1.Effect of beraprost sodium on HPH rats and expression of oxygen-sensitive Kv channels in pulmonary artery smooth muscle
Hua TIAN ; Jie GENG ; Jizhao DENG ; Fenling FAN ; Hongyan TIAN
Journal of Xi'an Jiaotong University(Medical Sciences) 2017;38(4):474-478,486
Objective To explore the effect of beraprost sodium (BPS) on hypoxia-induced pulmonary artery hypertension (HPH) in rats and the expression of oxygen-sensitive Kv channels in pulmonary artery smooth muscle (PASM).Methods The HPH model of rats was established by exposing rats to low-pressure and low-oxygen cabin which was auto-modulated for 8h every day.Rats in the BPS group were given an intragastric administration of BPS [300 μg/(kg · d)],while those in the control group and HPH group were given an intragastric administration of 3 ml/kg of 0.9% saline.After 4 weeks,the mean pulmonary artery pressure (mPAP) was measured and right heart ventricle hypertrophy index (RVHI) was calculated;pulmonary artery remodeling was evaluated by HE staining;the expressions of Kv 1.2,Kv 1.5 and Kv2.1 in the pulmonary artery were detected by Real-time PCR and Western blot.Results The HPH model was successfully established in rats exposed to chronic hypoxia for 4 weeks.Compared with those in HPH group,mPAP,RVHI and pulmonary artery remodeling were decreased in BPS group [mPAP:(13.48±2.18)mmHg vs.(23.87±2.23)mmHg vs.(17.09±1.20)mmHg;RVHI:0.28±0.02 vs.0.46±0.03 vs.0.36±0.04;% area of medial smooth muscle:35.72±6.58 vs.68.52±5.64 vs.46.58±8.43;P<0.05],and the mRNA and protein expressions of Kv 1.2,Kv 1.5 and Kv 2.1 were increased (relative protein expression level:Kv1.2,0.78±0.10 vs.0.15±0.03 vs.0.57±0.13;Kv1.5,0.61±0.10 vs.0.31±0.05 vs.0.59±0.13;Kv2.1,0.29±0.05 vs.0.10±0.02 vs.0.28±0.07;P<0.05).Conclusion BPS can improve pulmonary arterial hypertension induced by hypoxia,and upregulate the decreased mRNA and protein expressions of Kv 1.2,Kv 1.5 and Kv 2.1 in pulmonary artery.
2.The current situation and strategies of snake antivenomimmunoglobulins research and development.
Jing TIAN ; Quan-shui FAN ; Ying ZHENG
Acta Pharmaceutica Sinica 2015;50(12):1573-1580
Snake antivenomimmunoglobulins are considered to be the most efficient drugs in snake envenomings. Most snake antivenomimmunoglobulins all over the world are still prepared by fragmentation of polyclonal antibodies isolated from hyper-immunized horse serum till now. In this review, we retrospect the history of snake antivenomimmunoglobulins, analysis the present situation and pay the close attention on the key technological links in the process of research and manufacturing, such as properties of IgG and its fragments, selection and preparation of immunogen, optimization of immunization schedule and protein isolation and purification, which can be available for the reference in the research and development of snake antivenom.
Animals
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Antivenins
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pharmacology
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Humans
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Immunoglobulin G
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pharmacology
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Snake Bites
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drug therapy
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Snakes
3.Progresses in studies on allogeneic hematopoietic stem cell transplantation in children with acquired severe aplastic anemia.
Chinese Journal of Pediatrics 2013;51(5):345-348
Anemia, Aplastic
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mortality
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pathology
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therapy
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Child
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Disease-Free Survival
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Graft vs Host Disease
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prevention & control
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Hematopoietic Stem Cell Transplantation
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methods
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Humans
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Immunosuppressive Agents
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therapeutic use
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Risk Factors
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Severity of Illness Index
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Tissue Donors
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Transplantation Conditioning
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methods
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Transplantation, Homologous
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Unrelated Donors
5.Apoptosis and expression of PCNA and distribution of TUNEl positive cells in human vulnerable atherosclerotic plaque
Changqing FAN ; Ye TIAN ; Wei YANG
Chinese Journal of Practical Internal Medicine 2001;0(02):-
ObjectiveTo investigate the roles of apoptosis and express io n of PCNA in human vulnerable atherosclerotic plaque.MethodsAp optosis was detected by TUNEL.Expessions of PCNA were detected in samples of vul nerable plagues From 23 cases of coronary heart disease.by SP immunohistochemica l technique.ResultsThe expession level of PCNA in the smooth m uscle cells (SMCs) on fibrous cap region was significantly elevated than that of the other three regions.The expession level of PCNA in the inflammatory cells ( ICs) on fibrous cap and shoulder region were significantly elevated than that of the other two regions.The amount of apoptosis in the SMCs on lipid-rich core wa s much more than that of the other three regions.The amount of apoptosis in the ICs on lipid-rich core was more than the other three regions.The expression leve ls of PCNA in fibrous cap and shoulder were significantly higher than TUNEL posi tive cell (P
6.Synergistic Enhancement of Cytotoxicity of Mitomycin C on Gastric Cancer Cells in vitro by Tumor Necrosis Factor a and Interferon ?
Fengchi TIAN ; Xueyong ZHANG ; Daiming FAN
Chinese Journal of Cancer Biotherapy 1995;0(03):-
The aim is to investigate the effects of tumornecrosis factor ?(TNF?0 and interferon a(IFN?) on the cytotoxicity of mitomycin C(MMC) in vitro. The cytotoxicity of the three agents against gastric cancer cell line and freshly isolated gastric cancer cells were determined by the MTT assay. TNF? and/or IFN? (each 0~5000U/ml) showed cytotoxicity effects only on one of ten cell cultures dose-dependently; the combination resulted in a more significant effect. The combination of MMC with TNF? (30U/ml) or IFN?(l00U/ml) could enhance the cytotoxicity of MMC on two of three cell cultures synergisticly. When three agents were used together, the cytotoxicity of MMC against the ten cell cultures was enhanced synergisticly. TNF? and IFN? could enhance the cytotoxicity of MMC synergisticly, and this may make the cancer chemotherapy more effective.
7.Effects of Qi-Boosting Toxin-Resolving Formula on CD4+CD25+ Regulatory T Cells and Th17 Cells of Patients with Middle to Late Staged Nasopharyngeal Carcinoma
Zhichao JIANG ; Daofa TIAN ; Jingying FAN
Chinese Journal of Information on Traditional Chinese Medicine 2014;(2):23-26,31
Objective To investigate the effects of Qi-Boosting Toxin-Resolving Formula (QBTRF) on CD4+CD25+ regulatory T cells and Th17 cells of patients with middle to late staged nasopharyngeal carcinoma (NPC). Methods Flow cytometry was performed to detect the ratio of CD4+CD25+ regulatory T cells and Th17 cells in the peripheral blood mononuclear cells (PMBC) among 18 patients with middle to late stage of NPC treated by QBTRF added to conventional therapy (treatment group), Foxp3 mRNA and ROR-γt mRNA in PMBC was determined by RT-PCR technique. Furthermore, serum levels of IL-6 and TGF-β were assayed by ELISA. Meanwhile, 15 patients with NPC treated by conventional therapy were taken as the control group. Results The ratio of CD4+CD25+ regulatory T cells to the total CD4+ T cells and the transcriptional level of Foxp3 mRNA in PMBC were significantly lower in treatment group than that of control group (P<0.05), the ratio of Th17 cells to the total CD4+T cells and the transcriptional level of ROR-γt mRNA in PMBC were significantly higher in treatment group than that of control group (P<0.05). However, the serum level of IL-6 was obviously higher in treatment group than that of control group (P<0.05), and the serum leve of TGF-βwas obviously lower in treatment group than that of control group (P<0.05). Conclusion QBTRF can significantly affect the number ratio and functional activity of CD4+CD25+ regulatory T cells and enforce the differentiation of Th17 cells among patients with middle to late staged NPC, which it may be reversed the immune tolerance of NPC through regulating the level of IL-6 and TGF-β.
8.The short term follow-up study of shoulder hemi-arthroplasty
Fan LIU ; Liang TANG ; Tian MAO
Chinese Journal of Orthopaedics 1999;0(07):-
Objective To evaluate the short term results of shoulder hemi-arthroplasty. Methods Twenty-one patients with comminuted intra-articular fracture of proximal humerus underwent Neer type (Link, Germany) humeral head replacement from August 2000 to January 2004, which involved 5 males and 16 females,with an average age of 70 years (range, 64-76 years). According to AO M?ller classification, 3 cases were of C1 type, 8 C2 type, and 10 C3 type. The important steps of the procedure included insertion of the prosthesis in 40 degree retroversion and reconstruction of the rotary cuff of shoulder. The rehabilitation training course was divided into three stages. The patients were trained passively in the first stage (1 day-6 weeks); The muscles around the shoulder undertook isometric contraction once the bone fusion of greater and lesser tuberosity were obtained on X-ray for the second stage (7 -12 weeks); The improvement of the muscle strength and ROM of the shoulder were targeted throughout the third stage (13-24 weeks). All cases were followed up with Neer's scoring system comprising the symptoms, function, range of motion (ROM) of shoulder, and position of the prosthesis. Results The follow-up was performed twice a month within one year postoperatively, and one time three months after that.The mean duration of follow-up was 34.5 months (range, 14 to 55 months). Only slight pain was complained occasionally in 4 cases. No difficulty in active daily life was found in 21 cases .The average ROM of shoulder was flexion 115?(range, 65?-165?),extension 42.5?(15?-70?),abduction 85?(50?-120?), adduction 32.5?(20?-45?), external rotation 40?(20?-60?),and internal rotation 50?(10?-90?). No dislocation or subluxation of shoulder occurred in all cases. Assessed by Neer's scoring system, 8 cases were rated as excellent, 9 as good, 3 as fair and 1 as poor. The excellent and good rate was 80.9%. The patients' satisfactory rate was 90%. Conclusion The shoulder hemi-arthroplasty show good short term results for comminuted intra-articular fractures of the proximal humerus. The key points to improve the postoperative results are presented as follows: less invasive technique, appropriate retroversion angle for humeral head prosthesis, very solid reconstruction for rotary cuff of shoulder, and correct consecutive physical therapy after surgery.
9.The current situation and strategies of snake antivenomimmunoglobulins research and development.
Jing TIAN ; Quanshui FAN ; Ying ZHENG
Acta Pharmaceutica Sinica 2015;50(12):1573-80
Snake antivenomimmunoglobulins are considered to be the most efficient drugs in snake envenomings. Most snake antivenomimmunoglobulins all over the world are still prepared by fragmentation of polyclonal antibodies isolated from hyper-immunized horse serum till now. In this review, we retrospect the history of snake antivenomimmunoglobulins, analysis the present situation and pay the close attention on the key technological links in the process of research and manufacturing, such as properties of IgG and its fragments, selection and preparation of immunogen, optimization of immunization schedule and protein isolation and purification, which can be available for the reference in the research and development of snake antivenom.
10.Construction of specific interference vector and lentiviral packaging for SMAC gene in mice
Fan WU ; Zaifeng YI ; Deying TIAN
Journal of Clinical Hepatology 2014;31(5):442-445
Objective To construct and identify the specific interference vector for second mitochondria -derived activator of caspase (SMAC)gene in mice and perform lentiviral packaging.Methods According to the SMAC gene sequences in mice,three small interfering RNAs (siRNAs)(siRNA1,siRNA2,and siRNA3)and one negative control sequence (siRNAn)were designed and synthesized,and mouse hepatocytes were transfected with the above siRNAs using Lipofectamine 2000.The inhibitory effects of these siRNAs on SMAC mR-NA expression were evaluated by real-time PCR,and the optimal siRNA was screened out accordingly.Oligo DNA was synthesized based on the optimal siRNA and then connected to GV1 15 vector to obtain recombinant interference plasmid.The candidate clones were identified by PCR and sequenced.The recombinant interference plasmid,as well as pHelper 1.0 and pHelper 2.0,was used to transfect 293T cells for lentivirus packaging.Then,cell supernatants were collected and concentrated,and the lentiviral titer was determined by gradient dilution. Comparison between groups was made by analysis of variance,and multiple comparisons were made by SNK-q test.Results The three siRNAs had different inhibitory effects on SMAC mRNA expression in hepatocytes;siRNA1 showed the strongest inhibitory effect,with an inhibition efficiency of 70.3%.Based on siRNA1 ,the oligo DNA was successfully synthesized and correctly connected to the lentiviral vec-tor,as proved by sequencing.The lentiviral titer was 6 ×108 TU/ml,as determined by gradient dilution.Conclusion The recombinant lentivirus that inhibits SMAC expression in mice has been constructed successfully,laying the foundation for further studies on the role of SMAC gene in liver failure.