Effects of AG3340 on the migration and invasion of human retinal pigment epithelial cells induced by high glucose / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the effect and mechanism of matrix metalloproteinase(MMPs)inhibitor AG3340 on the migration and invasion ability of retinal pigment epithelial cells-19(ARPE-19)cultured in high glucose(CHG). METHODS: ARPE-19 cells cultured in vitro were divided into four groups: Control group, the glucose at the concentration of 5.6mmol/L in DMEM/F12 medium; HG group, the glucose at the concentration of 30mmol/L was cultured with DMEM/F12 medium; HG+AG3340 group, the cells were pretreated with AG3340 for 12h, and then cultured in DMEM/F12 medium containing 30mmol/L glucose; The mannitol(MA)group, cultured with DMEM/F12 medium of 5.6mmol/L glucose and 24.4mmol/L mannitol, which used as hypertonic control group. The migration ability of cells was detected by wound healing assay, the invasion ability of cells was detected by Transwell assay, and the relative expression levels of MMP-9, MMP-2, fibronectin and collagen Ⅳ were detected by Western blot.RESULTS: The results of wound healing assay showed that compared with the Control group, the cell migration rate of scratching after 24h and 48h in the HG group was significantly increased(all P<0.001).After pretreated by AG3340, the cell migration rate was significantly lower than that in the HG group(all P<0.01).Transwell assay showed that compared with the Control group, the number of cell invasion in the HG group was significantly higher than that in the Control group(all P<0.001). After pretreated by AG3340, the number of cell invasion was decreased than the HG group(all P<0.01). Western blot results showed that compared with the Control group, the relative expression levels of MMP-9 and MMP-2 of the cells in the HG group were increased, and the relative expression levels of Fibronectin and Collagen Ⅳ were decreased(all P<0.001). Compared with the HG group, the relative expression levels of MMP-9 and MMP-2 protein in AG3340 pretreatment group were decreased, and the relative expression levels of Fibronectin and Collagen Ⅳ were increased(all P<0.05). CONCLUSION: High glucose induced ARPE-19 cells with enhanced migration and invasion ability, and AG3340 partially reversed this effect, which was related to the inhibition of MMP-9 and MMP-2 expression and the stability of extra-cellular matrix components.

Assessment of outcomes of idiopathic macular epiretinal membrane based on preoperative classification of the epiretinal membrane with OCT / 眼科新进展

Objective To classify idiopathic macular epiretinal membrane (IMEM) on the basis of OCT and its application value in predicting visual acuity of patients after operation.Methods Totally 29 patients (30 eyes) with IMEM admitted in Shenzhen Eye Hospital from February 2013 to May 2016 were included in this study and divided into 4 levels based on the morphologic change of macular fovea by preoperative OCT examination.All the patients were given minimally invasive vitrectomy for stripping macular epiretinal membrane (MEM),and examined for best corrected visual acuity (BCVA) and central foveal thickness (CFT) 6 months after surgery.Results As the grading increased,the patient's macular foveal became disorganized,the appearance of the macular membrane was presented and the loss of macular foveal was more obvious.There was significant difference between preoperative and postoperative LogMAR BCVA (all P ＜ 0.01).Preoperative and postoperative LogMAR BCVA was increased with the grading,and the difference was statistically significant (all P ＜ 0.05),suggesting that patients with high-level epiretinal membranes suffered from poor visual acuity.The difference between preoperative and postoperative LogMAR BCVA was the largest in patients with grade Ⅱ idiopathic macular degeneration,and the difference was statistically significant compared with other levels (all P ＜ 0.001).It is suggested that BCVA of grade Ⅱ patients has the greatest improvement after operation.Apart from grade Ⅲ patients,there was no significant difference in CFT among the patients with other levels before and after operation (all P ＞ 0.05).The preoperative and postoperative CFTs between patients were increased with the grading,and the differences were statistically significant (all P ＜ 0.001),suggesting that the higher the level was,the larger the macular thickness.Conclusion The classification of IMEM based on OCT examination images not only helps to predict the postoperative visual acuity and CFT recovery but also the timing of operation in patients with idiopathic macular degeneration.

Research advances in the treatment for uveal melanoma / 眼科新进展

Uveal melanoma (UM) is the most common primary intraocular malignancy in adults.Despite the availability of local treatment modalities,the survival rates of patients with UM have not improved in 30 years.And the main reason is lack of therapeutic drugs for UM,especially agents that inhibit tumor metastasis.Foreign researchers have carried out extensive clinical trials on the agents,but traditional ocular enucleation has been performed for the treatment of UM without any clinical drug tests in China.This paper will give a review on these clinical trials abroad,hoping to provide insight in the treatment of UM for domestic ophthalmologists and oncologist.

Post-conditioning with gradually increased reperfusion provides better cardioprotection in rats / 世界急诊医学杂志（英文）

BACKGROUND:Rapid and complete reperfusion has been widely adopted in the treatment of patients with acute myocardial infarction (AMI), but this process sometimes can cause severe reperfusion injury. This study aimed to investigate different patterns of post-conditioning in acute myocardial ischemia-reperfusion injury, and to detect the role of mitogen activated protein kinase (MAPK) during the injury. METHODS:Rats were randomly divided into five groups:sham group, reperfusion injury (R/I) group, gradually decreased reperfusion group (GDR group, 30/10-25/15-15/25-10/30 seconds of reperfusion/ischemia), equal reperfusion group (ER group, 20/20 seconds reperfusion/ischemia, 4 cycles), and gradually increased reperfusion group (GIR group, 10/30-15/25-25/15-30/10 seconds of reperfusion/ischemia). Acute myocardial infarction and ischemic post-conditioning models were established in the rats. Six hours after reperfusion, 3 rats from each group were sacrificed and myocardial tissues were taken to measure the expressions of phosphorylation of extracellular signal-regulated protein kinase (P-ERK), phosphorylated c-Jun N-terminal kinase (P-JNK), mitogen-activated protein kinase p38 (p38 MAPK), tumor necrosis factor-α (TNF-α), caspases-8 in the myocardial tissue, and cytochrome c in the cytosol using Western blot. Hemodynamics was measured at 24 hours after reperfusion, the blood was drawn for the determination of cardiac enzymes, and the heart tissue was collected for the measurement of apoptosis using TUNEL. One-way analysis of variance and the Q test were employed to determine differences in individual variables between the 5 groups. RESULTS:Three post-conditioning patterns were found to provide cardioprotection (P<0.05) compared with R/I without postconditioning. GIR provided the best cardioprotection effect, followed by ER and then GDR. Apoptotic index and serum marker levels were reduced more significantly in GIR than in ER (P<0.05). The enhanced cardioprotection provided by GIR was accompanied with significantly increased levels of P-ERK 1/2 (1.82±0.22 vs. 1.54±0.32,P<0.05), and lower levels of p-JNK, p38 MAPK, TNF-α, caspase-8, caspase-9 and cytochrome in the cytoplasm (P<0.05), compared with ER. The infarct size was smaller in the GIR group than in the ER group, but this difference was not significant (16.30％±5.22％ vs. 20.57％±6.32％,P<0.05). Allthe measured variables were improved more significantly in the GIR group than in the GDR group (P<0.05). CONCLUSION:Gradually increased reperfusion in post-conditioning could attenuate reperfusion injury more significantly than routine method, thereby the MAPK pathway plays an important role in this process.

Change s of haemagglutination inhibitionan tibody level within one month after in fluenza A (H1N 1) vaccination / 中华微生物学和免疫学杂志

Objective To find the changes of haemagglutination inhibition ( HI ) antibody level against A/California/07/2009 (H1N1) within one month after pandemic A/H1N1 influenza vaccine (A/H1N1InfV) vaccination, and to provide data for drawing up immunization protocols against novel influenza . Methods The HI antibodies against A/California/07/2009 (H1N1) in sera from the inoculated subjects were tested by HI test .The geometric mean titer ( GMT) , geometric mean increase ( GMI) , seroconversion (SC) rate, seroprotection (SP) rate of HI antibodies were compared among the sera collected on day 3, 7, 14, 30 post vaccination .Results 961 participants were injected with A/H1N1InfV.In subjects aged 3 to 11 years, the antibody level peaked on day 14 post vaccination, but neither on day 14 nor on day 30, the lower bound of the two -sided 95%CI for the SP rate could fulfill the criteria of the FDA for influenza vac-cine.In subjects aged 12 to 60 years, the antibody level peaked on day 14 post vaccination and the SC rate , SP rate and GMI fulfilled the criteria of the European Medicines Agency ( EMEA) and the FDA for influenza vaccine. In subjects aged more than 60 years, the antibody level peaked on day 30 post vaccination , and the SC rate, SP rate and GMI on day 30 fulfilled the criteria of the EMEA and the FDA .Conclusion One dose A/H1N1InfV vaccination was able to induce enough protection on day 14 for subjects aged 12 to 60 years, on day 30 for subjects aged more than 60 years;however , for subjects aged 3 to 11 years who were antibody-negative at baseline , the lower bound of the two-sided 95%CI for the SP rate on day 14 and day 30 couldn′t fulfill the criteria of the FDA for influenza vaccine .

Pharmacological postconditioning with lactic acid and low dose edaravone could attenuate myocardial reperfusion injury through mitochondrial pathway / 中华心血管病杂志

<p><b>OBJECTIVE</b>To test the hypothesis that pharmacological postconditioning with lactic acid and low dose edaravone could mimic the upper trigger of mechanical postconditioning and relieve reperfusion injury through mitochondrial pathway.</p><p><b>METHODS</b>Rats were randomly divided into 6 groups (n = 18 each): sham, reperfusion/injury(I/R), postconditioning (IP), lactic acid (Lac, 60 µl), low dose edaravone (Eda, 3 µg/kg), and Lac+Eda. After 45 min myocardial ischemia, different drugs or saline were administrated around the infarct border according to different groups using micro syringe at the time of reperfusion. After 10 min reperfusion, right atrial plasma pH value was determined in all rats. Then the rats were sacrificed at 1, 6 and 24 h (n = 6 each), apoptotic index was measured by TUNEL, infarct area and ischemic area were measured through Evans blue-TTC double staining, mitochondrial absorbance, the contents of MDA and SOD in ischemic myocardium were detected by spectrophotometry, and the expression of apoptotic pathway molecules, such as Bcl-2, Bax and Cytochrome c (Cyt-c) , were detected by Western blot.</p><p><b>RESULTS</b>Right atrial plasma pH value was significantly lower, the content of MDA was significantly lower, and the content of SOD was significantly higher in IP and Lac+Eda groups than in I/R group (all P < 0.05). The mitochondrial absorbance in Lac+Eda group at all time points were all significantly higher than those in I/R group (all P < 0.05). The level of Bcl-2 in ischemic myocardium in Lac+Eda group was significantly higher than in I/R group (1.02 ± 0.19 vs.0.02 ± 0.01, P < 0.05), the level of Bax (0.38 ± 0.07 vs.2.40 ± 0.45, P < 0.05) and Cyt-c(0.78 ± 0.05 vs.6.54 ± 1.86, P < 0.05) were all lower than those in I/R group. The content of CK[(849 ± 228) vs.(1249 ± 211) U/L, P < 0.05] and CK-MB[(470 ± 266) vs. (966 ± 263) U/L, P < 0.05] in Lac+Eda group were all significantly lower than in I/R group, apoptotic index [(10.51 ± 1.52)% vs. (15.00 ± 1.90) %, P < 0.05] and infarct area [(27.12 ± 5.55)% vs. (45.66 ± 10.81)%, P < 0.05] in Lac+Eda group were all significantly lower than those in I/R group.</p><p><b>CONCLUSION</b>Pharmacological postconditioning with lactic acid and low dose edaravone could mimic the upper triggers of mechanical postconditioning and attenuate myocardial reperfusion injury through mitochondrial pathway.</p>

Effect of ischemic postconditioning on activation of p38 mitogen activated protein kinase and cardiac myocyte apoptosis in rats / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore the change of phospho-p38 (P-p38) mitogen activated protein kinase(MAPK) and its influence on myocardial apoptosis in reperfusion injury in postconditioning.</p><p><b>METHODS</b>Totally 60 rats were equally and randomly divided into six groups: Sham group,reperfusion injury (R/I) group, postconditioning (Post) group, SB203580 (I_p38) group, anisomycin plus postconditioning (Ani+post) group,and anisomycin (Ani) group. After the model of acute myocardial infarction was established,placebo solution (DMSO), SB203580 (1 mg/kg), or anisomycin (2 mg/kg) was injected through jugular vein 5 minutes before reperfusion. Six hours later, 3 rats in each group were executed and the hearts were separated to measure the signaling molecules including phospho-p38,tumor necrosis factor-alpha (TNF-α), Caspase-8, Bcl-2/Bax, and cytochrome-c (Cyt-c). Twenty-four hours later,the hemodynamic data were measured in the remaining rats,and then blood was collected to determine the serum markers of cardiac damage. After that,hearts were separated to measure the infarction area and apoptosis.</p><p><b>RESULTS</b>Six hours after reperfusion,the expressions of P-p38 in Post and I_p38 group were significantly lower than those in R/I group (P<0.05), significantly higher in Ani+post and Ani group than in Post group (P<0.05), and significantly lower in Ani+post group than in R/I group (P<0.05). The expressions of TNF-α and Caspase-8 were significantly lower in Post and I_p38 group than in R/I group (P<.05) and significantly higher in Ani+post and Ani group than in Post group (P<0.05). The expression of TNF-α was significantly lower in Ani+post group than in R/I group (P<0.05). The expression of Bcl-2 was significantly higher in Post and I_p38 groups than in R/I group (P<0.05) and significantly lower in Ani+post and Ani groups than in Post group (P<0.05). The expression of Bax was significantly lower in Post and I_p38 groups than in R/I group (P<0.05) and were significantly higher in Ani+post and Ani group than in Post group (P<0.05). The expression of Cyt-c after the removal of the cytoplasm mitochondria was significantly lower in Post and I_p38 group than in R/I group (P<0.05) and was significantly higher in Ani+post and Ani group than in Post group (P<0.05). Twenty-four hours after reperfusion,the values of rate-pressure product and ± delta pressure/delta time max were significantly lower in R/I group than in Post and I_p38 groups (P<0.05) and was significantly higher in Post group than in Ani+post and Ani group (P<0.05). The apoptotic index (AI) was significantly lower in Post and I_p38 groups than in R/I group (P<0.05) and was significantly higher in Ani+post and Ani groups than in Post group (P<0.05). The values of creatine kinase and creatine kinase-MB were significantly lower in Post,Ani+post, and I_p38 groups than in R/I group (P<0.05) and were significantly higher in Ani+post and Ani group than in Post group (P<0.05). The area of necrosis/area at risk ratio was significantly lower in Post and I_p38 groups than in R/I group (P<0.05) and was significantly higher in Ani+post and Ani groups than in Post group (P<0.05).</p><p><b>CONCLUSION</b>Postconditioning can inhibit the phosphorylation of p38 MAPK,through which it can attenuate cardiac myocyte apoptosis by both extrinsic and mitochondria pathways.</p>

Gradual algorithm of postconditioning reduced reperfusion injury through mitochondrion pathway in rats / 中华心血管病杂志

<p><b>OBJECTIVE</b>The effects of various postconditioning algorithm on reperfusion injury and the role of mitochondrion pathway were investigated in a rat model of reperfusion/injury.</p><p><b>METHODS</b>Rats were divided into 5 groups: sham, reperfusion/injury (R/I group), reverse algorithm of postconditioning (R-Post, 30/10-25/15-15/25-10/30 s of reperfusion/re-occlusion), standard algorithm of postconditioning (S-Post, 4 cycles of 20/20 s of reperfusion/re-occlusion), and gradual algorithm of postconditioning (G-Post, 10/30-15/25-25/15-30/10 s of reperfusion/re-occlusion).</p><p><b>RESULTS</b>The levels of Bax, Cytochrome-c, Caspase-9, serum marker of myocardium and apoptosis index were significantly lower while the level of Bcl-2 was significantly higher in the three postconditioning groups than those in R/I group (all P < 0.05). The levels of Bax (0.35 +/- 0.10 vs. 0.50 +/- 0.02, P < 0.05), Cytochrome-c (0.66 +/- 0.16 vs. 1.68 +/- 0.22, P < 0.05), Caspase-9 (0.61 +/- 0.17 vs. 1.66 +/- 0.55, P < 0.05), serum marker of myocardium [CK: (251.00 +/- 45.16) U/L vs. (388.56 +/- 75.01) U/L, P < 0.05; CK-MB: (146.00 +/- 60.12) U/L vs. (291.16 +/- 52.41) U/L, P < 0.05] and apoptosis index [(4.32 +/- 1.16)% vs. (8.58 +/- 1.12)% , P < 0.05] were all significantly lower while Bcl-2 level (2.00 +/- 0.34 vs. 1.40 +/- 0.18, P < 0.05) was significantly higher in G-Post group than those in S-Post group. Moreover, above mentioned cardiac protective effects were significantly stronger in the G-Post group compared to R-Post group (all P < 0.05).</p><p><b>CONCLUSION</b>In conclusion, gradual algorithm of postconditioning could attenuate reperfusion injury more significantly than standard algorithm, and mitochondrion pathway plays an important role in this cardioprotective process.</p>

Change of JNK MAPK and its influence on cardiocyte apoptosis in ischemic postconditioning / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To test whether postconditioning could inhibit the expression of phospho-JNK (P-JNK) mitogen activated protein kinase (MAPK) and study its relation to apoptosis of cardiocyte.</p><p><b>METHODS</b>Sixty rats were randomly divided into six groups: sham, reperfusion injury (R/I), postconditioning (Post), SP600125 (I_JNK), anisomycin and postconditioning (Ani+Post) and anisomycin (Ani) groups. After acute myocardial infarction was induced in rats, placebo solution (DMSO), SP600125 (6 mg/kg) or anisomycin (2 mg/kg) was injected through jugular vein 5 min before reperfusion; 6 h later 3 rats of each group were executed and the hearts were separated to measure the signaling molecules (phospho-JNK, TNF alpha, Caspase-8, Bcl-2/Bax, cytochrome-c). Twenty-two hours later hemodynamic data were measured in the left rats, and then blood samples were taken to determine serum markers of cardiac damage, and hearts were separated to measure the infarction area and cardiocyte apoptosis.</p><p><b>RESULT</b>Postconditioning improved +/-DP/DTmax of left ventricle, limited infarct area, relieved apoptosis and necrosis of cardiocytes, and inhibited the expression of P-JNK (1.12 +/-0.21 Compared with 1.90 +/-0.32, P<0.05). At the same time the levels of TNFalpha Caspase-8, Bax and Cyt-c were lower in Post group than those in R/I group, but Bcl-2 expression levels were higher. I_JNK group presented the similar protection effect of postconditioning [TUNEL index: (6.23 +/-2.43)% Compared with (18.22 +/-5.10)%, P<0.05; Infarct area: (23.44 +/-6.34)% Compared with (42.31 +/-8.21)%, P<0.05]. On the other hand, Ani+Post group partially lost cardioprotection effect [TUNEL index: (14.12 +/-2.00)% Compared with (18.22 +/-5.10)%,P>0.05; Infarct area: (35.27 +/-5.28)% Compared with (42.31+/-8.21)%,P>0.05], because of the activation of JNK MAPK.</p><p><b>CONCLUSION</b>Postconditioning can inhibit phosphorylation of JNK MAPK, which attenuates cardiocyte apoptosis by both extrinsic and mitochondria pathway.</p>

Investigation of Magnetic Resonance Imaging Features and Treatment of Sacrococcygeal Teratoma in Children / 实用儿科临床杂志

Objective To analyze the magnetic resonance imaging (MRI) features of sacrococcygeal teratoma in children,and explore the diagnostic value of MRI and the critical suggestion for treatment.Methods The MRI features,clinical features,pathological data and operation complication of sacrococcygeal teratoma in 26 children were retrospectively analyzed. Phenotype teratoma in 9 cases grew from apex of coccyx to fundament.They presented spherical or ellipsoidal,distinct boundary and septatus in cystic MRI.All pathological data presented benign tumors.Mixed teratoma in 16 cases growed from fundament to cavitas pelvis.They presented dumbbell,distinct boundary,mixed signal of cystiform and solid density in MRI.There were 12 cases in benign tumors and 4 cases in malignant in pathology,the invisible teratoma in 1 case which lay between rectum and sacrum,extended to cavitas pelvis.Its boundary was not intact and segmented,and the pathological data presented malignant tumors.Results All of the 26 cases were completely excised only once and 21 of them received sacrococcygeal incision,5 cases had abdominosacral combined incision.Twenty-two cases were followed up (benign 20 cases,malignant 2 cases),and 2 cases of them died. 8 cases had complications,of the 8 cases,2 cases with necrosis of flap,1 case with chronic sinus tract and 2 cases with recidivism occurrd.Conclusions MRI may detect discovering the location and appearance of sacrococcygeal teratoma in children in diagnosis and in differential diagnosis of tumor nature.It is a very significant to reconstruct cavities pelvis and eradicase focal dead space in prevention and treatment of post-operation complication in childrens sacrococcygeal teratoma.