The innate immune sensor NLRP3 inflammasome overactivation is involved in the pathogenesis of ulcerative colitis. PGAM5 is a mitochondrial phosphatase involved in NLRP3 inflammasome activation in macrophages. However, the role of PGAM5 in ulcerative colitis and the mechanisms underlying PGAM5 regulating NLRP3 activity remain unknown. Here, we show that PGAM5 deficiency ameliorates dextran sodium sulfate (DSS)-induced colitis in mice via suppressing NLRP3 inflammasome activation. By combining APEX2-based proximity labeling focused on PGAM5 with quantitative proteomics, we identify NEK7 as the new binding partner of PGAM5 to promote NLRP3 inflammasome assembly and activation in a PGAM5 phosphatase activity-independent manner upon inflammasome induction. Interfering with PGAM5-NEK7 interaction by punicalagin inhibits the activation of the NLRP3 inflammasome in macrophages and ameliorates DSS-induced colitis in mice. Altogether, our data demonstrate the PGAM5-NEK7 interaction in macrophages for NLRP3 inflammasome activation and further provide a promising therapeutic strategy for ulcerative colitis by blocking the PGAM5-NEK7 interaction.

【Objective】 To investigate the role of microRNA-218 (miR-218) in regulating prostate cancer (PCa) cell stemness and epithelial-mesenchymal transition (EMT). 【Methods】 PCa cell line stably overexpressing miR-218 was constructed with lentivirus transfection. The expression of miR-218 was detected with real-time fluorescence quantitative polymerase chain reaction (q-PCR). The migration ability was detected with Transwell assay. The expression of EMT related proteins were detected with Western blot. The properties of cells were determined with colony formation and tumor sphere formation assays. 【Results】 The results of q-PCR showed that the mRNA level of miR-218 was significantly lower in PCa cell lines LNCaP and C4-2 than in BPH-1. Transwell assay showed that miR-218 inhibited the migration of PCa cells. Western blot showed that the expression of EMT related proteins were inhibited by miR-218. Colony formation and tumor sphere formation assays showed that overexpression of miR-218 significantly inhibited the properties of cells. 【Conclusion】 The expression of miR-218 is downregulated in PCa cell lines. miR-28 can inhibit cell migration, EMT and cancer stem cell properties.

OBJECTIVE: To investigate the effects of melatonin (MT) on bone mass and serum inflammatory factors in rats received ovariectomy (OVX) and to investigate the effects of MT on the levels of inflammatory factors in culture medium and osteogenic ability of bone marrow mesenchymal stem cells (BMSCs) stimulated by lipopolysaccharide. METHODS: Fifteen 12-week-old Sprague Dawley (SD) rats were randomly divided into 3 groups. The rats in Sham group only received bilateral lateral abdominal incision and suture, the rats in OVX group received bilateral OVX, and the rats in OVX+MT group received 100 mg/(kg·d) MT oral intervention after bilateral OVX. After 8 weeks, the levels of serum inflammatory factors [interleukin-1β (IL-1β), IL-6, and tumor necrosis factor α (TNF-α)] were detected using ELISA assay. Besides, the distal femurs were detected by Micro-CT to observe changes in bone mass and microstructure, and quantitatively measured bone volume fraction, trabecular thickness, and trabecular number. The BMSCs were extracted from the femurs of three 3-week-old SD rats using whole bone marrow culture method and passaged. The 3rd-5th passage BMSCs were cultured with different concentrations of MT (0, 1, 10, 100, 1 000 µmol/L), and the cell viability was then detected using cell counting kit 8 (CCK-8) to select the optimal concentration of MT for subsequent experiments. Cells were devided into osteogenic induction group (group A) and osteogenic induction+1/5/10 μg/mL lipopolysaccharide group (group B-D). The levels of inflammatory factors (IL-1β, IL-6 and TNF-α) in cell culture medium were detected using ELISA assay after corresponding intervention. According to the results of CCK-8 method and ELISA detection, the cells were intervened with the most significant concentration of lipopolysaccharide for stimulating inflammation and the optimal concentration of MT with osteogenic induction, defining as group E, and the cell culture medium was collected to detect the levels of inflammatory factors by ELISA assay. After that, alkaline phosphatase (ALP) staining and alizarin red staining were performed respectively in groups A, D, and E, and the expression levels of osteogenic related genes [collagen type Ⅰ alpha 1 chain (Col1a1) and RUNX family transcription factor 2 (Runx2)] were also detected by real time fluorescence quantitative PCR (RT-qPCR). RESULTS: ELISA and Micro-CT assays showed that compared with Sham group, the bone mass of the rats in the OVX group significantly decreased, and the expression levels of serum inflammatory factors (IL-1β, IL-6, and TNF-α) in OVX group significantly increased (P<0.05). Significantly, the above indicators in OVX+MT group were all improved (P<0.05). Rat BMSCs were successfully extracted, and CCK-8 assay showed that 100 µmol/L was the maximum concentration of MT that did not cause a decrease in cell viability, and it was used in subsequent experiments. ELISA assays showed that compared with group A, the expression levels of inflammatory factors (IL-1β, IL-6, and TNF-α) in the cell culture medium of groups B-D were significantly increased after lipopolysaccharide stimulation (P<0.05), and in a concentration-dependent manner. Moreover, the expression levels of inflammatory factors in group D were significantly higher than those in groups B and C (P<0.05). After MT intervention, the expression levels of inflammatory factors in group E were significantly lower than those in group D (P<0.05). ALP staining, alizarin red staining, and RT-qPCR assays showed that compared with group A, the percentage of positive area of ALP and alizarin red and the relative mRNA expressions of Col1a1 and Runx2 in group D significantly decreased, while the above indicators in group E significantly improved after MT intervention (P<0.05). CONCLUSION MT may affect the bone mass of postmenopausal osteoporosis by reducing inflammation in rats; MT can reduce the inflammation of BMSCs stimulated by lipopolysaccharide and weaken its inhibition of osteogenic differentiation of BMSCs.
Female ; Rats ; Animals ; Tumor Necrosis Factor-alpha ; Osteogenesis ; Rats, Sprague-Dawley ; Core Binding Factor Alpha 1 Subunit ; Melatonin/pharmacology* ; Interleukin-6/genetics* ; Lipopolysaccharides/pharmacology* ; Coloring Agents ; Inflammation

This study aimed to obtain the first national estimate of the prevalence of autism spectrum disorder (ASD) in Chinese children. We targeted the population of 6 to 12-year-old children for this prevalence study by multistage convenient cluster sampling. The Modified Chinese Autism Spectrum Rating Scale was used for the screening process. Of the target population of 142,086 children, 88.5% (n = 125,806) participated in the study. A total of 363 children were confirmed as having ASD. The observed ASD prevalence rate was 0.29% (95% CI: 0.26%-0.32%) for the overall population. After adjustment for response rates, the estimated number of ASD cases was 867 in the target population sample, thereby achieving an estimated prevalence of 0.70% (95% CI: 0.64%-0.74%). The prevalence was significantly higher in boys than in girls (0.95%; 95% CI: 0.87%-1.02% versus 0.30%; 95% CI: 0.26%-0.34%; P < 0.001). Of the 363 confirmed ASD cases, 43.3% were newly diagnosed, and most of those (90.4%) were attending regular schools, and 68.8% of the children with ASD had at least one neuropsychiatric comorbidity. Our findings provide reliable data on the estimated ASD prevalence and comorbidities in Chinese children.

Objective To observe the expression change of melanoma-associated antigen(MAGE)-A9 in colorectal cancer (CRC)tissue and to explore its significance.Methods The samples in 23 cases of initially diagnosed CRC in the Affiliated Hospital of Nantong University from January 2006 to December 2008 were collected.The quantitative real-time(qRT)-PCR was adopted to detect MAGE-A9 mRNA expression in cancer tissue and corresponding paracancerous tissue.Its correlation with the clinicopatho-logical features and prognosis was analyzed.Results The positive rate of MAGE-A9 in CRC tissue was significantly higher than that in paracancerous normal tissue(P<0.05).MAGE-A9 protein expression in CRC was related to the clinicopathological features such as tumor differentiation degree(P=0.011),TNM stage(P=0.003),tumor infiltration depth(P=0.001)and lymph node me-tastasis(P=0.003).The survival analysis showed that the expression of MAGE-A9 was closely related to the prognosis of CRC pa-tients.Conclusion MAGE-A9 expression is increased in CRC tissue,suggesting the poor prognosis.

Objective To prepare quercetin ( QT )-loaded polylactic-co-glycolic acid-D-α-tocopheryl polyethylene glycol 1000 succinate ( PLGA-TPGS) nanoparticles ( QPTN) and QT-loaded polylactic-co-glycolic acid ( PLGA) nanoparticles ( QPN) by using QT as model drug and PLGA-TPGS or PLGA as carrier materials, and to investigate the quality of the two nanoparticles. Methods QPTN and QPN were prepared by using the ultrasonic emulsification-solvent evaporation method, and their surface morphology,size and surface charge were detected by using a transmission electron microscope ( TEM) and a Nano ZS90 light scattering and laser Doppler anemometry, respectively. Drug loading ( DL) , entrapment efficiency ( EE) and in vitro drug release of QT in the two nanoparticles were determined by using a reverse phase-high performance liquid chromatography (RP-HPLC) on Hypersil C18 column (4.6 mm×250 mm, 5 μm) with methanol and 0.03% phosphoric acid (3︰2) as mobile phase, and the detective wavelength was 370 nm. Results TEM images exhibited that two nanoparticles were all spherical and regular. The average sizes of QPTN and QPN were (155.4±2.7) nm and (363.8±3.2) nm, while DL and EE of QPTN were approximately (21.6±2.8)%, (93.7±2.9)% (n=6), and DL and EE of QPN were approximately (15.0±1.5)%, (64.6± 1.6)% (n=6), respectively. Both of nanoparticles exhibited sustained release, and the cumulative QT release of QPTN and QPN reached (85.8±2.8)% and (68.6±1.4)% (n=6) at day 30, respectively, with a significant difference between them (P<0.05) . Conclusion QPTN gets smaller size, higher DL and EE, and exhibits sustained release, and the in vitro cumulative QT release is faster and more complete than QPN relatively.

Objective To investigate the expressions of intercellular adhesion molecule (ICAM)-1 and matrix metalloproteinase (MMP)-2 in human gliomas and their correlation.Methods Sixty glioma samples,resected in our hospital from March 20,2010 to May 10,2014,were chosen in our study,and corroding to the WHO central nervous system tumor histological grading,they were divided into low-grade glioma group (grade Ⅰ-Ⅱ,n=26) and high-grade glioma group (grade Ⅲ-Ⅳ,n=34);other 10 normal brain tissues were used as controls.Reverse transcription-PCR and SP immumohistochemical staining were used to detect the ICAM-Ⅰ and MMP-2 mRNA and protein expressions.Results (1) The ICAM-1 and MMP-2 mRNA expressions in normal brain tissues,low-grade glioma group and high grade glioma group were increased in sequence,with significant difference (P＜0.05);the positive rate of ICAM-1 protein expression in normal brain tissues,low-grade glioma group and high grade glioma group was 20％,46.1％ and 88.2％,with significant differences (P＜0.05),and that in MMP-2 protein expression was 10％,50％ and 91.1％,with significant differences (P＜0.05).The mRNA expressions ofICAM-1 and MMP-2 in gliomas were positively correlated (r=0.702,P=0.001).Conclusions The expressions of ICAM-1 and MMP-2 are positively related to the malignant degrees of gliomas;the two may be useful reference factors to evaluate prognosis of human gliomas and potential targets for therapy.

Objective To observe any therapeutic effect of transcutaneous electric nerve stimulation (TENS) on patients with shoulder-hand syndrome after stroke (SHSAS) and to examine the influence of TENS on sympathetic skin response (SSR).Methods Sixty-eight patients with SHSAS were randomly divided into a treatment group (35 cases) treated with routine rehabilitation training and TENS therapy and a control group (33 cases)treated with routine rehabilitation training only.The therapy for both groups lasted 3 weeks.The severity of pain and edema of the affected upper limb was assessed with a visual analogue scale (VAS) while sympathetic skin response was recorded from the affected upper limb before and after treatment.Results VAS scores improved significantly in the treatment group,and significantly more than in the control group.There was no significant difference in the SSR latencies,amplitudes or abnormality rates between the two groups before treatment.The latencies and abnormality rates of both groups improved significantly after treatment,but the improvement in the treatment group was more obvious.The SSR amplitudes did not change significantly after treatment in either group.There was a positive correlation between the SSR latencies and abnormality rates and the VAS scores,but no significant correlation between SSR amplitude and the VAS scores.Conclusions TENS therapy combined with routine rehabilitation training showed not only good clinical results,but also significant changes in SSR among patients with SHSAS.This indicates that SSR could be used to evaluate therapeutic effects in SHSAS patients.

This paper introduced the implementing process, characteristics and effects of the web research learning of physiology. Research learning based on Web promoted reform of physiology teaching, enhanced ability of self-study, integration and innovative of students

Aim To observe the changes of diaphragm contractility and cytoskeletal proteins titin,nebulin and sarcoplasmic reticulum Ca~(2+)-ATPase gene expressions in adriamycin-induced cytotoxicity in rats.Methods The animal models of diaphragm damage were duplicated by injecting adriamycin into abdominal cavity one time.Forty male sprague-dawley rats were randomly divided into four groups(n=10):Three groups received adriamycin in low,middle and high dosage(10,20 and 40 mg·kg~(-1))respectively.Meanwhile,the normal saline was given to rats in control groups.Three days later,these rats were killed,and the diaphragm was removed by thoracotomy.The diaphragm contractility was assessed in isolated diaphragm strips perfusion by these paramemters including peak twitch tension(Pt),maximum tetanic tension(Po),time to peak contraction(CT),half relaxaion time(1/2RT),maximal rates of contraction(+dt/dt_(max))and maximal rates of relaxation(-dt/dt_(max)).The expressions of titin,nebulin and sarcoplasmic reticulum Ca~(2+)-ATPase(SERCA)at mRNA level were detected by RT-PCR analysis.Results In contrast to those in control group,Po,Pt,±dt/dt_(max) in the adriamycin group were lower(P<0.01);CT,1/2RT in the adriamycin group increased significantly(P<0.01).The levels of titin,nebulin and SERCA gene expressions in middle-dose group were lower than those in control group(P<0.01).Conclusions The mRNA levels of titin,nebulin and SERCA of diaphragm are down-regulated in adriamycin-induced cytotoxicity in rats.It may be associated with the decline of diaphragm contractility.