Objective To study the relationship of serum 25-hydroxyvitamin D3[25-(OH)D3],myostatin(MSTN),limb skeletal muscle index(ASMI)with osteopenia(OP)in patients with rheumatoid arthritis(RA)complicated with sarcopenia(SA).Methods A total of 244 RA patients selected as case group,and 123 healthy people who underwent physical examination in our hospital during the same period were selected as control group.The general data,serum levels of 25-(OH)D3,MSTN,ASMI and peripheral blood immune cells were compared among the two groups.The diagnostic value of serum 25-(OH)D3,MSTN and ASMI in RA patients with SA complicated with OP and the correlation between serum 25-(OH)D3,MSTN,ASMI and peripheral blood immune cells were analyzed.Results The levels of serum 25-(OH)D3,peripheral blood regulatory T lymphocyte(Treg)cells and ASMI in the case group were lower than those in the control group,while the levels of serum MSTN,peripheral blood helper T lymphocyte 17(Th17)cells and Th17/Treg were higher than those in the control group(P＜0.05).Among the 244 RA patients,171 had no SA,73 had SA,and the incidence of SA was 29.92％.The levels of serum 25-(OH)D3,peripheral blood Treg cells and ASMI in the SA group were lower than those in the non-SA group,and the levels of serum MSTN,peripheral blood Th17 cells and Th17/Treg were higher than those in the non-SA group(P＜0.05).The levels of serum 25-(OH)D3,peripheral blood Treg cells and ASMI in the OP group were lower than those in the non-OP group,while the levels of serum MSTN,peripheral blood Th17 cells and Th17/Treg were higher than those in the non-OP group(P＜0.05).The area under the curve(AUC)of combined detection of serum 25-(OH)D3,MSTN and ASMI for the diagnosis of OP in RA patients with SA was higher than that of the three alone(P＜0.05).Serum 25-(OH)D3 and ASMI in RA patients with SA complicated with or without OP were negatively correlated with peripheral blood Th17 cell level and Th17/Treg(P＜0.05),and positively correlated with peripheral blood Treg cell level(P＜0.05).The level of serum MSTN was positively correlated with the level of Th17 cells and Th17/Treg in peripheral blood(P＜0.05),and negatively correlated with the level of Treg cells in peripheral blood(P＜0.05).Conclusion The incidence of OP in RA patients with SA is high.At the same time,the occurrence of SA and OP can lead to the decrease of serum 25-(OH)D3,peripheral blood Th17 cell,Th17/Treg and ASMI,and the increase of serum MSTN and peripheral blood Treg cell.The combination detection of 25-(OH)D3,MSTN and ASMI can effectively improve the diagnosis of OP in RA patients with SA,and the levels of the three are significantly correlated with peripheral blood Th17 cells,Treg cells and Th17/Treg.

Objective To study the relationship of serum 25-hydroxyvitamin D3[25-(OH)D3],myostatin(MSTN),limb skeletal muscle index(ASMI)with osteopenia(OP)in patients with rheumatoid arthritis(RA)complicated with sarcopenia(SA).Methods A total of 244 RA patients selected as case group,and 123 healthy people who underwent physical examination in our hospital during the same period were selected as control group.The general data,serum levels of 25-(OH)D3,MSTN,ASMI and peripheral blood immune cells were compared among the two groups.The diagnostic value of serum 25-(OH)D3,MSTN and ASMI in RA patients with SA complicated with OP and the correlation between serum 25-(OH)D3,MSTN,ASMI and peripheral blood immune cells were analyzed.Results The levels of serum 25-(OH)D3,peripheral blood regulatory T lymphocyte(Treg)cells and ASMI in the case group were lower than those in the control group,while the levels of serum MSTN,peripheral blood helper T lymphocyte 17(Th17)cells and Th17/Treg were higher than those in the control group(P＜0.05).Among the 244 RA patients,171 had no SA,73 had SA,and the incidence of SA was 29.92％.The levels of serum 25-(OH)D3,peripheral blood Treg cells and ASMI in the SA group were lower than those in the non-SA group,and the levels of serum MSTN,peripheral blood Th17 cells and Th17/Treg were higher than those in the non-SA group(P＜0.05).The levels of serum 25-(OH)D3,peripheral blood Treg cells and ASMI in the OP group were lower than those in the non-OP group,while the levels of serum MSTN,peripheral blood Th17 cells and Th17/Treg were higher than those in the non-OP group(P＜0.05).The area under the curve(AUC)of combined detection of serum 25-(OH)D3,MSTN and ASMI for the diagnosis of OP in RA patients with SA was higher than that of the three alone(P＜0.05).Serum 25-(OH)D3 and ASMI in RA patients with SA complicated with or without OP were negatively correlated with peripheral blood Th17 cell level and Th17/Treg(P＜0.05),and positively correlated with peripheral blood Treg cell level(P＜0.05).The level of serum MSTN was positively correlated with the level of Th17 cells and Th17/Treg in peripheral blood(P＜0.05),and negatively correlated with the level of Treg cells in peripheral blood(P＜0.05).Conclusion The incidence of OP in RA patients with SA is high.At the same time,the occurrence of SA and OP can lead to the decrease of serum 25-(OH)D3,peripheral blood Th17 cell,Th17/Treg and ASMI,and the increase of serum MSTN and peripheral blood Treg cell.The combination detection of 25-(OH)D3,MSTN and ASMI can effectively improve the diagnosis of OP in RA patients with SA,and the levels of the three are significantly correlated with peripheral blood Th17 cells,Treg cells and Th17/Treg.

Objective:To evaluate the role of exosomes in neuronal injury induced by M1 microglia.Methods:Liposolysaccharide 100 ng/ml and interferon-γ (IFN-γ)20 ng/ml were added to well-growing BV2 microglia to induce the polarization of microglia into M1 phenotype.Cell supernatant of M1 microglia was collected and M1 microglia exosomes (M1-exo) were extracted with exosome kit.The well-growing N2a cells were divided into 4 groups ( n=24 each) using a random number table method: control group (group C), M1 microglia group (group M), exosome group (group E), and exosome inhibitor+ M1 microglia group (group G+ M). The cells in group C were conventionally cultured, the cells in group M were cultured with the supernatant of M1 microglia for 24 h, and the cells in group E were cultured with M1 microglia-derived exosomes for 24 h. In G+ M group, exosome inhibitor GW4869 was added, M1 microglia were incubated for 24 h, then the supernatant was collected and added to N2a cells, and the cells were incubated for 24 h. Cell viability of N2a cells was measured by the cell counting kit 8 assay, cell apoptosis rate was determined by flow cytometry.The expression of apoptosis-related genes Bcl-2 and Bax mRNA was detected by quantitative real-time-polymerase chain reaction, and the expression of apoptosis-related genes Bcl-2 and Bax protein was detected by Western blot. Results:Compared with group C, the cell viability was significantly decreased, the apoptosis rate was increased, the expression of Bcl-2 protein and mRNA was down-regulated, and the expression of Bax protein and mRNA was up-regulated in the other three groups ( P<0.05). Compared with group M, the cell viability was significantly increased, the apoptosis rate was decreased, the expression of Bcl-2 protein and mRNA was up-regulated, and the expression of Bax protein and mRNA was down-regulated in group G+ M ( P<0.05). There was no significant difference in the above indexes between group E and group M ( P>0.05). Conclusions:M1 microglia can mediate neuronal injury via exosomes.

Objective:To evaluate the role of miR-205-3p in oncosis in astrocytes subjected to oxygen-glucose deprivation and restoration (OGD/R) and the relationship with aquaporin4 (AQP4).Methods:Primary astrocytes were cultured in vitro to the logarithmic growth phase and divided into 5 groups ( n=16 each) using a random number table method: control group (C group), OGD/R group (O group), OGD/R+ miR-205-3p mimic group (M group), OGD/R+ miR-205-3p inhibitor group (I group), and OGD/R+ negative control group (NC group). Cells were cultured routinely in C group.Cells were subjected to 4 h of oxygen-glucose deprivation in a 37℃ anaerobic incubator (containing 94% N 2, 1% O 2 and 5% CO 2) followed by restoration of O 2-glucose supply for 24 h in O group.Cells in M, I and NC groups were transfected with miR-205-3p mimic, miR-205-3p inhibitor and miR-205-3p negative control for 48 h, respectively, and then cells were subjected to 4 h of oxygen-glucose deprivation followed by restoration of O 2-glucose supply for 24 h. The cell viability was evaluated by CCK-8 assay, the cell injury and oncosis were analyzed by flow cytometry, the expression of AQP4 mRNA was detected by quantitative reverse transcription-polymerase chain reaction, and the expression of AQP4 and porimin was detected by Western blot. Results:Compared with C group, the expression of miR-205-3p was significantly down-regulated, the cell viability was decreased, the rates of cell injury and oncosis were increased, and the expression of AQP4 protein and mRNA and porimin was up-regulated in O group ( P<0.05). Compared with O group, the expression of miR-205-3p was significantly up-regulated, the cell viability was increased, the rates of cell injury and oncosis were decreased, and the expression of AQP4 protein and mRNA and porimin was down-regulated in M group, the expression of miR-205-3p was significantly down-regulated, the cell viability was decreased, the rates of cell injury and oncosis were increased, and the expression of AQP4 protein and mRNA and porimin was up-regulated in I group ( P<0.05), and no significant changes were found in NC group( P>0.05). Conclusions:miR-205-3p is involved in oncosis in astrocytes subjected to OGD/R, which is associated with regulation of AQP4 expression.

OBJECTIVETo investigate the effect of aloin on apoptosis of human gastric cancer cells and explore the molecular mechanism.

METHODSGastric cancer MKN-28 and HGC-27 cells were cultured routinely in 1640 medium supplemented with 10% fetal bovine serum and 10% non-essential amino acids (for HGC-27 cells) and treated with different concentrations of aloin for different durations. The cell viability, cell nuclear morphology, and apoptotic rate of the cells were detected using CCK-8 assay, DAPI staining and AnnexinV-FITC/PI, respectively; Western blotting was used to detect the expression levels of PARP, procaspase 3 and the phosphorylation of p38, ERK and JNK. The cells were treated with specific inhibitors of p38, ERK and JNK, and the inhibitory effects on these pathways were detected with Western blotting; DAPI staining was used to detect the effects of inhibitors on apoptosis of gastric cancer cells.

RESULTSAloin dose-dependently inhibited the viability and induced apoptosis of HGC-27 and MKN-28 cells. Alion treatment obvious enhanced the phosphorylation of p38 and JNK but decreased ERK phosphorylation in the cells. Blocking ERK activation with the ERK inhibitor obviously enhanced aloin-induced cell apoptosis, where inhibiting p38 and JNK activation partly reversed alion-induced apoptosis in the cells.

CONCLUSIONSAloin induces apoptosis of human gastric cancer cells by activating p38 and JNK signaling pathways and inhibiting ERK signaling pathway.

Objective To evaluate the performance of fast Treponema pallidum(TP) detection in voluntary blood donors and optimize the strategy for pre-donation screening.Methods Before blood donation,the gold standard TP test strip was used to make a fast detection.After blood donation,the TP-ELISA was used to test the blood.Then,analyze the donors′ anti-TP positive rate,times and intervals of donating,false positive and negative of TP fast detection.Results From 2014 to 2015,among 73 990 donors who were tested by using fast TP detection,0.71% of them(529 donors) were positive.Among the positive donors,89.2% of them(472 donors) were first-time blood donors.35 donors′ donating intervals were more than 3 years,who accounted for 61.4% of the donors who had donated for more than once.The numbers of the false positive obtained from fast TP detection were 5 and the false negative was 15.By applying the fast TP detection before blood donation,the rate of anti-TP positive had been declined from 0.71% to 0.17%.Conclusion The rejection rate of TP positive can be significantly reduced by using fast TP detection before blood donation.The fast TP detection can be used to optimize the pre-donation screening and promote blood donation service efficiency and level,while donating times and intervals of the blood donors were also considered.

Objective To evaluate the effect of hypoxia-preconditioned bone marrow mesenchymal stem cells (BMSCs) on the expression of HO-1 during spinal ischemia-reperfusion (I/R) in rats.Methods Thirty healthy adult Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 5 groups with 6 animals in each group using a random number table:sham operation group (group S),I/R group,PBS group,BMSC transplantation group (BMSC group) and hypoxic preconditioning group (group HP).In group HP,BMSCs were exposed to 3％O2-5％CO2-92％N2 for 24 h for hypoxia preconditioning.In PBS,BMSC and HP groups,PBS,BMSCs and BMSCs for hypoxic preconditioning were injected intrathecally,respectively,and 60 min later spinal I/R was induced by clamping the aorta in the rats anesthetized with chloral hydrate.At 6,12,24 and 48 h and 7 days of reperfusion,the neurological function was evaluated and scored.At day 7 of reperfusion,the rats were sacrificed,and spinal cord tissues were obtained for determination of HO-1 expression (by Western blot) and HO-1 mRNA expression (using real-time PCR).Results Compared with group S,the neurological function score was significantly decreased at each time point of reperfusion,and the expression of HO-1 mRNA and protein in spinal cord tissues was up-regulated in the other groups.Compared with group I/R,the neurological function score was significantly increased at each time point of reperfusion,and the expression of HO-1 mRNA and protein in spinal cord tissues was upregulated in BMSC and HP groups,and no significant change was found in group PBS in the parameters mentioned above.Compared with group BMSC,the neurological function score was significantly increased at each time point of reperfusion,and the expression of HO-1 mRNA and protein in spinal cord tissues was up-regulated in group HP.Conclusion The mechanism by which hypoxic preconditioning enhances protection of spinal cord by BMSC transplantation may be related to up-regulation of HO-1 expression in rats.

Objective To investigate the protective effect of hypoxic-preconditioned bone marrow mesenchymal stem cells (BMSCs) on spinal cord tissue after ischemia reperfusion injury.Methods Healthy adult Sprague Dawley (SD) rats weighing 200 ～ 250 grams (g) were randomly divided into 3 groups with 6 animals in each group:The sham group received simple surgical manipulation without ischemia/reperfusion treatment;The spinal cord ischemia/reperfusion group (Control group) only received spinal cord ischemia/reperfusion surgery.The hypoxic preconditioned BMSC transplantation group (HP-MSCs group) was injected with hypoxic preconditioned BMSCs 2 days before ischemia/reperfusion.The control group,HP-MSCs group received spinal cord ischemia/reperfusion for 10 min and observed for 48 h.The permeability of the blood-spinal cord barrier was examined with Evans blue (EB),and the histomorphology changes were observed with hematoxylin and eosin (HE) staining.Results EB red fluorescence was significantly weakened in the HP-MSCs group than that in the Control group (P ＜ 0.05),and more intact motor neurons were found in the lumbar spinal cords in the HP-MSCs group than that in the Control group (P ＜0.05).Conclusions The hypoxic-preconditioned BMSCs could effectively attenuate spinal cord ischemia reperfusion injury,it may be associated with protective effect of the blood-spinal cord barrier integrity.

Objective To evualate the clinical efficacy of Minor Bluegreen Dragon Decoction in the treatment of chronic cough.Methods 106 cases of chronic cough were randomly divided into Minor Bluegreen Dragon Decoction in the treatment group of 59 cases, and Compound Liquorice Tablets treatment of 47 cases of the control group, the improvement de-gree of the symptoms (cough and expectoration) were observed before and after treatment.Results The treatment group and the control group the total effective rate were 86.4% and 36.2%, and the treatment group was significantly higher than that in control group (p<0.01).Two groups of cough and expectoration symptom improvement in varying degrees after treatment, but the treatment group was more effective(p<0.05).Conclusion Minor Bluegreen Dragon Decoction treatment of chronic cough patients (excluding other respiratory disease), is effective and safe, which is worthy of clinical promotion.

Objective To evaluate the efficacy of 10-day compound allantoin containing quadruple regimen in the treatment of chronic gastritis with Helicobacter pylori (H.pylori) infection,and to compare with the bismuth-containing quadruple therapy.Methods Altogether 173 patients with H.pylori positive chronic gastritis confirmed by gastric endoscope were divided into 10-day compound allantoin containing quadruple regimen group (n =43),24-day compound allantoin containing quadruple regimen group (n =46),10-day bismuth-containing quadruple regimen group (n =42) and 24-day bismuth-containing quadruple regimen group (n =42).After the treatment,the eradication rate of H.pylori,the rate of gastrointestinal symptoms (epigastric pain,bloating and belching) relief and the adverse effects of each group were observed.Intention-to-treat (ITT),per-protocol (PP) statistical analysis and chi-square analysis were performed for statistical analysis.Results H.pylori eradication rates of 10-day compound allantoin containing quadruple regimen group,24-day compound allantoin containing quadruple regimen group,10-day bismuth-containing quadruple regimen group and 24-day bismuth-containing quadruple regimen group analyzed by ITT were 90.7％ (39/43),91.3％ (42/46),90.5％ (38/42) and 88.1％ (37/42),respectively; while analyzed by PP were 90.7％ (39/43),93.3％ (42/45),90.5％ (38/42) and 90.2％ (37/41),respectively.And there were no statistical differences between groups (all P＞0.05).Ten days after the treatment,the rates of epigastric pain relief of 24-day compound allantoin containing quadruple regimen group and 24-day bismuth-containing quadruple regimen group were 81.1％ (30/37) and 78.8％ (26/33),respectively,the rates of bloating relief were 82.4％ (28/34) and 71.0％ (22/31),respectively,and the rates of belching relief were 76.9％ (20/26) and 75.0％ (21/28),respectively.There were no statistical differences between the two groups (all P＞ 0.05).However after 24-day treatment,the rates of epigastric pain relief of 24-day compound allantoin containing quadruple regimen group and 24-day bismuth-containing quadruple regimen group were 91.9 ％ (34/37) and 87.9％ (29/33),respectively,the rates of bloating relief were 94.1％ (32/34) and 87.1％ (27/31),respectively,and the rates of belching relief were 96.2％ (25/26) and 85.7％ (24/28),respectively.There were no statistical differences between the two groups (all P＞0.05).And the rates of epigastric pain and bloating relief increased after 24-day treatment compared with those of 10-day treatment,however the differences were not statistically significant between the two groups (all P＞ 0.05).In 24-day compound allantoin containing quadruple regimen group,the rate of belching relief was higher after 24-day treatment compared with that of 10-day treatment,and the difference was statistically significant (x2=4.127,P=0.042).No severe adverse effects were observed in each group,and there were no adverse effects such as oral metal odor,tongue black and melena in compound allantoin containing quadruple therapy.Conclusions Ten-day compound allantoin containing quadruple therapy as first-line approach in the treatment of chronic gastritis with H.pylori infection can get better H.pylori eradication and the efficacy is similar to bismuth quadruple therapy.Meanwhile the symptom relief rate is high and no obvious adverse effects were found.