1.Effect of hypoxia on expressions of stromal cell-derived factor-1 and integrin-linked kinase in retinal pigment epithelium cell in vitro
Ye-qing, WANG ; Xia, LI ; Yong, L(U) ; Jian, WANG ; Xiao-mei, ZHANG
Chinese Journal of Experimental Ophthalmology 2012;(11):1013-1017
Background Hypoxia is a crucial factor of neovascularization.Many researches found that stromal cell-derived factor-1 (SDF-1) and integrin-linked kinase (ILK) play an important role in the neovascular disease.However,effect of SDF-1 and ILK in eye neovascular disease is below understood.Objective The aim of this study was to investigate the effect of hypoxia on the expressions of SDF-1 and ILK in cultured retinal pigment epithelium(RPE) cells in vitro.Methods RPE tissue was isolated from 4-week-old C57BL/6 mouse and was digested and cultured in DMEM/F12 with 10% fetal bovine serum (FBS).The cells with 80% confluence were collected and passaged.The third generation of cells were identified with cytokeratin 18 (CK18) antibody by immunochemistry.The cells were inoculated at the density of 5×104 cells/ml to free-serum DMEM/F12 for 24 hours and then were cultured in regular medium in the normoxic control group.RPE cells were cultured for 1 hour and 3,6,12,24,48,72 hours with 200 μmol/L CoCl2 in the hypoxia group.Reverse transcription-PCR(RT-PCR) was used to evaluate the expressing change of SDF-1 mRNA and ILK mRNA in RPE cells,and Western blot was used to assay the expressing change of SDF-1 protein and ILK protein in RPE cells in different time points.The detected outcomes were represented as the ratio of target gene A value/β-actin A value.Results Cultured cells showed the polygon in shape with the black pigment granules in cytoplasm.Over 90% cells were positive response for CK18.Expressions of the SDF-1 mRNA and ILK mRNA were increased in different time points after CoCl2 co-cultured(SDF-1 mRNA:F=281.875,P=0.000 ;ILK mRNA: F=187.566,P=0.000),with the highest expressing value in hypoxia at 12 hours.No significant change in the expression of SDF-1 mRNA and protein was found 1 hour after CoCl2 co-cultured,but expressions of SDF-1 mRNA and ILK mRNA were significantly higher in 3,6,12,24,48 and 72 hours than the normoxic control group(P<0.01).The expressions of SDF-1 protein and ILK protein were gradually ascended with the time increase of CoCl2 co-culture,showing a significant difference among different time points(SDF-1: F=44.719,P =0.000 ; ILK: F =144.481,P =0.000),and the up-regulation of SDF-1 protein and ILK protein expression was seen mainly in 3,6,12,24,48 and 72 hours after CoCl2 co-cultured in comparison with the normoxic control group (P<0.01).Conclusions SDF-1 and ILK are involved in the hypoxic response of RPE cells and may play a potential role in ischemic/hypoxic retinopathy.
2.Diagnosis of hyperfunctioning ectopic parathyroid glands using ~(99)Tc~m-MIBI SPECT scintigraphy combined localizable CT
Xue-min, L(U) ; Shu-hong, YU ; Jian-kui, HAN ; Yan, DONG ; Fu-yong, YANG
Chinese Journal of Nuclear Medicine 2010;30(1):42-45
Objective To evaluate the diagnostic value of ~(99)Tc~m-methoxyisobutylisonitrile (MIBI) SPECT scintigraphy combined Iocalizable CT in the localization of ectopic parathyroid glands in hyperparathyroidism.Methods Retrospective data of surgery,pathology and imaging were collected from 28 patients with hyperfunctioning ectopic parathyroid glands.All cases underwent CT studies.Twenty-five patients had ~(99)Tc~m-MIBI planar imaging first:SPECT scintigraphy combined localizable CT was performed for the patients with abnormal radionuclide foci immediately.The fusion images obtained after reconstruction showed the exact location of the ectopic foci.Operative histopathologic results were regarded as "gold standards".Presuming 4 parathyroid glands as normal findings,findings confirmed by operation and pathology were regarded as positive,otherwise negative.The results of CT and radionuclide imaging were compared by X~2-test of four-foId table.Results Twenty-eight ectopic parathyroid glands were found in 28 patients,all pathologically confirmed as adenomss.CT found 22 foci,of which 17 were true positive,5 false positive,11 false negative,and 79 true negative.~(99)Tc~m-MIBI SPECT scintigraphy combined localizable CT found 23 foci,no false positive,2 false negative,and 75 true negative.The results showed that the sensitivities were 61% (17/28),92%(23/25),specificities 94%(79/84),100%(75/75),accuracies 86%(96/112),98% (98/100),positive predictive values 77%(17/22),100%(23/23),and negative predictive values 88% (79190),97%(75/77),respectively,for CT and radionuclide imaging.~(99)Tc~m-MIBI SPECT scintigraphy combined localizable CT was therefore significantly higher than CT in sensitivity(X~2=6.98,P<0.01),specificity (X~2=4.61,P<0.05),accuracy (X~2=10.30,P<0.01),positive predictive value(X~2=5.88,P<0.05) and negative predictive value (X~2=5.36,P<0.05).Conclusion ~(99)Tc~m-MIBI SPECT scintigraphy combined localizable CT is superior to CT alone in the localization of ectopic parathyroid glands in hyperparathyroidism,but false negative can be found in some patients.
3.Inhibitory effect of eicosapentaenoic acid on proliferation of human vascular endothelial cells induced by vascular endothelial growth factor
Yong-qin, BAO ; Jing-xue, MA ; Shu-fen, WANG ; Lan-cun, L(U) ; Ying-hua, DU
Chinese Journal of Experimental Ophthalmology 2011;29(1):38-42
Background Eicosapentaenoic acid(EPA)function as the critical lipid mediators involved in several biological events in human body and play important role in suppressing the genesis of vascular endothelial growth factor (VEGF),migration and proliferation of vascular endothelial cells.Many ocular diseases were proved to be associated with neovascularization.Objecfive The purpose of this study was to investigate the inhibitory effect of EPA on the proliferation of human umbilical vein endothelial cells (HUVEC) indueed by VEGF. Methods HUVEC strain was cultured and passaged,and difierent concentrations of EPA were added to the medium with and without VEGF.The cultured cells were identified by antiofactor Ⅷ polyclonal antibody.The suppressing role of different concentrations of EPA on the proliferation of VEGF-induced or-uninduced HUVEC was assessed by MTT method.The influence of difierent concentrations of EPA on the cellular cycle of VEGF-induced HUVEC was assayed using flow eytometry.The expression of Flk-1,a receptor of VEGF,in the HUVEC Was detected by immunohistochemistry. Results Cultured HUVEC showed the ftlsiform in shape and presented with the cobblestone-like arrangement with the positive response for Ⅷ factor-related antigen.Various concentrations of EPA showed obviously inhibitory effect on VEGF-induced or-unindueed HUVEC at a dose-dependent manner (F=23.072.P=0.000).The inhibitory ability of EPA on VEGF-induced HUVEC was stronger than VEGF-uninduced HUVEC(F=41.417,P=0.000).In 24,48 and 72 hours,the action of EPA on the proliferation of HUVEC was gradually enhanced with the prolong of time(F=1.495,P=0.236).Cell cycle analysis indicated that EPA arrested VEGF-induced HUVEC in G0/G1 phase.The ratio of HUVEC in G0/G1 phase in EPA group was(75.83±1.56)%,and that in control groups was(68.62±1.44)%,showing a significant difference between them(t=-5.88,P=0.00),and no apoptosis of HUVEC was found in both groups.Flk-1 was strongly expressed in the cellular nucleus and cytoplasm in control group.However,the positive expressing intensity of Flk-1 in the HUVEC weakened,and the positive cell number was evidently less in EPA group. Conclusion EPA can inhibit the proliferation of VEGF induced HUVEC through arresting the synthesis of DNA of HUVEC and downregulate the expression of Flk-1 in HUVEC.These results suggest that EPA might exert an antiangiogenic effect.
4.Effect of Thumb Pressing at Yangming Meridian on Function Rehabilitation After Total Knee Arthroplasty
Jun-Qing GU ; Yong-Ying LIANG ; Ying L(U) ; Lian-Bo XIAO
Shanghai Journal of Acupuncture and Moxibustion 2017;36(11):1348-1351
Objective To observe the effect of thumb pressing at Yangming Meridian on the function rehabilitation after total knee arthroplasty (TKA) for knee osteoarthritis (KOA).Method Sixty KOA patients after TKA were randomized into a treatment group (thumb pressing at Yangming Meridian plus conventional rehabilitation) and a control group (single conventional rehabilitation),30 cases each.The treatment group was intervened by thumb pressing and kneading manipulation at the acupoints of Foot Yangming Meridian and Foot Taiyin Meridian plus rehabilitation training of passive flexion and extension by CPM machine,while the control group was intervened by rehabilitation training of passive flexion and extension by CPM machine.The American Hospital for Special Surgery (HSS) for knee joint,the American Knee Society score (KSS) and the Western Ontario and the McMaster Universities Osteoarthritis Index (WOMAC) of KOA were used to evaluate the function.Result After the treatment,the HSS,KSS and WOMAC scores were changed significantly in both groups (P<0.05).The improvements of HSS,KSS and WOMAC scores in the treatment group were superior to those in the control group,and the differences were statistically significant (P<0.05).Conclusion The efficacy of thumb pressing at Yangming Meridian plus conventional rehabilitation for post-TKA KOA is superior to single conventional rehabilitation.
5.Analysis of the surveillance result of iodized salt at household level in Hebei province from 2007 to 2009
Li-hui, JIA ; Sheng-min, L(U) ; Jing, MA ; Dong-rui, MA ; Yong-gui, DU ; Jun, ZHAO ; Zhen-shui, CHONG
Chinese Journal of Endemiology 2010;29(6):656-658
Objective To find out the situation of household consumption of iodized salt in Hebei province so as to provide scientific basis for prevention and control of iodine deficiency disorders(IDD). Methods According to the "national iodine deficiency disorders surveillance program (Trial)", the county (city, district) was taken as a unit, township (town) and administrative villages were selected in accordance with the principle of systematic sampling, then households were chosen by random sampling to collect their edible salt in Hebei province from 2007 to 2009. Salt iodine content was detected by direct titration method. Results A total of 48 675, 48 448 and 48 756 salt samples were collected from 2007 to 2009, respectively. The consumption rate of qualified iodized salt from 2007 to 2009 was 91.16%, 91.96% and 96.17%, respectively. There were 24.6%(41/167)and 18.0%(30/167)counties with consumption rate of qualified iodized salt under 90% in 2007 and 2008, respectively. The percentage of counties with consumption rate of qualified iodized salt above 90% was 100.0%(167/167) in 2009. In general there was a significant differences in frequency distribution of consumption rates of qualified iodized salt among the three years(H = 10.778, P < 0.01 ), and the difference was found significant between 2007 and 2009 as well as between 2008 and 2009(all P < 0.05), but was not significant between 2007 and 2008(P > 0.05). Conclusions The consumption rate of qualified iodized salt at household level increases annually from 2007 to 2009. By 2009,the consumption rate of qualified iodized salt in each county is more than 90%, which has reached the national standard stipulated in "the evaluation programs for fulfilling the goal of eliminating IDD at county level".
6.Inhibitory effects of eicosapentaenoic acid on expression of nuclear factor-kB and cytokine in rat corneal neovascularization
Yong-qin, BAO ; Jing-xue, MA ; Gun-xi, YE ; Lan-cun, L(U) ; Bai-xia, DONG ; Ying, ZHAI
Chinese Journal of Experimental Ophthalmology 2011;29(8):707-712
Background Corneal neovascularization (CNV) is an important cause of visual impairment and graft rejection after allograft corneal transplantation in inflammatory corneal diseases. The mechanisms and therapy relating to CNV are intensely investigated at all times. Objective This study was to evaluate the effect of eicosapentaenoic acid (EPA) on CNV induced by alkali cauterization and its mechanism. Methods The animal models of corneal neovasculation were induced in the right eyes in 72 Sprayue-Dawley rats by putting a piece of 3 mmfilter paper with 1 mol/L NaOH at the center of the cornea for 30 seconds. The rats were then divided randomly into the 0.02 mg EPA treatment group (24 rats) ,0.03 mg EPA treatment group (24 rats) ,model group (24 rats) and normal group (6 rats). EPA of 0.04 ml with doses of 0.02 mg or 0. 03 mg or saline solution of 0. 04 ml was injected subconjunctivally in model rats and immediately after cauterization. The presence of CNV and corneal edema were observed daily by slit lamp biomicroscope. 1,4,7 and 14 days after operation, corneal histopathological examination was performed by hematoxylin and eosin staining. The vascular endothelial cells were stained with CD34 by immunohistochemistry,and the expression of IL-1α,IL-6 mRNA and the nuclear factor-κBp65 ( NF-κBp65 ) proteins was measured by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis. The use of animals complied with the Regulations for the Administration of Affair Concerning Experimental Animals by Hebei Province( version 1998 ). Results Under the slit lamp, CNV grew slowly from days 2-4 with obvious corneal edema and defect of epithelium. Larger CNV area and less edema were seen from days 7-10. Maximal vessel growth was observed 14 days after injury with thinner vessels in the model group. Histological examination showed that part of the corneal epithelium was damaged;serious corneal edema, more inflammatory cells and a lot of CNV in the stroma were presented in the model group. However, repairing of the corneal epithelium without CNV ,light corneal edema and less inflammatory cells were found in both the 0. 02 mg EPA and 0. 03 mg EPA treatment groups 7 days after alkali cauterization. The relative area of CNV in the 0. 02 mg EPA treatment group was ( 15.80±6.43 )% and ( 11.06±2. 14)% ,and that in the 0. 03 mg EPA treatment group was (16. 10±7.41 )% and (11.06±2. 51 )%, showing significant reduction in comparison with the model group [ (84. 74±7.77)% and (89.63±7.50) % ] 7 days and 14 days after operation ( P<0. 05 ). Stronger expression of CD34 in the vascular endothelial cells of the cornea stroma was observed in the model group and an absence of CD34 was observed in the EPA-treated groups on the 7th day. RT-PCR revealed that the expression of IL-1α mRNA and IL-6 mRNA was lower in the EPA treatment groups than the model group ( P<0. 05 ), and Western blot analysis showed that the expression of NF-κB/p65 in the corneas in the EPA treatment groups was significantly lower than that in the model group on the 4th day after operation (P<0.05).Conclusion Topical application of EPA suppresses CNV induced by alkali burn possibly by inhibiting the expression of NF-κB,IL-1α and IL-6.
7.Application of avastin in vitrectomy for proliferative diabetic retinopathy and its mechanism
Xiang-gui, WANG ; Yong-ping, LI ; Shi-yi, LI ; Yong-hao, LI ; Jing-lin, ZHANG ; Xin-hua, HUANG ; Lin, L(U)
Chinese Journal of Experimental Ophthalmology 2011;29(1):53-57
Background Researches demonstrated that avastin-assisted vitrectomy for serious proliferative diabetic retinopathy can decrease intra-operation complication and bring down difficulty of surgery.It is speculated that this is associated with suppression of avastin on neovascularization.However,the evidence of its pathology is lack.Objective The aim of this study was to evaluate the application and mechanism of avastin in vitrectomy for severe proliferative diabetic retinopathy. Methods Twenty-four eyes from consecutive 24 patients with vitrectomy for severe proliferative diabetic retinopathy were enrolled in this study.Fourteen eyes received all intravitreal injection of 0.06 ml avastin(1.5 mg)14 days prior to vitrectomy。And the other 10 eyes underwent only vitrectomy without avastin injection as controls,Preretinal membranes were collected during vitrectomy for histopathologic examination by hemotoxylin and eosin staining.The differences in the density of the neovessels and micro-neovessels between the two groups were observed by detecting the expression of CD34 in vesse]endothelial cells using immunohistochemistry.Written informed consent was obtained from each patient before surgery. Results No statistically significant differences were found in the demography and eye manifestations between only vitreetomy group and avastin+vitrectomy group(P>0.05).The neovessels with grade three was in 10 eyes in only vitrectomy group and 1 eye in avastin+vitrectomy group(P<0.01).More capillary-like neovascularization with single vascular endothelial cells,obvious hemorrhage and inflammatory cells infiltration were observed on preretinal membranes in vitrectomy group.However,there were less hemorrhage,ceUular components and few capillary-like neovascularization but more hyaline degeneration of fibrous tissue were observed in avastin+vitrectomy group under the light microscope.Immunochemistry revealed that CD34 was positively expressed in vascular endothelial cells on preretinal membrane in both two groups.The neovessels density and miero-neovessels density were 15.40±7.42/field and 1.88±1.70/field in avastin+vitrectomy group and those in vitreetomy group were 18.00±3.80/field and 0.45±0.56/field respectively,showing significant differences between these two groups(neovessels density:Z=-4.102,P<0.01;micro-neovessels density:Z=-4.137,P<0.01).Conclusion As an adjunct drug during the vitrectomy for proliferative diabetic retinopathy, avastin can improve the successful rate of surgery by inhibiting the neovascular formation and alleviating retinal edema.
8.Development of time resolved fluorescence microparticle-conjugated anti-human procalcitonin antibody for homogeneous immunoassay
jun Can WU ; yu Qing L(U) ; jie Huai HAO ; na Cheng ZHAO ; ling Yu ZHENG ; qiang Yong JIANG
Military Medical Sciences 2017;41(8):689-693
Objective To develop europium (Ⅲ) [Eu (Ⅲ)] chelated microparticles for homogeneous immunoassay.Methods Anti-human PCT antibodies were labeled with Eu (Ⅲ) chelated nanoscale microparticles as the detection antibody,and another anti-human PCT antibody was labeled with biotin as the solid-phase antibody.Magnetic microspheres labeled with streptavidin were used to separate the complexes of Eu-IgM-PCT-IgM-Biotin.Results In the homogeneous immunoassay,the standard curve fit was not linear.The quadratic curve was Y =19170.12 + 75493.74X-26.00X2(r =0.9986).According to the standard curve,the limit of detection for PCT was 0.04 ng/ml.Conclusion The homogeneous immunoassay which uses Eu (Ⅲ) chelated microparticles is highly sensitive for detection of PCT recombinant antigens and may serve as a promising method to measure serum PCT levels in the future.
9.Isokinetic dynamometry in anterior cruciate ligament injury and reconstruction.
Yong Hao PUA ; Adam L BRYANT ; Julie R STEELE ; Robert U NEWTON ; Tim V WRIGLEY
Annals of the Academy of Medicine, Singapore 2008;37(4):330-340
The use of isokinetic dynamometry has often been criticised based on the face-validity argument that isokinetic movements poorly resemble the everyday multi-segmented, dynamic activities of human movements. In the anterior cruciate ligament (ACL) reconstruction or deficiency population where muscle deficits are ubiquitous, this review paper has made a case for using isokinetic dynamometry to isolate and quantify these deficits in a safe and controlled manner. More importantly, the usefulness of isokinetic dynamometry, as applied in individuals with ACL reconstruction or deficiency, is attested by its established known-group and convergent validity. Known-group validity is demonstrated by the extent to which a given isokinetic measure is able to identify individuals who could and could not resume pre-morbid athletic or strenuous activities with minimal functional limitations following an ACL injury. Convergent validity is demonstrated by the extent to which a given isokinetic measure closely associates with self-report measures of knee function in individuals with ACL reconstruction. A basic understanding of the measurement properties of isokinetic dynamometry will guide the clinicians in providing reasoned interventions and advancing the clinical care of their clients.
Adolescent
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Adult
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Anterior Cruciate Ligament
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physiopathology
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surgery
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Anterior Cruciate Ligament Injuries
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Biomechanical Phenomena
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Female
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Humans
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Knee Injuries
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physiopathology
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surgery
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Male
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Middle Aged
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Muscle Contraction
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physiology
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Orthopedic Procedures
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Range of Motion, Articular
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physiology
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Torque
10.Analysis in the result of Keshan disease surveillance in Hebei province from 1990 to 2007
Dong-rui, MA ; Jing, MA ; Li-hui, JIA ; Yong-gui, DU ; Guang-jun, YAO ; Chao-hui, ZHOU ; Sheng-min, L(U)
Chinese Journal of Endemiology 2010;29(2):203-207
Objective To observe the dynamic changes of prevalence of Keshan disease (KD) in Hebei province from 1990 to 2007, to provide scientific basis for its prevention and treatment. Methods The surveillance data of KD was analyzed according to "the National Scheme of KD Surveillance and the Surveillance of KD" (W/T 78-1996) in Hebei province from 1990 to 2007 by the Institute for Prevention of Endemic Disease in Hebei Province Center for Disease Control and Prevention. The data included physical examination, electrocardiogram (ECG), the chest X-ray film of KD patients and the suspected patients, as well as selenium contents of hair collected in 1990, 1992 and 1999. Results No new cases of acute and subacute types of KD patients were found at the surveillance sites from 1990 to 2007. Thirty-five cases of new latent KD and one case of spontaneous chronic KD were identified respectively. Prevalence of chronic and latent KD ranged from 1.12% (8/713) to 8.18% (27/330) and 2.29% (19/831) to 8.20% (45/549) in Hebei province from 1990 to 2007, respectively. The prevalence of KD in children aged 3 - 14 years old and childbearing woman aged 20 - 45 years old decreased year by year, however population over 45 years old were more likely suffering from KD. The major abnormal changes of ECG in KD) patients were complete fight bundle branch block, ST-T change frequent premature ventricular contraction, and left anterior faseicular block. The prevalence of the heart enlargement in KD patients was 47.00% (211/449) averagely, and the prevalence of heart enlargement of medium grade increased remarkably after 2005 [28.57%(8/28) - 48.39%(15/31)]. The average mortality in chronic KD patients was 18.0%(18/100) from 1990 to 2007. Conclusions The prevalence of KD decreased slowly in Hebei province. Hebei province is still the region with higher prevalence of KD around the country, and the tasks of prevention and treatment of KD is still urgent. Enhancing the surveillance of of KD and carrying out management and treatment of KD patients should be emphasized in the future.