1.Parathyroid Hormone Promotes the Proliferation and Invasion of Chondrosarcoma Cells by Regulating the Expression of Primary Cilia
Kai XU ; Wei XIANG ; Weiting CHENG
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong 2017;46(2):122-126
Objective To investigate the effect of parathyroid hormone(PTH)on the proliferation and invasion of chondrosarcoma cells,and the relationship between PTH and the regulation of primary cilia expression.Methods After stimulation of the chondrosarcoma cell line SW1353 with different concentrations of PTH,induction of the expression of cilia with hypoxia and destruction of cilia structure with chloral hydrate,the cell viability was detected by CCK8 assay,the proliferation and invasion of SW1353 by Western blotting and Transwell method,the primary cilia expression by immunofluorescence assay and the GLI1,PTCH1 and IFT88 expression levels by real-time PCR.Results PTH could promote the proliferation of chondrosarcoma cells in a concentration-dependent manner and this effect was correlated with the structural integrity of the primary cilia.PTH could up-regulate the invasive ability of SW1353 cells and increase the expression levels of MMP9,which was suppressed when the primary cilia structure was damaged.Additionally,it was found that PTH could down-regulate the number of primary cilia,which was related to the structural integrity of the primary cilia.It could also regulate the expression levels of GLI1 and PTCH1,the target genes in Hedgehog pathway,and the expression levels of IFT88,the gene associated with the cilia function.Conclusion PTH can promote the proliferation and invasion of chondrosarcoma cells,down-regulate the expression of primary cilia and the downstream target genes.PTH may regulate the malignant biological features of chondrosarcoma by regulating the primary cilia expression.
2.Oxymatrine suppressed the activation of pancreatic stellate cells and p38-MAPK mRNA expression in vitro
Wei XU ; Kai CHEN ; Xiaohui XIANG ; Shihai XIA
Chinese Journal of Pancreatology 2016;16(3):185-188
Objective To clarify whether oxymatrine ( OM) could suppress the activation of pancreatic stellate cells ( PSC) and explore the potential molecular mechanism .Methods The proliferation of PSC line LTC 14 being activated by TGF-β1 with OM treatment at different concentrations (OM group) was measured. SOD level was determined by ELISA and p 38-MAPK mRNA was determined by real-time PCR.Results The proliferation of PSC in the control group , 0.1, 0.5, 1, 2, 5 g/L OM group was (1.51 ±0.08), (1.50 ± 0.07), (1.15 ±0.04), (1.15 ±0.04), (1.08 ±0.06), and (1.08 ±0.10), respectively.The level of the control group was lower than the groups where the concentration of OM reached or exceeded 0.5mg/ml ( all P=0.000).SOD level of LTC 14 cells in the control group, TGF-β1 group, 0.5 and 1 g/L OM group was (0.087 ±0.005), (0.073 ± 0.004), (0.085 ± 0.010), and (0.086 ± 0.007), respectively. No statistically significant difference existed among the groups (P=0.095).The p38-MAPK mRNA expression of PSC in the control group, TGF-β1 group, 0.5, and 1 g/L OM group was (1.000 ±0.000), (1.979 ± 0.505), (0.606 ±0.111), and (0.303 ±0.159), respectively.The p38-MAPK mRNA level of TGF-β1 group was higher than that of the control group (P=0.002), and that of 0.5 mg/ml OM group and 1 mg/ml OM group was lower that of TGF-β1 group ( P=0.000 ) , while no statistical difference was found between 0.5 mg/ml OM group and 1 mg/ml OM group.Conclusions OM could suppress the activation of PSC in vitro and the suppression of p38-MAPK mRNA expression may be involved .
3.Repair and functional reconstruction for compound defects of proximal phalanx and dorsal skin at multiple digits
Sheng-Li LI ; Wei-Gang CAO ; Kai-Xiang CHENG ;
Chinese Journal of Orthopaedic Trauma 2004;0(08):-
Objective To summarize our surgical experience in repair and functional reconstruction for compound defects of proximal phalanx and dorsal skin at multiple digits,Methods Six patients with multiple digital defects were treated in our department between June 1996 and March 2005.At the first stage,free iliac bone grafts were used to repair defects of proximal phalanges and temporary syndactyly between adjacent affected fingers was created through digital palmar skin sutures.The defects were covered with free flap transfer finally.Dorsalis pedis flaps were used in four patients,a lateral arm flap in one and a lateral thoracic flap in one respectively.At the second stage,a partial debulking procedure and division of syndaetyty followed three to six months later.Additional procedures were performed in three eases to reconstruct the digital extensor function through tendon transfer.The follow-ups ranged from six months to nine years.Results The flaps survived uneventfully in the six patients postoperatively.The dorsal aspects of reconstructed fingers demonstrated an aesthetically pleasing effect after the flap debulking procedures and division of syndactyly.Follow-up X-Ray examinations showed good lilac bone union and nearly normal structure of digital bone.The distal interphalangeal extension restored to normal in the three cases after extensor reconstruction.Conclusions Iliac bone graft to repair phalangeal defects and free flap transplantation to cover skin defects can be a good treatment for compound defects of proximal phalanx and dorsal skin at multiple digits.Secondary plastic procedure may greatly improve the appearance of a reconstructed digit,and extensor re- construction the function of distal interphalangeal extension too.
4.Resveratrol activates Notch signalling pathway via Sirtuin 1 in podocytes
Kewei XIE ; Kai WEI ; Mingying GU ; Peng XIANG ; Leyi GU
Chinese Journal of Nephrology 2015;31(4):296-302
Objective To explore the relationship between resveratrol and Notch 1 signalling pathway in podocytes.Methods Interference RNA (RNAi) and doxycycline (Dox) were used to inhibit the Sirtuin (SIRT) 1 expression in the wild-type and inducible SIRT1 shRNA (CAGGS) podocytes respectively.Recombinant mouse delta-like ligand 4 (DLL4) was used to activate Notch1 signalling.The message RNA of SIRT1,Notch1 downstream gene Hes1 and Hey2,as well as the key enzymes of Notch1 signalling pathway were detected by using real-time PCR.Western blotting was used to detect intracellular domain of Notch 1 (ICD1),SIRT1,and metalloprotease (ADAM) 10 and components of γ-secretase complex protein expression.Results In WT murine podoytes,resveratrol up-regulated ICD1 protein production,as well as the mRNA of Hes1 and Hey2 in a dose-dependent manner.Treatment with resveratrol resulted Nicastrin mRNA and protein increase in podocytes (P <0.05),as well as inhibit ADAM10 expression (P < 0.05),but all these changes were prevented after the use of SIRT1 RNAi(P < 0.05).DLL4 up-regulated the expression of mRNA of Hes1 and Hey2,as well as ICD1 protein production in a dose-dependent manner.Treatment with doxycycline resulted decrease of SIRT1 gene and protein expression in CAGGS podocytes after 24 h and 48 h respectively(P < 0.05),which weakend the role of DLL4 significantly(P < 0.05).Conclusion Resveratrol induces Nicastrin expression,as well as activation of Notch1 signalling pathway in a SIRT1-dependent manner.
5.Adenocarcinoma of non-pigmented ciliary epithelium: report of a case.
Mei-Fu GAN ; Zhen-Yang XIANG ; Chun-Kai YU ; Wei-Ming XU
Chinese Journal of Pathology 2008;37(3):213-214
Adenocarcinoma
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pathology
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Aged
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Ciliary Body
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pathology
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Epithelium
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pathology
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Female
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Humans
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Uveal Neoplasms
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pathology
6.Effects of xuefu zhuyu decoction on antioxidant and drug-metabolizing enzymes in liver of rats.
Xing-hua FAN ; Wei-Zhou SHI ; Yun-xiang CHENG ; Kai-jie ZOU ; Xiu-fen YANG
China Journal of Chinese Materia Medica 2014;39(22):4453-4458
Xuefu Zhuyu decoction (XFZYD) is a famous traditional Chinese medicine (TCM) formula, is widely used in the treatment of cardiovascular and cerebrovascular diseases in China over one hundred years. But its effect on antioxidant and drug-metabolizing enzymes are unknown. This study was to observe the effects of Xuefu Zhuyu decoction (XFZYD) on the activities of antioxidant and drug metabolism enzymes (DMEs) in liver of rats. Male SD rats, treated with XFZYD at the dosage of 3.51, 7.02 and 14.04 g x kg(-1) per day for 15 days, serum were collected, tissue fluid, cytosols and microsomes isolated from liver tissues were prepared by centrifugation according to the standard procedure, the activities of antioxidant enzymes and drug-Metabolizing Enzymes were determined by UV-V is spectrophotometer. In serum, the activities of AST was not significantly affected by the treatment with XFZYD, at the high- est dose, the levels of ALT, Cr and BUN were significantly decreased (P < 0.05). GPX were significantly increased at the dose of 7.02, 14.04 g x kg(-1) (P < 0.05), CAT were significantly increased at the highest dose (P < 0.05). T-SOD was not significantly af- fected by this treatment. In the liver tissue, GPX was significantly increased at the dose of 3.51, 7.02 g x kg(-1) (P < 0.05), GST, CAT and T-SOD were not significantly affected following this treatment. In cytosols, GST was significantly increased at the dose of 3.51 g x kg(-1) (P < 0.05), T-SOD was remarkable induced at the dose of 3.51 and 7.02 g x kg(-1) (P < 0.05). In microsomes, XFZYD had no significant effect on Cytochromeb5, NADPH-Cytochrome P450 reductase, CYP3A, CYP2E1 and UGT, XFZYD significantly in- duced GST at the dose of 3.51 and 7.02 g x kg(-1) (P < 0.05), and the level of GSH were significantly increased by XFZYD at the dose of 3.51, 7.02 and 14.04 g kg(-1) (P < 0.05). These findings suggest XFZYD can induce the activities of GPX, CAT, SOD, GST and increase GSH level in liver of rats, which indicate XFZYD may have detoxification and antioxidant functions.
Animals
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Antioxidants
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metabolism
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Drugs, Chinese Herbal
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pharmacology
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Inactivation, Metabolic
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drug effects
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Liver
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drug effects
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enzymology
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Male
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Rats
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Rats, Sprague-Dawley
7.The influence of LPS on the protein expression of related molecules in Smads and ERK1/2 signal pathway in LTC-14 cells
Qing ZHANG ; Kai CHEN ; Meili LU ; Wei XU ; Xiaohui XIANG ; Shihai XIA ; Runli JI
Chinese Journal of Pancreatology 2017;17(2):93-98
Objective To explore the influence of LPS treatment on related molecules in Smads and ERK1/2 signal pathway in pancreatic stellate cell line LTC-14.Methods LTC-14 cells were cultured in vitro, and were treated with LPS at different dose in different time points.Protein expressions of related molecules in Smads pathway and ERK1/2 pathway and α-SMA in LTC-14 Cells were examined by Western blot.Results On Treated LTC-14 cells by 0, 1, 5, 10, 20 and 50 mg/L LPS,protein expressions of Smad3 were 0.15±0.02, 0.37±0.02, 0.44±0.01, 0.46±0.02, 0.372±0.01 and 0.24±0.03;expressions of Smad7 were 0.79±0.05, 0.84±0.02, 0.55±0.03, 0.45±0.03, 0.34±0.02 and 0.92±0.07;p-ERK1/2 levels were 0.48±0.05, 0.74±0.03, 0.72±0.04, 0.89±0.02, 0.81±0.02 and 0.72±0.03;p-cPLA2 levels were 0.15±0.03, 0.30±0.01, 0.31±0.01, 0.30±0.02, 0.28±0.03 and 0.32±0.02;α-SMA levels were 0.56±0.06, 0.62±0.06, 0.54±0.04, 1.03±0.11, 1.39±0.08 and 1.28±0.10.The changes of protein expressions before and after LPS treatment were obvious (all P<0.01).The protein expressions of ERK1/2 were 0.56±0.03, 0.57±0.02, 0.53±0.02, 0.58±0.02, 0.59±0.05 and 0.55±0.04, which did not change obviously along with increased LPS dosages.LTC-14 cells treated with 10 mg/L LPS for 0, 1, 3, 6 and 9 h,the expressions of Smad3 were 0.69±0.05, 0.68±0.07, 1.02±0.14, 1.82±0.0 and 2.04±0.11,those of Smad7 were 2.77±0.10, 1.37±0.08, 1.45±0.14, 0.78±0.09 and 0.63±0.06,those of p-ERK1/2 were 0.16±0.03, 0.32±0.05, 0.79±0.03, 1.50±0.07 and 1.77±0.04,those of p-cPLA2 were 0.15±0.04, 0.32±0.06, 0.63±0.04, 0.95±0.04 and 1.49±0.10,those of α-SMA were 0.84±0.03, 1.26±0.21, 1.81±0.19, 4.28±0.26 and 4.37±0.15, all of which changed obviously as the treatment time increased (P<0.05 or 0.01).The expressions of ERK1/2 were 0.75±0.03, 0.72±0.02, 0.80±0.04, 0.74±0.03 and 0.85±0.09, which did not change obviously as the treatment time increased.Conclusions LPS could upregulate the expression of α-SMA in a time-and dose-dependent way, and activate intracellular Smads and ERK1/2 inflammatory pathways, which may be the potential molecular mechanism of the development of chronic pancreatitis.
8.A New Design of Impactor for Rat and Monkey's Spine Cord Injury Models
Xiang ZHANG ; Wei SONG ; Chaoyang YANG ; Kai LIAO ; Haijun NIU ; Xiaoguang LI
Chinese Journal of Rehabilitation Theory and Practice 2009;15(12):1138-1140
Spinal cord injury is very high morbidity trauma, seriously affecting the survival and health of patients. To establish a standard animal model has a great significance for studying SCI pathogenesis and pathological changes. A spinal cord injury device for experimental animal was presented in this paper. It can implement fixed-point and fixed-quantity impact by new design. It also can create many animal models of different levels injury. More importantly, it has many advantages such as simple to operate, free from human disturbance, high accuracy and reproducibility. Finally, some existing constraints and challenges about the animal spinal cord injury models made by the device were discussed.
9.Genetic diversity of Fritillaria from Sichuan province based on ISSR.
Kaiqiang LI ; Wei WU ; Youliang ZHENG ; Yong DAI ; Li XIANG ; Kai LIAO
China Journal of Chinese Materia Medica 2009;34(17):2149-2154
OBJECTIVETo provide more proofs for expounding the genetic relationships among the (varietal) species in genus Fritillaria from Sichuan province.
METHODThe ISSR marker technique was used to study relationships and genetic polymorphism of nineteen populations in ten species and one varietal species of genus Fritillaria. Genetic similarities were calculated by using NTSYS software and the dendrogram was constructed by using UPGMA method.
RESULTEleven primers were selected from 35 ISSR primers, and 179 DNA fragments were amplified from 19 populations. Of which, 179 fragments were polymorphic (percentage of polymorphic bands was 86.8%). The genetic similarity among all accessions ranged from 0.569 to 0.855. Clustering analysis showed that the 19 populations of Fritillaria could be distinctively classified into 4 groups. F. cirrhosa, F. przewalskii, F. cirrhosa var. logirnectarea and F. dajitensis were in the first group; The second group was the cluster of F. cirrhosa and F. mellea (wild and cultivated species); The third group was F. sulcisquamosa, F. thunbergii, wabunesis and F. delavayi; F. hupehensis alone formed the fourth group.
CONCLUSIONISSR marker technique is suitable for the genetic diversity of Fritillaria from Sichuan province. Interspecific identifications among the four original species of Bulhus Fritillariae Cirrhosae recorded by pharmacopoeia of China, and between them and the other species of genus Fritillaria from Sichuan province could not be gained by using ISSR markers technique. In addition, the cluster result of genus Fritillaria had some relationships with the geographical distribution.
China ; Fritillaria ; classification ; genetics ; Genetic Variation ; Microsatellite Repeats ; Phylogeny
10.Study on chemical constituents from seed of Oroxylum indicum.
Xiang-yu ZHAI ; Wei XIAO ; Biao YANG ; Zhao-qing MENG ; Zhen-zhong WANG ; Wen-zhe HUANG ; Kai-jin WANG
China Journal of Chinese Materia Medica 2015;40(15):3013-3016
Oroxylum indicum was a traditional Chinese medicine. In order to study the chemical constituents from the seed of O. indicum, the chemical constituents of 80% methanol extract of seeds of O. indicum were subjected to chromatography on silica gel, Sephadex LH-20, and preparative HPLC, leading to the isolation of eleven compounds. The structures were identified by various spectroscopic data including ESI-MS, 1H-NMR and 13C-NMR data as oroxin B (1), chrysin (2), baicalein (3), neglectein (4), quercetin-3-O-β-D-galactopy ranoside (5), quercetin-7-O-β-D-glucopyranoside (6), 2α,3β-dihydroxylluPeol (7), lupeol (8), rengyol (9), β-sitostero (10), and stigmasterol (11). Among them, compound 5 were firstly obtained from O. indicum.
Bignoniaceae
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chemistry
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Magnetic Resonance Spectroscopy
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Seeds
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chemistry