2.Effect of LY294002 on protein kinase B activation in human lens epithelial cell
Rui, GUO ; Hai-ke, GUO ; Qian-li, MENG
Chinese Journal of Experimental Ophthalmology 2013;32(11):1026-1030
Background Protein kinase B (Akt) is the center of multiple cellular signaling pathways,and it participates in the regulation of cell function,such as proliferation,migration,and metabolism of cells.Phosphoinositide 3-kinase (PI3K) promotes activation of Akt and therefore triggers many signal pathways.PI3K inhibitor can silent Akt,but whether it can affect the biological behavior of lens epithelial cells (LECs) during the posterior capsular opacity (PCO) is worthy of investigation.Objective This study was to explore the effect of LY294002,a PI3K inhibitor,on Akt activation in human LECs.Methods Human LECs strain,HLEC-B3 cells,were cultured and passaged.The cells were incubated to 96-well plate for 24 hours,then LY294002 was added with the final concentration at 0,10,20,30,40,50,60,70,80 μmol/L,respectively.After incubation for 24 hours,cell counting kit-8 (CCK-8) was used to detect the inhibitory rate of cell proliferation.Transforming growth factor-β2 (TGF-β2) of 10 μg/L was added in the medium of cells as TGF-β2 group,TGF-β2 + LY294002 (20 μmol/L) was used to coculture the other group of cells,and the cells without TGF-β2 and LY294002 served as the control group.Phosphorylation of Akt (p-Akt) in the cells was detected by laser scan confocal immunofluorescence microscope.The expression level of p-Akt was evaluated using Western blot.Results CCK-8 assay showed that the A value of the cells was gradually reduced with the increase of LY294002 concentration (Fgroup =9.72,P =0.00),but the A value was significant raised along with the lapse of time (Ftime =1737.54,P=0.00).Confocal immunofluorescence revealed that a little of p-Akt was expressed in the control cells.After induced by TGF-β2,lots of red fluorescence of p-Akt was seen in cells around the cell membranes.But in the TGF-β2+LY294002 co-culture cells,the fluorescence of Akt was much weaker.Western blot showed that the expression level of p-Akt was 0.91±0.08,1.48±0.13 and 0.95±0.19 in the control group,TGF-β2 group and TGF-β2 +LY294002 group,respectively,with a significant difference among the three groups (F =15.04,P =0.00).Conclusions LY294002 can inhibit the activation of Akt induced by TGF-β2.LY294002 may have utility in the prevention and treatment of PCO.
3.A study on the percutaneous penetration of artesunate from various reservoir vehicles.
Li-ling ZHOU ; Hai-xia HOU ; Rui LI
China Journal of Chinese Materia Medica 2002;27(9):662-665
OBJECTIVETo determine the main factors which affect the percutaneous penetration of artesunate and provide efficient data for the artesunate transdermal delivery system.
METHODTransdermal speed constant and accumulative amount of 12 hours were used for the estimations of various reservior vehicles, and the supplement orthodox design was used to study the effect of pH, various proportion of IPA/Water/IPM, and drug concentration.
RESULTDrug concentration and pH were the main factors which affected the percutaneous penetration of artesunate.
CONCLUSIONThe suitable reservior vehicle can prompt the percutaneous penetration of artesunate, and artesunate TTS will be made with further studies.
2-Propanol ; pharmacology ; Administration, Cutaneous ; Animals ; Antimalarials ; administration & dosage ; pharmacokinetics ; Artemisinins ; administration & dosage ; pharmacokinetics ; Drug Carriers ; Drug Delivery Systems ; Hydrogen-Ion Concentration ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Sesquiterpenes ; administration & dosage ; pharmacokinetics ; Skin Absorption ; drug effects
4.The Condition for Sporulation of Helminthosporium carposaprum
Ning-Hai LU ; Rui-Fu XU ; Li-Min WU ; Gang-Feng SHAO ; Gui-Yuan LI ;
Microbiology 1992;0(05):-
Caused by Helminthosporium carposaprum, tomato brown lea f spot was a serious disease in green house in Henan Province. The condition for promoting sporulation of fungi were tested in this paper. The results showed th at the number of sporulation were different on the different medium,the fungi c ould sporulate a lot on the PDA+tomato leaf and Czapek medium, but V8、PSA and t omato juice restrained sporulation.The best carbon source and nitrogen source f or the fungi promoting sporulation were fructose and ammonium chloride respectiv ely,mannitol and Peptone ammonium sulfate restrained sporulation. Light and ult raviolet radiation were in favor of sporulation , ultraviolet radiation irradiat ing for 60~80min promoted sporulation. The fungi were promoted sporulation on the condition of lower or higher temperature and alkalescence,which 15℃o r 30℃,pH 8~9.
5.Association of Increased Urine Brain Derived Neurotrophic Factor with Lower Urinary Tract Symptoms in Men with Benign Prostatic Hyperplasia
WANG LONG-WANG ; LI JIAN-LONG ; YU YI ; XIAO RUI-HAI ; HUANG HONG-WEI ; KUANG REN-RUI ; HAI BO
Journal of Huazhong University of Science and Technology (Medical Sciences) 2017;37(4):531-535
Urinary brain-derived neurotrophic factor (BDNF),an ubiquitous neurotrophin,was found to rise in patients with benign prostatic hyperplasia (BPH).We hypothesized that the urinary level of BDNF could be a potential biomarker for lower urinary tract symptoms (LUTS) in patients with BPH.Totally,76 patients with BPH-caused LUTS and 32 male control subjects without BPH were enrolled.International Prostate Symptom Score (IPSS) was applied to assess the symptom severity of LUTS.Urodynamic tests were performed for the diagnosis of underlying detrusor overactivity (DO) in the patients with BPH.Urine samples were collected from all subjects.Urinary BDNF levels were measured using enzyme-linked immunosorbent assays and normalized by urinary creatinine (Cr) levels.Seventy-six BPH patients were divided into moderate LUTS group (n=51,7<IPSS ≤ 20) and severe LUTS group (n=25,IPSS>20) according to the IPSS.Of the 76 BPH patients,DO was present in 34 (44.7%)according to the urodynamic test.The urinary BDNF/Cr levels were significantly higher in BPH patients with moderate LUTS (8.29±3.635,P<0.0001) and severe LUTS (11.8±6.44,P<0.0001) than normal controls (1.71±0.555).Patients with severe LUTS tended to have higher urinary BDNF/Cr levels than patients with moderate LUTS (11.8±6.44 vs.8.29±3.635,P=0.000).The conditions of BPH with LUTS correlated with elevated urinary BDNF levels,and urinary BDNF levels were even higher in BPH-DO patients.The results of this study have provided evidence to suggest that urinary BDNF level test could evaluate the severity of LUTS in BPH patients,and BDNF level can be used as a biornarker for the diagnosis of DO in BPH patients.
6.Application of various preoperative imaging examinations for peferator in anterolateral thigh flap
Yongjun RUI ; Yan ZHANG ; Hong YANG ; Hai HUANG ; Li QIANG ; Gang ZHAO ; Tong YANG
Chinese Journal of Microsurgery 2015;38(1):33-37
Objective To assess the diagnostic value of various preoperative imaging examinations for the point of the lateral femoral circumflex artery perforators through the deep fascia,in order to provide reasonable evidence for the design of location and protocol of anterolateral thigh perforator flap.Mehtods From September,2013 to December,2013,23 patients (including 1 patient with bilateral anterolateral thigh perforator flap) preparing anterolateral flap surgery were randomly divided into 4 groups(every group was 6 side).The patients in different groups were using different imaging techniques respectively,such as handheld doppler (HHD),Color doppler sonography (CDS),MDCT angiography (MDCTA),CDS and MDCTA united image location technology.The flap sizes varied from 8 cm × 5 cm to 28 cm × 12 cm.Moreover the coincidence rate between preoperative and intraoperative location of the perforator vessel was evaluated.Results Ali flaps were survived after the operation.The donor site were primarily closed while partial skin grafting was performed in 8 cases.The appearance and functional resuhs were satisfactory with following up for 3 to 9 months.The rates of coincidence between the preoperative and intraoperative location of perforating branches were HHD 61.53%,CDS 87.50%,MDCTA 85.71%,CDS and MDCTA united image location technology 93.33% respectively.The HHD group showed significant decrease compared with the other groups (x2 =7.92,P < 0.01; x2 =6.15,P < 0.05; x2 =12.54,P < 0.01,respectively).The CDS group had no statistically difference with the MDCTA group (P > 0.05).The united CDS and MDCTA group showed significant increase compared with the other groups (x2 =4.22,P < 0.05; x2 =3.94,P < 0.05).Conclusion CDS and MDCTA united image location technology are more accurate for perforator preoperative location in auterolateral thigh flap,and should be widely used.
7.Research status of photodynamic therapy combined anti - VEGF drugs to treat exudative age-related macular degeneration
Ling, LI ; Hai-Hui, QI ; Xue-Ying, MA ; Rong, ZHANG ; Rui-Juan, GUAN
International Eye Science 2015;(2):247-250
Photodynamic therapy ( PDT ) is a new technique to diagnose and treat diseases with photodynamic effect produced by photosensitizer and light, and is now a main method of treating exudative age - related macular degeneration ( AMD ) . ln recent years, with the development of science and technology, combinations of PDT have become a research hot spot. ln this paper, we reviewed the research status of treatments on exudative AMD with PDT combined anti-VEGF drugs.
8.Diagnostic Value of Computed Radiography on Neonatal Respiratory Distress Syndrome
ji-cheng, DU ; hai-bin, ZHOU ; fu-chun, LI ; rui-zhen, HONG ; man-hua, BAO
Journal of Applied Clinical Pediatrics 1986;0(02):-
Objective To improve the knowledge and diagnostic ability of imagiology on neonatal respiratory distress syndrome (NRDS) computed radiograph(CR).Methods The doubtful patients were done to photographs bedside using the high resolution imaging plate, 50 cases of newborn with NRDS were selected whose clinical diagnosed clearly and had been treated and had the complete CR image documents.The CR change and clinical characteristics were observed dynamically.Results Nine of 50 cases were combined with aspirated pneumonia,8 cases with infective pneumonia,3 cases with intra-alveolar hemorrage,and 2 cases with pneumothorax.Accoding to X-ray manifestations,all cases were divided into four stages:Ⅰ stage(n=5), Ⅱ stage(n=20),Ⅲ stage(n=22),Ⅳstage(n=3).Typical CR signs included:the pulmonary lucency decreasd,wide-ranging net and grain shadowes of high density, and in companing with a lot of air brunchus sing.Conclusions Computed radiography is the most important imaging method in diagnosis of NRDS bedside ,and shall be improved the ability of diagnosis and differential of NRDS combined with the clinic.
9.In vitro screening and in vivo identification of rat IκBα-siRNA
Rui, ZENG ; Yu-Qing, LAN ; Hai-Jun, GONG ; Chi, ZHANG ; Jin-Miao, LI
International Eye Science 2014;(6):986-991
AIM: To seek a small interfering RNA ( siRNA ) sequence targeting rat inhibitor of nuclear factor kappa Bα ( IκBα) that can specifically and effectively suppress IκBα mRNA expression of rat ciliary muscles in vivo.METHODS:Three IκBα specific double stranded siRNAs were designed and synthesized. They were transfected into rat A7r5 cells which express IκBα gene. Flow cytometry was used to assess transfected efficiency. The mRNA and protein levels of IκBα were examined by Real Time quantitative polymerase chain reaction ( Real Time-PCR ) and western blot to screen a candidate valid sequence with the highest inhibitory rate. The Cy3 labeled non-specific control siRNA or the candidate valid siRNA was then injected into rat anterior chamber. Distribution of Cy3- siRNA in rat ciliary muscles was viewed by fluorescence microscopy, and the inhibitory effect in vivo of the valid siRNA was identified via Real Time-PCR and immunofluorescence. RESULTS: The suppression effect of the siRNA targeting the CTACGATGACTGTGTGTTT of IκBα gene was most obvious by vitro screening. By anterior chamber injection, this valid siRNA could reach rat ciliary muscles and effectively suppress IκBα gene expression with the highest inhibitory rate of 59. 0% on mRNA level at 24h after RNAi, and 52. 3% on protein level at 72h after RNAi (P<0. 01).CONCLUSION: It proves that the siRNA targeting the CTACGATGACTGTGTGTTT of IκBα gene is the valid sequence to suppress rat IκBα expression of ciliary muscles by RNAi in vivo.
10.Effects of low-iodine diet on the expression of homeobox gene NKX-6.1 and NKX-6.2 in rat cerebrum tissue
Rui, ZHANG ; Hai-ze, GE ; Xiu-juan, ZHAO ; Yuan, LI ; Gang, GUO
Chinese Journal of Endemiology 2009;28(5):488-492
Objective To study the influence of low-iodine diet on the expression of homeobox gene Nkx-6.1 and Nkx-6.2 in rat cerebrum tissue, and to explore the possible molecular mechanism of cerebrum development retardation caused by low-iodine. Methods Twenty female Wistar rats were randomly equally divided into two groups: low-iodine group and control group, both fed with low-iodine diet as low as 13.66 μg/kg determinated by spectrophotometry in Tianjin Institute of Endocrinology and the former with deionized water, the later 200 μg/L potassium iodate. Thyroid hormone level was detected using chemiluminescence immunoassay 3 months later and they were mated with male rats normally fed. Rats of 16-day pregnancy, new-born and 20th days old were detected the content of Nkx-6.1 and Nkx-6.2 mRNA in the cerebrum tissue by real-time fluorescence quantitative PCR 0.61), (3.28±0.80)pmol/L] were lower than the control group[(1.04±0.06), (39.42±14.68)nmol/L, (4.83±0.33), day pregnancy, new-born and 20th days old of control group was (1.90±0.23)×10-3,(1.86±0.40)×10-4, (1.11± 0.27)×10-4(F=827.58, P<0.01), Nkx-6.1 mRNA expression level gradually decreased along with aging(all P<0.05). The intra-group difference was significant (F=297.25, P<0.01) and the Nkxr.1 mRNA expression level during 16 days of pregnancy was the highest(P<0.01). It was higher in the control group than in the low-iodine group during 16 days of pregnancy (t=10.14, P<0.01) as well as in the low-iodine group than in the in 16-day pregnancy, new-born and 20th days old of control group was respectively(1.03±0.19)×10-2, (1.33± 0.10)×10-3, (8.79±0,87)×10-3, and that of low-iodine group was (0.31±0.03)×10-2, (1.53±0.13)×10-3, (7.51±0.86)×10-2. The intra-group difference was significant(F=1293.02,1065.83, all P<0.01). Nkx-6.2 expression level during 20th days old was the highest(P<0.01) and that of newborn was the lowest(P<0.01). The Nkx6.2 mRNA expression level in control group were higher than the low-iodine group during 16-day pregnancy and 20th days old(t=14.35, 4.05, all P<0.01). It was higher in the low-iodine group than in the control group during newboru(t=4.78, P<0.01). Conclusions The difference in the expression of Nkx-6.1 and Nkx-62 is highly related to the brain development retardation caused by low-iodine.