2.Reconstruction of corneal surface layer using human amniotic epithelial cells modified by lentiviral vector-mediated EGFP gene
Ling, JIN ; Jian, CHEN ; Jing, WU ; Jin-tang, XU ; Qing, ZHOU ; Ru-shan, YE ; Hong, ZHANG
Chinese Journal of Experimental Ophthalmology 2011;29(8):685-689
Background Studies demonstrated that human amniotic epithelial cells (AECs) have some characteristics of embryonic stem cells and they were used to re-establish the surface of eyes. Human AECs may serve as new seed cells in tissue engineering for corneal epithelium reconstitution in the future. Objective The present study was to investigate the application value of human amniotic epithelium cells transfected by lentiviral vectormediated enhanced green fluorescent protein (EGFP) gene as new seed cell source for engineering the corneal surfacelayer. Methods Lentiviral vector carrying the objective gene EGFP was transfected into human amniotic epithelial cells (pLenti6/V5-DEST),and the transient expression of the transgene in the human amniotic epithelial cells was observed under the fluorescence microscope. Flow cytometry was used to detect the positive expression rates of EGFP in transfected cells. The transfected human amniotic epithelial cells were seeded onto the fresh corneal stromal surface of New Zealand white rabbit and cultured in vitro. The stem cell deficiency ( SCD ) models were established by cutting off the limbus of cornea in 20 eyes of New Zealand white rabbits, and the model rabbits were then divided into 2 groups randomly. The transplanted grafts carrying the pLenti6/V5-DEST-EGFP gene-transferred human amniotic epithelium cells were regarded as the pLenti6/V5-DEST-EGFP group, and the corneal stroma graft without any epithelial cell served as the control group. The opacity of stroma and corneal conjunctivalization and vascularization were observed daily. The rabbits' eyes were extracted one month after operation. The expression of EGFP in the cornea was detected under the fluorescence microscope, and the expression of CK8, CK18 and CK12 in cornea was detected by immunohistochemical staining. Results The shape of the transferred human amniotic epithelial cells resembled normal human amniotic epithelial cells. 48 hours after the transient transfection of EGFP presented with the highest expression level throughout the observation duration, with a positive expression rate of EGFP of 61.5% ,showing significant differences in comparison with that of 12 ( 5.24% ) , 24 ( 38.27% ) or 96 ( 39. 10% ) hours ( P <0. 05) post-transfection; but no obvious difference was found in the positive rate of transiently transfected EGFP between 48 hours and 72 hours ( 58.36% ) ( P>0. 05 ). Six cornea grafts were clear in 1 month and two corneas were rejected during the observation period in the pLenti6/V5-DEST-EGFP group. A few new blood vessels were seen around the graft. Ten corneas of the control group became opaque and cloudy with new blood vessels growth around the grafts. Imunohistochemistry revealed the positive expressions of CK8, CK1 8 and CK12 in the corneal epithelial layer in the pLenti6/V5-DEST-EGFP group. However,the expression of CK12 was absent in the control group. Conclusion Human amniotic epithelium cells transfected with the pLenti6/V5-DEST-EGFP gene is a new and ideal feed cell type to reconstruct the corneal surface layer. Lentivirus is a relatively safe gene transfection vector.
3.Scientific practice of Chengjiang Acupuncture School in the Republic of China.
Jian-Bin ZHANG ; Hong-Ru ZHANG ; Xun JIN ; You-Bing XIA
Chinese Acupuncture & Moxibustion 2014;34(2):199-202
Under the influence of Chinese medicine scientization, Chengjiang Acupuncture School, which was originated in the Republic of China (1912 - 1949), has also undergone a series of scientific practice. The Chengjiang School established scientific research, education and medical organizations such as the Research Society of China Acupuncture-Moxibustion, China Acupuncture-Moxibustion School, Training School of China Acupuncture-Moxibustion and Sanatorium of Acupuncture-Moxibustion. Zhenjiu Zazhi (Journal of Acupuncture-Moxibustion) was established, and monographs and textbooks such as Zhongguo Zhenjiu Zhiliaoxue (Therapeutics of Chinese Acupuncture-Moxibustion) and Zhongguo Zhenjiuxue Jiangyi (Lectures on Chinese Acupuncture-Moxibustion) were published. Meanwhile, based on clinical practice, large members of acupuncture personnel with scientific literacy emerged. Therefore, it is held that the scientization of Chengjiang School has performed a profound influence on the development of modern acupuncture-moxibustion.
Acupuncture
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education
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history
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Acupuncture Therapy
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history
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China
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History, 20th Century
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History, 21st Century
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Humans
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Moxibustion
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history
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Schools
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history
4.Study on Effect of Carvedilol on Children with Endocardial Fibroelastosis
rong, LI ; yong-ru, QIAN ; qi-jian, YI ; yong-hong, BAI
Journal of Applied Clinical Pediatrics 2006;0(13):-
Objective To observe the clinical effect and security of carvedilol on children with endocardial fibroelastosis(EFE).Methods Eighteen children with EFE treatment with carvedilol.The improvement of clinical symptom,heart rate,heart function,side effect and maximal tolerance dose after treatment with carvedilol were observed.Results The clinical symptom was obviously improved;eiection fraction(EF),fractional shortening(FS),mean velocity of circumferential fiber(Mvcf)were significantly increased,left ventricular end-systolic dimension(LVDS),left ventricular mass(LVmass),interventricular septal thickness(IVSs)were notably decreased after treatment with carvedilol.Conclusions The data indicates that carvedilol can significantly reduce left ventricular diastolic dimension(LVDD),IVSs,and LVmass,inhibit the remodeling of ventricle,significantly elevate the heart function of EFE.So carvedilol is benefit and security on children with EFE.
5.Melatonin Ameliorates Liver Fibrosis Induced by Carbon Tetrachloride in Rats via Inhibiting TGF-β1/Smad Signaling Pathway
Yu-Rong WANG ; Ru-Tao HONG ; Yuan-Yuan XIE ; Jian-Ming XU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2018;38(2):236-244
Melatonin has been reported to inhibit hepatic fibrosis and the mechanism may be correlated to its anti-oxidant effect.Nevertheless,the mechanism is not completely identified.This study was conducted to investigate the effects of melatonin on TGF-β1/Smad signaling pathway in liver fibrosis in rats.The liver fibrosis model was made by the subcutaneous injection of CCl4.The liver pathology changes were detected using hematoxylin and eosin (H&E) staining and Van Gieson (VG) staining.Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities were measured with an autoanalyzer.Glutathione peroxidase (GPx) activities and levels of malondialdehyde (MDA) and hydroxyproline (Hyp) in liver were evaluated by spectrophotometry.Expression levels of TGF-β1,Smad2/3,phosphorylated Smad2/3 (p-Smad2/3) and Smad7 in liver were detected by immunohistochemistry and Western blot analysis.Results showed that melatonin suppressed CC14-induced liver fibrosis,along with an improvement in histological changes,significant decreases in pathologic grading sores and obvious decreases in Hyp levels in liver.Melatonin improved liver function indicated by decreased serum ALT and AST activities.In addition,melatonin exerted its anti-oxidant effects,as supported by decreased MDA levels and increased GPx activities in liver.Furthermore,melatonin inhibited TGF-β1/Smad pathway,as evidenced by decreased TGF-β1,Smad2/3 and p-Smad2/3 expression and increased Smad7 expression in liver.In conclusion,melatonin may suppress CCl4-induced hepatic fibrosis in rats via inhibiting TGF-β1/Smad pathway.It is possible for melatonin to be a potential reagent to treat and cure liver fibrosis.
6.Inhibitory effects of total flavones of Metasequoia glyptostroboides on experimental cardiac hypertrophy and expression of c-Fos protein in ventricle
Wei-wan LIU ; WEI-WAN LIU ; Xiao-ru YANG ; Ming-jian SHI ; Hong-ying WANG ; Ying AO ; Zhi-ben TU ;
Chinese Traditional and Herbal Drugs 2001;32(4):329-332
To observe the effects of total flavones of Metasequoia glyptostroboides Hu et Cheng (TFM) on volume-overload cardiac hypertrophy and the expression of c-Fos protein in rat. Methods Volume-overload cardiac hypertrophy of rat was induced by aortocaval shunts. The rats were given ig TFM (400, 40 and 4 mg/kg/d). c-Fos protein in the ventricles were measured by immunocytochemical study. Results TFM at the above dosage decreased heart weight and contents of RNA and protein in the myocardium, inhibited the expression of c-Fos protein in the ventricles. Conclusion TFM can prevent volume-overload cardiac hypertrophy in rats. The inhibitory effects on the expression of c-Fos protein may be its mechanism in the molecular level.
7.Influence of nickel sulphate on lens in SD rat
Cai-cai, SHI ; Xing-ru, ZHANG ; Huan-ming, ZHOU ; Qing-song, LI ; Min-hong, XIANG ; Jian-min, TANG ; Long, ZHANG
Chinese Journal of Experimental Ophthalmology 2011;29(2):135-138
Background Researches showed that the incidence rate of cataract is high in the nickel mining area. Nickel sulphate can apparently inhibit the metabolism and proliferation of human lens epithelium cells. But the study on the injury mechanism of nickel on lens is still seldom. Objective Present study was to investigate the effect of nickel sulphate on the lens of SD rats. Methods Forty-five SPF SD rats aged from 7 to 14 days were grouped randomly into subcutaneous injection group, intraperitoneal injection group and blank group. Nickel sulphate of 2 g/L ( 10 mg/kg) was subcutaneously or intraperitonealy injected for 45 days. The opacity of rat lens was examined under the slit lamp at two-week interval and scored based on the criteria of LOCS II and LOCS III. The rats were sacrificed in 45 days after experiment and the lens were obtained for the pathological examination. Result The mean score of the anterior subcapsule opacity of rat lens was obviously higher in subcutaneous injection group compared with blank control group with a significant difference between them (t= 14. 311, P < 0. 05 ) , but no significant difference in the anterior subcapsule opacity between intraperitoneal injection group and blank control group (t = 4. 355 , P>0. 05 ). The score of posterior subcapsule opacity of lens were evidently higher in both subcutaneous injection group and intraperitoneal injection group than the blank control group (t = 9. 316,P = 0. 004;t = 7. 464, P = 0. 009) ,so was the mean score of the anterior +posterior subcapsule opacities(t = 23. 387,P=0. 000;t= 10. 533,P = 0. 002) and the total score of rat lens opacity ( t = 12. 358 , P = 0. 001; t = 10. 188 , P = 0. 003 ) . No significant differences were found in cortex opacity score and nuclear opacity score among three groups ( P > 0.05 ). Histopathology examination revealed that the degeneration of lens collagen protein was more serious in subcutaneous injection group and intraperitoneal injection group than the blank control group,and the injury degree of lens collagen protein was more dominant in subcutaneous injection group. Conclusion System administration of nickel sulphate induced the injury of anterior and posterior subcapsule of lens in SD rat.
8.Influence of mesenchymal stem cells implantation on ventricular remodeling and heart function after acute myocardial infarcion
ri-tai, HUANG ; hong-sheng, ZHU ; song, XUE ; feng, LIAN ; gang, HUANG ; jian-jun, LIU ; ru-qi, TAO
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(04):-
Objective To study the influence of mesenchymal stem cells(MSCs) implantation on ventricular remodeling and heart function after myocardial infarcion. Methods Bone marrow was aspirated from Gui-zhou Xiang swines.After being isolated,cultured and co-cultured with 5-azacytidine,either autologous MSCs(experiment group) or a comparable volume of physiologic saline(control group) were injected into the infarcted myocardium.Three and six weeks later,echocardiographic measurement was performed to assess the myocardial structure and heart function,and single photon emission computed tomography(SPECT) was employed for myocardial imaging.Implanted stem cells were detected by the anti-Brdv antibody DAB with HE staining. Results Left ventricular ejection fraction(EF),fractional shortening and wall thickening were higher in experiment group than control group.The thickness of the ventricular wall and septum was found increased while the left ventricular chamber size was smaller in experiment group.It was indicated by SPECT that three and six weeks after implantation,there was obvious image defect in control group while in experiment group there were some imaging areas in the infarcted area.Brdv-labelled cells were observed in the central part of and around the infarcted area.Conclusion Implantation of MSCs into the infarcted myocardium is believed to attenuate the remodeling process,inhibit the extent of wall thinning and dilatation of the ventricular chamber.MSCs implantation may also improve the contractile ability of the myocardium and heart function.
9.Effect of substrate of edible mushroom on continuously cropping obstacle of Rehmannia glutinosa.
Rui-Hong RU ; Xuan-Zhen LI ; Xiao-Shu HUNAG ; Feng GAO ; Jian-Ming WANG ; Ben-Yin LI ; Zhong-Yi ZHANG
China Journal of Chinese Materia Medica 2014;39(16):3036-3041
The continuous cultivation of Rehmannia glutinosa causes the accumulation of phenolic acids in soil. It is supposed to be the reason of the so called "continuously cropping obstacle". In this study, phenolic acids (hydroxybenzoic acid, vanillic acid, eugenol, vanillin and ferulic acid) were degraded by the extracta of all the tested spent mushroom substrate (SMS) and the maximal degradation rate was 75.3%, contributed by extraction of SMS of Pleurotus eryngii. Pot experiment indicated that hydroxybenzoic acid and vanillin in soil were also degraded effectively by SMS of P. eryngii. The employment of SMS enhanced ecophysiology index to near the normal levels, such as crown width, leaves number, leaf length, leaf width and height. At the same time, the fresh and dry weight and total catalpol concentration of tuberous root weight of R. glutinosa was increased to 2.70, 3.66, 2.25 times by employment of SMS, respectively. The increase of bacteria, fungi and actinomycetes numbers in rhizosphere soil were observed after the employment of SMS by microbial counts. The employment of SMS also enhanced the enzyme activity in soils, such as sucrase, cellulase, phosphalase, urease and catelase. These results indicated that the employment of SMS alleviated the continuously cropping obstacle of R. glutinosa in some extent.
Agaricales
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chemistry
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metabolism
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Agriculture
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methods
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Biodegradation, Environmental
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Hydroxybenzoates
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analysis
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metabolism
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Rehmannia
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growth & development
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metabolism
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Soil
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chemistry
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Soil Microbiology
10.Analysis of environmental fluoride of the coal-burning endemic fluorosis areas in Chongqing
Jing, CHEN ; Bang-zhong, XIAO ; Wei, YAN ; Qian-ru, ZHOU ; jie, ZHANG ; Zheng-hong, WANG ; Jian, ZHAO ; Xiao-ling, GUO ; Xing-jian, LUO
Chinese Journal of Endemiology 2009;28(5):541-544
Objective To find out the distribution pattern of environmental fluoride in the coal-burning endemic fluorosis areas to provide scientific evidence for establishing prevention and remedial measures in Chongqing. Methods According to historical data in Chongqing In 2008, 4 endemic villages(Lingyun and Lizi Villages of Wushan County, and Taiping and Daqing Villages of Pengshui County) and 2 non-endemic villages (ShuangLou and XianLong Villages of Yongchuan County) were investigated. Dental fluorosis of children in 8 to 12 year old and residents over 16 years of age for clinical skeletal fluorosis were examined in four endemic villages. Five households from each of 6 villages were taken, where 500 g of coal, mixed clay with coal, mixed coal of soil, coal cinder, soil were sampled; 15 people were taken in each village, each household gathering 500 g of corn, rice, potatoes, vegetables, grain and vegetable, 100 g of dried pepper and 250 ml of drinking water were sampled from 15 families of each village. Household drinking water samples were collected 1, each 250 ml. For those having tea-drinking habit, each household was collected 50 g of tea and 600 ml of drinking tea, the amount of fluoride were determined. Indoor and outdoor air was collected and measured in 5 households in each village. Results The detected rate of dental fluorosis of children in endemic areas was 74.65% (736/986). The detected rate of skeletal fluorosis of adult was 7.20%(736/986). The average fluoride content of coal, mixed clay with coal, mixed coal of soil, cinder coal, soil in the endemic villages was (310.56±209.46), (360.51±224.96), (293.62±65.15), (186.59±133.66), (497.54±294.70)mg/kg. The average fluoride content in non-endemic villages was (48.68±10.62), (275.66±62.69), (152.20±34.43), (209.14±188.66),269.98±58.21)mg/kg. The fluoride content level of endemic villages was significantly higher than that of non-endemic villages(t=7.67,31.54,5.82, 5.82, all P<0.05). The average fluoride content of drinking water, corn, pepper, flee, potato and vegetable in the endemic villages was (0.30±0.14)mg/L, (1.83±2.67), (23.50±91.80), (0.77±0.25), (0.44±0.11), (0.48±0.18)mg/kg, The average fluoride content in non-endemic village was (0.18±0.06)mg/L, (2.21±0.46), (2.82±2.51), (1.31±0.21), (0.64±0.41), (1.10±0.77)mg/kg. The fluoride content in drinking water and pepper in the endemic villages was significantly higher than that of the non-endemic villages(t=7.79, 2.33, all P<0.05). The fluoride content of rice, potato and vegetable in the non-endemic villages was significantly higher than that of the endemic villages(t=39.29,4.69,4.01, all P<0.05). There was no significant difference of fluoride content of tea and drinking tea between endemic villages[(99.41±55.83)mg/kg, (1.59±0.91)mg/L] and non-endemic villages[(79.95±43.78)mg/kg, (1.80±1.16)mg/L, t=1.01, 0.27, all P>0.05]. The amount of drinking tea in the endemic village[(1.45±0.68)L/d] was higher than that in non-endemic village[(1.00±0.47)L/d, t=4.27, P<0.05]. The average fluoride content of indoor air in the endemic village[(12.77±8.08)μg/m3] was higher than that in non-endemic village [(1.16±1.08)μg/m3, t=9.49, P<0.01]. There was no significant difference of fluoride content of outdoor air between endemic village and non-endemic village[(1.10±1.57), (0.39±0.31)μg/m3, t=2.01, P>0.05)]. Conclusions The fluoride source of coal-burning endemic fluorosis areas are coal and mixed coal of soil in Chongqing. Fluoride enters into human bodies mainly via respiratory, not from food. Although fluoride is rich in pepper, people don't eat it, so reducing the fluoride content in indoor air is the principle measure. Drinking tea may be was one factor of endemic fluorosis, which needs to be further studied.