1.Analysis of clinical and genetic features of one Chinese family with paroxysmal exercise-induced dyskinesia
Journal of Shanghai Jiaotong University(Medical Science) 2017;37(6):774-779
Objective· To study the clinical and genetic features of familial paroxysmal exercise-induced dyskinesia (PED) in a Chinese mainland family,and review the advances of clinical and genetic studies on PED.Methods· The clinical information of 7 family members in one Chinese pedigree,including 5 patients and 2 healthy people,was analyzed and the patients' response to treatment and prediction were followed up.The SLC2A1 gene in all 7 members of this family was sequenced.The clinical and genetic characteristics of 5 patients were analyzed.Advances of recent clinical and genetic studies related with PED were further reviewed.Results · Among the total 5 patients (male:female=1:4),four patients had pure form of PED,and one patient had PED plus epilepsy.Attacks of the proband and his daughter could not be well controlled by carbamazepine or sodium valproate.In addition,three patients showed a remission trend with age advancing.In this family,the SLC2A1 c.C284T (p.S95L) was identified in all 5 patients,but not in 2healthy members.According to the American College of Medical Genetics and Genomics (ACMG) criteria and guideline,the variant SLC2A1 c.C284T (p.S95L) was classified as pathogenic variant.Conclusion · PED is a rare paroxysmal movement disorder with highly phenorypic heterogeneity as well as a remission trend with age advancing.This paper reviews advances in clinical and genetic studies on PED recently,in order to contribute to the clinical diagnosis and appropriate treatment of PED.
3.Inhibitory effect of genistein on growth of hepatoma carcinoma cells SMMC-7721 and influence on apoptosis
Xiaofeng TIAN ; Hong CAO ; Li TIAN
Journal of Jilin University(Medicine Edition) 2006;0(03):-
Objective To investigate the influence of genistein(Gen) on the growth and apoptosis regulation of hepatoma carcinoma cells SMMC-7721.Methods SMMC-7721 cells were divided into three Gen groups treated with different concentrations(5.0,10.0,20.0 mg?L-1) and control group(without Gen).After SMMC-7721 cells were treated for 24 and 48 h,the changes of ultrastructure of cells were observed under electron microscope, MTT was used to detect the inhibitory rate of proliferation of SMMC-7721 cells,flow cytometry was performed to analyze cell phase;immunohistochemistry was used to detect the expression of Caspases-3 protein.Results Compared with control group,chromatin in cellular nucleus became conglomeration or lumping,with chondriosome swelling in Gen groups.With the increasing of Gen concentration,the cellular nucleus became condensed,heterochromatin agglutinating,ground cytoplasm obviously cavitating;with the increase of Gen concentration and prolongation of time,from 24 to 48 h,MTT showed the inhibitory rate of proliferation of SMMC-7721 cells increased in a time-and dose-dependent manner.Flow cytometry analysis showed that with the increase of Gen concentration,the number of cells at G2/M phase increased,the number of cells at S phase decreased and the ratio of G0/G1 also decreased.Immunohistochemistry results showed the expression of protein Caspases-3 increased with the Gen concentration in a dose-dependent manner(P
4.Study on the inlfuening faltors of altivity detemination of four coagulation factors in human prothrombin complex concentrates
Tian TIAN ; Haijun CAO ; Yao CAO ; Zechao HE ; Changqing LI
Chinese Journal of Biochemical Pharmaceutics 2014;(2):17-19
Objective To study the inlfuence factors on detection of activity of four coagulation factors in prothrombin complex concentrates (PCC) by several factors. Methods Using Pharmacopoeia of the People’s Republic of China (2010) as reference, the activity of four coagulation factors in PCC were investigated by choosing different pre-treatments, different diluents, different salt concentration, different standard human plasma and different company reagents. Results The activity of FII, FVII, FX were decreased and FIX was increased in the condition of adding protamine sulfate, and there were no differences of four coagulation factors whether warm bath in 37 for 15 min or not. However, the differences of four coagulation factors were significant by using deficient plasma, saline, distilled water and commercial dilution buffer(P<0.05). The activity of coagulation factor II, X in 1 mol/L salt concentration of PCC were significantly lower than in 0.25 mol/L(P<0.05), while coagulation factor VII, IX were not. The activity of FII, FVII, FIX, and FX were different by using different standard human plasma to make standard curve. The activity of four coagulation factors existed significant difference(P=0.00) by using SIEMENS company reagents and domestic reagents. Conclusion Choosing different pre-treatments, different dilution buffers, salt concentration, standard human plasma and commercial kits will inlfuence the detection result of coagulation factors.
5.Cashmere goat bacterial artificial chromosome recombination and cell transfection system.
Tian HUANG ; Zhongyang CAO ; Yaohui YANG ; Gengsheng CAO
Chinese Journal of Biotechnology 2016;32(3):317-328
The Cashmere goat is mainly used to produce cashmere, which is very popular for its delicate fiber, luscious softness and natural excellent warm property. Keratin associated protein (KAP) and bone morphogenetic protein (BMP) of the Cashmere goat play an important role in the proliferation and development of cashmere fiber follicle cells. Bacterial artificial chromosome containing kap6.3, kap8.1 and bmp4 genes were used to increase the production and quality of Cashmere. First, we constructed bacterial artificial chromosomes by homology recombination. Then Tol2 transposon was inserted into bacterial artificial chromosomes that were then transfected into Cashmere goat fibroblasts by Amaxa Nucleofector technology according to the manufacture's instructions. We successfully constructed the BAC-Tol2 vectors containing target genes. Each vector contained egfp report gene with UBC promoter, Neomycin resistant gene for cell screening and two loxp elements for resistance removing after transfected into cells. The bacterial artificial chromosome-Tol2 vectors showed a high efficiency of transfection that can reach 1% to 6% with a highest efficiency of 10%. We also obtained Cashmere goat fibroblasts integrated exogenous genes (kap6.3, kap8.1 and bmp4) preparing for the clone of Cashmere goat in the future. Our research demonstrates that the insertion of Tol2 transposons into bacterial artificial chromosomes improves the transfection efficiency and accuracy of bacterial artificial chromosome error-free recombination.
Animals
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Bone Morphogenetic Proteins
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genetics
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Chromosomes, Artificial, Bacterial
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DNA Transposable Elements
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Fibroblasts
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Goats
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genetics
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Keratins
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genetics
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Transfection
6.The exosome secreted by B16 cells promoted the proliferation and migration of mesenchymal stromal cell
Lei CAO ; Mifang YANG ; Fengfeng PING ; Tian TIAN ; Lei WANG
Chinese Journal of Microbiology and Immunology 2016;36(1):42-47
Objective To investigate the interactions between melanoma-derived exosomes and the microenvironment.Methods The exosomes were isolated from the culture medium of mouse melanoma cells and then co-cuhured with mesenchymal stromal cells (MSC) after identification.Immunofluorescence assay was performed to observe the exosomes engulfed by MSC.CCK-8 and transwell assays were used to evaluate the proliferation and migration of MSC.Effects of the exosomes on the expression of α-smooth muscle actin (α-SMA) in MSC were analyzed by Western blot.Results Co-culture of MSC with melanoma cell-derived exosomes enhanced the proliferation and migration of MSC as well as the expression of α-SMA.All of the changes mediated by the exosomes could be blocked by using the inhibitor of TGF-β receptor.Conclusion Melanoma cell-derived exosomes enhanced the proliferation and migration of MSC as well as the expression of α-SMA through TGF-β signaling pathway,which provided an advantageous microenvironment for melanoma progression.
7.Case of elephantiasis.
Chinese Acupuncture & Moxibustion 2012;32(11):994-994
8.Treatment of metastatic liver cancer in rat by hepatic artery injection of cytokine recombinant adenoviruses
Qiang HAO ; Jianming TIAN ; Xuetao CAO
Chinese Journal of Radiology 1999;0(10):-
Objective To investigate the therapeutic effects of TNF and IL 2 recombinant adenoviruses via intra arterial injection on metastatic liver cancer in rat model. Methods Recombinant adenoviruses harboring hTNF ? or hIL 2 gene were amplified in 293 cells and subjected to titration by the pathogenetic effects on 293 cell. The rats bearing metastatic liver cancer of Walker 256 breast carcinoma were randomly grouped and administered via gastra intestinal artery with hTNF ? recombinant adenoviruses alone, or hIL 2 recombinant adenoviruses alone, or at the dose of 1.0?10 9 pfu/rat. The therapeutic effects were observed including their survival time. Results The prepared recombinant adenoviruses of hTNF ? and hIL 2 were with the titers of 2.0?10 9 pfu/ml and 2.1?10 9 pfu/ml, respectively. 1.0 ?10 9 pfu hTNF was the proper dose. Administration of hTNF ? or hIL 2 recombinant adenoviruses via hepatic artery could extend the survival time of metastatic liver cancer bearing rats, with the better therapeutic effects achieved by combinatorial administration of these two adenoviruses. Conclusion Arterial administration of adenoviruses may be an effective approach to targeted immunogene therapy for cancer.
9.The in vivo gene transfer efficacy and expression patterns by hepatic artery administration of recombinant adenovirus
Qiang HAO ; Jianming TIAN ; Xuetao CAO
Chinese Journal of Radiology 1999;0(10):-
Objective To investigate the gene transfer efficiency and lasted time in rat organs by hepatic artery injection with LacZ reporter gene recombinant adenoviruses. Methods Seven groups of rats were injected with Ad.LacZ (2?10 9 pfu/ml) and two groups of rats were injected with PBS 1 ml as control separately through gastra intestinal artery, and liver, spleen, lung, and kidney were gotten at 12 hrs, 18 hrs, 72 hrs, 7 day, 14 day, 21 day, and 28day, respectively. X gal staining was used to check up expression of LacZ gene. Results Expression of LacZ gene was detected in liver 12 hrs after injection, but none were done in spleen, lung, and kidney. Up to 21days, LacZ gene expressed in liver, but the gene expression lasted for only 14 days in spleen, lung, and kidney LacZ gene was not detected in the two control groups in all organs at 7 day. Conclusion When recombinant adenovirus was administrated through hepatic artery, the introduced gene expressed preferentially in liver. This result was the basis of intraarterial administration of cytokines gene to treat liver tumor.
10.Comparison of clinical efficacy between continuous renal replacement therapy and intermittent haemodialysis for the treatment of sepsis-induced acute kidney injury
Tian DAI ; Shuhua CAO ; Xiaolong YANG
Chinese Critical Care Medicine 2016;28(3):277-280
Objective To compare the clinical effects between continuous renal replacement therapy (CRRT) and intermittent haemodialysis (IHD) for the treatment of sepsis-induced acute kidney injury (AKI). Methods A prospective study was conducted. Seventy-three patients with sepsis-induced AKI admitted to the intensive care units (ICUs) of Tianjin Hospital and Tianjin First Center Hospital from January to December in 2014 were enrolled. They were randomly divided into two groups: CRRT group (n = 35) and IHD group (n = 38). Data were recorded for the patients in two groups before treatment, including acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score, mean arterial pressure (MAP), urine volume, and the levels of C-reactive protein (CRP) and serum creatinine (SCr) before and 1 week after treatment, the time of recovery of urine volume, the length of ICU stay, the duration of organ support, and the incidence of cardiovascular events. Results There was no statistically significant difference in APACHE Ⅱ scores (21.63±2.46 vs. 21.34±2.46), MAP [mmHg (1 mmHg = 0.133 kPa): 71.26±10.70 vs. 75.74±15.17], urine volume (mL: 404.00±79.13 vs. 438.97±87.17), CRP (mg/L: 100.94±14.73 vs. 95.17±27.03), and SCr (μmol/L: 394.02± 50.26 vs. 390.47±54.42) before treatment between CRRT group and IHD group (all P > 0.05). One week after treatment, compared to the IHD group, CRRT could dramatically reduce the levels of CRP (mg/L: 41.05±10.15 vs. 60.21±14.78, t = 6.401, P < 0.001), SCr (μmol/L: 185.97±65.48 vs. 232.02±71.93, t = 2.862, P = 0.006), urine output recovery time (days: 7.94±3.06 vs. 11.08±3.71, t = 3.923, P < 0.001), the length of ICU stay (days: 9.54±3.39 vs. 13.42±3.89, t = 4.521, P < 0.001), organ support time (days: 3.23±2.70 vs. 6.34±3.36, t = 4.343, P < 0.001), and the incidence of cardiovascular events [23.53% (8/35) vs. 39.47% (15/38), χ2 = 5.509, P = 0.025]. Conclusion Compared to IHD, CRRT can more efficiently help patients with sepsis-induced AKI in removing excessive water, metabolic waste, and lower the levels of pro-inflammatory cytokines, maintain homeostasis of the internal environment, lower the adverse effects on cardiovascular system, so that it significantly improve the prognosis of patients, shorten the time of organ support and the length of ICU stay.