1.Protective effect of isorhamnetin on H9 C2 cell line against oxidative stress
Xi DONG ; Guibo SUN ; Yun LUO ; Xiaobo SUN ; Suhong CHEN
Chinese Pharmacological Bulletin 2015;(6):853-859,860
Aim To investigate the protective effect of isorhamnetin on H9 C2 myocardial cell line and its mechanisms. Methods The toxicity and optimal pro-tective concentration of isorhamnetin were determined by MTT assay. The experimental subjects were divided into four groups:group N ( normal ) , group M ( mod-el) , group M + ISO ( model + isorhamnetin ) , and group ISO ( isorhamnetin only ) . Group M +ISO and ISO were pre-incubated with isorhamnetin for 12 hours while other groups with plain DMEM. Group M and M+ ISO were treated with 300μmol · L-1 H2 O2 for 4 hours after pre-incubation. Mitochondrial membrane potential depolarization of H9 C2 was measured by fluo-rescence microscope. Apoptotic rate and ROS produc-tion of injured myocardial cell line were detected using
flow cytometry. The oxidative indictors were measured by spectrophotometry. The expressions of cytoplasmic cytochrome C, caspase-9, caspase-3, Bcl-2, Bax, Nrf2 and HO-1 were examined by Western blot. Result There was no difference in mitochondrial membrane potential depolarization, apoptotic rate, ROS produc-tion, oxidative indictors production and expressions of cytoplasmic cytochrome C, caspase-9,caspase-3, Bcl-2 , Bax between groups ISO and N ( P>0. 05 ) . Apop-totic rate, ROS production, expressions of cytoplasmic cytochrome C, caspase-9, caspase-3, Bax, MDA pro-duction of group M+ISO were significantly lower than those of group M ( P < 0. 01 ) . And mitochondrial membrane potential, Bcl-2, CAT, SOD and GSH-Px of group M + ISO were increased compared to group M .
Nuclear translocation of Nrf2 and expression of HO-1 in myocardial cell line were increased with the prolonged isorhamnetin incubation time. Conclusion Isorham-netin could protect myocardial cell line against H2 O2-induced oxidative injury and apoptosis through the in-
terruption of mitochondrial dependent apoptotic path-way and activation of Nrf2/ARE pathway.
2.The effect of high glucose on the expression of erythropoietin receptor in human umbilical vein endothelial cells
Huizhuo, XU ; Shuqian, DONG ; Xiaobo, XIA ; Siqi, XIONG
Chinese Ophthalmic Research 2010;28(1):45-49
Objective Although vascular endothelial growth factor(VEGF)is a primary mediaor in diabetic retinopathy(DR).VEGF inhibition alone is insufficient for preventing retinal neovascularization.Some studies showed that erythropoietin(EPO)is a potent retinal angiogenic factor of independent of VEGF in DR.The present study is to investigate the effect of high glucose on the expression of mRNA and protein of the erythropoietin receptor(EPOR)in cultured human umbilical vein endothelial cells(UVECs)in vitro.MethodsHuman UVECs from the cell center of the hospital were cultured in vitro and passaged in DMEM containing 10% neonatal bovine serum with 22mmol/L of glucose for 12,24,48 and 72 hours in the experimental group.Cells cultured in 5.5mmol/L glucose were used as control group Ⅰ and mannitol + 22mmol/L of glucose(isotope)as control group Ⅱ.The expression of EPOR mRNA in Human UVECs were detected by semi-quantitative reverse transcriptase(RT)-polymerase chain reaction(PCR)and detected at A_(260mm)/A_(280mm).The PCR product was calculated as the A value of EPOR mRNA amplification/the A value of GAPDH mRNA amplification.The expression of EPOR protein in Human UVECs was detected by immunocytochemistry.ResultsThe A_(260mm)/A_(280mm) value of EPOR mRNA receptor expression was 0.32±0.02 in the 5.5mmol/L glucose group,and 0.34±0.02 in the mannitol+22mmol/L glucose group(P>0.05).In 12 hours,24 hours and 72 hours after the experiment,the A260mm/A280mm value of EPOR mRNA was 0.82±0.01,0.96±0.02 and 1.02±0.01,respectively,indicating a significant increase in comparison with the 5.5mmol/L glucose group.The expression of Human UVECs protein was gradually increased with passage in the experimental group.Expression of Human UVECs protein was stronger in various time points in the 22mmol/L glucose group than in the 5.5mmol/L glucose group.ConclusionHigh glucose elevates the expression of EPOR mRNA and protein in Human UVECs in a time-dependent manner.The effect of high glucose(22mmol/L glucose)on the expression of EPOR mRNA and protein in Human UVECs is not related to osmotic pressure.
3.Role of micro RNA-106b-25 in cancer
Xi DONG ; Guibo SUN ; Xiaoyan XING ; Xiaobo SUN
Chinese Pharmacological Bulletin 2014;(12):1639-1641
Micro RNA (miRNA)were small RNAs encoded by the non-coding RNA genes,which functioned through degrada-tion of mRNA or inhibition of mRNA translation.The miRNA was involved in numerous biological processes,including cell proliferation,apoptosis,development and differentiation.And ectopic expression of miRNA could result in diseases,such as neoplastic hematologic disorder and solid tumor.miR-1 06b-25 cluster contained miR-1 06b,miR-93 and miR-25.These miR-NAs were correlated with the development of tumor.Therefore, we reviewed recent articles on the relationship of miR-1 06b-25 cluster with tumor.
4.Clinical study of autologous platelet rich plasma gel in arthroscopic rotator cuff repair:a randomized, controlled trial
Peilong DONG ; Xiaobo TANG ; Jian WANG ; Zhenan ZHU
Chinese Journal of Orthopaedics 2015;(9):942-947
Objective To investigate the effect of autologous platelet rich plasma (PRP) gel in arthroscopic rotator cuff re?pair. Methods All of 44 patients with rotator cuff tear undergent arthroscopic rotator cuff repair were randomly divided into two groups:PRP group (22 patients were received autologous PRP and hemocoagulase) and normal saline (NS) group (22 patients were received NS and hemocoagulase). All patients had the same accelerated rehabilitation protocol and were followed up in 1, 3, 12 months. Evaluation consisted of inflammatory reaction, wound healed, visual analogue scores (VAS), University of California at An?geles (UCLA) Shoulder Scores and American Shoulder and Elbow Surgeons (ASES) Scores. Results After operation, no inflam?matory reaction was in 20 cases of PRP group and 19 cases of NS group, mid inflammatory reaction 1 case in PRP group and 2 cas?es in NS group, moderate inflammatory reaction 1 case in PRP group and 1 case in NS group. Wound healed by first intention in all of PRP group and 21 patients of NS group. In PRP group, the preoperative, 3 months and 12 months postoperative VAS were 6.6±2.0, 3.4±1.8, 1.8±1.3, UCLA were 15.2±2.9, 24.3±2.7, 32.4±2.1, ASES were 35.6±12.4, 63.4±10.4, 92.3±7.5. In NS group, the preoperative, 3 months and 12 months postoperative VAS were 6.7 ± 1.9, 4.6 ± 1.9, 2.0 ± 1.2, UCLA were 14.8 ± 3.0, 21.2 ± 2.5, 31.7 ± 2.3, ASES were 32.7 ± 13.8, 55.8 ± 11.8, 90.7 ± 8.1. Three months postoperative VAS, UCLA, ASES were statistically signifi?cant differenece in PRP group and NS group. Twelve months postoperative VAS, UCLA, ASES were not statistically significant dif?ferenece in the two groups. Conclusion Using autologous PRP gel in arthroscopic rotator cuff repair can speed up the healing of operation incision with no adverse effect, reduce pain in the postoperation three months, accelerate the rotator cuff repair and re?covery of the function of shoulder joint. It has good short?term clinical effect.
5.Inhibitory effects of ostcopontin antisense oligodeoxynucleotide on osteopontin mRNA expression and adhensive ability of rat renal mcsangial cell
Xiaobo LIU ; Xueqing YU ; Xiaoyan LI ; Xiu-Qing DONG ;
Chinese Journal of Nephrology 1994;0(04):-
Objective To investigate the effects of osteopontin (OPN) antisense oligodeoxynucleotide (AS-ODN)on OPN mRNA expressions and adherence to collagen gel of rat renal mesangial cell line (1097). Methods One AS-ODN complementary chain to rat OPN cDNA sequences was designed and synthesized. All nucleotides of different kinds of oligodeoxynucleotide (including antisense, sense and mismatch sense) were modified with phosphrothioate. All the ODNs were mixed with cationic liposome(DOTAP) respectively. The cultured 1097 cells were incubated with different concentration of ODN at 37t for 48 hours, then total RNA was extracted from cultured cell line with Trizol reagent. OPN mRNA expression was detected with RNA dot blot and RT-PCR. The cell adhesion ability was measured with collagen gel attachment lest. Results OPN mRNA expression of mesangial cells was higherly upregulated in the medium with calf serum compare to that in the medium without serum. AS-ODN could suppress the expression of OPN mRNA in mesangial cells with dose-dependent and its lowest inhibitory concentration was 2. 5 ?mol/L, but mismatch ODN or sense ODN have no inhibitory effects on the OPN mRNA expression of mesangial cells even if at a high ODN concentration of 30 ?mmol/L. AS-ODN-treated cells weakly adhered to the collagen gel and could be easily detached off. The mesangial cells treated with mismatch ODN or sense ODN still had a good adherent ability to collagen gel. Conclusions Calf serum can induce rat renal mesangial cells higherly expressing OPN mRNA. Osteopontin antisense oligodeoxynucleotide can specifically suppress mesangial cells OPN mRNA expression and adhension to collagen gel.
6.The effect of platelet-rich plasma in anterior cruciate ligament reconstruction
Peilong DONG ; Xiaobo TANG ; Jian WANG ; Zhenan ZHU
Chinese Journal of Orthopaedics 2014;34(6):672-678
Objective To investigate the effect of platelet-rich plasma (PRP) in anterior cruciate ligament (ACL) reconstruction.Methods From January 2010 to January 2013,40 patients with ACL ruptures who underwent arthroscopic ACL reconstruction with gracilis and semitendinosus tendon were randomly divided into two groups:PRP group and normal saline group.20 patients received graft soaked with PRP and hemocoagulase while 20 patients received graft soaked with normal saline and hemocoagulase.All patients were followed up in 1,3 and 12 months.Evaluation consisted of postoperative drainage volume,inflammatory reaction,grade of wound healed,anterior drawer test,Lachman test,pivot shift,Lysholm knee score and KNEELAX3.Results The average follow-up period was 18 months.Postoperative drainage volume was 142±24 ml in PRP group and 324±22 ml in saline group.The difference was statistically significant.At 4 days after the operation,no inflammatory reaction was observed in 18 cases of PRP group and in 16 cases of saline group,mid inflammatory reaction in 1 case of PRP group and 2 cases of sa line group,and moderate inflammatory reaction in 1 case of PRP group and 2 cases of saline group.Wound healed by first intention in 20 patients of PRP group and in 19 patients of saline group.The preoperative results of anterior drawer test,Lachman test and pivot shift were positive,while postoperative results were negative in both two groups.In PRP group,the preoperative and postoperative Lysholm knee scores of patients in 12 months were 39.8±8.9 and 92.1±2.7 points respectively.In saline group,the preoperative and postoperative Lysholm knee scores of patients in 12 months were 38.7±9.8 and 89.9±4.1points respectively.The differences were not statistically significant.KNEELAX3 measuring results:in PRP group,preoperative measurement was 9.4±1.2 mm in average,while measurement in 12 months postoperatively was 1.2±1.1 mm.In saline group,preoperative measurement was 9.6±1.3 mm,while measurement in 12 months postoperatively was 2.2±1.2 mm.The differences were statistically significant.Conclusion Using graft soaked with PRP in ACL reconstruction could reduce postoperative drainage volume,accelerate the healing of tendon-bone interface in the bone tunnel and the recovery of knee joint function.
7.MSC-conditioned medium activates Nrf2/ARE pathway to protect H9c2 cells against oxidative stress
Xi DONG ; Guibo SUN ; Yun LUO ; Suhong CHEN ; Xiaobo SUN
Chinese Journal of Pathophysiology 2015;(6):961-966
[ ABSTRACT] AIM: To investigate the protective effect of mesenchymal stem cell ( MSC)-conditioned medium (MSCCM) on myocardial cell line H9c2 and its mechanism.METHODS:Verification of MSC was performed by flow cy-tometry analysis, followed by MTT assay to determine the optimal incubation time of MSCCM with myocardial cells.The cells were divided into 4 groups:normal ( N) group, model ( M) group, M+MSCCM group and MSCCM group.The cells in M+MSCCM group and MSCCM group were pre-incubated with MSCCM for 24 h.The cells in M group and M+MSCCM group were treated with 300 μmol/L H2 O2 for 4 h to imitate oxidative injury of myocardial cells.Mitochondrial membrane potential and apoptotic rate of injured myocardial cells were detected by flow cytometry.The ROS production was measured by fluorescence microscopy.The nuclear translocation of Nrf2 and expression of HO-1 was examined by Western blot.RE-SULTS:No difference of mitochondrial membrane potential, apoptotic rate or ROS production between MSCCM group and N group was observed (P>0.05).The mitochondrial membrane potential depolarization, apoptotic rate and ROS produc-tion in M+MSCCM group were significantly lower than those in M group ( P<0.01 ) .The nuclear translocation of Nrf2 and expression of HO-1 in the myocardial cells were increased with MSCCM incubation time prolonged.CONCLUSION:MSCCM protects the myocardial cells against oxidative injury induced by H2 O2 .The anti-oxidative mechanism would be as-sociated with the activation of Nrf2/ARE pathway.
8.Therapeutic effects of ultrasound biomicroscopy assisted intraocular lens suspension surgery with one loop suturing for residual lens capsule aphakia
Ruifeng SU ; Xiaobo TAN ; Litao GUO ; Weili DONG
Recent Advances in Ophthalmology 2017;37(2):175-178
Objective To evaluate the therapeutic effects of ultrasound biomicroscopy (UBM) assisted intraocular lens suspension surgery with one loop suturing for residual lens capsule aphakia.Methods Fifty aphakia patients (50 eyes) with residual lens capsular equal or greater than 180 degrees were chosen.The patients were randomly divided into 2 groups and underwent intraocular lens suspension surgery with one loop suturing:Trial group (25 cases,25 eyes) selected suture site according to the preoperative UBM results,and control group (25 cases,25 eyes) selected the suture site according to the clinical experience.The uncorrected visual acuity (UCVA),astigmatism at preoperative and postoperative 1 week,1 month and 3 months were observed,the eccentric and tilt value of the lens were calculated by UBM measurement at 3 month after operation,and the postoperative complications of the two groups were observed.Results UCVA at each time point after operation were higher than that before operation,and gradually increased with time.There was no significant difference between the two groups at 1 week before and after the operation of UCVA (P > 0.05).At 1 month and 3 months after operation,UCVA in trial group were significantly higher than those of control group (all P < 0.05).The astigmatism value at 1 week after operation in two groups was higher than that before operation and gradually decreased with time.The astigmatism value in trial group at 1 month and 3 months after operation had no significant difference with that of preoperative (P > 0.05),while the astigmatism value in control group increased (all P < 0.05).The astigmatism values had no significant difference between the two groups before and 1 week after operation (all P >0.05),which were significantly lower in trial group at 1 month and 3 months after operation (all P < 0.05).Three months after surgery,the proportion of lens accurately implanted in the ciliary sulcus was significantly higher than control group (P < 0.01),the eccentric and tilt values of the lens in trial group were significantly less than control group (P < 0.01).No serious complication was observed in all patients.Conclusion UBM assisted intraocular lens suspension surgery with one loop suturing can significantly improve the postoperative visual quality,safety and lens stability,which can be used as a routine preoperative examination of intraocular lens suspension surgery.In addition,UBM provides an alternative method for the calculation of the eccentricity and tile value of the intraocular lens after the implantation of the intraocular lens.
9.Early effect of tonsillotomy on children′s snoring with tonsil hypertrophy
Xiaoguang WANG ; Mingfu DONG ; Yaqin FAN ; Zhongsheng HAN ; Xiaobo SHUI
International Journal of Laboratory Medicine 2016;37(4):484-486
Objective To explore short-term effect of tonsillotomy on children′s obstructive sleep apnea hypopnea syndrome (OSAHS) with tonsil hypertrophy by Ellman double-frequency radio-frequency machine .Methods Totally 30 cases children′s OS-AHS with tonsil hypertrophy were had tonsillotomy by Ellman double-frequency radio-frequency machine .They were compared with 30 cases children′s OSAHS with tonsil hypertrophy by traditional tonsillectomy .Short-term effect of tonsillotomy were ana-lyzed by wound bleeding ,operation time ,postoperative pain ,difficulty eating ,PSG testing ,immune function ,growth and develop-ment .Results Postoperative symptom in 1 weeks and 6 months ,compared with the traditional tonsillectomy ,30 cases of OSAHS patients were found that there were less bleeding ,shorter operative time ,mild postoperative pain ,no difficulty in feeding ,improving sleep-disordered breathing ,no decline in immune function ,good growth and development .Conclusion Short-term effect of tonsillot-omy on children′s OSAHS with tonsil hypertrophy is good by Ellman double-frequency radio-frequency machine ,it is a minimally invasive ,safe and effective surgical method children OSA HS .
10.Association of a miRNA-137 rs1625579 polymorphism with neurocognitive function in patients with schizophrenia
Dong LYU ; Fang FANG ; Xiaobo ZOU ; Juda LIN ; Jingwen YIN
Tianjin Medical Journal 2016;44(5):613-616
Objective To investigate the aassociation of a microRNA-137 (miR-137) polymorphism, single nucleotide polymorphism (SNP) rs1625579, with neurocognitive function in patients with schizophrenia. Methods A total of 250 patients with schizophrenia were included in this study. The positive and negative syndrome scale (PANSS) was used to evaluate patients. The brief assessment of cognition in schizophrenia (BACS) scale was used to determine neurocongnitive functions in patients. Blood samples of patients were collected, and SNaPshot technique was used to compare the neurocognitive functions of different genotypes of rs1625579. Results The genotypes of TT, GT and GG were 221 (88.4%), 28 (11.2%) and 1(0.4%). There was no significant difference in PANSS score between TT genotype carriers and G allele (GG+GT) carriers. The detection of BACS showed that the digit sequencing score was significantly lower in patients with TT genotype than that of G allele (GG+GT) carrier ( P<0.05). There were no significant differences in other scores of BACS evaluation between two groups of patients. Conclusion The miR-137 polymorphism influences the working memory performance of schizophrenic patients in Chinese Han population.