1963 to August 2002, 33 patients with ACC were retrospectively reviewed.Among them,13 received multimedality therapy(S + R in 9 and R +S in 4) and the remaining 20 received radiotherapy alone.The interval between the first visit and clinical diagnosis was 12 months(1.0-36.4 months).The misdiagnosis rate was 55% (18/33),with the most common error of otitis media.The clinical symptoms of ACC were similar to those of squamous carcino ma in nasopharynx,though the cervical lymph node metastasis was rare(12%) and cranial nerve invasion was common (58%). ResultsAfter a median follow-up of 60.8 months (5.0-245.2 months),the 5-and 10-year overall survival rates,free-disease survival rates,local regional control rates and metastasis-free sur vival rates were 66% and 29% ,41% and 27% ,61% and 40% ,62% and 62% ,respectively.Among the 20 patients(61%) with recurrence,15 had local regional failure(13 in situ and 2 in cervical lymph node) ,and 9 had distant metastasis(5 in the lung,2 in the lung and bone,1 in the liver and I in the bone).Univariate analysis showed that clinical stage was a prognostic factor(Ⅰ + II vs Ⅲ + Ⅳ,P = 0.009),while treatment modality(radiotherapy alone vs multimodality therapy) was not. ConclusionsNasopharyngeal ACC is a locally aggressive disease with a long course.Either radiotherapy alone or muhimodality therapy(S + R/R + S) is effective in the treatment.