Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 17 de 17
Filter
1.
Journal of Experimental Hematology ; (6): 433-438, 2019.
Article in Chinese | WPRIM | ID: wpr-774297

ABSTRACT

OBJECTIVE@#To investigate the effect of steadily down-regulating the expression of calreticulin (CALR) on the invasion of natural killer/T-cell lymphoma SNK6 cells, and explore its possible mechanism.@*METHODS@#The sequences of specific short hairpin RNA (shRNA) targeting on human CALR were designed, and were inserted into pLKO.1-puro lentivirus vector, and the reconbinant lentivirus vector was obtained; the lentivirus particles were backed by three-plasmid system and transfected into SNK6 cells, the SNK6 cells stably down-regulating the CALR expression were sercened by puromytain, the CALR-silencing effect was verified by real-time PCR and Western blot. CCK-8 assay was used to evaluate the cell viability, The transwell invasion assays was used to analyse invasion of SNK6 cells. The mRNA expression of Calreticulin, MMP2, MMP9 and VEGF was determined by real time PCR, the protein expression of Calreticulin and GAPDH was analyzed by Western blot.@*RESULTS@#The recombinant lentiviral vector pLKO.1-puro-shCALR was successfully constructed, packed into the lentivirus, then the SNK6 cells stably down-regulating Calreticulin expression was obtained. When Calreticulin was down-rengulated in SNK6 cells, the proliferation rate was reduced and the invasion ability was decreased; the mRNA levels of VEGF and MMP-2/9 also were reduced.@*CONCLUSION@#The stable down-regnlation of CALR expression in SNK6 cells can attenuate the imvasiveness of SNK6 cells, which maybe related with transcriptional decrease of MMP2, MMP9 and VEGF.


Subject(s)
Humans , Calreticulin , Genetics , Metabolism , Cell Line, Tumor , Cell Proliferation , Down-Regulation , Genetic Vectors , Lentivirus , Matrix Metalloproteinase 2 , Matrix Metalloproteinase 9 , RNA Interference , RNA, Small Interfering , Transfection , Vascular Endothelial Growth Factor A
2.
Journal of Experimental Hematology ; (6): 1397-1405, 2017.
Article in Chinese | WPRIM | ID: wpr-301717

ABSTRACT

<p><b>OBJECTIVE</b>To explore the effects of mTOR inhibitor rapamycin on proliferation, cell cycle and apoptosis of Burkitt's lymphoma cell line Raji and CA46 cells and its mechanism, so as to provide the experimental evidence for a therapeutic target of Burkitt's lymphoma.</p><p><b>METHODS</b>3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide(MTT) assay was performed to assess the inhibitory effect of rapamycin on proliferation of Burkitt's lymphoma cell line Raji and CA46 cells. The cell cycle distribution of Raji and CA46 cells was analyzed by flow cytometry with propidium iodide(PI) single staining. The cell apoptosis of Raji and CA46 cells was analyzed by flow cytometry with FITC Annexin V+PI double staining. The expressions of RPS6, p-RPS6, survivin and caspase-3 proteins were detected by Western blot after treating with rapamycin.</p><p><b>RESULTS</b>Rapamycin markedly inhibited the proliferation of both Raji and CA46 cells in a time- and concentration-dependent manners, showing good biological activity, the cell proliferation inhibition rate reached about 20% after treatment with 1 nmol/L rapamycin. After treatment with different concentrations of rapamycin for 24 and 48 hours, the proportion of both cells in G/Gphase in the treated groups was significantly increased in a time- and concentration-dependent manners in comparison with the solvent control group. With regard to the cells in S and G/M phase, the decreased population was accompanied by the increase of G/Gphase cells. After treatment with 100 nmol/L rapamycin for 48 hours, both Raji and CA46 cells demonstrated an apparent apoptosis,especially late apoptosis by flow cytometry with Annexin V+PI staining. After treatment with rapamycin, the expression of p-RPS6 and survivin of Raji and CA46 cells was obviously down-regulated, the expression of caspase-3 was obviously up-regulated in a time- and dose-dependent manners. However, rapamycin did not obviously affect the expression of RPS6.</p><p><b>CONCLUSION</b>The rapamycin can effectively inhibit cell proliferation, arrest Raji and CA46 cells in G/Gphase, and this effect associates with inhibiting the activation of mTOR/RPS6 signal pathway through down-regulating the expression of phosphorylated RPS6, i.e. mTOR downstream signal pathway. It also can induce apoptosis by down-regulating the expression of anti-apoptotic protein survivin and activating the intrinsic pro-apoptotic protein caspase-3.</p>

3.
Journal of Experimental Hematology ; (6): 921-925, 2017.
Article in Chinese | WPRIM | ID: wpr-271894

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effect of 2 kinds of red blood cells (RBC) on the laboratorial indexes and therapeutic efficacy of patients with autoimmune hemolytic anemia (AIHA).</p><p><b>METHODS</b>The clinical data of 120 patients with AIHA from June 2015 to June 2016 were analyzed retrospectively. These 120 patients were divided into A goup and B group. The patients in A group (60 cases) were infused with washed RBC, while the patients in B group (60 cases) were infused with WBC-deplated RBC. The changes of laboratotial indexes, clinical symptoms and incidence of adverse reactions in 2 groups before treatment and at 24 hourse after treatment as well as the therapeutic efficacy and improvement status of clinical symptoms were compared.</p><p><b>RESULTS</b>The RBC count and Hb level in 2 groups after treating for 1 day were significantly higher than those before treatment (P<0.01), while the TBIL level and reticulocyte (Ret) count were significantly lower than those before treatment (P<0.01). However, the RBC, Hb, TBIL levels and Ret count in 2 groups before and after treatment showed no statistical difference (P>0.05); the improvement of clinical symptoms, complex therapeutic efficacy and incidence of adverse reactions in 2 groups after treatment also showed no significantly difference (P>0.05).</p><p><b>CONCLUSION</b>Both washed and leukocyte-deplated RBC can alleviate the anemia in patients with AIHA in short-time, but the leucocyte-deplated RBC is more expensive, suggesting that the wased RBC is more practical for treatment of AIHA patients.</p>

4.
Journal of Central South University(Medical Sciences) ; (12): 1163-1168, 2017.
Article in Chinese | WPRIM | ID: wpr-669234

ABSTRACT

Objective:To explore the risk factors,preventive measures and therapeutic methods for bronchopleural fistula (BPF) after lung resections.Methods:A restrospective analysis for 11 patients with BPF after pneumonectomy from April 2012 to June 2016 in Department of Thoracic Surgery,Xiangya Hospital,Central South University was performed.Their clinical characteristics,treatment and prognosis were analyzed,and the risk factors and effective therapeutic strategies were summarized.Results:Among the 11 patients with BPF,10 cases were cured finally,and 1 case with conservative treatment was dead.The total mortality rate was 9.09%.The 10 patients treated with positive measures were all cured,including 5 cases with pulmonary lobectomy and pneumonectomy,4 cases with amplatzer and covered stent,and 1 case with fibrin glue.One case with conservative treatment was dead because of respiratory failure.Conclusion:It is important to intervene BPF as early as possible.Fibrin glue via bronchoscope for tiny BPF after lung resection is preferred to be considered.We recommend to take early positive operation (pulmonary lobectomy and pneumonectomy) after pulmonary resection if the BPF cannot be cured via bronchoscope whereas the patients' condition is allowed.The amplatzer or covered stent should be considered first for the patient with BPF after pneumonectomy.

5.
Journal of Shanghai Jiaotong University(Medical Science) ; (12): 1658-1663, 2017.
Article in Chinese | WPRIM | ID: wpr-665379

ABSTRACT

Objective·To explore the difference between multi-artery-graft off-pump coronary artery bypass grafting (OPCABG) and single-artery-graft OPCABG on left main coronary artery or multivessel disease with propensity score matching. Methods·A total of 1578 patients with left main coronary artery or multivessel disease underwent isolated OPCABG were selected in Ruijin Hospital from January 2012 to September 2016. The propensity score methodology was used to obtain risk-adjusted outcome. Kaplan-Meier analysis was applied for estimation of freedom from major adverse cardiac and cerebrovascular events (MACCE) and readmission for heart disease. Independent predictor of MACCE were assessed by Cox regression analysis. Results·The average follow-up time was 28 months (7-55 months). There was no statistical difference in short-term clinical endpoints in hospital. In the follow-up results, multi-artery-graft OPCABG patients had statistical improvement in readmission for heart disease (2.7% vs 12.7%, P=0.023), CCS class (1.2±0.4 vs 1.4±0.6, P=0.020) and patency rate of grafts in 1 year after operation (95.8% vs 85.9%, P=0.025), compared with single-artery-graft OPCABG. There was no statistical difference in other endpoints. There was statistical improvement for multi-artery-graft OPCABG patients in freedom from readmission for heart disease (P=0.028). Female was an independent predictor of MACCE (95% CI 0.117-0.906, P=0.032). Conclusion·Multi-artery-graft OPCABG appears to be safe and with good patency of grafts and clinical outcomes in treating left main coronary artery or multivessel disease. The follow up of female patients should be paid more attention.

6.
Chinese Journal of Interventional Cardiology ; (4): 549-552, 2014.
Article in Chinese | WPRIM | ID: wpr-453802

ABSTRACT

Objective This study intends to explore the impacts of the establishment of chest pain center(CPC) on the door-to-balloon(D-to-B) time in patients with ST-elevation myocardial infarction (STEMI) by different transfer ways to hospital. Methods A regular CPC and a regional cooperative network were established based on the pre-hospital transmitted real-time 12-lead electrocardiogram system. The STEMI patients were divided into the following three groups by the different transfer ways to hospital before and after the establishment of chest pain center:self-referral groups (group A1, n=52, and group A2, n=65), EMS (emergency medical service ) groups (group B1, n=31, and group B2, n=92) and transfer PCI groups (group C1, n=23, and group C2, n=552). The mean D-to-B time and the rate of D-to-B below 90 minutes were compared between before and after the establishment of CPC and the reasons of reperfusion delay were analyzed. Results There were no statistical differences of the average D-to-B time [(123±78) min vs.(140±123)min, P > 0.05] and the rate of D-to-B time below 90 min (44.2%vs. 46.2%) between group A1 and group A2. The average D-to-B time was significantly shortened in group B2 [(89±66)min] while compared with that in group B1 [(155±115)min, P<0.05] and the rate of D-to-B time below 90 min was remarkably elevated in group B2 compared with that of group B1 (69.6%vs. 32.3%, P<0.05). The average D-to-B time was significant shorter in group C2 than in group C1 [(77±43)min vs. (337±662)min, P<0.05] and the rate of D-to-B time below 90 min was remarkable higher in group C2 than in group C1 (75.7%vs. 21.7%, P<0.05). The longer D-to-B time in self-referral groups was mainly due to the delay of getting informed consent before PCI when occupied catheterization laboratory was the major cause of reperfusion delay in EMS groups and transfer PCI groups. Conclusions The establishment of CPC may significantly shorten the D-to-B time and increase the rate of D-to-B time below 90 min for these patients admitted by EMS and transferred from non-PCI hospitals. However, the pathway for the self-referral patients should be further modified.

7.
Chinese Journal of Emergency Medicine ; (12): 1147-1152, 2013.
Article in Chinese | WPRIM | ID: wpr-442311

ABSTRACT

Objective To study the efficiency of tele-consultation on Internet with transmitting realtime 12-lead ECG carried out by the Chest Pain Center evaluated by the length of time required for the emergency percutaneous coronary intervention (PCI) in patients with ST segment elevation myocardial infarction (STEMI).Methods A total of 435 STEMI patients treated by emergency PCI were divided into the group A (n =98,admitted in 2010),group B (n =114,admitted in 2011) and group C (n =223,admitted in 2012).Data were collected before (2010) and after establishment of the Chest Pain Center (2011 to 2012) including the length of time elapsed from onset of symptoms to the first medical contact (FMC),the length of time required from FMC to the intra-aortic balloon inflated (FMC-2B) and the length of time required from entering the gate of hospital to the intra-aortic balloon inflated (D-2B).Measure data were described with non-normal median and interquartile intervals.Comparisons were made among groups with rank sum test.Results The median time of D2B of three groups were 107,78 and 59 mins in groups A,B and C,respectively.The differences in D2B among three groups were significant (P =0.000).The time of the D2B was shortened significantly because of the patients transferred to the hospital with a variety of ways (P =0.008).However,the length of D2B time was not significantly changed (P =0.846) when patients came to the hospital all on themselves.The median times from symptom onset to FMC in the group A,group B and group C were 112,62 and 78 mins.and the differences among three groups were not statistically significant (P =0.368).The median times of FMC2B in three groups were 287.0,313.5 and 421.8 mins,respectively,and there were no significant differences (P =0.135).Conclusions The establishment of the Chest Pain Center and Internet of things can effectively shorten the duration of D2B in STEMI patients.However,the reduction of time length from the symptom onset to reperfusion must rely on the coordination between communities and health care system.

8.
Chinese Journal of Emergency Medicine ; (12): 669-673, 2013.
Article in Chinese | WPRIM | ID: wpr-437919

ABSTRACT

Objective To evaluate the pre-hospital diagnostic reliability of real-time tele-transmission of 12-lead electrocardiogram of patients with ST-segment elevated acute myocardial infarction (STEMI).Methods The 12-lead electrocardiogram was simultaneously recorded with real-time tele-transmission system and a conventional electrocardiograph in 40 STEMI cases.The width and amplitude of each wave,the deviated amplitude of ST-segment in the same leads were compared by t-test and rank-sum test.Results There were no statistical differences in the width and amplitude of P wave,QRS wave and t wave as well as the deviated altitude of ST-segment between the two separate electrocardiographs (P >0.05).There was a significant positive correlation between the two ECG devices in respect of ST-segment elevated altitude (r =0.912,P =0.000).The differential ability of ST-segment elevation between two separate ECG devices kept highly consistent (Kappa value:0.976).Conclusions Real-time tele-transmission of 12-lead electrocardiogram is reliable for the pre-hospital diagnosis of STEMI.

9.
Chinese Journal of Surgery ; (12): 83-85, 2009.
Article in Chinese | WPRIM | ID: wpr-238953

ABSTRACT

<p><b>OBJECTIVE</b>To study the therapeutic effect of splenic autotransplantation combined with lower esophagus transaction anastomosis in the treatment of liver cirrhosis induced portal hypertension.</p><p><b>METHODS</b>Thirty-six patients admitted from January 2003 to December 2006 were randomly divided into splenic autotransplantation group undergoing splenic autotransplantation after splenectomy combined with lower esophagus transaction anastomosis, and splenectomy group only undergoing splenectomy combined with lower esophagus transaction anastomosis. The general conduction, splenic scanning, liver function, and the level of serum Tuftsin and IgM of each patient were observed before and after operation.</p><p><b>RESULTS</b>The levels of Tuftsin and IgM in splenic autotransplantation group were significant higher than that of splenectomy group 2 months after the operation, and the liver function showed no significant difference between these two groups. Splenic tissue was detected in the retroperitoneal space by 99mTc-DRBC 2 months after operation.</p><p><b>CONCLUSIONS</b>Splenic autotransplantation combined with lower esophagus transaction anastomosis is a safe and effective treatment strategy for patients with liver cirrhosis induced portal hypertension, and the spleen tissue transplanted into the retroperitoneal space can partially preserve the immune function.</p>


Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Young Adult , Anastomosis, Surgical , Esophagus , General Surgery , Hypertension, Portal , General Surgery , Liver Cirrhosis , Spleen , Transplantation , Transplantation, Autologous , Treatment Outcome
10.
Journal of Experimental Hematology ; (6): 929-932, 2009.
Article in Chinese | WPRIM | ID: wpr-333994

ABSTRACT

The purpose of this study was to investigate the lethal effect of cytotoxic lymphocytes against U266 cells induced by DCs pulsed with multiple myeloma (MM) U266 lysate and transfected with GM-CSF recombinant adenovirus. The cytotoxic lymphocytes against U266 cells were induced by culturing with DCs, which pulsed with MM U266 antigens and transfected with GM-CSF recombinant adenovirus. The effect of cytotoxic lymphocytes against U266 cells were measured by LDH release detection. Experiments were divided into 3 groups: N-DC group as control in which DCs were normal; U-DC group in which DCs were pulsed by U266 soluble antigen, and G-U-DC group in which DCs were stimulated by U266 soluble antigen and GM-CSF transfected with Ad-CMV. The results showed that there was significant difference on killing rate against U266 cells between 3 groups (F = 10.939, p < 0.05). The killing rate of G-U-DC group was the highest (p < 0.001), and killing rate of U-DC group was higher than that of N-DC group (p < 0.001). It is concluded that the cytotoxic lymphocytes against U266 cells can be induced by DCs pulsed with U266 lysate, and the lethal effect of CTLs can be enhanced when DCs transfected by recombinant adenovirus with exogenous gene GM-CSF.


Subject(s)
Humans , Adenoviridae , Genetics , Antigens, Neoplasm , Genetics , Allergy and Immunology , Cancer Vaccines , Allergy and Immunology , Cell Line, Tumor , Cytotoxicity, Immunologic , Dendritic Cells , Allergy and Immunology , Granulocyte-Macrophage Colony-Stimulating Factor , Genetics , Allergy and Immunology , Multiple Myeloma , Recombinant Proteins , T-Lymphocytes, Cytotoxic , Allergy and Immunology , Transfection
11.
China Journal of Orthopaedics and Traumatology ; (12): 264-266, 2008.
Article in Chinese | WPRIM | ID: wpr-307037

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the clinical value of T shape approach in the treatment of proximal tibial fractures.</p><p><b>METHODS</b>One handrend and thirteen patients of proximal tibial fractures were randomly divided into two groups. Group A: 62 cases underwent the traditional exposure approach. According to Schatzker classification,the cases of II to VI type was 25, 10, 16, 6, 5 respectively. Group B:51 cases underwent T shape approach ahead of knee joint, the cases of II to VI type was 21, 8, 13, 5, 4 respectively. All data were analyzed by SPSS 10.0 to compare operation time, blood loss, duration of hospitalization, healing time, the time of osseous union and complications after operation.</p><p><b>RESULTS</b>Sixty patients in group A and 50 patients in group B were followed-up from 12 to 24 months. (1) Operation time:group B was longer than A (P < 0.01). (2) Mean blood loss and duration of hospitalization was the same. (3) Clinical healing time:group B was shorter. (4) Mean time of osseous union: 48 group B was shorter. Function of knee: group B was better than group A. (Complication: group B was less than group A.</p><p><b>CONCLUSION</b>As compared with traditional exposure approach, T shape approach of knee joint had advantages of small scar, fewer complications, faster union of fracture and earlier recovery of joint function. The approach is valuable for the treatment of proximal tibial fractures.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Fracture Fixation , Methods , Knee Joint , Postoperative Complications , Epidemiology , Tibial Fractures , General Surgery
12.
Chinese Medical Journal ; (24): 2180-2184, 2008.
Article in English | WPRIM | ID: wpr-350778

ABSTRACT

<p><b>BACKGROUND</b>Survivin is a rather specific gene in tumor tissue. We transfected dendritic cells (DCs) with recombinant adenovirus (Ad) containing survivin gene and granulocyte-macrophage colony-stimulating factor (GM-CSF) gene and tested the inducing effect of the transfected DCs on cytotoxic T lymphocytes (CTL) to kill leukemic cells.</p><p><b>METHODS</b>After derived from the peripheral, DCs was assayed by mixed leukocyte reaction (MLR) tests. Lactate dehydrogenase (LDH) release test was used to evaluate cytotoxicity of CTL.</p><p><b>RESULTS</b>Expression of survivin in transfected DCs was confirmed by Western blotting analysis. GM-CSF expression was confirmed by enzyme-linked immunosorbent assay (ELISA). In MLR assay, DCs coinfected with Ad-survivin and Ad-GM-CSF induced higher allogeneic lymphocyte reaction than control DCs at ratios of 1:5, 1:10, 1:50 and 1:100. DCs coinfected with Ad-survivin and Ad-GM-CSF had much higher activity of CTL to HL-60 cells than DCs infected with Ad-survivin only, Ad-GM-CSF only, or control DCs. Levels of interleukin-12 (IL-12) and interferon gamma (IFN-gamma) in lymphocyte supernatants containing DCs coinfected with Ad-survivin and Ad-GM-CSF were significantly higher than those in the control group.</p><p><b>CONCLUSION</b>DCs coinfected with Ad-survivin and Ad-GM-CSF induce much higher anti-leukemic response in vitro than those infected with either factor. Therefore, adenovirus vectors containing survivin and GM-CSF genes may be promising vaccine candidates for leukemia therapy.</p>


Subject(s)
Humans , Adenoviridae , Genetics , Cytotoxicity, Immunologic , Dendritic Cells , Physiology , Genetic Therapy , Granulocyte-Macrophage Colony-Stimulating Factor , Genetics , HL-60 Cells , Inhibitor of Apoptosis Proteins , Interferon-gamma , Interleukin-12 , Leukemia , Therapeutics , Lymphocyte Activation , Microtubule-Associated Proteins , Genetics , Neoplasm Proteins , Genetics , T-Lymphocytes, Cytotoxic , Allergy and Immunology , Transfection
13.
Acta Academiae Medicinae Sinicae ; (6): 469-473, 2008.
Article in Chinese | WPRIM | ID: wpr-270667

ABSTRACT

<p><b>OBJECTIVE</b>To explore the expression profile of microRNAs during the course of embryonic stem cells differentiation towards hepatocytes induced by sodium butyrate.</p><p><b>METHODS</b>Total RNA was extracted from embryonic stem cells on day 0, 6, and 9 during cell differentiation, and microRNA was isolated from the total RNA. Microarray analysis of microRNA expression was performed to detect the different expression levels of microRNA among the indicated time points (day 0, 6, and 9).</p><p><b>RESULTS</b>Compared with the microRNA expression level on day 0 of cell differentiation, 17 different microRNAs exhibited higher expressions both on day 6 and day 9. Twenty-two and 27 microRNA demonstrated lower expressions on day 6 and day 9, respectively. Further analysis revealed that 15 microRNA among the above microRNAs with significant differential expression may keep close interation with histone deacetylase.</p><p><b>CONCLUSION</b>During the course of embryonic stem cells differentiation towards hepatocytes induced by sodium butyrate, histone deacetylase and its relevant microRNAs may play important roles in cell differentiation.</p>


Subject(s)
Animals , Mice , Butyrates , Pharmacology , Cell Differentiation , Cells, Cultured , Embryonic Stem Cells , Cell Biology , Metabolism , Gene Expression , Hepatocytes , Cell Biology , Metabolism , MicroRNAs , Genetics , Metabolism
14.
Journal of Forensic Medicine ; (6): 330-332, 2008.
Article in Chinese | WPRIM | ID: wpr-983404

ABSTRACT

OBJECTIVE@#To screen the AFLP primers with good diversity to distinguish various species of Cannabis.@*METHODS@#The AFLP was used to analyze the genetic diversity of 12 species of Cannabis using 55 primer combinations.@*RESULTS@#A total of 285 AFLP bands were obtained using five primer combinations with better diversity, among which 99 bands were polymorphic and 10 bands were special, with 47-76 bands amplified in each pair of primers.@*CONCLUSION@#AFLP may has good resolution in the diversity study of Cannabis. It may provide an essential basis for further study of the genetic diversity of Cannabis.


Subject(s)
Amplified Fragment Length Polymorphism Analysis/methods , Cannabis/genetics , DNA, Plant/genetics , Forensic Genetics , Genetic Variation , Polymorphism, Genetic
15.
Journal of Experimental Hematology ; (6): 591-593, 2007.
Article in Chinese | WPRIM | ID: wpr-276866

ABSTRACT

This study was purposed to investigate the immunological effects of modified dendritic cells (DCs) in inducing cytotoxic T cells (CTLs) effect against lymphoma cells. The DCs were derived from human peripheral blood and transfected with recombined adenovirus vector carrying survivin gene, Western blot was used to detect the expression of survivin, the lactate dehydrogenase (LDH) release test was used to determine the cytotoxicity of CTLs, the mixed lymphocyte reaction (MLR) was used to measure the ability to proleferate allo-lymphocyte by DCs, ELISA was used to assay IL-12 level in supernatant. The results showed that the expression of survivin in transfected dentritic cells was confirmed by Western blot analysis. In MLR assay, DCs transfected with Ad-survivin could induce higher allogeneic lymphocytes reaction at the ratios of 1:5, 1:10, 1:50 and 1:100. DCs transfected with Ad-survivin had much higher activity of CTL to CA46 cells than control DCs. The levels of IL-12 of supernatants containing DCs transfected with Ad-survivin were significantly higher than that in the control group. It is concluded that DCs transfected with Ad-survivin can induce CTL response in vitro against lymphoma cells.


Subject(s)
Humans , Adenoviridae , Genetics , Metabolism , Apoptosis , Allergy and Immunology , Dendritic Cells , Allergy and Immunology , Genetic Vectors , Genetics , Inhibitor of Apoptosis Proteins , Interleukin-12 , Metabolism , Lymphoma , Allergy and Immunology , Microtubule-Associated Proteins , Genetics , Allergy and Immunology , Recombination, Genetic , T-Lymphocytes, Cytotoxic , Allergy and Immunology , Transfection , Tumor Cells, Cultured
16.
Journal of Experimental Hematology ; (6): 791-794, 2006.
Article in Chinese | WPRIM | ID: wpr-233494

ABSTRACT

The study was aimed to construct the recombinant adenovirus vectors containing human survivin gene, and to investigate their expression in transfected dendritic cells. Full length cDNA encoding survivin was obtained by PCR amplification from plasmid pcDNA3.0-survivin. The PCR product was restricted, and then inserted into pShuttle-CMV. The plasmids of pShuttle-CMV-survivin were linearized with PmeI, and the fragment containing survivin was ligated with pShuttle-CMV and transfected into E. coli BJ5183. After homologous recombination in bacteria, the extracted plasmid from the positive bacteria were linearized with PacI, transfected into HEK293 cells with liposome Lipofectamine 2000. Then, the harvested adenovirus supernatants were transfected into dendritic cells. The results showed that the recombinant adenovirus-survivin was constructed successfully and its titer was about 2.65 x 10(9) pfu/ml. The expression of survivin in transfected dendritic cells was confirmed by Western blot analysis. It is concluded that the recombinant adenovirus vector containing human survivin was constructed successfully, which may provide preliminary laboratory evidence for anti-leukemia immunotherapy.


Subject(s)
Humans , Adenoviridae , Genetics , Metabolism , Dendritic Cells , Allergy and Immunology , Metabolism , Escherichia coli , Genetics , Metabolism , Gene Expression Regulation, Viral , Genetic Vectors , Inhibitor of Apoptosis Proteins , Microtubule-Associated Proteins , Genetics , Neoplasm Proteins , Genetics , Recombinant Proteins , Genetics , Transfection
17.
Chinese Journal of Clinical Pharmacology and Therapeutics ; (12)2002.
Article in Chinese | WPRIM | ID: wpr-561703

ABSTRACT

AIM: To explore the effect and mechanism of endothelin receptor antagonist on local angiotensin system of hypertrophic myocardial tissue. METHODS: Twenty four rats were randomly divided into 3 groups, control group (CG), hypertrophy group (HG) and endothelin receptor antagonist group (EG). The ratios of left ventricle weight to body weight (LVW/BW) and left ventricle weight to heart weight (LVW/HW) were measured to reflect myocardial hypertrophy. Protein expression of AT1R was detected by immunohistochemistry. The concentration of Ang Ⅱ was determined by ria. Activity of ACE was examined by spectrophotometry. A piece of myocardial tissue was observed in light microscope after HE or VG dyeing. RESULTS:(1) In comparison to CG, LVW/BW, LVW/HW, ACE activity, the concentration of Ang Ⅱ and the expression of AT1R of HG were increased significantly (P

SELECTION OF CITATIONS
SEARCH DETAIL