1.Hydroxycamptothecin-induced apoptosis of human Tenon capsule fibroblast and its mechanism
Xue, YIN ; Yu-xuan, FU ; Zhi-lan, YUAN
Chinese Journal of Experimental Ophthalmology 2013;(3):221-225
Background The hyperplasia of human Tenon capsule fibroblasts (HTFs) is a common cause of filtering surgery failure in glaucomous eye.Researches demonstrated that hydroxycamptothecin is a cell cycle arresting drug and induce apoptosis of cancer and fibroblasts.However,its mechanism is currently less understood.Objective This study was to investigate whether hydroxycamptothecin induce the apoptosis of HTFs and explore the possible mechanism.Methods Human Tenon capsule tissue was obtained from EyeBank of Jiangsu Province Hospital.HTFs were cultured using explant method in vivo and passaged in DMEM containing 10% FBS.The cells were identified using vimentin and keratin by immunochemistry,and the cells of generation 3-6 in the logarithmic growth phase were used in the experiment.The cells were incubated with 0.01,0.05 or 0.10 g/L hydroxycamptothecin for 5 minutes respectively,and the cells without any hydroxycamptothecin were served as the control group.Cell viability then was assessed by cell counting kit-8 (CCK8) for the optimal inhibition concentration.The cells were treated by 0.10 g/L hydroxycamptothecin for 24 hours,and the apoptotic rate of the cells were assayed with annexin V/PI double-staining.Mitochondrial membrane potential of HTFs was assessed using JC-1 staining.The expressions of caspase-3,caspase-9 and cytochrome C (cyt C) in mitochondria and cytoplasm of HTFs were detected by Western blot.Results The proliferative value (A450) of the HTFs 0,0.01,0.05,0.10 g/L was 0.9716±0.0608,0.8035 ± 0.0346,0.7048 ±0.0446,0.6265 ±0.0286,with a significant difference (F =26.372,P =0.002).A450 of HTFs in the 0.01,0.05,0.10 g/L groups was significantly lower than the control group (P<0.05),with the lowest A450 value in the 0.10 g/L group.The apoptotic percentage of HTFs was (18.72±1.41)%,in the 0.10 g/L hydroxycamptothecin group and that of the control group was (3.67 ±0.36)%,showing a significant difference between them (t =-10.374,P=0.001).The expression intensity of caspase-3 and caspase-9 protein in HTFs was higher in the 0.10 g/L hydroxycamptothecin group than that in the control group.JC-1 staining showed that the green fluorescence of the monomer JC-1 in cytoplasm was stronger in the 0.10 g/L hydroxycamptothecin group than that in the control group,but the red fluorescence of the polymer JC-1 in the 0.10 g/L hydroxycamptothecin group was weaker than that in the control group.The grey scale of cyt C protein in HTFs in mitochondrion was 0.0605±0.0022 in the 0.10 g/L hydroxycamptothecin group,showing a significant increase in comparison with 0.0301 ±0.0016 of the control group (t=4.865,P=0.014).However,the grey scale of cyt C protein in cytoplasm was declined in the 0.10 g/L hydroxycamptothecin group than that in the control group (0.0605 ±0.0022 vs.0.0301 ±0.0016) (t =-11.177,P =0.001).Conclusions Hydroxycamptothecin can induce the apoptosis of HTFs through activating the mitochondrial apoptosis pathway.
2.Morphologic Localization and Purified Cultivation of Sinoatrial Nodes Isolated from Newborn Rabbits
qing-zhi, WANG ; guang-lan, MAO ; lei, YU ; xiao-zhong, QIU ; lin, YUAN
Journal of Applied Clinical Pediatrics 1993;0(03):-
Objective To localize the sinoatrial node (SAN) of the newborn rabbits in vivo and cut it for purifying cultivation and study the morophologic characters of primary cultured pacemaker cells of SAN under light microscope and transmissional electron microscope. Methods Hearts of the newborn rabbits were embedded in paraffin for HE-staining and observed the location, form of SAN under optical microscope; SAN cells isolated from neonatal rabbits cultured and purified with the method of differential attachment and BrdU-treatment.Results SAN localized in the anterior wall of the superior vena cava and the posterior-lateral atrial wall.There was about 0.32 mm between its lowest point and sulcus terminalis. Three distinctly different types of cells were observed among the cultured cells of SAN: spindle, araneiform and polygon. The spindle cells covered the greatest proportion of the cultured cells of SAN (59.6%?7.3%). The frequency of spontaneous contraction of spindle cells was the highest among the constrcting cells (145 ?9)time/min. The results of ultrastructure observation showed that myofibrils and other organelles in spindle cells were poorly organized and significantly decreased in number compared with araneiform cells. There was no significant difference between araneiform cells isolated from SAN and from atrial muscle.Conclusion Among the cultured cells from neonatal rabbits SAN, the spindle cells are the pacemaker cells of SAN.
3.Systemic blood pressure,intraocular pressure and primary open-glaucoma: A populationbased study in Shaanxi Province of China
Zhi-Lan, BAI ; Bai-Chao, REN ; Jian-Gang, YANG ; Yuan, HE ; Li, CHEN ; Nai-Xue, SUN
International Eye Science 2005;5(6):1122-1127
AIM :To investigate the association of primary open-glaucoma (POAG), intraocular pressure (IOP) and systemic blood pressure in a rural population aged 50 years old or above in Shaanxi Province, China.METHODS: In the population-based, cross-sectional study, 1 775 (83.53%) residents, aged 50 years old or above, from 3 counties of Shaanxi Province, China, undertook an interview with a standard questionnaire and a detailed eye examination, including logarithm of the minimum angle of resolution visual acuity, slit-lamp biomicroscopy, applanation tonometry and dilated fundus examination. Two blood pressure reading in the sitting position were taken. Gonioscopy was performed if a narrower peripheral anterior chamber (less than one fourth of the corneal thickness) was indicated. Automated visual field testing was performed by participants assessed to have suspicion of glaucomatous disc damage or if IOP was 22mmHg or higher.RESULTS: IOP significantly correlated with systemic blood pressure, and both IOP and systolic blood pressure increased significantly with increasing age. No association between POAG and hypertension was found. The frequency of POAG increased significantly with lower diastolic perfusion pressure.CONCLUSION: Data in our study are accordance with those reported in other population-based studies, and confirm that lower diastolic perfusion pressure is a significant risk factor for primary open angle glaucoma.
4.Epidemiological investigation on age-related macular degeneration in rural area of Shaanxi Province,China
Zhi-Lan, BAI ; Bai-Chao, REN ; Jian-Gang, YANG ; Yuan, HE ; Li, CHEN ; Nai-Xue, SUN
International Eye Science 2005;5(6):1114-1121
AIM: To assess the prevalence and risk factors for age-related macular degeneration (AMD) in a rural population in Shaanxi Province of China.METHODS: A total of 2 835 (81.00%) people aged 40 years old or more, from Fuping county, Jingbian county and Yang county of Shaanxi Province, China, underwent a comprehensive interview and a relative eye examination. The present of AMD was classified into neovascular AMD (NV) and pure geographic atrophy (GA) by using direct ophthalmoscopy for fundus examination according to International Classification System.RESULTS: The prevalence (95% CI) of AMD was 3.00% (2.42, 3.71) in this population, of which NV accounted for 1.45% (1.05, 1.98) and 1.55% (1.14, 2.10)for GA. The prevalence of AMD increased significantly with increasing age (P <0.001). AMD was present in 0.47% of participants aged 40 to 49 years, rising to 11.90% of participants older than 80 years, of which the corresponding data increased from 0.28% to 4.76% for NV and from 0.19% to 7.14% for GA. No significant difference was found in the prevalence of NA and GA between genders in this population. With multiple logistic analyses, apart from advancing age, only smoking was found to have a strong association with any type of AMD.CONCLUSION: The prevalence of AMD in the rural population of Shaanxi Province of China is lower than that reported from other population-based studies in different provinces of China, less than that reported in whites, more than that reported in blacks. Except increasing age, smoking is also a significant well-known risk factor for AMD.
5.Epidemiology of primary angle-closure glaucoma in a rural population in Shaanxi Province of China
Zhi-Lan, BAI ; Bai-Chao, REN ; Jian-Gang, YANG ; Yuan, HE ; Li, CHEN ; Nai-Xue, SUN
International Eye Science 2005;5(5):872-880
· AIM: To assess the prevalence and related risk factors for primary angle-closure glaucoma in a rural population for 40 years of age or older in Shaanxi Province.· METHODS: By using a stratified, cluster-based, random sampling technique, 8 500 persons of all ages were selected randomly, including 3 500 those aged 40 years or more, from the North, the South and the Middle of Shaanxi Province in Western China from July to December in 2003. All participants had an interview with a standard questionnaire and those questions related to glaucoma of previous diagnosis and treatment, family history and outbreak history. Then a detailed and relative eye examination was performed, including logarithm of the minimum angle of resolution visual acuity, external eye examination with slit-lamp biomicroscopy and fundus examination. The intraocular pressure was measured with Perkins applanation tonometry to those persons aged 50 years old or more and those with suspected increased IOP. A further examination was performed to those persons with suspicious glaucoma, including repeated tonometric examination, gonioscopy, dark room test, automated visual field testing, et al.·RESULTS: 6 815 of the eligible 8 500 persons of all ages were interviewed and examined from July to December in 2003, a response rate of 80.18%, including 2835 of the eligible 3 500 persons of 40 years old or more with a response rate of 81.00%. According to Van Herick method, the percentage of shallow peripheral anterior chamber was 13.6%, of which 10.4% was in grade 2, 3.0% in grade 1, and 0.2% in grade 0. In this rural population for 40 years of age or older 31 participants were found to have primary angle-closure glaucoma, with the prevalence of 1.09%. With multiple logistic analyses, the prevalence increased significantly with age (P=0.008), whereas no significant difference (Pgender=0.180, Peducation=0.199) was found in the rate with gender and education, although women and illiteracy seemed to be at increased risk of the disease (OR: 1.77, 95%CI,0.77-4.10; OR: 1.71, 95%CT, 0.76-3.87). Of 31 participants, 21 persons (67.74%) had not been previously diagnosed and treated. Of 10 participants previously diagnosed, 6 (60%) had been received peripheral iridectomy; nevertheless, no any other treatments were performed. 48.39% participants suffered from visual impairment in various degrees, of which the percentage of blindness in either eye accounted for 29.03%. Of those with primary angle-closure glaucoma, 22 persons (70.97%) were classified as having chronic from of the disease.· CONCLUSION: In this rural population in Shaanxi Province of China, the rate of occludable angles is 13.6%, more common than that reported previously in other Chinese populations. The prevalence of primary angle-closure glaucoma is close to that from other Asian populations, increasing with age. A majority of glaucoma was undiagnosed and untreated previously.
6.Epidemiology of primary open angle glaucoma in a rural population in Shaanxi Province of China
Zhi-Lan, BAI ; Bai-Chao, REN ; Yuan, HE ; Jian-Gang, YANG ; Li, CHEN ; Nai-Xue, SUN
International Eye Science 2005;5(5):864-871
· AIM: To assess the prevalence and related risk factors of primary open angle glaucoma (POAG) in a rural population in Shaanxi Province.· METHODS: By using a stratified, duster-based, random sampling technique, 8 500 persons of all ages were selected randomly, from the North, the South and the Middle of Shaanxi Province from July to December in 2003. All participants had an interview with a standard questionnaire and those questions related to glaucoma of previous diagnosis and treatment, family history and outbreak history. Then a detailed and relative eye examination was performed, including logarithm of the minimum angle of resolution visual acuity, external eye examination with slit-lamp biomicroscopy and fundus examination. The intraocular pressure was measured with Perkins applanation tonometry to those persons aged 50years old or more and those with suspected increased IOP. A further examination was performed to those persons with suspicious glaucoma, including repeated tonometric examination, gonioscopy, dark room test, automated visual field testing, et al.· RESULTS: 6 815 of the eligible 8 500 persons of all ages were interviewed and examined from July to December in 2003, a response rate of 80.18%. 9 participants were found to have POAG, with the prevalence of 0.13%, age ranging from 38 to 80 years (mean year:62.0). In those aged 30 years old or more, the prevalence of the disease was 0.23%, 0.28% and 0.39% for those more than 40 and 50 years old, respectively. The rate of suspicious POAG was 0.18% (12 cases) with age ranging from 35 to 77 years (mean year: 54.7). With multiple logistic analyses, the prevalence increased significantly with age (P=0.023). Other than increasing age, myopia was also a strong risk factor for POAG. Of 9participants with POAG, only 2 cases (22.22%) had been previously diagnosed. No one with POAG was received any treatment previously. 66.67% (6 cases) participants with POAG suffered from visual impairment in various degrees secondary to POAG. The percentage of blindness in either eye was 33.33%.· CONCLUSION: The prevalence of POAG is close to that from other Chinese populations, increasing with age. A majority of glaucoma was undiagnosed and untreated previously.
7.Spectrum-effect relationship between HPLC fingerprints and bioactive components of Radix Hedysari on increasing the peak bone mass of rat
Xin-Yue CHEN ; San-Hu GOU ; Zhi-Qiang SHI ; Zhi-Yuan XUE ; Shi-Lan FENG
Journal of Pharmaceutical Analysis 2019;9(4):266-273
The traditional Chinese medicine of Radix Hedysari plays an important role in invigorating gas for as-cending, benefiting blood for promoting production of fluid, and promoting circulation for removing obstruction in collaterals, which is consistent with the principle of treatment for osteoporosis. This study is designed to investigate the bioactive components on increasing peak bone mass (PBM) by exploring the spectrum-effect relationship between chromatography fingerprints and effect. Multiple indicators are selected to evaluate the pharmacological activity. In fingerprints, 21 common peaks are obtained, five of which are identified. Furthermore, gray relational analysis (GRA) is a quantitative method of gray system theory and is used to describe the correlation degree of common peaks and pharmacological activities with relational value. 21 components are then divided into three different regions, of which ononin and calycosin play an extremely significant role in increasing PBM. In addition, factor analysis and hierarchical cluster analysis (HCA) are used to screen the optimal producing area for Radix Hedysari. This provides a comprehensive and efficient method to improve the quality evaluation of Radix Hedysari, confirming the bioactive components for PBM-enhancement and further develop its medicinal value.
8.Comparison of peg-interferon monotherapy to peg-interferon and nucleoside analogue combination therapy for hepatitis B: a meta-analysis of randomized controlled trials.
Mao-ying LI ; Xue-lan YUAN ; Da-zhi ZHANG
Chinese Journal of Hepatology 2012;20(6):442-447
To evaluate the efficacy and safety of pegylated-interferon (Peg-IFN) treatment as monotherapy or in combination with nucleoside analogues (NAs) for treating chronic hepatis B (CHB) infection.Searches of PubMed, OVID, EMBASE, and the Chinese Medical (WanFang, CNKI, and VIP) databases were conducted to identify all relevant randomized controlled trials published since January 1990. Twelve studies comparing Peg-IFN monotherapy to NA combination therapy (lamivudine (LAM), n =8); adefovir (ADV), n = 4) met the inclusion criteria (treatment duration, range: 48-52 weeks; follow-up, range: 24 weeks to three years). Meta-analysis was performed with RevMan 5.0 using the fixed-effects and random-effects models. Patients who had received combination therapy had a higher biochemical response rate at the end of treatment than those who had received monotherapy (51.1% vs. 38.9%, odds ratio (OR) = 1.63, 95% confidence interval (CI): 1.33-2.01, P less than 0.01). Subgroup analysis of Peg-IFN combination therapies with LAM or ADV indicated that neither NA type significantly enhanced the increased efficacy of combination therapy compared to monotherapy. The combination therapy subgroups also had higher virologic response rates at the end of treatment than the monotherapy subgroups (LAM: 65.9% vs. 34.9%, OR = 3.57, 95% CI: 1.83-6.95, P less than 0.01; ADV: 74.6% vs. 46.2%, OR = 3.66, 95% CI: 2.13-6.30, P less than 0.01). Moreover, the combination therapy group had a higher sustained biochemical response rate at the end of follow-up than the monotherapy group (47.6% vs. 42.1%, OR = 1.28, 95% CI: 1.05-1.55, P less than 0.05). The LAM combination therapy subgroup had a significantly higher biochemical response rate than the monotherapy subgroup, but there was no significant difference between the LAM and ADV combination therapy subgroups. At the end of follow-up, the ADV combination therapy subgroup had a significantly lower rate of hepatitis B e antigen (HBeAg) than the monotherapy subgroup, but there was no significant difference between the ADV and LAM combination therapy subgroups for HbeAg reduction. The combination therapy group and monotherapy group showed no statistically significant differences in HBsAg reduction or occurrence of severe adverse events. Peg-IFN/NA combination therapy produces a higher biochemical response rate in CHB patients than PEG-IFN monotherapy. Moreover, Peg-IFN/ADV combination therapy produces a greater reduction in HBeAg than Peg-IFN monotherapy.
Adenine
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administration & dosage
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analogs & derivatives
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Antiviral Agents
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administration & dosage
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adverse effects
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therapeutic use
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Drug Therapy, Combination
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Hepatitis B, Chronic
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drug therapy
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Humans
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Interferons
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administration & dosage
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adverse effects
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therapeutic use
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Lamivudine
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administration & dosage
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Nucleosides
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administration & dosage
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adverse effects
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therapeutic use
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Organophosphonates
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administration & dosage
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Polyethylene Glycols
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administration & dosage
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adverse effects
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therapeutic use
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Randomized Controlled Trials as Topic
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Treatment Outcome
9.Activity Study of Ciliary Neurotrophic Factor(CNTF)Mutant
Hua BI ; Li-Yong YUAN ; Xin-Chang SHI ; Yang ZHAO ; Lan LIU ; Chun-Ming RAO ; Jun-Zhi WANG ;
China Biotechnology 2006;0(01):-
To study still further the activity of CNTF mutant designed by computer molecular modling,the methods of dissociated cultures of chick dorsal root ganglion、TF-1 prolification and the normal mice'weight loss tests weve used.The results indicated that the mutant protein promoted the survival of dorsal root ganglion、induced TF-1 prolification and made the normal mice lose weight,decrease appetite and reduce fat index.The weight loss effect was dependant with its administration dosage,ED50 was 150.986?g/kg/d.To TF-1,the specific activity reached 2.0?106U/mg against international reference reagent.In a word,CNTF mutant had excel bioactivity.So it provided clues for its development and application.
10.Identification of moutan cortex and its adulterants by ITS2 sequence.
Meng WEI ; Lan WU ; Yuan TU ; Wei-Chao REN ; Li XIANG ; Wei SUN ; Lin-Bi ZHANG ; Zhi-Gang HU
China Journal of Chinese Materia Medica 2014;39(12):2180-2183
To explore a new method to identify Moutan Cortex to guarantee its safe use, internal transcribed spacer 2 (ITS2) sequence was used to identify Moutan Cortex and its adulterants. DNA was extracted and target fragments were amplified. Sequences were analyzed and assembled by CodonCode Aligner V3.7.1. Genetic distances were computed and phylogenetic tree was constructed based on kimura 2-parameter (K2P) model by MEGA 5.0. The length of the 20 ITS2 sequences of Moutan Cortex from nine different places is 227 bp, and no variation site was detected. The maximum inter-specificK2P distance of Moutan Cortex is 0, the minimum intra-specific K2P distance is 0.041, the average intra-specific K2P distance is 0.222. According to NJ analysis, Moutan Cortex from different places can get together as one branch with bootstrap support values 99%, which indicates Moutan Cortex can be easily distinguished from its adulterants. Using ITS2 sequence can accurately identify Moutan Cortex and its adulterants, it is an effective supplementary to traditional identification methods.
Base Sequence
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China
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DNA Barcoding, Taxonomic
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methods
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DNA, Plant
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genetics
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DNA, Ribosomal Spacer
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genetics
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Drug Contamination
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prevention & control
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Drugs, Chinese Herbal
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chemistry
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classification
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Molecular Sequence Data
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Paeonia
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classification
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genetics
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Phylogeny
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Quality Control