1.Research method of psoriasis based on HaCaT cells
Jia-Feng WANG ; Jie LI ; Jin-Qi LI
Chinese Journal of Pharmacology and Toxicology 2018;32(4):302-303
Psoriasis is a chronic,refractory,inflammatory skin disease that occurs in young adults.The traditional animal model cannot simulate the skin characteristics of patients with psoriasis effectively, so it is difficult to be used for in-depth study of psoriasis mechanism. Immortalized human epidermal cells (HaCaT)is a non-tumor,immortalized human epidermal cell which is widely used in the study of dermatosis.HaCaT cells are the best choice for the study of psoriasis mechanism because their immu-nological characteristics and reproductive ability are coincide with the pathological features of psoriasis. This article reviews the specific methods such as establishment of cell method, cytokine and chemo-kine analysisin the pathogenesis study of psoriasis based on HaCaT cells, hoping to provide some thoughts for drug′s pharmacological activity research.
2.Effect of AAVC-I on the migration of human umbilical vein endothelial cells.
Shu LI ; Xin JIN ; Xue-feng LONG ; Jin-li JIA ; Gen-bao ZHANG ; Yun HONG
Chinese Journal of Applied Physiology 2015;31(5):407-410
OBJECTIVETo investigate the effect of component I from agkistrodon acutus venomon (AAVC-I) the migration of human umbilical vein endothelial cells (HUVECs), and to elucidate the possible anti-angiogenic mechanism of AAVC-I.
METHODSThe effect of AAVC-I on the migration of HUVECs which was cultivated in vitro and treated with AAVC-1 at four concentrations: 0, 20, 40, 80 microg/ml, was observed by methods of scratch wound-healing and Transwell assay. The expression level of mRNA and protein of P-selectin and intercellular cell adhension molecule-I (ICAM-1) were examined by RT-PCR and Western blot assay.
RESULTSCompared with the blank group, the migration ability of HUVECs in each AAVE-I treated group was reduced in a dose-dependent manner, and the expression level of the mRNA and protein of P-selectin and ICAM-1 were decreased.
CONCLUSIONAAVC-I inhibits the migration of endothelial cell, which is acted by down-regulation of the expression content of mRNA and protein of P-selectin and ICAM-1.
Cell Movement ; drug effects ; Cells, Cultured ; Crotalid Venoms ; pharmacology ; Down-Regulation ; Human Umbilical Vein Endothelial Cells ; drug effects ; Humans ; Intercellular Adhesion Molecule-1 ; metabolism ; P-Selectin ; metabolism ; RNA, Messenger
3.Analysis of the characteristics of juvenile primary Sj(?)gren's syndrome with renal damage
Jin-Li RU ; Xiao-Feng ZENG ; Li-Min ZHANG ; Jin-Chang HE ; Jia ZHU ;
Chinese Journal of Rheumatology 2001;0(05):-
Objective To study the renal damage of patients with juvenile primary Sjgren's syndrome (pSS)and its clinical manifestations,pathologic characteristics with biopsy,treatment and prognosis.Methods Ten patients with juvenile pSS complicated with renal impairments were retrospectively analyzed.Data of these 10 patients were compared with those without renal impairments.Results Ten patients complicated with renal impairments in 24 patients with juvenile pSS,9 of them presented with type I renal tubular acidosis(RTA), 5 with hypokalemia paralysis,3 with calcification of the renal tissue,3 with positive urine protein.1 with dia- betes insipidus.There was no significant difference between patients with renal impairments and those without. Three patients underwent kidney biopsy that showed chronic interstitial nephritis(CIN)with extensive lymp- hoplasmie cell infiltration.Two patients had glomerular lesions and one of them was diagnosed as pSS over- laped with systemic lupus erythematosus(SLE).Steroid and immunosuppressive agents had significantly alle- viated symptoms and the hypergammaglobulinemia was significantly improved.Conclusion Renal impairment may be the major complication in juvenile pSS.The major clinical manifestations are RTA and the glomeruli are involved occasionally.Treatment with steroid anti immunnsuppressive agents should be given to those who have evidence of systemic involvement.
4.Risk stratification and analysis of prognostic factors of acute myeloid leukemia with t(8;21)
Jia FENG ; Jin LIU ; Haichan XU ; Wenli ZHANG ; Qian ZHANG ; Fengluan ZHONG ; Qingxiang MENG
Journal of Leukemia & Lymphoma 2013;22(7):431-435
Objective To investigate the cytogenetic and immunological phenotypes of acute myeloid leukemia (AML) with t(8;21),and explore the risk stratification and risk-adapted treatments.Methods The chromosomal karyotype of bone marrow was detected and analyzed in 22 newly diagnosed patients with t(8;21) AML by direct culture and G banding technique.Patients were divided into two groups according to the chromosomal karyotypes.Clinical characteristics and immunological phenotypes were compared between patients with isolated t(8;21) and those with additional aberrations.A follow-up study with median time 30 months (4-68 months) was conducted to analyze prognostic factors.Results 13 cases (59.1%) were isolated t(8;21) AML,while 9 (40.9 %) had additional aberrations.Loss of sex chromosome was found in 3 cases and complex variant translocation in 2.The 10q-,9q-,-18 and +10 were found in single cases.Overall survival of patients with additional aberrations was significantly poorer than those with isolated t (8;21) (P =0.0176).Analysis of prognostic factors showed that t(8;21) chromosomal karyotype,initial white blood cells at diagnosis,and treatment regimen (chemotherapy alone or plus hematopoietic stem cell transplantation) had effects on overall survival.Conclusion Patients with t (8;21) AML are frequently associated with additional chromosomal aberrations.The latter indicates a poorer outcome and can be one of the bases of risk stratification.Hematopoietic stem cell transplantation might help to improve the overall survival.
5.Mutation (Thr704Met) of the SCN4A gene causes severe familial hypokalemic periodic paralysis
Dan YANG ; Haijiang LI ; Jia GENG ; Tao CHEN ; Jianping SONG ; Lipin LIANG ; Fangfang LI ; Feng JIN
Chinese Journal of Neurology 2016;(2):98-101
Objective To investigate the clinical features and pathogenic genes of a familial hypokalemic periodic paralysis ( HOKPP).Methods PCR amplification and DNA sequencing were used to screen candidate genes of the HOKPP family members (CACNA1S, SCN4A, KCNE3), and the clinical features were carefully analyzed at the same time.Results The sequencing analyses of the SCN4A gene in the proband identified three nucleotide sequence mutations, which influenced the amino acid sequence of the skeletal sodium channel.One of the mutations was identified as a C/T heterozygous pattern at the 2111th nucleotide position in exon 13, resulting in a change from Thr to Met at the 704th amino acid position of the sodium channel protein.All affected patients carried the Thr704Met mutation, whereas unaffected family members did not.Clinical symptoms in this family followed an autosomal dominant inheritance pattern.Muscles weakness, pain and hypokalemia in the period between attacks were seen in all patients.Paralytic symptoms occurred early, lasted longer and recurred frequently, while cold was the main predisposing factor.With the progress of the disease, patients represented persistent weakness and atrophy in proximal muscles.Conclusions Mutation (Thr704Met) in the SCN4A gene should be responsible for this family.This mutation causes severe HOKPP and progressive muscle atrophy.
6.The optimized culture and identification of astrocytes from rat cortical tissue
Hui JIN ; Gaifeng FENG ; Pengbo YANG ; Ning JIA ; Weina YANG ; Yihua QIAN ; Weixi WANG
Journal of Xi'an Jiaotong University(Medical Sciences) 2015;(6):849-853
Objective To obtain highly purified astrocytes and identify the cells in each stage to support further studies.Methods The cerebral cortex of a neonatal SD rat was isolated and prepared into single cell suspension.The obtained cells were purified by differential adherence and shook at a constant temperature.By inverted phase contrast microscopy and HE staining,cell morphology was observed.The immunofluorescence staining with anti-mouse GFAP was used to identify the cells.Results The primary cortical cells developed rapidly at 3 d after culture and covered the flasks at 9-12 d.At this time,the cells showed stratification and the astrocytes lay at the lower layer.GFAP positive rate was only about (67.2 ±7.1)%.After the first passage,GFAP positive rate increased obviously (84.0±6.0)%. However, oligodendrocytes and microglias could not be removed completely,and the cells also showed stratification.Through 3 times of passages,we obtained many single species of astrocytes showing satellite shape with 2 or 3 processes,big cell body and round or oval-shaped nuclei leaned to one side.Immunofluorescence staining showed that nearly all of the cells were strong positive and the positive rate reached as high as (97.6 ± 2.4 )%.Conclusion Through differential adherence and shaking at a constant temperature,more astrocytes of high purity and in good state can be obtained.
7.WGCNA Reveals Key Roles of IL8 and MMP-9 in Progression of Involvement Area in Colon of Patients with Ulcerative Colitis
Xue LIN ; Jin LI ; Qiu ZHAO ; Jue-Rong FENG ; Qian GAO ; Jia-Yan NIE
Journal of Huazhong University of Science and Technology (Medical Sciences) 2018;38(2):252-258
Ulcerative colitis (UC) is a chronic inflammatory disease and its involvement area in colon is influenced by a complex network of gene interactions.We analyzed the weighted gene co-expression networks in microarray dataset from colonic mucosa of patients with UC and identified one gene co-expression module that was highly associated with the progression of involved area in UC colon (Pearson coefficient=0.81,P<0.0001).In total,523 hub genes in this module were found to be involved in immune system process after enrichment analysis in Gene Ontology.By the STRING and Cytoscape analysis,we observed that interleukin-8 (IL-8) and matrix metalloproteinase-9 (MMP-9) were centered in the network of hub genes.We then detected the expression of IL-8 and MMP-9 in mucosa from left-sided colon of patients using quantitative PCR and immunofluorescence assay respectively.Both quantitative PCR and immunofluorescence assay revealed the expression levels of IL-8 and MMP-9 were significantly different among the healthy controls,left-sided colitis group and pancolitis group (P<0.05).IL-8 and MMP-9 were detected with an enhanced expression in pancolitis as compared with left-sided colitis and healthy controls,respectively (P<0.05).This study demonstrates that immune system process is indispensable in the progression of disease in colon,and identifies that IL-8 and MMP-9 play potential critical roles for the progression.
8.Clinic research of video-assisted thoracoscopic solitary pulmonary nodule resection after CT-guided hardening agent localization
Ninglei QIU ; Zhi ZHANG ; Yiping ZHUANG ; Yong YU ; Jin ZHANG ; Hui JIA ; Dongjie FENG ; Lin XU
Chinese Journal of Thoracic and Cardiovascular Surgery 2012;28(7):398-400
Objective The aim of this study is to evaluate the efficacy and safety of preoperative CT-guided hardening agent localization.Methods From December 2010 to January 2012,27 patients with 29 solitary pulmonary nodules who had undergone CT-guided hardening agent localization and video-assisted thoracoscopic surgery (VATS) were studied.Results All cases were underwent CT-guided hardening agent localization successfully,and no patient had serious complication that required any intervention.The diameter of nodules ranged from 3 to 21 mm as measured by CT[mean (11.27 ± 6.32) mm].The distance between the center of nodule and visceral pleural ranged from 4 to 38 mm[mean (14.45 ± 4.32) mm].Conversion from VATS to thoracotomies was not necessary during the diagnostic resection procedure nodules.29 solitary pulmonary nodules underwent thoracoscopic wedge resection,and no intra-or postoperative mortality or morbidity was recorded.Conclusion CT-guided hardening agent localization before video-assisted thoracoscopic solitary pulmonary nodule resection is a safe and effective procedure for accurate diagnosis and resection of indeterminate solitary pulmonary nodules.
9.Heterogeneous acellular dermal matrix patch for repair of oral mucosal defects in 71 patients
Lingfa XUE ; Wei SHANG ; Yuanyong FENG ; Xiaoming JIN ; Fengtong LIU ; Muyun JIA ; Rongtao YUAN ; Lingxue BU
Chinese Journal of Tissue Engineering Research 2010;14(16):3015-3018
BACKGROUND:Recently,acellular dermal matrix allograft has been widely used in the repair of oral mucosal defects.But little information is about the heterogeneous acellular dermal matrix (HADM) patch for repair of oral mucosal defects.OBJECTIVE:To investigate the efficacy and biosafety of HADM in the repair of oral mucosal defects.METHODS:In total 71 patients with oral benign or malignant tumors who had oral mucosal or soft tissue defects following tumorectomy were included in this study.These patients comprised 37 males and 34 females,and were averaged 45 years (range,20-70 years old).Of them,42 suffered from benign tumors and 29 from malignant tumors.HADM patches were used for repair of oral mucosal defects.The survival,color,and texture of HADM patches were observed.Shrinkage rate of HADM patches was compared between regions without supports from hard tissues (cheeks,tongue,and mouth floor) and with supports from hard tissues (gingiva,hard palate).RESULTS AND CONCLUSION:All 71 HADM completely survived.No necrosis and infection occurred.At 2 weeks after transplantation,(98.20±5.20) % of patch area survived.At 3 months after transplantation,patches showed similar color to surrounding oral mucosa and most patients had sense of tension to different extents.At 6 months after transplantation,cell creeping substitution and vasculadzation were successfully accomplished in the region of patch transplantation.Patches grew stably,with smooth pink appearance and good elasticity,and no further shdnkage.Patients felt normal.HADM patch shrank primarily at 2 weeks-1 month after transplantation,and tended to be stable at 3 months.There was no significant difference in tissue morphology between surgical region and normal tissue.The HADM shdnkage rate was significantly higher in regions without supports from hard tissues than regions with supports from hard tissues.These findings indicate that HADM patches have advantages in repair of oral mucosal defects including good histocompatibility,wide source,simple manipulation,and able to cover the wound surface in the early state,promote wound surface healing,and reduce scar formation,and can be used as an ideal matedal for repair of oral mucosal defects.
10.Research on bone marrow mesenchymal stem cells in the treatment of severe acute pancreatitis
Yuqiang SHAN ; Liang LUO ; Qi XIE ; Yi WU ; Zhong JIA ; Huicheng JIN ; Guanghua FENG
Chinese Journal of Hepatobiliary Surgery 2013;19(9):711-715
Objective To investigate the mechanism of acute inflammatory response and tissue repair when rats accepted transplanted bone marrow mesenchymal stem cells (MSC) in severe acute pancreatitis (SAP).Methods A total of 40 rats were randomly divided into 4 groups which included the normal group (n=10),the model severe acute pancreatitis group (n=10),the transplanted bone marrow mesenchymal stem cells group (n =10),and the combination of bone marrow mesenchymal stem cells and granulocyte colony-stimulating factor (G-CSF) group (n=10).To cure the acute severe pancreatic injury caused by SAP,rats were injected with EDU-labeled MSCs and granulocyte colonystimulating factor (Gr-CSF).The conversion rate of MSCs to pancreatic cells or MSCs to endothelial cells was detected to assess the role of MSCs in tissue regeneration and repair.The expression of amylase,interleukin-6 (IL-6),and interleukin-10 (IL-10) in serum was detected to assess the role of MSCs in an acute inflammatory response.Results The concentrations of amylase and IL-6 were reduced and the concentration of IL-10 was increased in MSC and MSC+G-CSF groups after the onset of SAP.Flow cytometry showed a small amount of MSCs converting to endothelial or pancreatic cells,but the conversion rate was relatively low.Conclusions MSCs can play an important role in the antipre-release of inflammatory mediators,reducing acute immune response to control the acute inflammatory response in SAP.Moreover,MSCs can repair a damaged pancreas by converting into both pancreatic and endothelial cells.