1.Correlation Analysis of Wnt/β-catenin Signaling Pathway, OPN and MMP-13 in Varying Degrees of Osteoarthritis
Jiyuan SHI ; Zhi YI ; Zongzhi LIU ; Le JI ; Huitong LIU
Progress in Modern Biomedicine 2017;17(24):4639-4644
Objective:To analysis the correlation of Wnt/β-catenin signaling pathway,OPN and MMP-13 in rabbit model with different degree of osteoarthritis (OA).Methods:Forty New Zealand white rabbits were randomly divided into four groups of 10 each,then we constructed the rabbit model with different degree of osteoarthritis (OA) by different concentrations of papain.The mRNA and protein levels of β-catenin,OPN,MMP-13,type Ⅱ collagen and proteoglycan were detected by Real-Time PCR and ELISA,respectively.Results:We have successfully established OA rabbit model by different concentrations of papain,and these OA rabbits model could be divided into mild,moderate and severe three level by Mankin scoring system,chondrocytes.Compared with the normal control group,the protein levels of β-catenin,OPN,and MMP-13 in the osteoarthritis were significantly increased,while the protein levels of type Ⅱ collagen and proteoglycan were significantly decreased (P<0.05).With the increasing severity of OA,the results were consistent (P<0.05).Conclusions:These results indicated that Wnt/β-catenin signaling pathway might regulate the expression of OPN,thereby affecting the expression of MMP-13,and ultimately have an impact on the occurrence ofosteoarthritis.
2.lnfluence of dexamethasone on lL-1β and TNF - α expression in rabbit corneal neovascularization
Rui, SHI ; Yu-Shun, XUE ; Le, YANG ; Ji-Min, WANG ; Feng, WANG ; Yi-Ning, SHI
International Eye Science 2014;(9):1574-1577
To discuss the influence of dexamethasion on lL-1β and TNF - α expression in suture - induced rabbit corneal neovascularization ( CNV ) and analyze the potential mechanism.
●METHODS: For 43 healthy rabbits, 40 were randomly selected for establishing CNV model in corneal stroma. The right eyes (group A) were received no medicine and the left eyes ( group B) were injected dexamethasone after successfully establishing the model. The no modeling 3 rabbits were normal control group. The morphologic change of corneal was observed with slit lamp microscope and the areas of CNV was calculated every day, then 8 rabbits were randomly chosen for sacrificing at 1, 4, 7, 14, 21d respectively. The pathological characteristics of CNV were observed after HE staining, and lL - 1β and TNF - α expression was detected by immunohistochemistry.
●RESULTS: CNV was grown at the 4d after suture, and the 7-14d was vigorous growth period. inflammatory cell infiltration appeared after HE staining, and CNV was located at the superficial stroma of cornea. lmmunohistochemistry results showed that lL - 1β and TNF - α expression was gradually increased with prolonged suture time. Compared with corneal stitch group, the rabbits cured by dexamethasone were found with less inflammatory cells infiltrating and neovescularization, moreover, the expression of lL - 1βand TNF-α decreased. There were statistical significance between the two groups (P<0. 05).
● CONCLUSlON: Dexamethasone can inhibit the CNV growth by controlling the inflammation of corneal and restraining lL-1β and TNF-α expression.
3.Application of cosmetic suture technique for reparation and reconstruction in facial emergency surgery
Xiaoge LI ; Hongxia WANG ; Shupeng HUANG ; Le LI ; Haishan SHI ; Yi JI ; Liang CHEN
Journal of Regional Anatomy and Operative Surgery 2015;(3):285-286,287
Objective To investigate the clinical effect of cosmetic suture technique applied in organs of facial emergency surgery. Methods There were 25 patients who were admitted in our hospital from August 2009 to December 2013. After anatomical reduction and functional restoration, they were given surgery with cosmetic suture technique on the base of debridement. Results All of the patients a-chieved good surgical result, the satisfaction is 100%. After the first stage of operation, 23 patients of them were of no obvious scar and good function recovery. The other 2 patiens also got good effects after secondary surgery. Conclusion Cosmetic suture technique pay attaintion to anatomical and functional restoration. It is of equisite technique, slight injury, less scar after operation and it can maximumly get close to the normal tissue structures and achieve the objective of beauty.
4.The mediating effect of personality between schizophrenia and prospective memory
Zhikai LIAN ; Zhanbiao SHI ; Yushan LIU ; Liu YANG ; He LI ; Yi LE ; Ling HE
Chinese Journal of Behavioral Medicine and Brain Science 2015;24(3):233-236
Objective To explore the differences between schizophrenic patients and normal at prospective memory,and investigate the effect of personality intermediary between schizophrenia and prospective memory.Methods A case-control study was utilized in this research.It used the color matching task to test prospective memory and MMPI-2 scale to test personality differences between the two groups.Results Schizophrenia prospective memory score(19.29±2.30),lower than the normal group(44.74±2.06),the difference was significant(P<0.01).In mediating effect on the personality inspection,the subscales within MMPI-2 of Social Scale(Si) and Anxiety and tension Scale(ANX) regression coefficients were tested(P<0.05),which showed significant effect.Further analysis with sobel statistics showed that only one factor tested was significant.The test coefficients of depression scale (D),mental weakness scale (Pt) and schizophrenia scale (Sc) were significantly different (P< 0.05),which showed that these subscales also had significant mediating effect.Conclusion The prospective memory performance is low in schizophrenia.The personality characteristics that depression,mental weakness,schizo phrenia,social anxiety and tension play an intermediary role in schizophrenia and prospective memory.
5.Procedures and application of Dubian-Azan stainning.
Yu-lan JIN ; Le LIANG ; Shao-hui SHI ; Yi-ding HAN ; Hideaki ENZAN ; Eriko MIYAZAKI
Chinese Journal of Pathology 2006;35(5):310-311
Animals
;
Liver
;
pathology
;
Liver Cirrhosis
;
pathology
;
Male
;
Rats
;
Rats, Wistar
;
Reproducibility of Results
;
Staining and Labeling
;
methods
6.Roles of adenosine and cytokines in the prostate tissue of rats with acute bacterial prostatitis.
Zhi LONG ; Xia-Ming PEI ; Le-Ye HE ; Ying-Bo DAI ; Dong-Yi PENG ; Yi-Chuan ZHANG ; Xuan-Yan SHI ; Jing-Liang HE
National Journal of Andrology 2014;20(4):315-319
OBJECTIVETo investigate the possible roles of adenosine and the cytokines TNF-alpha and IL-10 in the pathogenesis of acute bacterial prostatitis (ABP) in rats.
METHODSForty-eight male Wistar rats were randomly divided into groups A (ABP), B (ABP + theophylline intervention), C (sham) and D (blank control). ABP models were established by injecting Escherichia coli 0157 into the prostate, and those in group B were treated by intraperitoneal injection of theophylline immediately after modeling. At 4 and 14 days, the prostate tissues of the rats were collected for detection of the expressions of TNF-alpha and IL-10 by immunohistochemistry and the concentration of adenosine by high-performance liquid chromatography.
RESULTSAt 4 and 14 days, the concentrations of adenosine were significantly higher in group A ([48.38 +/- 17.27] and [26.54 +/- 11.22] microg/g) than in C ([0.45 +/- 0.25] and [0.46 +/- 0.29] microg/g) and D ([0.41 +/- 0.23] and [0.43 +/- 0.27] microg/g) (P < 0.05), and so were the expressions of TNF-alpha in A (0.23 +/- 0.08 and 0.21 +/- 0.03) than in C (0.07 +/- 0.03 and 0.07 +/- 0.01) and D (0.07 +/- 0.06 and 0.07 +/- 0.06) (P < 0.05), and those of IL-10 in A (0.13 +/- 0.03 and 0.25 +/- 0.01) than in C (0.07 +/- 0.03 and 0.07 +/- 0.03) and D (0.07 +/- 0.01 and 0.07 +/- 0.02) (P < 0.05). Compared with group A, the rats in group B showed significant increases at 4 and 14 days in the severity of inflammation, concentration of adenosine ([86.64 +/- 32.87] and [51.17 +/- 22.96] microg/g, P < 0.05) and expression of TNF-alpha (0.37 +/- 0.08 and 0.32 +/- 0.06, P < 0.05), but exhibited no remarkable difference in the expression of IL-10 (0.12 +/- 0.06 and 0.15 +/- 0.06, P > 0.05).
CONCLUSIONAdenosine may affect the progression of inflammation by regulating the expressions of the cytokines TNF-alpha and IL-10 in ABP rats through the adenosine receptor signaling pathway.
Adenosine ; physiology ; Animals ; Escherichia coli O157 ; Interleukin-10 ; metabolism ; Male ; Prostate ; drug effects ; metabolism ; Prostatitis ; metabolism ; microbiology ; Random Allocation ; Rats ; Rats, Wistar ; Theophylline ; pharmacology ; Tumor Necrosis Factor-alpha ; metabolism
7.Gold-immunochromatography assay for detection of Yersinia pestis F1 antigen
He-zhi, LIU ; Xue-wei, BAI ; Hai-feng, WANG ; Le-le, HU ; Song, ZHOU ; Xiao-yan, YANG ; Guo-yi, DU ; Shun-lin, YANG ; Xian-ming, SHI ; Yu-gui, LI
Chinese Journal of Endemiology 2010;29(6):678-681
Objective To study the sensitivity and specificity of gold-immunochromatography assay (GICA) for detection of Yersiniapestis(Y. pestis ) F1 antigen. Methods Viscera organ(liver and spleen) specimens of 308 mice with virulent Y. pestis infection and 225 control specimens of rats(217 Spermophilus dauricus, 5 mice,3 guinea pigs) were detected by GICA dipstick with monoclonal antibody against plague F1 antigen (F1MAb).Meanwhile, micro-method of reverse indirect hemagglutination assay(RIHA) and bacteria culture were carried out for parallel testing. Results Bacteriological examination of 225 control specimens, and F1 antigen detected with GICA and RIHA were all negative. No cross-reaction with related Yersinia pseudotuberculosis at 1 x 108 cfu/ml level was found in GICA and RIHA. Detection sensitivity of Y. pestis by GICA and RIHA were 2.5 × 105 cfu/ml and 2.0 × 105 cfu/ml, respectively, and of F1 antigen were 1μg/L and 10 μg/L, respectively. Coincidence was 97.94% (522/533) between GICA and bacteriological test, Kappa = 0.959, and the difference was statistically insignificant(x2 = 0.36, P > 0.05); and 97.94%(522/533) between GICA and RIHA, Kappa = 0.959, with statistically significant difference in the positive detection rates(x2 = 9.09, P < 0.05). Out of the 308 infected mice, 284 were positive of plague bacterial cultured, In 284 samples with positive bacterial culture, there were 280 of positive detected by GICA for F1 antigen, positive rate of F1 antigen was 98.59%, higher than that by RIHA[the positive rate of 96.13%(522/533)], with statistically significant difference(x2 = 5.14, P < 0.05). Sensitivity of GICA was 98.59% (280/284), specificity was 97.19% (242/249), positive predictive value (PPV) was 97.56% (280/287),negative predictive value ( NPV ) was 98.37% (242/246), and Youden index was 0.9578. Conclusions GICA is sensitive and specific, fast and simple in detection of F1 antigen of the plague. It's a valuable detection technique for early and rapid diagnosis of plague.
8.Microtuber induction in vitro from Rehmannia glutinosa.
Jian-ping XUE ; Le-yi SHI ; Ai-min ZHANG
China Journal of Chinese Materia Medica 2002;27(11):824-827
OBJECTIVETo set up the fittest medium and optimum condition for plantlets to form microtubers.
METHODPlant hormones, concentration of sucrose, active carbon and light time influenced formation of adventitious root and microtuber in vitro from plantlet of Rehmannia glutinosa (85-5).
RESULT AND CONCLUSIONThe plantlets were cultivated on the medium of microtuber induction after roots were induced on the 1/2MS medium with IBA 1 mg.L-1. The best medium was MS + 6-BA 2 mg.L-1 + NAA 0.1 mg.L-1 + Sucrose 5%. The fittest incubation temperature was 25 degrees C and the light length was (2,000-3,000 lx) 12 h.d-1. Active carbon and GA3 should not be added to the medium, which was not suitable for the induction of microtuber.
Culture Media ; Culture Techniques ; Light ; Plant Growth Regulators ; pharmacology ; Plant Roots ; growth & development ; Plants, Medicinal ; growth & development ; Rehmannia ; growth & development ; Sucrose ; pharmacology ; Temperature
9.The species traceability of the ultrafine powder and the cell wall-broken powder of herbal medicine based on DNA barcoding.
Li XIANG ; Huan TANG ; Jin-le CHENG ; Yi-long CHEN ; Wen DENG ; Xia-sheng ZHENG ; Zhi-tian LAI ; Shi-lin CHEN
Acta Pharmaceutica Sinica 2015;50(12):1660-1667
Ultrafine powder and cell wall-broken powder of herbal medicine lack of the morphological characters and microscopic identification features. This makes it hard to identify herb's authenticity with traditional methods. We tested ITS2 sequence as DNA barcode in identification of herbal medicine in ultrafine powder and cell wall-broken powder in this study. We extracted genomic DNAs of 93 samples of 31 representative herbal medicines (28 species), which include whole plant, roots and bulbs, stems, leaves, flowers, fruits and seeds. The ITS2 sequences were amplified and sequenced bidirectionally. The ITS2 sequences were identified using Basic Local Alignment Search Tool (BLAST) method in the GenBank database and DNA barcoding system to identify the herbal medicine. The genetic distance was analyzed using the Kimura 2-parameter (K2P) model and the Neighbor-joining (NJ) phylogenetic tree was constructed using MEGA 6.0. The results showed that DNA can be extracted successfully from 93 samples and high quality ITS2 sequences can be amplified. All 31 herbal medicines can get correct identification via BLAST method. The ITS2 sequences of raw material medicines, ultrafine powder and cell wall-broken powder have same sequence in 26 herbal medicines, while the ITS2 sequences in other 5 herbal medicines exhibited variation. The maximum intraspecific genetic-distances of each species were all less than the minimum interspecific genetic distances. ITS2 sequences of each species are all converged to their standard DNA barcodes using NJ method. Therefore, using ITS2 barcode can accurately and effectively distinguish ultrafine powder and cell wall-broken powder of herbal medicine. It provides a new molecular method to identify ultrafine powder and cell wall-broken powder of herbal medicine in the quality control and market supervision.
Cell Wall
;
DNA Barcoding, Taxonomic
;
DNA, Plant
;
genetics
;
DNA, Ribosomal Spacer
;
genetics
;
Drugs, Chinese Herbal
;
analysis
;
Phylogeny
;
Plants, Medicinal
;
classification
;
genetics
;
Powders
;
Quality Control
10.Antidiabetic activity of Callicarpa nudiflora extract in type 2 diabetic rats via activation of the AMPK-ACC pathway
Wen-Yu MA ; Le-Ping MA ; Bo YI ; Min ZHANG ; Shi-Xiu FENG ; Li-Ping TIAN
Asian Pacific Journal of Tropical Biomedicine 2019;9(11):456-466
Objective: To evaluate the antidiabetic effect of Callicarpa nudiflora extract in streptozotocin-induced diabetic rats. Methods: The chemical constituents in Callicarpa nudiflora extract were identified by UPLC-Q-TOF-MS. Callicarpa nudiflora extract (0.15 and 0.3 g/kg) was orally administered to streptozotocin-induced diabetic rats for 42 d. The effects of Callicarpa nudiflora extract on body weight, blood glucose, serum insulin, total cholesterol, triglyceride, LDL-C and HDL-C were investigated, and its effect on liver and pancreatic pathology was assessed by histopathological analysis. Moreover, the expression levels of adenosine 5'-monophosphate-activated protein kinase (AMPK), glucose transporter type 4 (GLUT4), phospho-AMPK/AMPK, and p-acetyl-coA carboxylase (P-ACC)/ACC in the skeletal muscles and liver were determined by reverse transcription-polymerase chain reaction, Western blotting, and immunohistochemistry. Results: A total of 34 compounds, including 8 iridoids, 14 phenylpropanoids, and 12 flavonoids, were identified from Callicarpa nudiflora extract. Callicarpa nudiflora extract significantly reduced blood glucose and significantly restored all other biochemical parameters to near normal levels, including serum insulin, total cholesterol, triglyceride, LDL-C, and HDL-C. Callicarpa nudiflora extract improved insulin resistance and reversed the damage in the liver and pancreas caused by diabetes. Furthermore, Callicarpa nudiflora extract increased the expression levels of phospho-AMPK, GLUT4, P-ACC, and insulin receptor substrate-1 and decreased the expression level of PPAR毭 in diabetic rats.Conclusions: Callicarpa nudiflora extract improved oral glucose tolerance, lipid metabolism, insulin resistance, and reversed the diabetes-related damage in the liver and pancreas by activating the AMPK-ACC pathway.