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Objective:To investigate the effect of rapamycin on the proliferation and apoptosis of rheumatoid arthritis synovial fibroblasts (RA-FLS) and its mechanism.Methods:Synovial tissues were collected from patients with RA during joint replacement surgery, and primary synovial fibroblasts were extracted by trypsin digestion. The effect of rapamycin on the proliferation of RA-FLS was detected by cell counting kit (CCK-8) method. RA-FLS were divided into the control group and the rapamycin group (10 nmol/L). The effect of rapamycin on apoptosis of RA-FLS cells was detected by flow cytometry. The mRNA expres-sion levels of mammalian target of rapamycin (mTOR), serine/threonine-protein kinase AKT, B lymphocy-toma-2 (Bcl-2) associated X gene (Bax) and Bcl-2 were detected by RT-PCR. The protein expression levels of Bax, Bcl-2, mTOR, p-mTOR (2448), AKT, p-AKT and mTORC1 downstream related molecules protein S6 kinase 1(S6K1), p-S6K1, eukaryotic translation initiation factor-binding protein 1 (4EBP1) and p-4EBP1 were detected by Western blot. Differences between the two groups were compared using two independent samples t-test. Results:The results showed that the proliferation efficiency of RA-FLS treated with rapamycin was significantly weaker than that of the control group, and the drug inhibition rate of rapamycin increased with the increase of rapamycin concentration. The apoptosis rate of rapamycin group was significantly higher than that of the control group (5.31±0.59)% vs (3.49±0.40)%, t=7.83, P=0.001). The expression of Bax mRNA in rapamycin group was significantly increased (1.35±0.04 vs 1.00±0.00, t=15.60, P=0.004), while the expression of Bcl-2 mRNA (0.790±0.003 vs 1.000±0.000, t=85.50, P=0.007), mTOR mRNA (0.41±0.08 vs 1.00±0.00, t=14.37, P=0.044) and AKT mRNA (0.59±0.08 vs 1.00±0.00, t=7.54, P=0.017) were decreased, and the differences were statistically significant when compared with the control group. Compared with the control group, the protein expression of Bax in rapamycin group was significantly increased (0.75±0.10 vs 0.48±0.09, t=4.04, P=0.007), and the expression levels of Bcl-2 (0.632±0.055 vs 0.758±0.020, t=7.35, P=0.002), p-AKT/AKT(0.61±0.07 vs 0.88±0.04, t=5.61, P=0.005), p-mTOR/mTOR(0.92±0.12 vs 1.28±0.09, t=5.05, P=0.002), p-S6K1/S6K1(0.884±0.020 vs 1.023±0.058, t=4.52, P=0.004) and p-4EBP1/4EBP1 were decreased(0.86±0.05 vs 1.11±0.05, t=6.00, P=0.004). Conclusion:Rapamycin may inhibit the proliferation and induce apoptosis of RA-FLS cells by inhibiting AKT/mTORC1 pathway.
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Objective:To explore the link between the differentially expressed long non-coding RNAs (lncRNAs) and the number of regulatory T cells (Tregs) by detecting the lncRNAs expression profiles in patients with systemic lupus erythematosus (SLE), then analyze the correlation between Tregs and lncRNAs and the clinical features of SLE patients. We also predict the mechanism by which lncRNAs regulate the differentiation and development of Tregs, and provid new approach for the treatment of SLE.Methods:Peripheral blood of 9 active SLE patients was collected and mononuclear cells (PBMCs) were extracted. The lncRNAs expression profiles of PBMCs was analyzed by whole transcriptome sequencing. Nine healthy people served as controls to screen the differentially expressed lncRNAs, and to analyze the correlation between lncRNAs and Tregs number. Pearson test was used to analyze the correlation between lncRNA and the number of Tregs, and the correlation between Treg-associated lncRNAs and systemic lupus erythematosus disease activity index(SLEDAI) score, erythrocyte sedimentation rate (ESR), C3, C4 in SLE patients. The targeted genes of Treg asso-ciated lncRNAs were predicted with miRcode and Targetscan databases and co-expression network.Results:There were 240 differentially expressed lncRNAs in SLE patients compared with healthy controls, including 134 highly expressed lncRNAs ( P<0.05) and 106 low expressed lncRNAs ( P<0.05). The expression of ANKRD44-AS1 ( r=0.74, P=0.022), LINC00200 ( r=0.70, P=0.037), AP001363.2 ( r=0.78, P=0.014) and LINC02824 (r=0.79, P=0.011) were positively correlated with the number of Tregs, and the expression of AP000640.1 ( r=-0.72, P=0.028), AC124248.1 ( r=-0.77, P=0.016), LINC00482 ( r=-0.83, P=0.005) and MIR503HG ( r=-0.96, P<0.001) were negatively correlated with the number of Tregs. Among these eight Tregs associated lncRNAs, the expression of LINC00482 ( r=-0.73, P<0.001) and MIR503HG ( r=-0.76, P<0.001) were negatively correlated with C3. LINC00200, ANKRD44-AS1 and AP000640.1 related to Tregs regulated the expression of STAT5, PLD1, HOPX and RUNX3 through competitively binding of miRNA or transregulatory mechanism, thereby regulating the differentiation and development of Tregs. Conclusion:The lncRNAs expression profiles are changed in SLE patients, the differentially expressed lncRNAs are associated with abnormal number and function of Tregs in SLE patients, and Treg associated lncRNAs are associated with SLE disease activity, which may affect the expression of STAT5, PLD1, HOPX, RUNX3 and regulate Tregs function and participate in the pathogenesis and progression of SLE by competitively binding to miRNAs or trans-regulatory mechanism.
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This paper summarized identification and reposition strategies of catheter malposition in 113 cancer patients with peripherally inserted central catheters(PICCs) during catheterization.In the process of PICC catheterization,catheter malposition was identified by ultrasound and ECG in a real-time manner.A series of timely and effective reposition procedures were performed by internal jugular vein blocking,breathing combined with anterior segment wire withdrawal Overall,111 cases were successfully repositioned,the success rate was 98.23%,and 2 cases failed because of other reasons such as diseases.
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Objective To investigate the protective mechanism of Interleukin 33 (IL-33) in preventing myocardial ischemia and reperfusion injury in rats. Methods A rat model with myocardial I/R with 32the adult male SD rats , which were randomly divided into 4 groups: SO group , I/R group , IL-33 + I/R group , SB230580 + IL-33 + I/R group. The levels of LDH, CK, TNF-α, IL-6, HMGB1, Bcl-2, total caspase-3, cleaved caspase-3 and P-P38 were detected. Results After reperfusion, IL-33 significantly decreased the levels of serum LDH and CK and the expression of TNF-α, IL-6 , cleaved caspase-3 , but significantly increased the expressions of Bcl-2, p-P38 (P < 0.05). SB230580 attenuated the protective role of IL-33 on myocardial I/R in a certain degree. Conclusions IL-33 may prevent myocardial I/R injury via inhibiting inflammation and cardiocyteapoptosis by way of P38 MAPK signaling pathway.
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<p><b>OBJECTIVE</b>To investigate whether selectively stimulating β1-adrenergic receptor could inhibit high mobility group box 1 protein and attenuate myocardial ischemia/reperfusion(I/R) injury in rats.</p><p><b>METHODS</b>Eighty healthy male Sprague-Dawley (SD) rats were randomly divided into seven groups: (1) Sham operated group (SO); (2) Ischemia/reperfusion (I/R) group; (3) Dobutamine1 (5 µg×kg⁻¹ · min⁻¹) + I/R group; (4) Dobutamine2 (10 µg·kg⁻¹ × min⁻¹) + I/R group; (5) LY294002 (0.3 mg/kg) + Dobutamine2 + I/R group; (6) SB203580 (1 mg/kg) + Dobutamine2 + I/R group; (7) ZnPPIX (10 mg/kg) + Dobutamine2+I/R group. Rats were pretreated by saline, dobutamine, LY294002, SB203580 and ZnPPIX, respectively, then underwent myocardial I/R. Myocardial I/R injury and oxidative stress were assessed, and myocardial HO-1, NF-κB and HMGB1 expressions were measured by Western blot analysis.</p><p><b>RESULTS</b>Dobutamine significantly reduced the myocardial infarct size (P < 0.05), myocardial enzymes (LDH and CK) (P < 0.05) and proinfiammation cytokines (TNF-α and IL-6), reduced oxidative stress (MDA and SOD) in a dose-dependent manner (all P < 0.05). Meanwhile, dobutamine significantly and dose-dependently mediated the induction of HO-1 (P < 0.05), the expression of NF-κB (P < 0.05) and HMGB1 (P < 0.05). However, all the effects could be significantly reversed by co-treatment with LY294002, SB203580 and ZnPPIX (all P < 0.05).</p><p><b>CONCLUSIONS</b>Current study demonstrates that selectively stimulating β1-adrenergic receptor by dobutasmine could reduce rat myocardial I/R injury in vivo through promoting the induction of HO-1 and inhibiting HMGB1 release.</p>
Subject(s)
Animals , Male , Rats , Chromones , Dobutamine , HMGB1 Protein , Metabolism , Imidazoles , Interleukin-6 , Morpholines , Myocardial Infarction , Myocardial Ischemia , Myocardial Reperfusion Injury , Drug Therapy , Metabolism , Myocardium , NF-kappa B , Oxidative Stress , Pyridines , Rats, Sprague-Dawley , Receptors, Adrenergic , Reperfusion Injury , Superoxide Dismutase , Tumor Necrosis Factor-alphaABSTRACT
Objective To investigate the effect of preconditioning with sodium butyrate on myocardial I/R injury. Methods Anesthetized rats were treated with sodium butyrate (100 or 300 mg/kg, i.p.) 30 mins before ischemia, and then subjected to ischemia for 30 min followed by reperfusion for 4 h. LDH, CK, TNF-α, IL-6, HMGB1, infarct size, MDA and SOD activity were measured. The infracted size was tested by TTC assay; The expression of HMGB1 was observed by western blot. Results After 4 h reperfusion, pretreatment of sodium butyrate (300 mg/kg) could significantly reduce the infarct size and the levels of LDH and CK (P<0.05)comparing to the control group; inhibit the increase of the MDA level and the decrease of the SOD level(P<0.05), also inhibit the expression of TNF-α, IL-6 and HMGB1 (all P < 0.05) induced by I/R. Conclusion Preconditioning of sodium butyrate can attenuate myocardial I/R injury by inhibiting inflammation response.
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Objective To investigate factors related to chronic pain in those injured with fractures 27 months after the Sichuan earthquake.The correlation between intensity of pain and quality of life was also analyzed.Methods A total of 705 victims were investigated on site.Their residual pain was categorized using a visual analogue scale (VAS) score as no pain,mild pain,moderate pain or severe pain.The pain-related biological,psychological and social factors were analyzed using the Barthel Index ( BI ),Life Satisfaction Questionaire-11 ( LiSat-11 )and the SF-36 health questionnaire. Rehabilitation and surgical interventions,employment,income and emotional status were also investigated. Results The incidence of chronic pain was 88.5% in this population,of which mild pain and moderate pain were 35.7% and 33.3%,respectively.The percentage of the victims who had received fracture surgery was 65.8% ; the percentage of those who had recovered was 96.9%.BI scores for the victims without pain,with mild,moderate and severe pain were 92.7 + 10.2,92.8 + 8.4,91.2 ± 9.9 and 90.4 + 14.7,respectively ; the differences between these groups were all statistically insignificant.The influence of pain intensity on life satisfaction showed a significant linear trend.The percentages of the victims with restricted occupational ability in the four groups were 38.3%,61.5%,75.7% and 62.8% respectively.The median of personal annual income were ¥ 3550,¥ 2500,¥ 2000 and ¥ 2500.The VAS scores were significantly related to abnormal emotions,life satisfaction,employment and annual income.The subjects with different levels of residual pain also showed significant differences in the physical functioning,role-physical,bodily pain,general health,vitality,social functioning,role-emotional and mental health sub-scales of the SF-36.The total SF-36 scores were highest among victims without pain (70.6 + 17.5) and declined significantly in those with mild (61.3 + 14.3 ),moderate (52.7 + 14.3 ) and severe pain (52.3 + 14.7 ).This negative correlation between pain intensity and SF-36 total score was statistically significant. Conclusions Chronic pain remains common among fracture victims 27 months after the earthquake.Its intensity is correlated with psychological and social factors as well as quality of life.
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Objective To analyze factors contributing to the quality of life of earthquake victims with nonunion fracture in limbs after medical rehabilitation and to provide a clinical reference for future disasters.Methods Thirty-one wounded who sustained their injuries in the Sichuan earthquake completed the short form-36(SF-36)questionnaire 2 weeks before their amputation,and after 3,6 and 12 months of medical rehabilitation.The SF-36scores were compared with Sichuan population norms.Stepwise multiple linear regression was used to highlight variables which influenced their functional recovery and their physical and mental health.Results The physical and mental health composite scores of the 31 victims were significantly lower than Sichuan population norms before medical rehabilitation,but they had a significant increasing trend after 3,6 and 12 months of treatment.Their scores on all the domains remained significantly lower than Sichuan population norms at a later follow-up.Smoking,external fixations and chronic diseases were related to their recovery of physical function.Age,permanent housing,family income and casualties among family members were related to their mental health recovery.Conclusion The quality of life of patients with nonunion fracture caused by earthquake can be improved significantly through medical rehabilitation.But social factors should be considered in addition to medical rehabilitation after an earthquake.
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Objective:To increase the solubility of the campher and oleum anisi stellati in the syrup of cough and reduce the precipitation of syrup.Methods: The technology of ? cyclodextrin inclusion was used in the experiment. Results: The experiment shows the method may meet the demand of production. Conclusions: The method is very good in the production of cough syrup.
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Objective To explore the relationship between the expression of thrombospondin-2 (TSP-2), the changes of MVD and angiogenesis in non-small cell lung cancer (NSCLC). Methods By using SP immunohistochemical staining, the expression of TSP-2 and microvessel counting were evaluated in surgically resected specimens from 55 patients with NSCLC and 30 patients' pancancerous tissues, using the anti-TSP-2 and anti-CD34 monoclonal antibodies respectively. Results Active angiogenesis took place in NSCLC tumor tissues. Microvessel density (MVD) was associated with metastasis and clinical stage of NSCLC. Expression level of TSP-2 in the tumor tissues of 55 NSCLC patients was significantly higher than that in the pancancerous tissues from other 30 patients (149.10?2.94 vs. 145.70?4.74, P