Objective Researches showed that matrix metalloproteinase-2 (MMP-2) has a critical role in the neovascularization of tumor,and vascular endothelial growth factor(VEGF)is a promoting factor of new blood vessel formation.There still is little literature about the effect of MMP-2 in retinal neovascularization up to now.This study tried to explore the expression and significance of MMP-2 and VEGF in retinal neovascularization.MethodsA retinal neovascularization model was established in 30 7-day-old cleaning C57BL/6J mice exposed to an environment of high concentration of oxygen for 5 days,and 30 matched mice were raised in normal air environment.Fifteen mice from hyperoxic group and control group were sacrificed in 10 days after treatment and the eyeballs were enucleated to make retinal stretched preparation.Adenosine diphosphate-ase (ADPase) stained retina flat-mounts was performed to assess the retinal vascular profiles,and H&E staining was applied to count the number of new vascular cell nuclei.The expression of MMP-2 and VEGF was detected using immunohistochemistry by calculating the intergrated value of positive cells.ResultsThe retinal stretched preparation presented more neovascularization in mice from the hyperoxic group compared with control group.The number of nuclei from the vascular endothelial cells in the new vessels breaking through the internal limiting membrane in the hyperoxic group was 33.51±2.55,indicating a significant increase in comparison with control group (7.27±0.20)(t=9.345,P＜0.05).There were stronger expression of MMP-2 protein and the VEGF protein in the ganglion cell layer,inner plexiform layer and inner nuclear layer,and neovascularization breaking through the internal limiting membrane in the hyperoxic group compared with control group (t=4.25,P＜0.05;t=6.38,P＜0.05).Expression of MMP-2 showed the positive correlation with the expression of VEGF(r=0.825,P＜0.05).ConclusionBoth MMP-2 and VEGF promote retinal neovascularization.The overexpression of MMP-2 and VEGF play a synergistic role during the formation of neovascularization.

AIM: To study the inhibitory effect of captopril on retinal neovascularization (RNV).METHODS: Sixty seven-day-old mice were randomly divided into treated group and control group with thirty mice in each group. These mice were exposed to 750 50mL/L oxygen for 5 days and then to room air.The treated group had been injected captopril (2.7mL/kg), while control group had been injected 9g/L sodium chloride (2.7mL/kg) by intravitreal for 5 days.The mice were sacrificed at the 17th day after birth and the eyes were enucleated. Adenosine diphosphate-ase(ADPase) stained retina flat-mounts was performed to assess the retinal vascular profiles, Hematoxylin Eosin (HE)staining method was applied to count the number of new vascular cell nuclei and the expression of matrix metalloproteinase-2(MMP-2)and pigment epithelium derived factor (PEDF)was detected by immunohistochemical method.RESULTS: Comparing with control group,regular distributions and good branch and reduced density of RNV were observed in the treated group. The number of nucleus of new vessels vascular endothelial cells breaking through the internal limiting membrane was less in the treated group than in the control group (P<0.05). Stain of retinal MMP-2 was weaker in the treated group than in the control group and stain of retinal PEDF was stronger in the treated group than in the control group.CONCLUSION: Intravitreal injection of captopril (2.7mL/kg) may block the RNV in the oxygen-induced mouse model and may provide an effective method for prevent-ing RNV.

Sperm DNA fragmentation index (DFI) refers to the percentage of DNA strand breaks in the total sperm. Many studies suggest that elevated DFI can lead to male infertility and early spontaneous abortion. High-DFI patients are more likely to fail in assisted reproduction and preliminary treatment or prevention methods have been developed for this population. This review focuses on the impact of DFI on clinical pregnancy outcomes and progress in the studies of its treatment.
Abortion, Spontaneous ; Chromatin ; DNA Fragmentation ; Female ; Humans ; Infertility, Male ; Male ; Pregnancy ; Pregnancy Outcome ; Sperm Injections, Intracytoplasmic ; Spermatozoa

Objective To explore the inhibition effect of Cysteine-rich 61 (CCN1; Cyr61) specific siRNA expression vector on RNV in a mouse model of oxygen-induced retinopathy (OIR).Methods One hundred and twenty healthy C57BL/6J mice were chosen and randomly divided into the experimental group and control group,with 60 mice in each group.The experimental group was intravitreously injected with CCN1siRNA recombinant plasmids.The control group was injected with vector plasmids.Adenosine diphosphate-ase stained retina flat-mounts was performed to assess the retinal vascular profiles,retinal section with HE staining was applied to count the number of new vascular cell nuclei and the protein and mRNA expression of CCN1 and vascular endothelial growth factor (VEGF) were detected by immunohistochemistry,Western blot and Real-time RT-PCR.Results Compared with control group,regular distributions,good branches and reduced density of retinal neovascularization were observed in the experimental group.The number of nucleus of vascular endothelial cells breaking through the inner limiting membrane was obviously less in the experimental group than that in the control group (t=8.756,P＜ 0.05).The expression of CCN1 and VEGF were obviously decreased in the experimental group compared with the control group (all P＜0.05).Conclusion The development of RNV of ROP can be markedly inhibited by RNA interference targeting CCN1,and CCNlsiRNA may provide an effective method for preventing vascular proliferative retinopathy.

Objective To explore the efficacy of GM6001,tissue inhibitor expression and significance of matrix metalloproteinase?9(MMP?9)in mice model of oxygen?induced retinal neovascularization(RNV)and evaluate the inhibition effect of MMP?9 inhibitor(GM6001)on RNV. Meth?ods Mice were placed in oxygen boxes to establish oxygen?induced RNV animal models. The GM6001 treated or hyperxia control groups received an intravitreal injection of 1μL GM6001(100μmol/L)or PBS at day 11 after birth. The normal control and hyperxia group were not treated. HE staining was used to detect RNV in retinal whole mounts,the mRNA level and protein expression of MMP?9 were measured by RT?PCR,Western blot and immunohistochemistry,respectively. Results RNV in the GM6001 treated group was decreased significantly compared with the hyperxia group and hyperxia control group. Compared with the normal control group,higher protein and mRNA expression of MMP?9 were observed in the hy?perxia group and hyperxia control group. The expression of MMP?9 protein and mRNA were decreased in the GM6001 treated group compared with the hyperxia control group(P<0.05). Conclusion The abnormal expression of MMP?9 was closely correlated with RNV. The development of RNV can be markedly inhibited by MMP?9 inhibitor(GM6001),which,we believe,will provide new molecular targets and therapeutic strategy for retinopathy of prematurity treatment.

Objective: To investigate the relationship between Bilirubin blood lipid comprehensive index and ifbrinogen (FIB) to severity of coronary lesions in patients with coronary artery disease (CAD).
Methods: A total of 324 patients with angiography (CAG) conifrmed diagnosis were divided into 2 sets of groups.①By CAG examination, the patients were divided into 2 groups: CAD group,n=262 and Non-CAD group,n=62.②By Gensini scoring system, the patients were divided into 4 quartile groups: 1st quartile group,n=58, 2nd quartile group,n=110, 3rd group, n=80 and 4th quartile group,n=76. The blood levels of total bilirubin (TBIL), total cholesterol (TC), LDL-C, HDL-C, TG and ifbrinogen were measured and bilirubin blood lipid comprehensive index, TC/(HDL-C+TBIL) and LDL-C/(HDL-C+TBIL) were calculated respectively.
Results:①By CAG examination, compared with Non-CAD group, CAD group had increased TC, LDL-C, ratios of TC/(HDL-C+TBIL), LDL-C/(HDL-C+TBIL) and FIB,P<0.05; decreased TBIL and indirect bilirubin (IBIL),P<0.05.②By Gensini scoring system, the level of TBIL, IBIL, ratios of LDL-C/(HDL-C+TBIL), TC/(HDL-C+TBIL), HDL-C and FIB were different among 4 groups,P<0.05.③ Logistic regression analysis indicated that with excluded interactional factors, the bilirubin blood lipid comprehensive index was the independent risk factor for CAD[TC/(HDL-C+TBIL): OR: 1.668, 95% CI:1.065~2.784,P=0.028; LDL-C/(HDL-C+TBIL ): OR: 1.786, 95% CI: 1.021~2.879,P=0.030].④Correlation analysis presented that TC/(HDL-C+TBIL), LDL-C/(HDL-C+TBIL) and FIB were positively related to Gensini scoring system (r=0.423,P<0.01), (r=0.417,P<0.01) and (r= 0.293,P <0.01) respectively.
Conclusion: Bilirubin blood lipid comprehensive index and ifbrinogen were positively related to severity of coronary lesions in CAD patients.

[Abstract ] Objective High-mobility group box-1 (HMGB1) is abundantly released in the epileptogenic brain tissue , but few reports are seen about the effect of HMGB 1 on the expression of P-glycoprotein ( P-gp) in the vascular endothelial cells of the epi-leptogenic tissue .This study is to explore whether HMGB 1 can regulate P-gp expression in the brain microvascular endothelial cells of the mouse in vitro . Methods Immortalized brain microvascular endothelial bEnd .3 cells of the mouse were cultured in vitro and al-located to different concentration groups ( treated with culture medium containing 10 , 100 , 500 , and 1000 ng/mL HMGB1 for 8 hours), treatment duration groups (treated with culture medium containing 100 ng/mL HMGB1 for 4, 8, 16, 24, and 32 hours), and a control group ( treated with culture medium without HMGB 1 ) .The mRNA expression of P-gp-encoding gene-multidrug resistance gene 1a (mdr1a) was detected by real-time qPCR, and its protein expression determined by Western blot and immunocytochemistry . Results The results of qPCR manifested that the expressions of mdr 1a mRNA were 1.646 ±0.176, 1.777 ±0.135, 1.617 ±0.043, and 1.398 ±0.182 in the 10, 100, 500, and 1000 ng/mL HMGB1 groups, respectively, significantly higher than 1.030 ±0.284 in the control group (P<0.05), and so were those in the 4, 8, 16, 24 h, and 32 h groups (2.655 ±0.112, 2.168 ±0.212, 1.823 ± 0.232, 1.418 ±0.376, and 1.445 ±0.123) than in the control (1.010 ±0.164) (P <0.05).Western blot showed a significant increase in the P-gp protein expression in all the concentration groups (P<0.05) as well as in the 8 h and 16 h treatment duration groups as compared with the control group (P<0.05).Immunocytochemis-try also revealed a higher P-gp expression in the HMGB1-treated than in the control cells (P<0.01). Conclusion HMGB1 can upregu-late the expressions of mdr1a mRNA and P-gp protein in the brain microvascular endothelial cells of the mouse , which may associated with drug resistance of central nervous system diseases , especially that of epilepsy .

Objective To evaluate the effects of dexmedetomidine combined with subanesthetic dose of ketamine on the emergence agitation in the patients undergoing thoracotomy.Methods Eighty ASA physical status Ⅱ or Ⅲ patients,aged 55-75 yr,weighing 50-75 kg,scheduled for elective esophageal cancer resection,were randomly divided into 4 groups (n =20 each) using a random number table:normal saline group (NS group),dexmedetomidine group (group D),subanesthetic dose of ketamine group (group K),and dexmedetomidine combined with ketamine group (group DK).In DK and K groups,ketamine 0.5 mg/kg was injected intravenously (within 1 min) at 10 min before the end of the operation.In DK and D groups,dexmedetomidine 0.5 μg/kg was infused intravenously over 10 min starting from 10 min before the end of operation.In group NS,the equal volume of normal saline was infused intravenously over 10 min starting from l0 min before the end of operation.The emergence time,extubation time,duration of ICU stay,occurrence and degree of agitation,and development of cardiovascular events and hypoxemia within 24 h after operation were recorded.Ramsay sedation scores were recorded before induction of anesthesia (T1),immediately after completion of administration at the end of surgery (T2),and at 0,5,10 and 30 min after extubation (T3-6).Results There was no significant difference in the emergence time,extubation time,and duration of ICU stay between the four groups.Compared with group NS,Ramsay sedation scores were significantly increased at T3-6,the incidence and degree of agitation were decreased,and the incidence of cardiovascular events and hyoxemia was decreased in D,K and DK groups.Compared with D or K group,Ramsay sedation scores were significantly increased at T3-6,the incidence and degree of agitation were decreased,and the incidence of cardiovascular events and hyoxemia was decreased in group DK.Conclusion Dexmedetomidine combined with subanesthetic dose of ketamine can prevent the emergence agitation in the patients undergoing thoracotomy,which provides better efficacy than either alone.

Objective To analyze the clinical data of recipients over 15 years after renal transplantation,and to find the factors that affect the long-term survival of recipients after renal transplantation.Method Before June 30,2000,326 renal transplant recipients in our hospital were collected retrospectively.The risk factors which affect the survival of kidney transplant recipients and kidney were analyzed from four dimensions.A Cox model was established to analyze these multi factors.Result Cox hazard model indicated that advanced age (P=0.010,RR =1.052),AMR (P＜0.001,RR =18.311),nonadherence (P =0.001,RR =2.854),smoking (P =0.025,RR =2.097)were the risk factors for recipients' survival.Using immunosuppressive regimen FK506 + MMF+ Pred (P =0.019,RR =0.433),or CsA + MMF + Pred (P =0.019,RR =0.413) was the protective factor for recipients' survival.Nonadherence (P＜0.001,RR =5.645),and diabetes (P＜0.001,RR =3.310) were the risk factors of grafts' survival.Using immunosuppressive regimen FK506 + MMF + Pred (P＜0.001,RR =0.236),or CsA + MMF + Pred (P =0.002,RR =0.317) was the protective factor of grafts' survival.Conclusion To enhance the long-term outcome of recipients and grafts,the individualization of immunosuppressive regiments and controlling of the chronic diseases progress by changing the unhealthy life style are cutting on edge.

Objective To explore the protective effect and mechanism of astaxanthin (AST) on the acute kidney injury induced by iohexol in rats.Method Thirty rats were randomly divided into five groups:control group (Ctrl);iohexol group (CM);astaxanthin group (AST,100 mg/kg),low astaxanthin dose group (LAST+CM,50 mg/kg) and high astaxanthin dose group (HAST+CM,100 mg/kg),6 in each group.The rats in AST,LAST+CM,HAST+CM groups were administrated with AST by oral gavages using an intubation needle for 10 consecutive days.The rats in Ctrl and CM groups rats in Ctrl,CM groups were given with dissolvant instead in equal volume.Except for the Ctrl and AST groups,on day 8,rats were given indomethacin,L-NAME and iohexol in their femoral vein under chloral hydrate anesthesia to build a contrast induced-nephropathy (CIN) model.At the end of the experiment (72 h after CIN induction),all rats were sacrificed.The Scr level,BUN level,renal histology,renal tissue activities in superoxide dismutase (SOD),catalase (CAT),glutathione peroxidase (GPx),Glutathione (GSH) and level of malondialdehyde (MDA) were performed.Apoptosis of renal cells was detected by Bcl-2,Bax and Caspase-3 p17 with Western blot.Results Compared with Ctrl group,the levels of Scr,BUN were significantly increased in CM group (all P ＜ 0.01);while compared with CM group,the indicators were decreased in treatment groups (P ＜ 0.01).Renal tubular structure damage,medulla congestion,loss of brush border,vacuolar degeneration,apoptosis and proteinaceous casts were observed in the CM group,and the renal injury scores were higher compared with Ctrl group (P ＜ 0.05),however,administrated with AST could significantly improve the changes (P ＜ 0.05).Oxidative stress indicators showed that MDA level were increased while SOD,GPx,GSH activities were significantly decreased at CM group (all P ＜ 0.05),and the indicators above were ameliorated in treatment groups (all P ＜ 0.05).Western blot showed that the expression of Bcl-2 was down-regulated while the Bax,Caspase 3 p17 was up-regulated respectively at CM group (P ＜ 0.05),while the HAST+CM group could prevent the changes.Conclusions Iohexol can results in oxidative stress increased in kidney,which activate Caspase-3 p17 signal path,down-regulated Bcl-2 expression,up-regulated Bax expression respectively,and lead to cell apoptosis.AST can ameliorate the changes,especially with high AST dose,which suggest that the possible protection mechanism is by ameliorating oxidative stress and inhibiting apoptosis pathways.