1.Effects of microRNA-16 on proliferation, invasion and cytokine secretion of synovial fibroblasts from rheumatoid arthritis patients
Chinese Journal of Pathophysiology 2014;(10):1868-1872
AIM:To investigate the effect of microRNA-16 ( miR-16) on the proliferation, invasion and cyto-kine secretion of rheumatoid arthritis ( RA) synovial fibroblasts ( RASFs) from the RA patients.METHODS: miR-16 mimic and miR-16 inhibitor were synthesized, and then Transfected into RASFs isolated from RA patients with lipo-fectamine.MTT assay, Transwell chamber and flow cytometry were used to determine the effect of miR-16 on proliferation, invasion and apoptosis of RASFs.The expression of matrix metalloproteinase 3/13 ( MMP3/13) and interleukin 1β( IL-1β) was measured by RT-PCR and Western blotting.RESULTS: The proliferation and invasion of RASFs were signifi-cantly inhibited by miR-16 mimic.The result of flow cytometry demonstrated that miR-16 had no effect on apoptosis of RASFs.Furthermore, miR-16 down-regulated the expression of MMP3/13 and IL-1β.CONCLUSION:miR-16 plays an important role in the development of RA and may inhibit the proliferation and invasion of RASFs through down-regulating the expression of MMP3/13 and IL-1β.
2.Effect of SREBP-2 silencing on tunicamycin-induced endoplasmic reticu-lum stress in chondrocytes
Jing XIE ; Dongliang SHI ; Huiqing GUO
Chinese Journal of Pathophysiology 2016;32(7):1291-1296
[ ABSTRACT] AIM:To explore the effect of sterol regulatory element-binding protein 2 ( SREBP-2) on tunicamy-cin-induced endoplasmic reticulum stress ( ERS) in chondrocytes.METHODS:After isolation of human normal chondro-cytes and osteoarthritis ( OA) chondrocytes, the normal cells were cultured and treated with tunicamycin and SREBP-2 siR-NA.After 24 h treatment, fluorescent quantitative RT-PCR ( RT-qPCR) was applied to quantify microRNA-185 ( miR-185) levels.The cell apoptotic rate was determined by flow cytometry.The expression of SREBP-2 and ERS-related pro-teins, C/EBP homologous protein (CHOP), phosphorylated eukaryotic initiation factor-2α(p-eIF2α) and activating tran-scription factor 4 (ATF4), and the expression of apoptosis-related proteins, Bcl-2, Bax and caspase-3, were determined by Western blot.The caspase-3 activity kit was used to determine the caspase-3 activity.RESULTS: Compared with hu-man normal chondrocytes, both SREBP-2 up-regulation and miR-185 down-regulation were observed in OA chondrocytes (P<0.05).SREBP-2 siRNA transfection enhanced tunicamycin-inhibited miR-185 level (P<0.05).miR-185 overex-pression reduced tunicamycin-induced SREBP-2 expression ( P <0.05 ) .OA control group and tunicamycin treatment group consistently resulted in ERS and cell apoptosis with concomitant enhancement of CHOP, p-eIF2αand ATF4 proteins, increases in Bax and caspase-3 proteins, and reduction of Bcl-2 (P<0.05).However, SREBP-2 silencing significantly re-versed these effects ( P<0.05) .The apoptotic rates were consistent with the expression tendency of apoptosis-related pro-teins (P<0.05).SREBP-2 siRNA transfection markedly down-regulated tunicamycin-induced caspase-3 activity, which was notably blocked by miR-185 inhibition (P<0.05).CONCLUSION:SREBP-2 silencing may inhibit tunicamycin-in-duced ERS and cell apoptosis via up-regulating miR-185 expression.
3.Activation Characteristics of Vastus Medialis, Rectus Femoris and Vastus Lateralis Muscles in Patients with Knee Osteoarthritis and Health Subjects
Dongliang SHI ; Ninghua WANG ; Bin XIE
Chinese Journal of Rehabilitation Theory and Practice 2009;15(6):508-513
Objective To investigate the activation characteristics (contraction velocity, force and anti-fatigue) of the vastus medialis (VM), rectus femoris (RF) and vastus lateralis (VL) by using surface electromyogram (sEMG) during isometric and isokinetic quadriceps femoris force tests in knee osteoarthritis. Methods The participants included an observation group (n=34) and a control group (n=34).The age, height, body weight, gender, dominant side and Q angle were matched between the two groups. The participants' quadriceps femoris force and sEMG of VM, RF and VL were tested in isometric knee extension at knee joint angles of 10°, 60°, 100° and isokinetic knee extension at velocities of 60°/s and 180°/s. Comparisons were made for recruitment rate, average amplitude, peak torque/bodyweight ratio, index of fatigue by independent-group t-test, with the alpha level set at 0.05 for significant differences between groups.Results ①Recruitment rate of VM in isometric contraction at 60° and isokinetic contraction at 60°/s and 180°/s, RF in isometric contraction at 60° and isokinetic contraction at 60°/s and VL in isometric contraction at 10°and 100° and isokinetic contraction at 60°/s were all significantly slower in the observation group than that in the control group(P<0.05). ②The average amplitudes of VM, RF in isokinetic contraction at 60°/s and 180°/s and RF, VL in isometric contraction at 10° were significantly higher in the observation group than that in the control group(P<0.05). ③Peak torque/bodyweight ratio of quadriceps was significantly lower in the observation group than that in the control group in isometric contraction at 10°, 60°, 100° and isokinetic contraction at 60°/s and 180°/s(P<0.05). ④In isokinetic contraction at 180°/s the index of fatigue was significantly higher in the observation group than that in the control group (P<0.05).Conclusion A significant decreasing contraction velocity and force, increasing anti-fatigue of the vastus medialis, rectus femoris and vastus lateralis were found in knee osteoarthritis patients when compared with healthy subjects.
4.Coordination of Vastus Medialis, Rectus Femoris and Vastus Lateralis in Patients with Knee Osteoarthritis
Dongliang SHI ; Ninghua WANG ; Bin XIE
Chinese Journal of Rehabilitation Theory and Practice 2010;16(5):473-477
ObjectiveTo compare the coordination characteristic of the vastus medialis (VM), rectus femoris (RF) and vastus lateralis (VL) between patients with knee osteoarthritis and healthy subjects. Methods34 patients with knee osteoarthritis and 34 controls with normal knee were tested in isometric knee extension at knee joint angles of 10°, 60°, 100° flexion and isokinetic knee extension at velocities of 60°/s and 180°/s. Surface electromyogram (EMG) was recorded in the VM, RF and VL. The activation sequence of muscles and VM/VL EMG ratio were analyzed. ResultsThe VM started significantly later than the VL in isokinetic contraction at 180°/s in patients compared with the controls (P<0.05), and the VM/VL EMG ratio decreased in isometric contraction with knee extension at 10° (P<0.05). ConclusionThe coordination of the vastus medialis, rectus femoris and vastus lateralis decreases in the knee osteoarthritis patients.
5.Involvement of neural mechanism in cardioprotection provided by renal ischemic preconditioniong in anesthetized rabbits
Yanfeng DING ; Mingzhu SHI ; Xinqing NIU ; Dongliang LI ; Ruirong HE
Journal of Xinxiang Medical College 2002;19(5):345-349
Objective To investigate the role of renal nerve in cardioprotection provided by renal ischemic preconditioning(RIP).Methods The effects of ischemia-reperfusion and RIP on the hemodynamics, myocardial oxygen consumption, epicardial electrography and infarct size were examined in anesthetized rabbit.Results During the 45 min of myocardial ischemia and 180 min of reperfusion, all hemodynamic parameters and myocardial oxygen consumption decreased progressively significantly. Epicardial electrographic ST-segment was elevated significantly during myocardial ischemia and return to baseline progressively in the course of reperfusion. The myocardial infarct size occupied 55.80±1.25% of area at risk,and RIP significantly reduced the myocardial infarct size to 36.51±2.80%(P<0.01). The renal nerve section (RNS) per se didn't affect myocardial infarct size produced by ischemia-reperfusion, while cardioprotection afforded by RIP was completely abolished by RNS.Conlusion RIP have the protective effect on heart, and activation of renal afferents by transient ischemia-reperfusion play an important role in such a cardioprotection.
6.Expression of Gli1 protein and vascular endothelial growth factor-C in papillary thyroid carcinoma and their relationship with cervical lymph node metastasis
Dongliang SHI ; Suyuan ZHAO ; Qianqian CHEN ; Qinghuai LI
Chinese Journal of Postgraduates of Medicine 2013;(20):30-34
Objective To study the expression of Gli1 protein and vascular endothelial growth factor (VEGF)-C in papillary thyroid carcinoma and their relationship with cervical lymph node metastasis.Methods The expression of Gli1 protein and VEGF-C were investigated by immunohistochemical EnVision method in 67 cases of papillary thyroid carcinoma and 32 cases of nodular goiter pathological specimen tissues.Nascent microvascular and micro-lymphatic of all pathological speimen tissues were examined by immunohistochemical monoclonal antibody CD34 and D2-40 staining assay respectively,and the microvascular density (MVD) and micro-lymphatic vessel density (MLVD) were calculated.The relationship between the positive expression of Gli1 protein and MVD,between the positive expression of VEGF-C and MLVD,and their relationship with cervical lymph node metastasis were analyzed.Results The positive rates of Gli1 protein,VEGF-C and MVD,MLVD were significantly higher in papillary thyroid carcinoma than those in nodular goiter [67.16%(45/67) vs.43.75%(14/32),70.15%(47/67) vs.31.25%(10/32),23.14 ± 2.06 vs.2.14 ± 0.31,13.36 ± 1.32 vs.3.53 ± 0.65,P < 0.05].The positive expression of Gli1 protein was related to the patient' s age and the tumor size (P < 0.05),while the positive expression of VEGF-C was not related to the patient' s age and the tumor size (P > 0.05).The positive expression of Gli1 protein and VEGFC were significantly higher in TNM stage Ⅲ-Ⅳ of papillary thyroid carcinoma than those in TNM stage Ⅰ-Ⅱ (P < 0.01) and also was significantly higher with cervical lymph node metastasis than without cervical lymph node metastasis (P < 0.01).MVD and MLVD were significantly higher with cervical lymph node metastasis than without cervical lymph node metastasis (P < 0.05).The positive expression of Gli1 protein was positively correlated with MVD (r =0.784,P< 0.01),the positive expression of VEGF-C was positively correlated with MLVD (r =0.529,P < 0.01),the positive expression of Gli 1 protein was positively correlated with the positive expression of VEGF-C (r =0.586,P <0.01).Conclusions Gli1 protein which may participate in the formation of nascent microvascular is abnormally activated by the Hedgehog signaling pathway to express,VEGF-C which may be participate in the formation of nascent micro-lymphatic is mediated and started by the MAPK signaling pathways to express.Expressions of Gli1 protein is positively correlated with expressions of VEGF-C,therefore the Hedgehog signaling pathway may be associated with the MAPK signal pathway.Suppressing nascent microvascular and micro-lymphatic may become new target to blockingup papillary thyroid carcinoma cervical lymph node metastasis.
7.Clinical application of primary bile duct closure improvement after choledochotomy
Bin ZHU ; Weibin SHAO ; Dongliang YAN ; Haiyan SHI
Chinese Journal of Primary Medicine and Pharmacy 2010;17(19):2649-2650
Objective To determine the value and reasonable application of slowly coated vicryl plus and albumin gel in primary suture after common BD exploration. Methods The operation was successfully performed in all 45 patients. The incision of common bile duct was directly sewed up by slowly coated vicryl plus and spurted by albumin gel after the exploration. Results There were no complications such as bile leakage and bile duct stricture etc.The mean payment of hospitalization was low 10.5%. Conclusion This method was safe,feasible and effective.
8.Analysis of risk factors to acute lung injury based on multiple trauma *
Yaqin YANG ; Yonghua XIA ; Bing SUN ; Jinhe SHI ; Dongliang LIANG
Chongqing Medicine 2013;(27):3255-3256,3259
Objective To investigate the risk factors to acute lung injury based on multiple trauma to provide a theoretical basis for early intervention .Methods The emergency surgical patients with multiple trauma in our hospital from March 2006 to March 2011 were selected .The patients meeting the diagnostic criteria for acute lung injury were taken as the study group and the others as the control group .All patients were enrolled for evaluating the injury severity score (ISS) ,acute physiology and chronic health Ⅱ(APACHE Ⅱ) score and recording smoking ,alcohol abuse ,diabetes mellitus ,number of organ damage ,gastrointestinal bleeding , pulmonary contusion ,diffuse intravascular coagulation (DIC ) ,vomiting ,traumatic shock ,time to correct shock ,blood transfusion . The polymorphism of rs3788853 ,rs13306087 ,rs12709426 of angiotensin-converting enzyme(ACE) gene were analyzed .Results In the study group and the control group ,there were statistical differences in 6 influencing factors of the ISS ,APACHE Ⅱ score ,blood transfusion ,DIC ,traumatic shock ,time to correct shock>6 h(P<0 .05);the gene and genotype frequencies of ACE between the two groups had no statistical difference (P>0 .05);the 6 kinds of influencing factors were risk to acute lung injury based on multi-ple trauma by Logistic regression analysis .Conclusion The ISS ,APACHE Ⅱ score ,blood transfusion ,DIC ,traumatic shock ,long time to correct shock are the risk factors to acute lung injury based on multiple trauma .
9.Best time of mini-traumatic operation to treat hypertensive intracerebral haemorrhage in internal capsule
Dongliang LIANG ; Pingfa LI ; Jinhe SHI ; Feiyun YANG ; Jianxin ZHANG
Chinese Journal of Postgraduates of Medicine 2006;0(26):-
Objective To investigate the best time of mini-traumatic operation to treat hypertensive intracerebral haemorrhage in internal capsule. Methods Sixty-four patients were divided into treatment group and control group according to operative time. Patients who were treated with mini-traumatic operation within 6 hours were as treatment group, those treated with mini-traumatic operation beyond 6 hours were as control group. Results Twelve of 40 patients in treatment group were cured, 16 became better, 5 had no effect, 7 were dead, and the effective rate was 70.00%. Five of 24 patients in control group were cured, 6 became better, 5 had no effect, 8 were dead, and the effective rate was 45.83%. Conclusion The effect of mini-traumatic operation to treat hypertensive intracerebral haemorrhage in internal capsule within 6 hours is clearly better than that beyond 6 hours.
10.Expression of miRNA-1247-3P in acute respiratory distress syndrome cell model and its function
Dongliang CHENG ; Yuan LIANG ; Yanchen CHEN ; Di QING ; Changsong SHI
The Journal of Practical Medicine 2015;(17):2802-2804
Objective To detect the expression pattern of microRNA in A549 cells treated by lipopolysaccharide, study the expression of miRNA-1247-3P in A549 cells under LPS treatment and explore the possible mechanism of miRNA-1247-3P in A549 cells under LPS treatment. Methods A549 cells were divided into experimental and control groups. Immunocytochemical method and RT-PCR were used to detect the changes of SP-A and SP-C. The expression of miRNAs were detected using miRNAs array in different groups. The key miR-1247-3P was collected to detect the changes of miR-1247-3P in all groups using quantitative real-time polymerase chain reaction. Results Compared with control group, the expressions of SP-A and SP-C were significantly decreased in the experimental groups (P < 0.05). MiRNA array showed that 31 miRNAs were up-regulated and 3 miRNAs were down-regulated. Compared with control group, the expression of miR-1247-3P was significantly increased in the experimental groups (P < 0.05). Conclusion The increased expression of miR-1247-3P may play an important role in the pathogenesis of ARDS.