Hydrogels are promising materials for electronic skin due to their flexibility and modifiability. Reported hydrogel electronic skins can recognize stimulations and output signals, but the single output signal and the requirement of external power source limit their further applications. In this study, inspired by the neuron system, the self‐powered neuron system‐like hydrogels based on gelatin, water/glycerin and ionic liquid modified metal organic frameworks (MOFs) are prepared. The optimized hydrogel exhibits excellent adhesion (40 kPa), stretchability (0%–100%), water retention (>92% at 0% relative humidity (RH) atmosphere), ionic conductivity (>10 −3 S m −1 ) and stability (>30 days). Besides, the neuron system‐like hydrogels are highly sensitive to pressure (0—10 N) and humidity (0%–75% RH) with dual‐modal output, without external power source. Finally, the optimized hydrogel ionic skin is applied in human motion detection, energy harvesting, and low humidity sensing. This study provides a preliminary exploration of self‐powered ionic skin for multi‐application scenarios.

Objective To explore the expressions and significance of NF-κB and PTEN in endometriosis. Methods The expressions of NF-κB and PTEN were detected by immunohistochemical SP method in 40 cases of ectopic endometrium, 40 cases of eutopic endometrium, and 30 cases of normal endometrium. Results The expressions of NF-κB was the highest in ectopic endometrium , followed by eutopic endometrium , and was the lowest in normal endometrium ( P < 0 . 01 ) . While the expressions of PTEN was the lowest in ectopic endometrium, followed by eutopic endometrium, and was the highest in normal endometrium (P < 0.01). Conclusion The expressions of NF-κB and PTEN were positively associated with the development and progress of endometriosis.

目的:构建基于Tat细胞穿透肽和核定位信号NLS的重组酶Cre蛋白表达载体,引导Cre内化并入核实现细胞水平的基因敲除.方法:在带His标记的细胞穿透肽Tat和增强型绿色荧光蛋白(EGFP)表达载体pET14b-SBP-Tat-EGFP基础上,利用酶切连接的方法将NLS-Cre片段插入带上述表达载体中,构建新的融合蛋白表达载体pET14b-SBP-Tat-NLS-Cre-EGFP;经酶切、测序鉴定载体构建正确后,将重组质粒转化BL21 (DE3)宿主菌,经异丙基硫代-β-D-半乳糖苷(IPTG)诱导表达后,用Ni2+亲和层析纯化得到融合蛋白;将融合蛋白透析、过滤除菌后加入到培养的cdc42基因两端带Loxp位点的C57小鼠腹腔巨噬细胞中,在Zeiss荧光显微镜下观察蛋白转导效率,并用Western blot方法在蛋白水平检测并验证细胞水平目的基因敲除效果.结果:经酶切、基因测序证实重组载体构建成功,融合蛋白在大肠杆菌中可有效表达;Zeiss荧光显微镜下观察融合蛋白穿透细胞膜并进入细胞,细胞在经融合蛋白内化处理后目的基因蛋白的表达量与对照组没加Tat融合蛋白比较有明显降低.结论:利用Tat细胞穿透肽成功构建了基于细胞穿透肽的带有NLS-Cre的Tat蛋白表达运输载体,建立了可携带Cre进入细胞核进行基因敲除的系统,说明利用该系统可以实现细胞水平的基因敲除.

Objective:To investigate the expression of FAS receptor on idiosyncratic T cell subsets,including TCRαβ+cells,CD4-CD8-cells and TCRαβ+CD4-CD8-T cells,and to discuss the relationship between FAS-mediated apoptosis and peripheral TCRαβ+CD4-CD8-T cells in patients with rheumatoid arthritis(RA).Methods:Totally 24 RA patients and 24 normal controls were included.Flow cytometry was used to detect the percentages of T cell subsets and the expression of FAS receptor.The relationships between the percentages of T cell subsets and the expression of FAS receptor with RF,WBC,CRP and ESR were analyzed respectively by spearman relativity analysis.Results: The percentages of peripheral TCRαβ+ cells and CD4-CD8-cells from RA patients were similar to those in normal controls(P0.05).Strikingly higher levels of TCRαβ+CD4-CD8-T cells in the peripheral blood of RA patients were detected compared with those in normal controls(0.82±0.38 vs.0.57±0.26,P=0.013).The expressions of FAS receptor on TCRαβ+CD4-CD8-T cells were significantly increased in RA patients compared with those in normal controls(1.23±0.69 vs.0.80±0.45,P=0.016).Positively relationship was found between TCRαβ+CD4-CD8-T cells and TCRαβ+CD4-CD8-FAS+cells(r=0.809,P=0.000).There was no significant relationship between the percentages of TCRαβ+CD4-CD8-T cells,the level of FAS receptor expression and RF,WBC,CRP or ESR,respectively.TCRαβ+ cells as well as CD4-CD8-cells had no significant association with RF,WBC,CRP and ESR,respectively.Conclusion: The preliminary results suggest that the significantly upregulated percentages of DNT cells and its FAS receptor expression may increase the sensitivity to FAS-mediated apoptosis.It may lead to the imbalance of immune respective T cells in RA patients.There is no significant relationship between the percentages of TCRαβ+CD4-CD8-T cells,FAS receptor and the indicators of disease activity,though the percentages of them are markedly increased in the Chinese RA patients.The role of TCRαβ+CD4-CD8-T cells in the pathogenesy of RA patients remains to be further studied.

OBJECTIVE:To investigate the effect of NK-22 cells isolated from the synovial fluid (SF) of patients with rheumatoid arthritis (RA) on the proliferation of fibroblast-like synoviocytes and explore its possible mechanism.METHODS:The proportions of NK-22 cells in the peripheral blood (PB) and the SF of 20 RA patients and 20 healthy individuals were determined by flow cytometry. NK-22 cells in the SF sorted by flow cytometry were cultured for two weeks followed by a 4-h stimulation with 20 ng/ml phorbol 12-myristate 13-acetate and 0.5 µ mol/L ionomycin. The culture supernatant of NK-22 cells was then harvested, in which the levels of IL-22 and TNF-α were measured by ELISA. The fibroblast-like synoviocytes were exposed to the culture supernatant for 24, 48, 72, and 96 h, and the changes in the cell proliferation were detected by MTT assay.RESULTS:RA patients showed a significantly greater proportion of NK-22 cells in both the SF and PB than the normal control subjects (P<0.05). NK-22 cells sorted by flow cytometry reached a purity exceeding 90%, and the levels of IL-22 and TNF-α in the culture supernatant of NK-22 cells cultured for two weeks were 941.16 pg/ml and 368.1 pg/ml, respectively. The culture supernatant of NK-22 cells caused a rapid proliferation of the fibroblast-like synoviocytes at 24, 48, 72, and 96 h after the exposure.CONCLUSION:NK-22 cells in the SF of RA patients can promote the proliferation of fibroblast-like synoviocytes possibly due to the capacity of NK-22 cells to produce IL-22 and TNF-α.

We have developed a potential radiation-sensitive drug-delivery system using active diselenide-containing block co-polymer aggregates in aqueous solution that can load and release anticancer drugs. These aggregates were sensitive to even a low dose of γ-radiation, such as 5 Gy, which is close to the radiation dose received by patients during a single radiotherapy treatment. This line of research may open an avenue for the combination of radiotherapy and chemotherapy.

OBJECTIVE:To investigate the expression levels of miR-146a and miR -16 in the peripheral blood mononuclear cells (PBMCs) of patients with rheumatoid arthritis (RA) and their correlation to the disease activity. METHODS:The expression levels of miR-146a and miR-16 in the in PBMCs were measured using real-time RT-PCR in 40 RA patients and 16 healthy individuals. The correlations of miR-146a and miR-16 expressions to the disease activity of RA were analyzed. RESULTS:The expression levels of miR-146a and miR-16 in the PBMCs of RA patients increased significantly compared to those in the healthy individuals (P<0.05). Their expression levels was significantly higher in active RA patients than in patients with disease remission (P<0.05) and healthy individuals (P<0.01). The expression levels of miR-146a and miR-16 were positively correlated to ESR, CRP and the disease activity scores in 28 joints (DAS28) (P<0.01), but not to RF (P>0.05). CONCLUSION:The elevated expression levels of miR-146a and miR-16 are correlated to RA disease activity, suggesting their value in assessment of the clinical disease activity of RA.

OBJECTIVE:To explore the association of the expressions of human leukocyte antigen (HLA)-DR4, peptidyl arginine deiminase type4(PAD4), and signal transducer and activator of transcription 4 (STAT4) in the peripheral blood with the disease activity in patients with rheumatoid arthritis (RA).METHODS:Twenty-four RA patients in active stage (DAS28 score>or=2.6) and 14 RA patients in remission stage (DAS28 score<2.6) were enrolled in this study, with 12 healthy volunteers as the control. The QuantiGene Plex method was used to measure the expression level of HLA-DR4, PAD4, and STAT4 mRNA, and the relationship between the expressions of these genes and the DAS28 score, levels of anti-cyclic citrullinated peptide antibody (anti-CCP antibody) and rheumatoid factor (RF) was analyzed.RESULTS:The expressions of HLA-DR4, PAD4, and STAT4 were significantly higher in RA patients than in the healthy controls (P<0.05). The level of HLA-DR4 mRNA in the two RA groups showed no significant difference, but was significantly higher than that in the healthy controls. HLA-DR4 expression was not found to correlated to DAS28 score, anti-CCP antibody level or RF in the RA patients. The expressions of PAD4 and STAT4 were significantly different between the two RA groups (P<0.05). In the RA patients, PAD4 mRNA expression was positively correlated to DAS28 and anti-CCP antibody level (P<0.05), and STAT4 expression showed positive correlations to DAS28 and RF levels (P<0.05).CONCLUSION:HLA-DR4, PAD4 and STAT4 are overexpressed in RA patients and may be involved in the pathogenesis of RA. The expressions of PAD4 and STAT4, but not HLA-DR4, are closely related to the disease activity of RA. Detection of peripheral blood PAD4 and STAT4 expressions can be helpful for evaluating the disease activity of RA.

Objective To evaluated bone metabolism,radiology and OPG / RANKL serum levels in patients with ankylosing spondylitis (AS) treated with Gu Ling tang and rh TNFR:Fc. Methods Fifty two AS patients were randomly divided into 2 groups:the therapy group (26 cases) treated with Gu Ling tang and rh TNFR:Fc,the control group (26 cases) treated with rh TNFR:Fc only. Serum samples from the patients were obtained at baseline and 24 weeks after treatment. Disease activity indexes and serum levels of Osteocalcin(OC),C-terminal telopeptides (CTX),receptor activator of nuclear factor-κB ligand (RANKL),and osteoprotegerin (OPG) were measured before and after treatment respectively. And the X-ray images of patient's pelvis were graded based on BASRI before and after treatment. Results After 24 weeks of treatment,the clinical efficacy indexes were significantly improved in patients of both groups(P0.05),and no significant difference was found in the indexes between the two groups. Bone metabolism:Serum levels of OC were significantly increased(P0.05),while serum levels of CTX were significantly decreased(P0.05) in the two groups after 24 weeks. Between two groups,serum levels of OC and CTX were significantly(P0.05)after 24 weeks of treatment. BASRI:Improvement of BASRI-SIJ and BASRI-h were not significant(P 0.05) after 24 weeks and improvement in two groups. Between two groups,changes of BASRI-SIJ and BASRI-h were not significant(P 0.05). Serum levels of OPG and RANKL:Serum levels of OPG were significantly elevated (P 0.05) and serum levels of RANKL were significantly decreased after treatment at week 24 (P0.05),in two groups. Between two groups,serum levels of RANKL/OPG were significantly(P0.05)after 24 weeks of treatment. Conclusion Gu Ling tang with rh TNFR:Fc may be effective for reducing disease activity and improving bone metabolism. It is indicated that Gu Ling tang may interrupt the osteoclasia and radiological improvement through adjusting OPG system in patients with AS.

Objective To investigate the growth inhibition and apoptosis of fibroblast-like synoviocytes (FLSs) induced by the proteins extracted from Zaocys dhumnades, and to explore the underlying molecular mechanisms. Method Proteins from Zaocys dhumnades were obtained using the traditional protein extraction methods. Primary FLSs were obtained from synovium in patients with RA. Various doses (low, medium and high doses) of snake proteins were used to treat FLSs. Cell growth was determined by the MTT assay. Flow cytometry was employed to determine the apoptosis of FLS. Expression of wt-p53 and bcl-2 mRNA were determined using RT-PCR. Result The proliferation of FLS was significantly inhibited by the medium and high doses of proteins extracted from Zaocys dhumnades(P0.01), and the apoptotic rate of FLS was also increased(P0.01). The expression levels of wt-p53 in FLS treated with medium and high doses of the snake proteins were significantly higher than their corresponding negative control groups (P 0.01), and the expression levels of bcl2-mRNA were also significantly decreased (P0.01). Conclusion The growth inhibition and induction of apoptosis were probably achieved via the regulation of expression of the wt-p53 and bcl-2 genes.

Objective To observe the effect of sinomenine on the proliferation of fibroblast-like synoviocytes (FLS) and expression level of MMP-3 secreted by FLS in rheumatoid arthritis in vitro. Methods FLS were obtained by digesting synovial tissues with collagens and were divided into four groups:sinomenine high, middle, low concentration and control group. The proliferation of FLS was assessed by methyl- thiazolyl-tetrazolium (MTT) assay and the mRNA expression of MMP-3 was measured by semi-quantitative RT-PCR respectively after treated by sinomenine. Results The rate of FLS proliferation was significantly reduced in groups treated by sinomenine. Compared with control group, the mRNA expression of MMP-3 markly decreased in sinomenine groups (P <0.05) and especially in sinomenine high group (P <0.01). Conclusion Sinomenine can effectively inhibit the proliferation of FLS and reduce the mRNA expression of MMP-3 of FLS in rheumatoid arthritis, that may be a mechanism in therapy of rheumatoid arthritis by sinomenine.

Objective To observe the histomorphology effects and biomechanical abilities of Chinese herbal medicine invigorating the kidney(Guling Pian) on osteoporotic(OP) ovariectomized(OVX) rats model,and explore its therapeutic mechanism.Methods 36 femal 3-month-old SD rats were randomized into sham-operated,OVX,Guling Pian low,medium,high dosage and ethynylestradiol groups,6 in each group.The dynamic and static parameters of lumbar vertebrae were tested by bone morphometry,the maximum load and rigidity of lumbar vertebrae were determined by biomechanical indentation testing 4 months later.Results ①The bone trabecula resolution,bone formation rate(BFR)/BS and BFR/BV,the number of osteoclast per mm in different dosages of Guling Pian and ethynylestradiol groups were significantly decreased than those in OVX group(P<0.01).The BFR/TV in moderate dosage Guling Pian group and bone mineral apposition rate(MAR) in middle and high dosages of Guling Pian groups were significantly decreased(P<0.01).The bone trabecula area percentage,thickness and number in moderate and high dosages of Guling Pian and ethynylestradiol groups obviously increased,the bone trabecula thickness of moderate dosage of Guling Pian group was higher than that in ethynylestradiol group(P<0.05).②The rigidity of ethynylestradiol and different dosages of Guling Pian groups and the maximum load in moderate and high dosages of Guling Pian groups were significantly higher than those in OVX group(P<0.01).The maximum load of high dosage of Guling Pian group was significantly different from that of ethynylestradiol group(P<0.05).Conclusions Guling Pian has double effects on restraining bone absorption and promoting bone formation,may decrease bone turnover to improve bone quality and to increase bone biomechanics intensity.

Objective To investigate the effect of kdney-tonifying tranditional Chinese medicine,Gu Ling Pian(GLP)on regulating human osteoblasts function and the osteoprotegerin(OPG)/receptor activator of nuclear factor kappa B ligand(RANKL)expression via p38 MAPK pathway in vitro.Methods Human osteoblast were cultured with serum obtained from rat fed with high,medium,or low dose GLP.The expression levels of the OPG and RANKL mRNA were determined by real-time RT-PCR.Total p38 and phosphorylated p38 were detected by western blot.Mineralized bone nodule was detected by alizarin red staining method.Results GLP serum promoted osteoblasts differentiation,increased the formation of mineralized bone nodule,up-regulated the OPG expression while RANKL expression decreased and increased the total protein of phosphorylated p38.Conclusion Kdney-tonifying traditional Chinese medicine,Gu Ling Pian,induced osteoblasts differentiation and synthesis of OPG/RANKL probably through p38 MAPK pathway.

目的 :观察中药单体青藤碱(SIN)对脂多糖(LPS)刺激的 THP- 1细胞增殖、凋亡的影响.方法:培养的 THP- 1细胞经 LPS 1 mg/L刺激 2 h后,分为 SIN高(0.3 mmol/L)、中(0.15 mmol/L)、低(0.075 mmol/L)剂量组及空白对照组,干预 24 h.采用四甲基偶氮唑蓝法检测细胞的增殖情况,碘化丙啶染色流式细胞检测凋亡指数,吖啶橙(AO) /溴乙锭(EB)荧光显微镜观察细胞凋亡水平.结果 :与空白对照组相比,SIN高(P＜ 0.01)、中(P＜ 0.05)剂量组细胞增殖明显降低,流式细胞检测 SIN 3个剂量组细胞凋亡率增高,AO/EB染色示 SIN处理的细胞出现不同程度的核裂解和胞膜受损等凋亡现象.结论 :SIN可能通过抑制单核细胞的增殖,促进其凋亡来达到免疫抑制和治疗类风湿关节炎的作用.

AIM To explore the expression of avian reticuloendotheliosis viral(v-rel) oncogene-related B (RelB) mRNA in vitro in murine mature and immature myeloid dendritic cells (DCs). METHODS The bone marrow was collected from the femur and tibias of C57BL/6 mice in sterile condition. Bone marrow precursors were cultured with recombinant mouse granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and interleukin-4 (rmIL-4) to produce immature DCs. Immature DCs were stimulated by LPS 18 h before the end of culture to become mature. DCs' phenotype was detected by flow cytometry (FCM). RelB expression and protein level were determined by RT-PCR and immunofluorescence staining. RESULTS The expression level of co-stimulatory molecules (CD86 and CD40) and MHC-II class molecule were low in immature DCs, whereas high in mature DCs. RelB expression was significantly higher in mature DCs than in immature DCs (P<0.01). CONCLUSION RelB expression is closely associated with the maturative situation of DCs. Inhibition of RelB expression in DCs may induce tolerogenic DCs.

目的观察类风湿关节炎(rheumatoidarthritis,RA)患者血清巨噬细胞移动抑制因子(macrophagemigrationinhibitoryfactor,MIF)水平与疾病活动程度的关系。方法选取58例RA患者,根据疾病活动评分分为低度活动组14例、中度活动组33例和高度活动组11例,用ELISA法检测血清中MIF的表达,以15名正常人作对照,并检测RA患者血清中的RF,抗环瓜氨酸肽(cycliccitrullinatedpep-tide,CCP)抗体及C-反应蛋白,观察其与MIF的关系。结果与对照组比较,MIF在RA低、中、高度活动组的表达均有所增高(P<0.05~0.01),MIF与RA疾病活动评分及RF、C-反应蛋白均呈正相关关系(r=0.72~0.86,P=0~0.014;r=0.91,P<0.01;r=0.83,P<0.01),与抗CCP抗体则无相关关系(r=0.56,P>0.05)。结论在RA疾病活动时MIF水平增高,并能反映疾病活动的程度,可以作为临床观察RA疾病活动的指标之一。

OBJECTIVE: To observe the effect of sinomenine, a traditional Chinese medicine monomer, on the expression of in vitro cultured CD80mRNA and CD86mRNA in dendritic cells in rheumatoid arthritis (RA). METHODS: The peripheral blood mononuclear cells isolated from 10 patients with RA were divided into sinomenine (high, medium and low concentrations) groups and blank control group and costimulated by recombinant human GM-CSF and recombinant human interleukin-4 (rhIL-4). The expressions of CD80 mRNA and CD86mRNA were detected by semi-quantitation reverse transcription polymerase chain reaction (RT-PCR). RESULTS: As compared to the control group, the expressions of CD80 mRNA and CD86mRNA in dendritic cells intervened by sinomenine (high, medium and low concentrations, respectively) (P0.01 or P0.05),there were differences among the 3 subgroups of sinomenine(P0.05). CONCLUSION: The therapeutical effect of sinomenine on rheumatoid arthritis is probably achieved through its inhibition on the expressions of CD80mRNA and CD86mRNA of dendritic cells, thus blocking the costimulation of which on T cells.

Objective To explore the effects of Chinese herbs on the gene expression of osteopontin(OPN) in rat cultured in vitro. Methods Osteoblasts were from the skull of newborn SD rats and treated with high, middle or low concentration of the Chinese herbs. RT-PCR and western blot were used to detect the expression of OPN. Results At both high and middle concentration of the Chinese herb could up regulate the levels of OPN RNA and protein. Conclusion Kidney-tonifying traditional Chinese herb can promote the function of bone formation. Increase in mineral nodes may be due to the up regulation of both OPN RNA and protein.

Objective To observe the effect of sinomenine on the activity of nuclear transcription factor kappa B in dendritic cells in rheumatoid arthritis in vitro.Methods Peripheral blood mononuclear cells isolated from 12 rheumatoid arthritis patients were divided into four groups:sinomenine high,middle,low concentration groups and control group and were cultured with GM-CSF,(IL-4).The sinomenine group was intervened with different concentration of sinomenine after 6 days and harvested 48 housr later.The activity of nuclear factor kappa B was examined by electrophoretic mobility shift assay and IL-12 released by dendritic cells was measured by enzyme linked immunosorbent assay.Results Nuclear factor kappa B activity and expression of IL-12 were inhibited by sinomenine at concentration ranging from 0.05～0.3 mmol/L.Positive correlation was found between the activity of nuclear factor kappa B and the expression of IL12.Conclusion Sinomenine inhibit antigen presenting function of dendritic cells by decreasing its nuclear factor kappa B activity.