1.Effects and mechanism of protein tyrosine kinase inhibitor on blood-retinal barrier breakdown
Shi-jie, ZHANG ; Xi, ZHANG ; Qi, ZHU
Chinese Journal of Experimental Ophthalmology 2011;29(7):610-614
Background Several cytokines,especially interleukin-1β (IL-β) involve in the breakdown of blood-retina barrier,and the signal of cytokine is transduced through protein tyrosine kinase (PTK) pathway.Objective This study was to investigate the effects of PTK inhibitor,Genistein,on IL-1β-induced blood-retinal barrier breakdown and possible mechanism.Methods The animal models of blood-retinal barrier breakdown were induced through intravitreal injection of IL-1β(10ng) in 24 clean healthy SD rats and assigned to IL-1β group and Genistein group.5μl IL-1β+1μl Genistein with 0.2,1,5μg were intravitreally injected in 12 model rats and 5μl IL-1β (2mg/L)+1μl DMSO was used at the same way in other 12 models.Evans Blue was injected in rats via jugular vein in 1 hour before sacrifice of animals and the arterial blood was collected for the detect of serum Evans Blue.The retinas of the rats were obtained in 4 and 48 hours after injection of vitreous cavity to assay the content of Evans Blue in retina.The changes of vessels and infiltration of inflammatory cells were observed with hematoxylin-eosin stain.RT-PCR was employed to determine the expression of IL-8 and MCP-1mRNA in neuroretina after intravitreal injection.Expression of MCP-1 protein was localized by immunohistochemistry.Results The ratio of retinal Evans Blue and plasma Evans Blue was significantly decreased after intravitreal injection of different doses of Genistein among Genistein groups and IL-8 group with a statistical difference (4 hours:F=7.510,P=0.010;48 hours:F=5.960,P=0.019).With the increase of time after injection of Evans Blue,the ratio of retinal Evans Blue and plasma Evans Blue was gradually reduced in comparison to IL-1β group (P<0.05).After injection of IL-1β,the dilation of retinal vessel and adhesion of leukocyte to vessel wall were seen under the light microscope,but infiltration of less inflammatory cells was found in Genistein group.The expressions of IL-8 and MCP-1mRNA were obviously declined in retina of rats in Genistein groups compared with IL-8 group (P<0.05).Immunochemistry indicated that the expression of MCP-1 protein in neuroretina tissue was weaker in Genistein group compared with IL-8 group.Conclusion PTK inhibitor,Genistein,can decrease IL-1β-induced permeability of vessel and maintain the integrity of blood-retinal barrier by downregulating the expression of chemokines and infiltration of leukostasis in retinal vessels.This study imply that PTK pathway plays an important role in IL-1β-induced blood-retinal barrier breakdown.
2.Correlation of serum homocysteine,vitamin B12 and folate levels and subclinical hypothyroidism in pregnancy
Ling ZHANG ; Liang SHI ; Jie ZHANG
Chongqing Medicine 2016;45(27):3785-3787
Objective To analysis the correlation of Serum homocysteine ,vitamin B12 and folate levels and subclinical hypot‐hyroidism in pregnancy .Methods 150 cases of subclinical hypothyroidism during pregnancy were selected as the experimental group ,and 150 cases of pregnant women with normal thyroid function were collected as the control group .Compared the serum thy‐roid‐stimulating hormone ,free thyroid hormones ,three free triiodothyronine ,homocysteine ,serum vitamin B12 ,folic acid levels of pregnant women in the two groups ,and analyzed the related serum markers of thyroid‐stimulating hormone and serum homocys‐teine .Results The homocysteine levels in experimental group was higher than that of control group(P<0 .05) ,the vitamin B12 and folate levels in experimental group were lower than that of control group (P<0 .05) ,the thyroid‐stimulating hormone levels in ex‐perimental group was significantly higher than that of control group (P<0 .05) .Serum thyroid‐stimulating hormone and homocys‐teine levels were positively correlated (P<0 .05) ,serum thyroid‐stimulating hormone and vitamin B12 and folic acid had no signifi‐cant correlation (P>0 .05) .Serum homocysteine and thyroid stimulating hormone levels were positively correlated (P<0 .05) ,ser‐um homocysteine and folate levels were negatively correlated (P<0 .05) ,serum homocysteine levels and vitamin B12 were not corre‐lated (P> 0 .05) .Conclusion Subclinical hypothyroidism during pregnancy have some relevance with homocysteine ,vitamin B12 and folate levels .
3.Influence of angiotensin-(1-7) on inducible nitric oxide synthase expression after focal cerebral ischemia-reperfusion in rats
Jie LU ; Yingdong ZHANG ; Jingping SHI
Chinese Journal of Neurology 2011;44(2):128-131
Objective To investigate influence of Ang-(1-7) on the inducible nitric oxide synthase (iNOS) activity and gene expression following focal cerebral ischemia/reperfusion in rats. Methods Cerebral ischemia/reperfusion injury was induced by intraluminal thread occlusion of middle cerebral artery in the adult male Sprague-Dawley (SD) rats. Ang-(1-7) or artificial cerebrospinal fluid (aCSF) was continuous administrated by implanted Alzet osmotic minipumps into lateral cerebral ventricle after reperfusion. Experimental animals were divided into sham-operated group ( sham operation + aCSF), aCSF treatment group(MCAO+aCSF)and ang-(1-7)treatment groups(MCAO+Ang-(1-7))at low(1 pmol·0.5 μl-1·h-1),medium (100 pmol·0.5 μl-1·h-1)or hith(10 nmol·0.5 μl-1·h-1)dose levels.The activity of iNOS in ischemic tissues were measured by iNOS detection kits. Reverse transcription( RT)-PCR was used to determine messenger RNA (mRNA) of the iNOS in ischemic tissues. Results The cerebral ischemic lesion resulted in a significant increase of iNOS expression compared with sham operation group. The high-dose Ang-(1-7) markedly enhanced (iNOS) activity ( 160. 83 vs 116. 75 U/mg, F = 19. 22,P<0.01; 151.87 vs 113.07 U/mg, F=63.52,P<0. 01) and gene expression(0.43 vs 0.38, F=21.83,P < 0. 01; 0. 40 vs 0. 35, F = 19.49, P < 0. 01 ) compared with aCSF treatment group at 24 hours and 48hours after reperfusion, whereas medium and low-dose Ang-( 1-7 ) didn't stimulate iNOS activation.Conclusions The obtained results suggest that high-dose Ang-(1-7) upregulate iNOS expression following ischemic stroke.Moreover,overdose Ang-(1-7)(10 nmol·0.5 μl-1·h-1)may have Ang Ⅱ-like effects in iNOS expression increase.
4.Effect of Angong Niuhuang Wan, cinnabar and realgar against lipopolysaccharide-mediated neurotoxicity
Feng ZHANG ; Jie LIU ; Jingshan SHI
Chinese Journal of Pharmacology and Toxicology 2010;24(3):161-167
OBJECTIVE To investigate the role of cinnabar and realgar in Angong Niuhuang Wan (AGNH) -produced neuroprotection against lipopolysaccharide ( LPS) -mediated neuronal damage and further explore the corresponding mechanisms. METHODS Primary rat midbrain neuron-glia cultures were used as an in vitro model to investigate effects of AGNH on LPS-mediated degeneration of dopamine (DA) neurons. The experiment was divided into normal control group, LPS model group, LPS + cinnabar (4 and 40 mg·L-1) groups, LPS + realgar (4 and 40 mg·L-1 ) groups and LPS + AGNH (40 and 400 mg·L-1 ) group. Drugs were added 30 min before LPS treatment. After 7 d, dopaminergic neurotoxicity was assessed through the quantification of tyrosine hydroxylase (TH)-positive neurons and morphological analysis of TH-positive neurons; the activation of microglia was evaluated using OX-42 antibody; the gene expression of tumor necrosis factor-α (TNF-α) and induced nitric oxide synthase (iNOS) mRNA in microglia was performed by real-time RT-PCR analysis, and the release of TNF-α and nitric oxide (NO) in the supernatant of neuron-glia cultures was determined respectively by the ELISA and Griess reagent. RESULTS Compared with normal control group, DA neurons in LPS model group decreased by 40% (P <0.05) , microglial activation was induced, the expression of TNF-α mRNA and iNOS mRNA in microglia increased 9 and 2 times, respectively ( P < 0. 05 ) , and subsequent production of TNF-α and NO in the supernatant of neuron-glia cultures increased 20 and 30 times, respectively (P<0.05). Compared with LPS model group, AGNH 400 mg·L-1 and realgar 40 mg·L-1 significantly attenuated LPS-mediated DA neuronal loss by 40% and 30% , respectively (P<0.05) and inhibited activation of microglia and expression of TNF-α mRNA by 61% and 52% (P <0.05). iNOS mRNA was reduced by 58% and 51% (P <0.05 ) in microglia. The subsequent release of TNF-α was reduced by 55% and 43% (P<0.05) and NO reduced by 53% and 34% (P<0.05) in the supernatant of neuron-glia cultures. Cinnabar had no inhibitory effect on LPS-induced changes. CONCLUSION AGNH protects LPS-induced neurotoxicity through its anti-inflammatory properties and realgar might be the key contributor to the neuroprotective action of AGNH, while cinnabar fails to show any neuroprotection.
5.Relevance study of renal hemodynamic change and endothelia dysfunction in patients with hypertensive disorder complication pregnancy
Jie ZUO ; Lijun ZHANG ; Qiduo SHI
Chinese Journal of Ultrasonography 2013;(5):406-409
Objective To investigate if the endothelin-1 (ET-1),nitric oxide(NO) can affect renal artery blood flow and explore the clinical value of ultrasonography in hypertensive disorder complication pregnancy.Methods 18 patients of gestational hypertension,18 patients of gently preeclampsia,20 patients of severe preeclampsia and 20 normal pregnancy women were chosen in random.Color Doppler ultrasonography was used to detect the peak systolic velocity (PSV),end diastolic velocity (EDV),resistive index (RI),pulsatility index (PI) of renal interlobar artery.The levels of serum ET-1 and NO were measured by enzyme-linked immunosorbent assay (ELISA) and nitric acid recovery respectively.Results The level of EDV in preeclampsia patients were significant lower than the control group (P <0.05).The levels of RI,PI,S/D in preeclampsia patients were significant higher than the control group (P <0.05).The level of ET-1 in study group was higher than control group (P <0.01).The level of NO in study group was lower than control group (P <0.01).There was positive correlation between the RI,PI,S/D of renal interlobar artery and the level of serum ET-1.There was inverse correlation between the RI,PI,S/D of renal interlobar artery and the level of serum NO.Conclusions Endothelial dysfunction can cause unbalance of renal vessel systolic-diastolic function.The level of renal artery blood flow parameters can reflect the state of vessel function and have clinical value in hypertensive disorder complication pregnancy.
6.The comparision between two international QOL questionnaire of lung cancer:EORTC QLQ-LC43 and FACT-L
Jie YOU ; Zhiming SHI ; Xianjing ZHANG
China Oncology 2001;0(05):-
Background and purpose:Quality of life (QOL) questionnaires are tools to assess the quality of life of patients. How to choose a proper questionnaire is the fi rst problem of QOL assessment in clinic research. However, there is few report on it. Our current research compares the data obtained from two world-wide used QOL questionnaire of lung cancer patients EORTC QLQ-LC43 (European Organisation for the Research and Treatment of Cancer Quality of Life Questionnaire-Lung Cancer43) and FACT-L (Functional Assessment of Cancer Treatment-Lung) by various statistic methods in order to show differences between the two questionnaires and to provide some suggestions for choosing questionnaires to assess patients’ QOL in clinic research. Methods:The clinic data were obtained from 125 lung cancer patients by two different international QOL questionnaires of lung cancer patients EORTC QLQ-LC43 and FACT-L, and analyzed by various statistic methods such as Pearson correlation analysis, canonical correlation analysis and multiple regression. In addition, we also compare the general modality such as the frame, item numbers and the contents of the subscales, etc. Results:Of the two QOL there are some parts similar in length, ranking type and time construction, even in items. They all contain items in measuring the physical domain, emotion domain, function domain, society domain and lung domain. Correlations between corresponding subscales of the FACT-L and the EORTC QLQ-LC43 are all signifi cant ranging from r=0.331 for the social domain to r=0.664 for the emotional domain. Canonical correlation analysis for the two sets of subscales revealed there are four signifi cant canonical variables (canonical correlation coeffi cient r=0.87 to 0.26 and overall redundancy about 41% ). The eight EORTC QLQ-LC43 subscales are well represented by the fi ve FACT-L subscales (multiple linear regression, R 2=0.531 to 0.766) and the fi ve FACT-L subscales (except relationship with doctors) are also well represented by EORTC QLQ- LC43 subscales (R 2=0.537 to 0.823). The proportion of the corresponding subscale in the total explanation is 50% to 90%. Conclusion:These two questionnaires cover some common parts, but they measure different contents of QOL. Though have the same name, some domains cann’t be compared directly. The two questionnaires cann’t be alternative and the results of them shouldn’t be compaired directly. Both questionnaires are suited for clinic study (e.g compare the effect of two different theropies). But each has its own characters and should be chosen according to reseach goals.
7.Clinical significance of platelets activation in patients with acute cerebral hemorrhage
Jie LIN ; Guohua ZHANG ; Fuping SHI
Journal of Clinical Neurology 2001;0(05):-
Objective To study the platelets activation in patients with acute cerebral hemorrhage and its significance in perihematoma edema.Methods 33 patients with cerebral hemorrhage were treated with minimally invasive surgery.The volume of edema perihematoma was determined by CT technique before operation.The expressions of platelets CD62p and TSP in intracerebral hematoma fluid and venous blood were detected by flow cytometry(FCM).The correlation between the two variables was also analyzed.Results The expressions of platelets CD62p and TSP were higher in patients with acute cerebral hemorrhage than those in healthy individuals,and the expressions of platelets CD62p and TSP in intracerebral hematoma fluid were higher than those in venous blood.There were significant positive correlations between the expression levels of CD62p and TSP from the surface of platelets in hematoma and the edema size perihematoma(CD62p:r=0.4781,TSP:r=0.5183,all P
9.Rapid eye movement sleep behavior disorder as a predicting phenomenon of ?-synucleinopathies(report of 1 case)
Yingdong ZHANG ; Jie LU ; Jingping SHI
Journal of Clinical Neurology 1995;0(04):-
Objective To improve the recognition of rapid eye movement(REM) sleep behavior disorder(RBD) as an early marker for ?-synucleinopathies.Methods By studying a typical case of RBD followed with multiple system atrophy-P,the clinical features,pathogenesis and its correlation with ?-synucleinopathies of RBD were elucidated.Results This case manifested a serial of paroxysmal increased activities of the limbs and behavioral disturbances during his REM sleep,and parkinsonism features appeared 9 years later.His cranial MRI showed the abnormal long T1 and T2 signals at bilateral centrum ovale,corona radiate and basal ganglia area of the cerebral hemisphere.Conclusions RBD is clinically characterized with paroxysmal behavioral disorder in the REM sleep,the changes of the brain stem,striatum and cortical perfusion are attributed to the RBD pathogenesis.Closely linked to a-synucleinopathies,RBD may be clinical harbinger of those disorders.