Aim To study the action mechanism of hyperin(Hyp) on neonatal rat's neuron with anoxia/reoxygenation(A/R).Methods The dissociated neonatal rat brain cells were subjected to 30 min of anoxia or followed 40 min of reoxygenation.Lactate dehydrogenase(LDH),malondialdehyde(MDA)and nitric oxide(NO)in the supernatant were measured.The intracellular free calcium concentration([Ca~(2+)]_i)in brain cells was assayed with Fura 2-AM method.Results Anoxia induced a significant increase of LDH in the supernatant from (62.0±13.0) U·L~(-1)(Sham group)to (116.0±16.6) U·L~(-1)(Control group,P<0.01),and reoxygenation markedly increased LDH and MDA in the supernatant from (45.6±9.2) U·L~(-1) and (9.1±0.9) μmol·L~(-1)(Sham group)to (106.0±17.4) U·L~(-1) and (16.4±2.7) μmol·L~(-1)(Control group,P<0.01),respectively.In the range of 1.0 ～ 16.0 μmol·L~(-1),Hyp markedly and concentration-dependently inhibited anoxia-or reoxygenation-evoked increases of LDH and MDA.1.0～16.0 μmol·L~(-1) Hyp not only inhibited anoxia-induced increase of NO in the supernatant and rise of [Ca~(2+)]_i in brain cells(P<0.05 or P<0.01),but also attenuated reoxygenation-evoked increases of NO and[Ca~(2+)]_i(P<0.05 or P<0.01),Hyp 16.0 μmol·L~(-1) significantly reduced NO and[Ca~(2+)]_i from (34.4±6.3) μmol·L~(-1) and (640±94) nmol·L~(-1) to (25.0±5.1) μmol·L~(-1) and (331±56) nmol·L~(-1),respectively.Conclusion The protective effect of Hyp on A/R-injured neurons may be related to the inhibition of overload of[Ca~(2+)]_i,NO release and lipid peroxidation.

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.

Objective To explore the effect of oxygen inhalation on the retinae of newborn rats and its mechanism. Methods We mimicked the retinopathy of prematurity(ROP) by putting the newborn rats in high concentrated oxygen. One-day-old rats were put into the oxygen box with the oxygen concentration of 80% for continuous 7 days; then in air condition for 7 days. The arterial blood oxygen pressure, retinal superoxide dismutase (SOD), and malondialdehyde (MDA) of the rats (1,2,4,7,8,9,11,14 days old) were examined. The diameter of retinal vessels′ main branch and the coverage rate of peripheral vessels were measured in 7- and 14-day-old rats by ink perfusion. The retinal neovascularization of rats (8,9,11,14 days old) were observed by HE staining. The rats of the same age fed in air condition were in the control group. Results The differential pressures of blood oxygen of rats (1,2,4,7 days old) in study group were significantly higher than those in the control group (P0.05). The contents of SOD of the retinae in the rats (1,2,4,7,8 days old) were significantly lower than those in the control group(P

Split spinal cord malformation(SSCM) is a rare neurological abnormality that spinal cord was separated in the sagittal plane.Based on the tissue element and type of thecal shvac,SSCM is classified into Ⅰ,Ⅱ and mixed type.Congenital spinal deformity(CSD) is caused by the vertebral body formed or segmented obstacles.CSD can be divided into 3 types:Ⅰ,Ⅱ and mixed type.Both of SSCM and CSD are diagnosed depending on clinical symptoms,X rays,computed tomography (CT),magnetic resonance imaging(MRI) and other examine.Genitourinary and cardiac ultrasound should be performed at the same time.Progression of spinal deformity and nerve injury are accompanied with the growth process of children with CSD and SSCM.Early surgery can avoid further injury of nervous system,prevent spinal deformity progress and obtain good correction.Segmental tissue removal,tethered cord release and spinal deformity correction are generally consisted into an individual operative plan at present.Traditional surgical option included 2 steps:tissue removal and tethered cord release are conducted at Ⅰ phase surgery;then conducted the spinal deformity correction at Ⅱ phase.However,some reports suggested that completed the process of tissue removal,tethered cord release and spinal deformity correction at one surgery also obtain a satisfied effictiveness.The aim of this research is to review and discuss the diagnosis of the SSCM with CSD and evaluate the treatment effectiveness of Ⅰ phase surgery instead of the traditional surgical option.

Objective To analyse the economic loss from nosocomial infection(NI) in severe brain injury patients in neurosurgery.Methods The economic loss and hospitalization days of the severe brain injured patients in our hospital from January 2003 to December 2005 were analyzed by case control study.Results The hospitalization days and total cost of the NI group were higher than the controlled group,there was statistical significance(P

Objective To investigate the relationship between histopathological changes of donor liver and those after liver transplantation and explore its effects on prognosis.Methods The histopathological changes of donor liver and post-transplantation live biopsy were observed in 87 cases.Results The percentage of hepatic degeneration at immediate biopsy of donor liver was 100%,the injury of sinusoid endothelial cell accounted for 41.3%,the hepatic steatosis for 18.3%and the necrosis of liver cells for 10.3%.Conclusion Most of the pathological changes of the donor liver during and after liver transplantation are reversible.The damage of the hepatic sinusoidal cells is the important criterion for the observation of ischemia reperfusion injury.Moderate and heavy hepatic steatosis can be used in the liver transplantation. After liver transplantation,if the necrosis of liver cells gradually gets serious,the risk of transplanted liver losing its function gets bigger,so the comparative observation has very important value in diagnosis.

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.

BACKGROUND: There are currently few studies regarding the pharmacokinetics of vancomycin via intravitreous injection.OBJECTIVE: To determine the concentration of vancomycin injected into the vitreous chamber of conscious rabbits.METHODS: A microdialysis probe was implanted into vitreous chamber of normal rabbit eyes and rabbit eyes infected withbacterial endophthalmitis for 24 hours, and 10 g/L vancomycin 0.1 mL was administered intravitreally. The drug concentration inthe vitreous chamber of rabbit eyes was determined at 0.5, 1, 2, 4, 6, 12, 24, 48, 72 and 84 hours after injection, through themicrodialysis and high performance liquid chromatogram-ultraviolet detection.RESULTS AND CONCLUSION: The metabolism of vancomycin showed an open two-compartment model in normal rabbit eyes.Its half-life was 51.66 hours and the peak concentration was 695.92 mg/L. The metabolism of vancomycin in the infected vitreouschamber showed a one-compartment model. Its half-life was 11.91 hours and the peak concentration was 713.35 mg/L. All rabbitswere injected with drugs for 84 hours and the intravitreous concentration of vancomycin was higher than minimal inhibitoryconcentration. The experimental findings indicate that microdialysis coupled to high performance liquid chromatography is apowerful tool to investigate the ocular pharmacokinetics of vancomycin, and the samples are harvested in a real-time, continuousand dynamic fashion when the experimental animals are conscious.

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 investigate the expression of c-erbB-2 in breast cancer and its relationship with prognosis.Methods The expression of c-erbB-2 in breast cancer was detected by immunohistochemistry assays in 60 cases of breast cancer.Results The positive expression level of c-erbB-2 in invasive breast cancer was signifi-cantly higher than that in other kinds of breast cancer(P<0.01).The positive expression level of c-erbB-2 in the metastatic breast cancer with four lymph nodes was higher than in the early cancer with no lymph node involvement (P<0.01).The 10 years survival rate in negative expression of c-erbB-2 was higher than in positive expression(P<0.05).Conclusion c-erbB-2 is important for physicians to instruct the treatment of the breast cancer and judge the prognosis.