ObjectiveTo investigate the effects and mechanisms of IL-32γ-shRNA-mediated gene silencing on the proliferation and apoptosis of fibroblast-like synoviocytes in patients with rheumatoid arthritis (RA).MethodsA eukaryotic expression plasmid of shRNA targeting IL-32γ was transfected into fibroblastlike synoviocytes by liposome in patients with rheumatoid arthritis.RT-PCR was used to determine the expression level of IL-32γ.Western blotting was used to detect the levels of cyclin D1 and p-Akt.The proliferation of RA-FLS was examined by MTT.Cell cycles were analyzed by flow-cytometry.The apoptosis of cells were measured by TUNNEL.Comparisons between groups were tested by t test.Results ① The expression of IL-32γwas significantly inhibited by shRNA-IL-32γ-expressing plamid PGCsi 3.0 targeting sequence 1,2 and 3,and the inhibition rate had reached 75.6％,66.2％ and 64.1％,respectively.② The absorbance value of proliferation of RA-FLS in EASY-shRNA-IL-32γ group was significantly lower than that in the shRNA-control group and normal group at day 3 [(0.23±0.03) vs (0.35±0.03) and (0.36±0.04),P＜0.05] and 5 [(0.27±0.03) vs (0.52±0.05) and (0.53±0.04),P＜0.01 ] after transfection.③ The rate of RA-FLS at phase G1 in the EASY-shRNA-IL-32γ group was significantly higher than that in the shRNA-control group and normal group respectively [(88±6)％ vs (69±5)％ and (68±4)％,P＜0.05],while those at phase S+G2 in the EASY-shRNA-IL-32γgroup was significantly lower than that in the shRNA-control group and normal group [ ( 13.6±3.0)％ vs (30.2±4.1)％ and(32.1±4.3)％,P＜0.01].④The rate of RA-FLS apoptosis in the EASY-shRNA-IL-32γ group was significantly higher than that in the shRNA-control group and normal group[(20.50±3.21 )％ vs (9.20±0.32)％ and (8.60±0.22)％,P＜0.01].⑤ The expression of cyclin D1(0.36±0.04) and p-Akt(0.31±0.03) in the EASY-shRNA-IL-32γ group was significantly lower than that in the shRNA-control group [ (0.59±0.08) and (0.53±0.06)] and normal group [(0.61±0.07) and (0.52±0.06),P＜0.01].ConclusionEASYshRNA-IL-32γ can inhibit RA-FLS proliferation by down-regulating the expression of cyclin D1 and induce RA-FLS apoptosis by down-regulating the expression of p-Akt.

Objective To determine the contents of four effective constituents in Euphorbiae Semen decoction;To provide evidence for Euphorbiae Semen decoction into clinical application. Methods Established quantitative analysis multi-components by single marker method was used to determine the contents of diterpenoids constituents, such as euphorbia storoid, euphorbia factor L2, and euphorbia factor L3. HPLC method was used to determine the contents of aesculetin. Results Contents of euphorbia storoid, euphorbia factor L2, and euphorbia factor L3 in smashed Euphorbiae Semen decoction were 0.015 9%, 0.005 9% and 0.024 1%, respectively. However, the contents of the above three constituents could not be detected in whole Euphorbiae Semen decoction. The content of aesculetin (0.693 6%) in smashed Euphorbiae Semen decoction was more than that in whole Euphorbiae Semen decoction (0.288 2%). Conclusion Decoction digestion effect of diterpenoids constituents in Euphorbiae Semen decoction is not good. Decocting with water is not suitable for the clinical application of Euphorbiae Semen for purgation and diuresis. Aesculetin in smashed Euphorbiae Semen decoction has good decoction digestion effect, in which clinical use for antisepsis and anti-inflammation is effective.

The present study examined the functional profile of dendritic cells (DCs) in patients with rheumatoid arthritis (RA) and the effects of simvastatin on the function of DCs. A total of 40 patients who was recently diagnosed as having RA were equally assigned to two groups: the routine treatment group (group R) and the routine treatment plus simvastatin group (group R+S). Twenty healthy individuals served as control. The peripheral blood mononuclear cells (PBMCs) were isolated before and 4 weeks after the treatment and then cultured with interleukin-4 (IL-4) and granulocyte-macrophage colony stimulatory factor (GM-CSF) to prepare mature DCs. The expression of co-stimulating factor CD86 on the surface of DCs was assessed by flow cytometry. And the stimulating capacity of DCs was measured by mixed lymphocyte reaction (MLR). The contents of cytokines in culture supernatants of DCs in MLR were detected by ELISA. Blood lipids and high-sensitivity C-reactive protein (hs-CRP) were detected. The relationship between the expression of CD86 and the blood CRP level was also investigated. The results showed that, as compared with the control group, the CD86 expression and the level of cytokines secreted by DCs were significantly increased in RA patients and greater stimulating capacity of DCs in MLR was demonstrated in RA patients. T lymphocytes in MLR secreted higher levels of proinflammatory cytokines (IL-2, IL-17, TNF-α and INF-γ) and lower level of anti-inflammation cytokine (IL-10). The function of DCs was markedly weakened and the level of hs-CRP and low-density lipoprotein was substantially lowered in group R+S in comparison to group R. The CD86 expression was positively correlated with hs-CRP. It was concluded that DCs in RA are highly activated and DC-initiated immune reaction may play an important role in the pathogenesis of RA. Simvastatin administration can significantly inhibit the DCs function and reduce the level of hs-CRP, indicating the suppression on inflammatory reaction may be one of the mechanisms by which simvastatin exerts its effect in treating RA.

Objective To investigate the correlation of oxygen extraction rate (ERO2) with blood lactate clearance rate and cardiac output (CO) in the early stage of post-restoration of spontaneous circulation (ROSC) in patients resuscitated from cardiogenic cardiac arrest,and to analyze the relationship between the ERO2 and prognosis.Methods Fourteen patients successfully resuscitated from in-hospital cardiogenic cardiac arrest in the emergency ICU from October 2012 to January 2014 were retrospectively analyzed.These patients were assigned to survival group (n =5) or death group (n =9) as per the outcome at 72 h after ROSC.At admission (0 h),3,6 and 12 h after ROSC,arterial blood and venous blood were drawn to detect ERO2 and lactate clearance rate.Cardiac output (CO) was measured by thoracic impedance method,APACHE Ⅱ scores were assessed,and survival time was recorded.Results The patients in the death group died during the period of 12-72 hours after ROSC.The significantly decreased ERO2 at 6 h and 12 h after ROSC,and decreased blood lactate clearance rate and decreased CO at 3 h,6 h and 12 h after ROSC were found in the death group compared with the survival group (all P ＜ 0.05).The ERO2 at 6 h and 12 h after ROSC was significantly positively related to blood lactate clearance rate (r =0.857,r =0.947,both P ＜ 0.05) and CO (r =0.968,r =0.936,both P ＜ 0.05) at 3 h,6 h and 12 h after ROSC.The ERO2 at 6 h and 12 h after ROSC was significantly negatively related to APPACHE Ⅱ score (r =-0.970,r =-0.973,both P ＜ 0.05);APPACHE Ⅱ scores were significantly negatively correlated with blood lactate clearancerates (r=-0.880,r=-0.899,r=-0.850,all P＜0.05) and CO (r=-0.876,r=-0.922,r=-0.916,all P＜0.05) at 3 h,6 h and 12 h after ROSC.Conclusions The ERO2 at 6h after ROSC may be used to assess the severity and prognosis of patients resuscitated from cardiogenic cardiac arrest.

Objective To build learning evaluation system and indicators connotation of problem-based in small-group teaching model.Methods Delphi technique was used to build learning evaluation system of problem-based for course of nursing education through a two-round expert consultation among 30 experts in Zhejiang province.Results The experts' authority coefficient was 0.89,determination coefficient was 0.90,and familiar confficient was 0.88.The finally learning evaluation system of problem-based for course of nursing education consisted of 3 first-level indicators,14t second-level indicators and 41 third-level indicators.The coordination coefficients of first-level indicators and second-level indicators were 0.37 and 0.31,respectively,there was significant difference,P＜0.01.Conclusion This project has a rigorous and reliable construction process with strong theoretical and practical significance,and could be used as a guidance and reference for problem-based in small-group teaching for course of nursing education.

Objective To investigate the effects of recombinant adeno-associated virus-mediated myostatin propeptide (MPRO) on uptake and oxidation of glucose,and glycogen synthesis in C2C12 myotubes,as well as the associated molecular mechanism.Methods Mature C2C12 myotubes were assigned to the following 6 groups:control,insulin,green fluorescent protein (GFP),insulin + GFP,MPRO,and insulin + MPRO groups.Glucose uptake,glucose oxidation,and glycogen synthesis were detected by counting radioactivity of 14CO2 or 14C labeled glycogen derived from 2-deoxy-[1-14 C] glucose.The activity of insulin signal pathway was evaluated by Western blot.Results Compared with control group,glucose uptake and glycogen synthesis were significantly increased in insulin and insulin+GFP groups,and further increased in insulin+MPRO group as compared with insulin alone(all P＜ O.05).However,MPRO and insulin had no effect on glucose oxidation.The phosphorylations of insulin receptor (IR) β,insulin receptor substrate 1 (IRS-1),protein kinase B (Akt),glycogen synthase kinase-3 β (GSK-3β),and the expressions of phosphatidylinositol 3-kinase (PI3K) and glucose transporter 4 (Glut4) in membrane were significantly increased in insulin and insulin+GFP groups compared with control group(all P＜0.05),and were further increased after MPRO transfection (all P ＜ 0.05).Conclusion MPRO may increase insulin-stimulated glucose uptake and glycogen synthesis in C2C12 cells by activating the IRS/PI3K/Akt signal pathway.

Objective To observe any effect of low intensity pulsed ultrasound (LIPUS) on the expression of integrin-focal adhesion kinase (FAK)-mitogen-activated protein kinase (MAPK) mechanochemical transduction pathway-related proteins in the chondrocytes of rabbits with knee osteoarthritis (OA).Methods Of the 18 New Zealand white rabbits selected for the study,twelve received knee anterior cruciate ligament transection to model OA.The remaining 6 rabbits served as normal controls.At the 4th week after modeling the rabbits were sacrificed and chondrocytes were isolated and cultured in vitro.All the cultured cells were randomly divided into three groups:a normal control group (NC),an OA model group (OA) and an OA model plus LIPUS group (OA + LIPUS).When the cells had been cultured to the 2nd passage,the NC group and OA group cells had received no treatment.The OA + LIPUS group cells were exposed to 40 mW/cm2 of LIPUS for 20 min,once a day for 6 days.The expression of collagen protein type Ⅱ,aggrecan,MMP-13,integrin β1 p-FAK and p-p38,p-ERK1/2 and p-JNK Mapks were detected by Western blotting.Results Compared with the NC group,the expression of collagen type Ⅱ and aggrecan was significantly lower in the OA and OA + LIPUS groups,with more significantly lower expression of collagen type Ⅱ and aggrecan in the OA group than in the OA + LIPUS group.Compared with the NC group,the expression of MMP-13 was significantly higher in the OA and OA + LIPUS groups,with a significantly larger increase in the OA group.Compared with the NC group,the expression of integrin β1 and p-FAK was also significantly higher in the OA and OA + LIPUS groups,with a significantly larger increase in the OA + LIPUS group.Compared with the NC group,the expression of p-p38,p-ERK1/2 and p-JNK was also significantly higher in the OA group,but compared with the OA group,the expression of those kinases was,on average,significantly lower in the OA + LIPUS group.Conclusions LIPUS can inhibit the degradation of collagen type Ⅱ and aggrecan,and inhibit the expression of MMP-13,p-p38,p-ERK1/2 and p-JNK in OA chondrocytes,at least in vitro.At the same time,LIPUS can increase the expression of integrin β1 and p-FAK.The results show that LIPUS may activate an integrin-FAK-MAPK mechanochemical transduction pathway to induce changes in the extracellular matrix of chondrocytes.

Objective To study the optimization process of matching conditions using glutaraldehyde (GDA) as crosslinking agent of hemoglobin based oxygen carriers (HBOCs),to further reduce the average molecular weight and the content of super-weight molecular,and improve the conversion ratio of polymerization.Methods The orthogonal designs were done on the basis of the previous single influencing factor research of human placenta hemoglobin crosslinking GDA.Three factors were selected including molar ratio of GDA to hemoglobin,mass concentration of hemoglobin and the rate of the feeding GDA.Results The molar ratio of GDA to hemoglobin is the most important influencing factor on the molecular weight distribution of polymerized hemoglobin,followed by the mass concentration of hemoglobin and the rate of feeding GDA.When analyzing the impact on the mean molecular weight,there were significant differences between mean molecular weight corresponding to different molar ratios of GDA to hemoglobin (P＜0.05),while there was no statistical significance between mean molecular weight corresponding to different mass concentrations of hemoglobin and the rates of feeding GDA (P＞0.05).When analyzing the impact on the effective conversion ratio,there were significant differences between effective conversion ratios corresponding to different molar ratios of GDA to hemoglobin and different mass concentrations of hemoglobin (P＜0.05),while there were no statistical significances between effective conversion ratios corresponding to different rates of feeding GDA (P＞0.05).When analyzing the impact on the content of super-weight molecular,there were significant differences between content of super-weight molecular corresponding to different molar ratios of GDA to hemoglobin,while there were no statistical significances between content of super-weight molecular corresponding to different mass concentrations of hemoglobin and different rates of feeding GDA.Conclusions The optimal matching conditions of hemoglobin polymerization process were determined by orthogonal designs.

Objective To study the level of serum 25 (OH) D3in children in suzhou area, and to provide scientific basis for the rational supplement of vitamin D for children aged 0-6years.Methods From September2015to September 2016, 15 010children underwent routine physical examination in the Children′s Health Clinic of Suzhou Municipal Hospital were selected, of whom 7 905were male and 7 105were female.The serum 25 (OH) D3was detected by collecting their fingerling blood.Results (1) The mean serum 25 (OH) D3of15 010children aged 0to 6in Suzhou was (35.83±13.23) μg/L, and the mean serum 25 (OH) D3of male and female were (36.48±13.25) and (35.11±13.16) μg/L respectively, and the differences were statistically significant (P＜0.01). (2) The mean level of serum 25 (OH) D3of 0-＜3, 3-＜6, 6-＜12, 12-＜36, 36-＜48and≥48months old children were (34.49±11.53), (41.15±13.86), (48.03±17.25), (46.12±17.69), (28.49±16.55) and (42.28±17.59) μg/L.The detection levels of serum 25 (OH) D3between the age groups were statistically significant (P＜0.05) except the children 3-＜6months and≥48months. (3) From January to December, the detection levels of serum 25 (OH) D3 were statistically significant between different months (P＜0.01) except in January, February, March and November, as well as July and August.The serum25 (OH) D3in each month was graded according to the vitamin D level, and the detection levels of serum 25 (OH) D3between different months were statistically significant (P＜0.01).The proportion of serum 25 (OH) D3over 30μg/L was less than 50％in January, March and November.The ratio ranged from 50％to 60％in February, June and December.The ratio ranged from 60％to 70％in the July, August and September, while the proportion was over 70％in April, May and October.Conclusion The level of serum 25 (OH) D3in children in Suzhou area was decreased obviously, and health education should be strengthened, and attention should be paid to intaking of vitamin D in children.

The present study examined the functional profile of dendritic cells (DCs) in patients with rheumatoid arthritis (RA) and the effects of simvastatin on the function of DCs. A total of 40 patients who was recently diagnosed as having RA were equally assigned to two groups: the routine treatment group (group R) and the routine treatment plus simvastatin group (group R+S). Twenty healthy individuals served as control. The peripheral blood mononuclear cells (PBMCs) were isolated before and 4 weeks after the treatment and then cultured with interleukin-4 (IL-4) and granulocyte-macrophage colony stimulatory factor (GM-CSF) to prepare mature DCs. The expression of co-stimulating factor CD86 on the surface of DCs was assessed by flow cytometry. And the stimulating capacity of DCs was measured by mixed lymphocyte reaction (MLR). The contents of cytokines in culture supernatants of DCs in MLR were detected by ELISA. Blood lipids and high-sensitivity C-reactive protein (hs-CRP) were detected. The relationship between the expression of CD86 and the blood CRP level was also investigated. The results showed that, as compared with the control group, the CD86 expression and the level of cytokines secreted by DCs were significantly increased in RA patients and greater stimulating capacity of DCs in MLR was demonstrated in RA patients. T lymphocytes in MLR secreted higher levels of proinflammatory cytokines (IL-2, IL-17, TNF-α and INF-γ) and lower level of anti-inflammation cytokine (IL-10). The function of DCs was markedly weakened and the level of hs-CRP and low-density lipoprotein was substantially lowered in group R+S in comparison to group R. The CD86 expression was positively correlated with hs-CRP. It was concluded that DCs in RA are highly activated and DC-initiated immune reaction may play an important role in the pathogenesis of RA. Simvastatin administration can significantly inhibit the DCs function and reduce the level of hs-CRP, indicating the suppression on inflammatory reaction may be one of the mechanisms by which simvastatin exerts its effect in treating RA.
Adult ; Anti-Inflammatory Agents ; pharmacology ; therapeutic use ; Arthritis, Rheumatoid ; drug therapy ; immunology ; C-Reactive Protein ; metabolism ; Cytokines ; metabolism ; Dendritic Cells ; drug effects ; immunology ; Female ; Humans ; Male ; Middle Aged ; Simvastatin ; pharmacology ; therapeutic use