1.Dynamic changes in ultrastructural morphology and serum levels of hyaluronic acid, laminin and collagen in rats with dimethylnitrosamine-induced liver fibrosis
Chunhui LI ; Lihui PAN ; Dongming PIAO ; Zhengri YIN ; Wenxie XU
Chinese Journal of Tissue Engineering Research 2007;11(4):780-784,封4
BACKGROUND: Development of liver fibrosis accompanies many morphological and functional changes. The pathological alterations of dimethylnitrosamine (DMN)-induced liver fibrosis in rats are similar to those of human liver fibrosis.OBJECTIVE: To observe the dynamic changes in morphology and serum hyaluronic acid (HA), laminin (LN), and type Ⅳ collagen in rats with liver fibrosis induced by DMN.DESIGN: Randomized controlled animal trial.SETTING: Laboratory of Teaching and Research Section of Pathology, College of Medicine, Yanbian University.MATERIALS: Eighty 3-month-old male rats of clean grade with 175-200 g body mass were selected, which were provided by the animal center of College of Medicine, Yanbian University. Agents: Dimethylnitrosamine provided by Sigma company, α smooth muscle actin by Dako company, Sirius red by Aldrich chem company, serum hyaluronic acid,laminin and type Ⅳ collagen kit by Sino-American Biotechnology Company, rabbit-anti-rat I g by Dako, Denmak company. Devices: JEM-1200EX transmission electron microscope made in Japan; enzyme linked immuno analyzer made in Japan; and CMTAS multifunction true color pathological image analysis system developed by Beijing University of Aeronautics and Astronautics.METHODS: The experiment was conducted in the College of Medicine, Yanbian University from June 2004 to December 2005. The rats were divided into 2 groups by lot: model group (n =40): The rats were intraperioneally injected with 10 g/L DMN (10 μL/kg) once daily, 3 days a week for 4 weeks; control group (n =40): The matching normal saline was injected at the same period; the blood from the left ventricle was collected and frozen in refrigerator at -70 ℃ before the rats were killed at days 7, 14, 21, and 28 and the liver tissue was removed for electron and light microscope observation. ①The dynamic changes in the content of serum HA, LN and type Ⅳ collagen were measured by enzyme-linked immunoabsorbent assay (ELISA). ②The morphological changes and liver fibrosis grading were examined by HE staining,immunohistochemical Sirius-red staining (liver fibrosis degree was classified into 5 grades: grade 0: no fibrosis; grade 1:fibrosis in portal area; grade 2: fibrotic septa between portal tracts; grade 3: fibrosis septa and structure disturbance of hepatic lobule; grade 4: cirrhosis), meanwhile, the area-density percentage of collagen fibrosis was calculated. ③The hepatic stellate cells were detected with transmission electron microscope and immunohistochemical alpha smooth muscle actin (SMA) staining. ④The correlation between area-density percentage of collagen fibrosis during liver fibrosis formation and the serum levels of HA, LN and type Ⅳ collagen was analyzed.MAIN OUTCOME MEASURES: ①The changes in the serum levels of HA, type Ⅳ collagen and LN during liver fibrosis formation; ②The morphological changes and liver fibrosis grading and area-density percentage of collagen fibrosis; ③Transformation and distribution characteristics of hepatic stellate cells; ④The correlation between area-density percentage of collagen fibrosis and the serum levels of HA, LN and type Ⅳ collagen.RESULTS: Among the 80 rats, 34 of the experimental group were modeled successfully, which were involved in the result analysis with the 40 rats in the control group. ①The levels of serum HA, type Ⅳ collagen and LN of the model group were significantly higher compared with the control group from day 7 to 28 (P < 0.05), especially that on the 28th day. ②In the model group, the portal area of the rats showed hemorrhagic necrosis at day 7 after injection of DMN; at day 14,hemorrhage, necrosis and thin fibrotic septa joining central areas of liver were found; at days 21 and 28, thick septa was found; The area-density percentage of collagen fibrosis of the model group was significantly higher compared with the control group at days 7, 14, 21 and 28 (P < 0.05), especially that on the 28th day. There were significant differences in the liver pathologic grading between the two groups at each time point (P < 0.01); the pathologic grading of the model group at day 7 differed from those at days 14 and 28 (P < 0.01). ③The α-SMA positive cells and a transitional hepatic stellate cell were found under the electron microscopy; typical myofibroblast was observed in the model group at day 21 and 28 under the electron microscopy. ④The area-density percentage of collagen fibrosis was positively correlated with the content of serum HA, LN and type Ⅳ collagen (r=0.707, 0.675, 0.662, P< 0.01).CONCLUSTON: There are significantly progressional changes in morphological and serum levels of HA, type Ⅳ collagen and LN in different stages of DMN-induced liver fibrosis in rats, moreover, the area-density percentage of collagen fibrosis is positively correlated with the serum levels of HA, LN and type Ⅳ collagen at different stages.
2.Effect of phacoemulsification with different corneal micro-incision sizes on postoperative corneal healing
Ning REN ; Lixia SUN ; Hong CUI ; Zhengri LI ; Chenglin LI ; Haiyan JIN ; Yingjun LI
Chinese Journal of Experimental Ophthalmology 2021;39(4):319-326
Objective:To investigate the effect of phacoemulsification using different corneal micro-incision sizes on the postoperative corneal healing.Methods:A non-random controlled study was performed.Seventy-six patients (76 eyes) with age-related cataract who underwent phacoemulsification and cataract extraction combined with intraocular lens (IOL) implantation in Yanbian University Hospital from May 2016 to May 2017 were enrolled.The subjects were divided into 2.2 mm incision group (37 eyes) and 1.8 mm incision group (39 eyes) according to corneal incision size.The intraoperative cumulative dissipated energy (CDE) and ultrasound time of the two groups were measured and compared.The corneal incision structure and corneal thickness of operative eyes were measured by anterior segment optical coherence tomography before surgery and at postoperative 1 day, 1 week and 1 month.The corneal endothelial cells count, the central corneal thickness (CCT), and the corneal volume in the central corneal diameters of 3.0 mm (CV3) and 10.0 mm (CV10) were measured by Pentacam.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of the Yanbian University Hospital (No.2015153), and written informed consent was obtained from all subjects before surgery.Results:There were statistically significant differences in the corneal endothelial cells count, CCT, CV3, CV10, and corneal thickness at epithelial side of the incision between pre-operation and post-operation in each group ( Ftime=17.717, 67.356, 17.577, 13.559, 80.076; all at P<0.01).But there was no statistically significant difference in the indexes between the two groups ( Fgroup=0.788, 0.706, 3.692, 4.341, 4.182; all at P>0.05).The number of corneal endothelial cells in the two groups was gradually decreased after surgery, and was significantly reduced at one month after surgery in comparison with the pre-operation, and the differences were statistically significant (both at P<0.05).At 1 day after surgery, the CCT, CV3, CV10, and corneal thickness at epithelial side of the incision in the two groups were increased significantly in comparison with the pre-operation and the differences were statistically significant (all at P<0.05), and the differences were not statistically significant between preoperative and postoperative 1 week or 1 month (all at P>0.05).With time going by, the indexes were back to normal gradually.The incidence of endothelial gaping and dislocation at the corneal incision in the 1.8 mm incision group was 12.8% (5/39) and 5.1% (2/39), which were significantly higher than 0.0% (0/37) and 2.7% (1/37) in the 2.2 mm incision group, and the differences were statistically significant ( χ2=5.078, P=0.024; χ2=0.295, P=0.590).At 1 day after surgery, corneal thickness at the endothelial side in the 1.8 mm incision group was significantly thicker than that in the 2.2 mm incision group, and the difference was statistically significant ( P=0.042), and the corneal thickness at endothelial side of the incision was positively correlated with CDE in the two groups ( r=0.231, P=0.025; r=0.347, P=0.003). Conclusions:Compared with 2.2 mm incision, 1.8 mm corneal incision results in higher incidence of endothelial gaping and dislocation at the corneal incision, greater corneal thickness at endothelial side of the incision and slower recovery.
3.Application of Spider Web map in assessment of CSCD-covered journals of pediactrics
Jinlong LI ; Fangyu SHI ; Zhengri LU ; Jichun MA ; Long GE ; Jinhui TIAN
Chinese Journal of Medical Library and Information Science 2014;(10):75-79
Objective To assess the CSCD-covered 7 journals of pediactrics.Methods The average number of refe rences,cited half life, H index, citation half life, self citation, IF, immediate index, self-cited rate of each paper pub-lished in 2013 in 7 journals of pediactric, covered in China Science and Technology Journal Citation Index Database, were divided into positive correlation group and negative correlation group.Their Spider Web maps were plotted.The papers published in the 7 journals of pediactrics were assessed.Results The top 3 journals, assessed according to the average number of references, IF, and immediate index, were Chinese Journal of Pediatrics,Chinese Journal of Prac-tical Pediatrics, and Chinese Journal of Evidence-based Pediatrics while the top 3 journals, assessed according to their citation half life , self citation, cited half life, and self cited rate, were Chinese Journal of Evidence-based Pe-diatrics,Chinese Journal of Pediatrics, and Journal of Clinical Pediatrics.Conclusion The academic level of the 7 journals of pediatrics is rather high.Chinese Journal of Evidence-based Pediatrics and Chinese Journal of Pediatrics have the highest assessment score.There is a large room for the other 4 journals to improve their academic level.
4.Determination of harpagide and harpagoside in Scrophulariae Radix by HPLC-UV.
Yun'e BAI ; Pengfei YUAN ; Qinghui WANG ; Suli WANG ; Yuewei GE ; Zhengri NIU ; Mingying SHANG ; Guangxue LIU ; Chen LI ; Shaoqing CAI
China Journal of Chinese Materia Medica 2011;36(19):2697-2702
OBJECTIVETo develop a method for the determination of harpagide and harpagoside in Scrophulariae Radix (Xuanshen) by HPLC-UV under double wavelength, and to study the changes of these two constituents during processing, and to set the limitation of harpagide and harpagoside contents in crude drug and sliced pieces of Xuanshen.
METHODThe analyses were performed on an Agilent Technologies ZORBAX SB-C18 (4.6 mm x 250 mm, 5 microm) eluted with acetonitrile-water (containing 0.03% phosphoric acid) in gradient model. The flow rate was 1.0 mL x min(-1) . The column temperature was 25 degrees C. The UV detector wavelength was set at 210 nm before 13 min and then changed to 280 nm.
RESULTHarpagide and harpagoside were separated well. The linear calibration curves were obtained over of 0.0549 - 1.46 microg for harpagide (r = 0.9999, n =7) ,0.0225 - 0.900 microg for harpagoside (r = 0.9998, n = 9). The recoveries ( +/- RSD)% were 98.1 (+/- 2.4)% for harpagide and 98.8 (+/- 4.3)% for harpagoside. The contents of harpagide were 0. 277% - 0.620%, harpagoside were 0.078% - 0.362% in Xuanshen, and harpagide were 0.276% - 1.059%, harpagoside were 0. 059% - 0.183% in sliced Xuanshen, respectively. After the processing of Scrophulariae Radix, the content of harpagide increases 13.7% - 96.0%, while harpagoside decreases 11.0%-73.9%.
CONCLUSIONThis method is simple, accurate, and can be used for the quality control of Scrophulariae Radix. We propose that the total content of harpagide and harpagoside in either crude drug or sliced pieces of Scrophulariae Radix should not be less than 0.45%.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Glycosides ; analysis ; isolation & purification ; Iridoid Glycosides ; Magnoliopsida ; chemistry ; Plant Roots ; chemistry ; Pyrans ; analysis ; isolation & purification ; Spectrophotometry, Ultraviolet ; methods
5.Efficacy of the Mineral Oil and Hyaluronic Acid Mixture Eye Drops in Murine Dry Eye.
Jung Han CHOI ; Jung Han KIM ; Zhengri LI ; Han Jin OH ; Kyu Youn AHN ; Kyung Chul YOON
Korean Journal of Ophthalmology 2015;29(2):131-137
PURPOSE: To investigate the therapeutic effects of mineral oil (MO) and hyaluronic acid (HA) mixture eye drops on the tear film and ocular surface in a mouse model of experimental dry eye (EDE). METHODS: Eye drops consisting of 0.1% HA alone or mixed with 0.1%, 0.5%, or 5.0% MO were applied to desiccating stress-induced murine dry eyes. Tear volume, corneal irregularity score, tear film break-up time (TBUT), and corneal fluorescein staining scores were measured at 5 and 10 days after treatment. Ten days after treatment, goblet cells in the conjunctiva were counted after Periodic acid-Schiff staining. RESULTS: There was no significant difference in the tear volume between desiccating stress-induced groups. The corneal irregularity score was lower in the 0.5% MO group compared with the EDE and HA groups. The 0.5% and 5.0% MO groups showed a significant improvement in TBUT compared with the EDE group. Mice treated with 0.1% and 0.5% MO mixture eye drops showed a significant improvement in fluorescein staining scores compared with the EDE group and the HA group. The conjunctival goblet cell count was higher in the 0.5% MO group compared with the EDE group and HA group. CONCLUSIONS: The MO and HA mixture eye drops had a beneficial effect on the tear films and ocular surface of murine dry eye. The application of 0.5% MO and 0.1% HA mixture eye drops could improve corneal irregularity, the corneal fluorescein staining score, and conjunctival goblet cell count compared with 0.1% HA eye drops in the treatment of EDE.
Animals
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Conjunctiva/*drug effects/pathology
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Cornea/metabolism
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Disease Models, Animal
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Drug Combinations
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Dry Eye Syndromes/*drug therapy/metabolism
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Emollients/administration & dosage
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Female
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Goblet Cells/drug effects/metabolism/pathology
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Hyaluronic Acid/*administration & dosage
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Mice
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Mice, Inbred C57BL
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Mineral Oil/*administration & dosage
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Ophthalmic Solutions
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Tears/*metabolism
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Viscosupplements/administration & dosage
6. Tangshen formula improves cholesterol uptake and efflux of macrophages induced by high lipid via activating PGC-1α/LXR/ABCA1 pathway
Ke XU ; Junwei GAO ; Chenguang WU ; Feng TIAN ; Lifan WANG ; Peng LIU ; Zhengri SHEN ; Hui JIA ; Ping LI
Chinese Journal of Clinical Pharmacology and Therapeutics 2021;26(9):978-985
AIM: To observe the effects of Tangshen formula (TSF) treatment on lipid efflux and uptake in sodium palmitate (PA) induced RAW264.7 macrophages. METHODS: After 200 μmol/L PA induced RAW264.7 macrophages, TSF and PGC-1α-siRNA were given to intervene respectively. The lipid content in the cells was detected by ELISA kit; intracellular lipid droplet deposition was detected by BODIPY 493/503 and Filipin staining. Western blot and Real-time PCR were used to detect the expression of PGC-1α, LXR, ABCA1 and CD36. RESULTS: TSF diminished the levels of TC, TG and intracellular lipid droplet deposition in PA-induced RAW264.7 macrophages. Western blot and Real-time PCR analysis showed that TSF could up-regulate the expression of PGC-1α, LXR, ABCA1 and down-regulate the expression of CD36. Furthermore, silencing PCG-1α by SiRNA significantly suppressed the effects of upregulating the expression of PGC-1α, LXR and ABCA1, and downregulating the CD36 expression with TSF treatment. CONCLUSION: TSF may extenuate intracellular lipid droplet deposition in macrophages by upregulating cholesterol efflux through activating the PGC-1α/LXR/ABCA1 pathway and inhibiting lipid uptake through down-regulateing the expression of CD36.
7. Tangshen formula promotes cellular cholesterol efflux in HG-induced mTECs by suppression of TGF-β1/Smad3 pathway
Peng LIU ; Zhengri SHEN ; Chen WANG ; Yun WANG ; Xinping QIU ; Hailing ZHAO ; Ping LI
Chinese Journal of Clinical Pharmacology and Therapeutics 2021;26(1):1-9
AIM: To observe the effect of Tangshen formula (TSF) treatment on TGF-β1/Smad3 pathway and cellular cholesterol efflux and explore its potential mechanism in HG-induced mouse tubular epithelial cells (mTECs). METHODS: After 25 mmol/L high glucose induced mTECs, TSF and Smad3 inhibitor SIS3 were given to intervene respectively. The lipid content in the cells was detected by ELISA kit; TGF-β1/Smad3 pathway and PGC-1α, LXR, ABCA1, ABCG1 were detected by Western blot and real-time PCR. RESULTS: TSF diminished the levels of TC, TG, LDL-C and increased the levels of HDL-C in HG-induced mTECs. Western blot and real-time PCR analysis showed that expression levels of TGF-β1, Smad3, Collagen and Fibronectin were significantly downregulated in the HG-induced mTECs with TSF and SIS3 treatment. And PGC-1α, LXR, ABCA1, ABCG1 expression levels were significantly upregulated in the HG-induced mTECs with TSF and SIS3 treatment. CONCLUSION: TSF can promote the cellular cholesterol efflux in HG-induced mTECs vis suppression of TGF-β1/Smad3 pathway.