Background In vitro study showed that chemotaxis consist of chemotaxis factor 4(CXCR4)and stromal cells derived factor-1(SDF-1)and may play a role in the orientation and migration of retinal progenitor cells (RPCs)toward lesion.Overexpression of CXCR4 in RPCs can enhance the chemotaxis activity.Objective This work was to explore the feasibility and underlying mechanism of up-regulation of CXCR4 on RPCs induced by hypoxia.Methods RPCs were retained in an incubator with normal O2volume(16％)or hypoxia condition(10％ O2)for 12 hours and 24 hours respectively.Flow cytometer cell analysis screening(FACS)was conduced to measure the proportion of CXCR4-expressing cells,and CXCR4,HIF-1 mRNA were analyzed by reverse transcription-polymerse chain reaction(RT-PCR).The chemotical effect of 30 mg/L SDF-1 to RPCs cultured under the hypoxia condition was assessed using Boyden chamber.Results The expression level of CXCR4(CXCR4 mRNA/β-actin mRNA)inRPCs cultured by 10％ O2 for 12 and 24 hours were 0.28+0.07and 0.48+0.17 and increased by 1.75 and 3.00 fold more than that of 16％ O2 culture group(0.16+0.02)(P＜0.01).The expression level of HIF-1 mRNA(HIF-1 mRNA/β-actin mRNA)in RPCs cultured by 10％ O2 for 12 and 24 hours were 0.18 ±0.07and 0.38 ±0.13 and increased by 3.00 and 6.30 fold more than that of 16％ O2 culture group(0.06±0.01)(P＜0.01).The chemotical effect of 30 μg/L SDF-1 to RPCs increased from 13.00％ in 16％ O2 culture group to 36.00％ and 46.00％ in the cells cultured by 10％ O2for 12 and 24 hours.FACS revealed that the proportion of CXCR4+ cells in hypoxia-exposure for 12 and 24 hours were 26.90％ and 46.10％,respectively,but that in 16％ O2 culture group was 9.10％,showing a statistically significant difference(P ＜ 0.01).Conclusions RPCs induced by hypoxia can enhance the expression of CXCR4 in RPE cells and the chemotaxia to SDF-1.The overexpression of H1F-1 in RPCs may be involved in the up-regulation of CXCR4 expression.

AIM: To investigate the angiogenesis effect and protective mechanism of cordycepin on rhesus macaque choroid- retinal endothelial ( RF/ 6A) cell line cultured in high glucose condition.
METHODS: Cultured RF/ 6A cells were divided into normal control group, high glucose group and high glucose (HG) + different concentration cordycepin groups (HG+ 10μ g/ mL group, HG+ 50μ g/ mL group, HG+ 100μ g/mL group). The cell proliferation was assessed using cholecystokinin octapeptide dye after treated for 48h. The cell migration was investigated by a Transwell assay. The tube formation was measured on Matrigel. Furthermore, the impact of cordycepin on high glucose - induced activation of VEGF and VEGF receptor 2 (VEGFR-2) was tested by Western blot analysis.
RESULTS: Compared with normal control group, cell viability markedly increased in high glucose group ( P <0. 05). Cordycepin inhibited RF/ 6A cell proliferation in a dose- dependent fashion: 10. 2 ± 0. 9%, 23. 4 ± 1. 5% and 31. 1±1. 2% inhibition as the concentrations of cordycepin were 10, 50 and 100μ g/ mL, respectively. The difference had statistically significant (P<0. 05) compared with high glucose group. The number of cell migration were 55. 6±2. 70, 87. 4 ± 2. 40, 65. 4 ± 2. 7, 57. 8 ± 2. 38, 62. 4 ± 2. 77 in normal control group, high glucose group and HG+10μ g/mL group, HG + 50μ g/ mL group, HG + 100μ g/ mL group respectively. Migration of RF/ 6A conspicuously increased in high glucose group ( P < 0. 05) compared with normal control group; while showing a gradually reducing trend with the increase of cordycepin dose and a statistically significant difference compared with high glucose group (P<0. 05). The number of tube formation were 18. 7±2. 08, 25. 7 ± 1. 52, 19. 9 ± 1. 57, 16. 3 ± 2. 51, 5. 67 ± 1. 72 in the abovementioned group. Similarly showing a gradually reducing trend with the increase of cordycepin dose and a statistically significant difference with high glucose group (P< 0. 05). In addition, the number of tube formation of RF/ 6A in high glucose group significant increased compared with normal control group ( P < 0. 05 ). The expression of VEGF and VEGFR-2 dramaticlly increased in high glucose group vs normal control group, oppositely gradually decreased with the increase of cordycepin concentrations, and had a statistically significant difference vs high glucose group (P<0. 05).
CONCLUSION: Cordycepin can suppress the proliferation, migration and tubu formation of RF/ 6A in high glucose condition, might via inhibiting expression of VEGF and VEGFR-2.

BackgroundResearch demonstrated that vitamin D3 mediated by its receptor has the potent nonclassical effects,including immunomodulatory,antiinflammatory,and neuroprotective properties,and it can enhance the secretion and sensitivity of insulin and therefore down-regulate hyperglycemia and attenuate the corneal edema.ObjectiveThe present study was to investigate the protective effect of vitamin D3on ocular structure in experimental diabetic rat.Methods Twenty-two healthy SPF C57BL/6 rats were randomly divided into vitamine D3 group (8 rabbits),diabetic control group ( 11 rabbits) and normal control group ( 3 rabbits).2％ streptozotocin ( STZ,175 mg/kg)was intraperitoneally injected to create the diabetic models in the rats of the vitamine D3 group and diabetic control group.Blood glucose was examined for 3 times in the third day after STZ injection,and the rats with the blood glucose concentration ＞16.7 mmol/L was identified as the successful diabetic models.After modeling,the rat tail blood was collected for the monitoring of blood glucose.Two weeks after modeling,vitamine D3 was intraperitoneally injected in each week for 5 times.The fundus was examined using direct ophtalmoscope,and the eyeballs were obtained under the excessive anesthesia for the measurement of thickness of the central cornea,retina and choroids by histopathological examination once a week for 7 weeks after administration of vitamin D3.The administration of the animals complied with the Statement of ARVO.ResultsThe corneal edema appeared with the corneal thickness of (339.14± 11.13) μm in the first week and gradually attenuated with time elapse after modeling in the diabetic group ( F =382.446,P =0.000).The corneal thickness values were significantly decreased from the second week through the seventh week in the vitamin D3 group compared with diabetic control group(P＜0.05).The atrophy of the corneal epithelium was found from the fifth week to the seventh week in diabetic control group,but that in vitamin D3 group was slight (P＜0.05).The gradually thinning of the choroids was seen from the first week to the seventh week in the diabetic control group ( F =437.411,P =0.000 ),however,the thickness values in the vitamin D3 group were significantly increased in comparison with the diabetic control group in various time points (P＜0.05).The retina thickness was gradually reduced during the seven-week duration in the diabetic control group (F =91.859,P =0.000),but no significant change was identified in retina thickness in the vitamin D3 group(P＞0.05).ConclusionsVitamin D3 has prevent and therapeutic effects on experimental diabetic oculopathy.

Child, Preschool ; Female ; Humans ; Sarcoma ; diagnosis ; surgery ; Stomach Neoplasms ; diagnosis ; surgery ; Stromal Cells ; pathology

Withanolide A is a biologically active secondary metabolite occuring in roots and leaves of Withania somnifera. In the present study, adventitious roots from leaf explants of W. somnifera were induced for the production of withanolide-A by Agrobacterium tumefaciens strain C58C1 to obtain hair roots. Hair roots induction rate reached 30%. The withanolide A was determined by HPLC in different hair roots lines and different parts of W. somnifera. The average content of withanolide A in all hair roots lines were 1.96 times as high as that in wild-plant, the concentration of withanolide A in hair roots (1.783 mg x g(-1) dry weight) were 1.51 times as high as the roots of wild W. somnifera (1.180 mg x g(-1) dry weight), respectively. It is possible to obtain withanolide A from hair roots culture of W. somnifera.
Agrobacterium tumefaciens ; physiology ; Plant Extracts ; analysis ; biosynthesis ; Plant Roots ; chemistry ; growth & development ; metabolism ; microbiology ; Withania ; chemistry ; growth & development ; metabolism ; microbiology ; Withanolides ; analysis ; metabolism

Genomic DNA of two fungi Thamnidium elegans and Umbelopsis isabellina were extracted with an amended Cetyltrimethyl Ammonium Bromide (CTAB) method. This modified method uses repeated freezing in liquid nitrogen and thawing with combination of shocking with glass beads to replace of the tra- ditional method. Quality and concentration of DNA extracted by the modified methodwere tested. Compared with the traditional method, higher yield and purity of genomic DNA were obtained with less amount ofmy- celium. The result indicted that this is a simple and highly efficient method, which is suitable to treat many samples at one time and for basic molecular experiments, such as restriction endonuclease reaction and PCR.

Background Glaucoma is primarily characterized by the damage of retinal ganglion cells in the inner layer.Whether the outer retinal layers are involved is controversial.Although the functional abnormality of the outer retinal layers have been determined,structural studies present difierent outcomes.Objective This study measured and compared photoreceptor layer thickness between normal and glaucomatous eyes using spectral domain optical coherence tomography(OCT). Methods A case-control study was designed.The macular area of 38 eyes from 38 normal volunteers and 48 primary open-angle glaucomatous eyes(POAG)were imaged by spectral domain OCT(SD OCT).The outer nuclear layer(ONL)and inner and outer segments(IS/OS)layer thicknesses in fovea and parafovea(1.5 mm from the fovea) were measured by a single masked observer using an image analysis software (SigmaScan Pro version 5.0).Retinal nerve fiber layer(RNFL)thicknesses of the 86 eyes were measured by Stratus OCT.The photoreceptor layer thicknesses between normal and glaucomatous eyes were compared.The correlation of photoreceptor with RNFL thicknesses was evaluated with linear and non-linear regression models. Results The outer nuclear layer(ONL)thickness in the fovea in normal and glaucomatous eyes were 96.7±10.7μm and 103.7±13.3 μm,respectively,with a significant difference(P=0.011).The inner and outer segments(IS/OS)layer thicknesses in the fovea in the two groups were 59.3±5.5 μm and 59.5±5.5μm,respectively,without significant difference(P=0.890).The outer nuclear layer(ONL)thicknesses in the parafovea in normal and glaucomatous eyes were 70.9±14.0μm and 68.7 ±10.7μm,respectively,(P=0.410).The inner and outer segments(IS/OS)layer thicknesses in the parafovea in the two groups were 45.2±6.4 μm and 43.6±5.5μm,respectively(P=0.228).The relationship between ONL and RNFL thickness was best described with a second order polynomial regression model assoeiation(Y=-0.019X2+2.73X+10.34,R2=0.211,P=0.005). Conclusion The foveal ONL thickness is increased in glaucomatous eyes.The alteration of foveal ON L thickness is associated with the severity of the disease.

Background Fundus photography is a traditional method for detecting local retinal nerve fiber layer (RNFL) defects,but the evaluation of its result depends on the observer's experience.The spectral domain optical coherence tomography (OCT) exhibit the defects of RNFL very clearly.Objective This study was to evaluate the diagnosis value and correlation between topographic profiles of localized RNFL defects determined by spectral domain and time domain OCT with fundus photography.Methods Forty-one normal eyes of 41 subjects and 55 eyes of 55 glaucomatous patients with localized,wedge-shaped RNFL defects identified by two glaucoma specialists in fundus photographs were enrolled in the clinical study.The angular location and width of RNFL defects determined on the images of fundus photography,Cirrus HD-OCT and Stratus OCT were analyzed respectively using Pearson's correlation coefficient and linear regression analysis.This study followed the Helsinki declaration and was approved by Ethic Committee of Shenzhen Eye Hospital.Written informed consent was obtained from each individual before the clinical examination.Results Seventy-five RNFL defects were identified in 55 eyes by two glaucoma specialists unanimously with the defect position at superior-temporal and inferior-temporal quadrants.The sensitivity of Cirrus HD-OCT and Stratus OCT to determining RNFL defects were 88.0% and 69.3% respectively and their specificities were 92.7% and 97.6% respectively.The angular locations of RNFL defects by Cirrus HD-OCT and Stratus OCT were highly correlated with those by fundus photography(r=0.993,r=0.992,P＜0.001);while the angular widths of RNFL defects by Cirrus HD-OCT and Stratus OCT were moderately correlated with those by fundus photography(r=0.420,r=0.432,P=0.019,P=0.002).No significant differences were found in the defect width of RNFL between Cirrus HD-OCT or Stratus OCT and fundus photography(Cirrus HD-OCT:P=0.114;Stratus OCT:P=0.074),and significant difference was found in that between Cirrus HD-OCT and Stratus OCT(P=0.002).Conclusion Spectral domain OCT and time domain OCT can localize RNFL defects with high sensitivity and specificity.The measure value of Cirrus HD-OCT and Stratus OCT for RNFL defects shows a good diagnostic agreement with fundus photography.

0.05).The duration of pupil contraction in experimental group(54.78?4.52 hours)was longer than that in control group(21.33?2.28 hours)(P

Objective To analyze the physical agent detection ability of occupational health technical service institutions in Methods Guangdong Province. A total of 92 institutions in Guangdong Province that participated in the inter laboratory comparison of physical agents organized in 2020 were taken as the research subjects using purposive sampling method. The ， Results certified physical factor items number of measuring instruments and measurement standards were investigated. The ， ， certification and accreditation project on physical agents in 92 institutions with certification and accreditation rate >80.0% ， ， ， - were noise high temperature ultraviolet radiation hand transmitted vibration and illumination. The certification approval rates ， ， ， ， of power frequency magnetic field 1 Hz to 100 kHz electric field 1 Hz to 100 kHz magnetic field laser radiation physical ， - < （ ） labor intensity and whole body vibration were 60.0%. The certification and accreditation rate of noise 98.9% was the ， （ ） （P< ） ， highest and that of the physical labor intensity 3.3% was the lowest 0.01 . As for the number of measuring instruments （ ） the number of noise measuring instruments accounted for the highest proportion 52.6% in 92 institutions. The number of ， （P< ） measuring instruments from the most to the least was grade A grade B and unqualified institutions 0.01 . The number of - （P< ） measuring instruments in private institutions was greater than that in non private institutions 0.01 . In the measurement ， ， standards some institutions that have certified power frequency electric field power frequency magnetic field and microclimate ， - projects still used the abandoned standard. Some institutions that have certified microclimate illuminance and whole body Conclusion ， vibration used the wrong measurement standards. Among the physical agents the certification and accreditation ， rate of noise is the highest and the number of noise related measuring instruments is the largest. The qualified institutions and private institutions have greater number of certification and accreditation projects on physical agents and measuring instruments.