ABSTRACT
Objective:To investigate the therapeutic effects of a small interfering RNA (siRNA) targeting hypoxia inducible factor-1α (HIF-1α) on collagen-induced arthritis (CIA) in mice and to analyze the possible mechanism at the macrophage level.Methods:DBA/1 mice were injected with bovine type Ⅱ collagen to establish the CIA model. Then, they were injected with HIF-1α-siRNA adenovirus or negative control adenovirus through tail vein once a week for four weeks. This study included four groups: control group, CIA model group, negative control group and HIF-1α-siRNA group. Mouse bone marrow-derived macrophages (BMDMs) were isolated and cultured. The relative expression of CD206 and arginine (Arg) at mRNA level in mouse BMDMs was detected by RT-PCR. The proportions of F4/80 + CD16/32 + M1 and F4/80 + CD206 + M2 macrophages in spleen and thymus were detected by flow cytometry. Pathological changes in the ankle joint of mice were observed using HE staining. Immunohistochemistry was used to detect the expression of macrophages and the subsets in mouse synovial tissues. Results:(1) Compared with the control group, the CIA model group showed decreased expression of CD206 at mRNA level in BMDMs, but increased expression of Arg at mRNA level ( P<0.01). HIF-1α-siRNA increased the expression of CD206 at mRNA level ( P<0.05) and reduced the expression of Arg at mRNA level in BMDMs of mice with CIA ( P<0.01). (2) Compared with the control group, the mice in the CIA model group had increased proportions of F4/80 + CD16/32 + M1 macrophages in splenocytes and thymocytes ( P<0.05), but decreased proportions of F4/80 + CD206 + M2 macrophages in thymocytes ( P<0.05). HIF-1α-siRNA could down-regulate the proportions of F4/80 + CD16/32 + M1 macrophages in splenocytes and thymocytes ( P<0.05), and up-regulate the proportions of F4/80 + CD206 + M2 macrophages in thymocytes of CIA mice ( P<0.01). (3) CIA mice had synovial hyperplasia and macrophages infiltration, especially M1 macrophages, in the ankle joint. HIF-1α-siRNA could alleviate the synovial hyperplasia and macrophage infiltration. Conclusions:HIF-1α-siRNA could alleviate macrophage infiltration and synovial hyperplasia in CIA mice through reducing the proportions of M1 macrophages in thymocytes, BMDMs and synovial tissues and increasing the proportion of M2 macrophages, suggesting that HIF-1α-siRNA could treat CIA mice by regulating the differentiation of M1 and M2 macrophages.
ABSTRACT
Objective To investigate the clinical efficacy of neoadjuvant chemoradiotherapy in the treatment of locally advanced low and middle rectal cancer,and evaluate the effect of related clinical factors upon the long-term survival. Methods Clinical data of 101 patients with locally advanced low and middle rectal cancer admitted to our hospital from January 1,2010 to December 31,2014 were collected. All patients completed the preoperative intensity-modulated radiation therapy DT45-50. 4 Gy,synchronized with oxaliplatin+capecitabine/5-fluorouracil or single drug capecitabine chemotherapy,and total mesorectal excision) was performed 4-13 weeks after the end of the neoadjuvant therapy. The short-term efficacy and long-term prognosis of these patients were evaluated. Kaplan-Meier method was used for survival analysis,and Cox’s regression model for multivariate analysis. Results The total sphincter preservation rate was 53. 5%.The decrease rates of T,N staging and TNM total staging were 73. 26%,67. 32% and 72. 3%,respectively. The pathological complete response ( pCR) rate was 16. 8%.The median follow-up time was 41 months. The 3-year overall survival (OS), desease-free survival (DFS),local recurrence and distant metastases rates were 82. 2%,80. 7%,7. 2% and 12. 1%,respectively. The single factor analysis demonstrated that ypT and ypN stages were the risk factors affecting the 3-year OS,DFS anddistant metastases ( all P＜0. 05).Multivariate analysis revealed that ypT stage was an independent factor affecting the 3-year OS,and ypT and ypN stages were the independent factors of the 3-year DFS ( all P＜ 0. 05 ). ConclusionsNeoadjuvant chemoradiotherapy combined with TME in the treatment of locally advanced middle and low rectal cancer can partially decrease the tumor staging,enhance the sphincter preservation rate and improve long-term clinical prognosis. Both ypT and ypN stages are correlated with the clinical prognosis of patients.
ABSTRACT
Objective To investigate the expressions of AGGF1 in esophageal squamous cell carcinoma (ESCC) and their relationships with clinical features and prognosis of ESCC. Methods The expressions of AGGF1 in 70 cases of ESCC and 30 cases of normal esophageal tissue were examined using SP immunohistochemical staining and were analyzed according to the clinical features and follow-up data. Results The expressions of AGGF1 in 70 cases of ESCC was significantly higher than those in 30 cases of normal esophageal tissue [54.29%(38/70) vs. 23.33%(7/30)](P=0.004). The expressions of AGGF1 in ESCC were significantly related to the TNM stage, clinical stage and prognosis (P all<0.05). The OS was shorter in the positive teams of AGGF1 than that in the negative teams [(19.7 ± 3.5) months vs. (33.2 ± 4.0) months] (P=0.015). Cox- proportional multivariate analysis showed that positive expressions of AGGF1 and VEGF (P=0.043, 0.024) and clinical stage (P=0.035) were significant prognostic factors in overall survival. Conclusions AGGF1 has high expressions in ESCC, and it is closely related to the clinical features and prognosis of ESCC.
ABSTRACT
Objective To investigate the expression of miRNA-140 in chondrocytes and synovial fluid of osteoarthritis (OA) patients, and explore the relationship between the miRNA-140 expression and OA severity.Methods This study enrolled 30 OA patients who underwent total knee arthroplasty for chondrocytes sampling and 30 OA patients who underwent intra-articular injection for synovial fluid sampling. All OA patients were grouped into mild [Kellgren and Lawrence (KL) grade 1-2], moderate (KL grade 3) and severe (KL grade 4), with 10 in each subgroups for each sampling purposes. 7 non-OA patients and 10 patients with knee injury were collected for cartilage and synovial fluid sampling respectively as control groups. Chondrocytes were isolated from the cartilage tissue and cultured in vitro. Quantitative real time PCR for miRNA-140 in chondrocytes and synovial fluid were performed, and the U6 snRNA was used as internal control. The expression difference of miRNA-140 among groups and correlation between the expression and the KL grade of OA were analysed using one-way ANOVA and Spearman test respectively. Results The expression of miRNA-140 in chondrocytes of knees in OA patients was reduced than that in normal knees, and the between-group difference was statistically significant (F=305.464, P<0.001). miRNA-140 could be detected in synovial fluid of both normal knees and OA knees, its relative expression level was reduced in synovial fluid of OA group compared with normal group, and the between-group difference was statistically significant as well (F=314.245, P<0.001). The relative expression level of miRNA-140 in both chondrocytes and synovial fluid were negatively correlated with the KL grade of OA(r=-0.969, P<0.001; r=-0.970, P<0.001). Conclusion miRNA-140 could be detected in chondrocytes and synovial fluid of OA patients, and its expression was negatively correlated with the severity of OA.
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Cells, Cultured , Chondrocytes , Metabolism , Knee Joint , Metabolism , MicroRNAs , Osteoarthritis, Knee , Metabolism , Real-Time Polymerase Chain Reaction , Synovial Fluid , MetabolismABSTRACT
Objective To study the CBCT image registration of PTV enlarging distance and IMRT planning(CT-1) for patients with lung cancer,and evaluate their characters.Methods Ten patients with lung cancer were included in the study.Two sets image,before and after radiotherapy,were acquired every week.Then delineated the targeted volume and made the planning (CT-2) according the enlarging distance data.To comparize the parameters of DVH for lung and spinal cord,volumes and dose of PTV and NTCP with CT-1 and CT-2.The difference of two plan was analyzed by covariance analysis or Wilcoxson's z-test.Results The max,min and mean dose of PTV,the lung V5,V10,V20,V30,V50 were similar in both plans (P =0.242-0.663).There was superiority in CT-2 of PTV and lung's mean dose(P =0.049,0,035).The NTCP had the decent tendency followed by the increasing of lung Vs,V10,V20(P =0.146,0.053,0.000).Conclusions CBCT based image registration can reduce PTV,the mean dose of lung,NTCP,and increase PTV dose.This provides a tool for exploring acurate radiotherapy strategies.
ABSTRACT
Objective To analysis the influence factors of standardization in the hierarchical chain management of type 2 diabetes and to enhance the hierarchical chain management of type 2 diabetes.Methods ( 1 ) Six hundred and ninty patients with type 2 diabetes completed 1 years management were divided into well-controlled glycosylated hemoglobin ( HbAlc ) group (＜7.0％ ) and bad-controlled glycosylated hemoglobin (HbAlc) group ( ≥ 7.0％ ).The conditions of diet,physical activity,medication,self-blood sugar monitoring and participation in health seminars were investigated and analyzed.(2) The patients were divided into standardized management group and not standardized management group.Their age,sex,educational background,occupation,monthly income per person,medical security,the course,cognition for glycuresis,two-way transfer,and chronic complications were investigated and statistically analyzed.Results ( 1 ) The proportions of physical activity (70.1％ vs 54.2％,x2=6.163,P=0.018),self-blood sugar monitoring(60.4％ vs 43.8％,x2=6.268,P=0.016) and participation in health seminars (56.0％ vs 41.7％,x2=4.577,P=0.045) in the well-controlled HbAlc group were significantly higher than those in the bad-controlled HbAlc group.(2) Their age [(61.08 ±10.04) years old vs ( 57.75 ± 9.89 ) years old,t=2.539,P=0.012],educational background ( ratio of low educational attainment:8.3 ％ vs 17.2％,x2=6.426,P=0.041 ),medical security (own expense ratios:4.6％ vs 11.5％,x2=3.543,P=0.048 ),awareness of diabetes ( ratio of poor awareness of diabetes:19.4％ vs 41.0％,x2=17.518,P=0.000 ),two-way transfer ( ratio of not transfer treatment:4.6％ vs 14.8％,x2=7.662,P=0.022) and chronic complications ( ratio of chronic complication:41.7 ％ vs 26.2％,x2=6.130,P=0.017) were significantly different between the standardized management group and not standardized management group.(3) Logistic regression analyses indicated that the age ( OR=0.954,P=0.006),monthly income per person ( OR=4.101,P=0.018 ),medical security ( OR=7.617,P=0.003 ),cognition for glycuresis ( OR=0.030,P=0.000),two-way transfer ( OR=9.079,P=0.000) and chronic complications ( OR=0.456,P=0.031 ) were the risk factors of standardized management.Conclusion We should focus on the impact factors affecting the standardized management of patients including age,monthly income per person,medical security,awareness of diabetes,ratio of not transfer treatment,positive strategies for chronic complications,improve the hierarchical chain management of type 2 diabetes,and then make the diabetic patients to early participate in standardization management of diabetes mellitus and delay the appearance of complications.
ABSTRACT
Objective To explore the hierarchical chain management model in blood glucose control and its influence factors in patients with diabetes mellitus.Methods Health management database of diabetic patients was established in 2007 and managed by hierarchical chain management.The number of the patients reached to 1010 till 2011.The blood glucose control of diabetic patients was analyzed and its influence factors were analyzed by multivariate unconditional Logistic regression method.Results The concentration of glycosylated hemoglobin( HbA1c ) of 1010 patients with type 2 diabetes was (8.21 ±:2.70)％.Four hundred and eighty-seven cases (48.22％) reached the blood glucose standard,303 cases (30.00％)reached the blood pressure standard,245 cases (24.26％) reached the blood lipids standard,and 76 cases (7.52％) reached all three standards.Multivariate analysis showed that occupation (OR =2.521,95％ CI:1.871 - 3.397),education level (OR =1.890,95％ CI:1.642 - 2.174),disease course (OR =1.035,95％CI:1.016 -1.055),systolic pressure (OR =1.016,95％ CI:1.007 -1.025) and triglyceride (OR =1.204,95％CI:1.063 - 1.365) were the risk factors of blood glucose control (P ＜0.01).Conclusions Hierarchical chain management model is helpful for the blood glucose control in patients with type 2 diabetes.The comprehensive control and treatment of type 2 diabetes should be taken combined with related risk factors,such as blood pressure,blood lipids and diabetes disease course.
ABSTRACT
Objective To investigate the relationship and mechanism of the heme oxygenase-1 expression in peripheral blood mononuclear cells (PBMC) and the oxidative stress in diabetic nephropathy. Methods Two groups of diabetic patients with or without diabetic nephropathy and a normal control group were enrolled in this study. Fasting blood glucose (FBG), HbA1c, serum MDA level, ROS level, HO-1 mRNA level and HO-1 protein expression in PBMC were determined. Results In control group, diabetic group and diabetic nephropathy group , the MDA levels significantly increased[(14.23±5.07)nmol/ml vs (24.90±7.12)nmol/ml vs (43.83±16.97)nmol/ml](F＝37.022,P＜0.01), the ROS levels significantly increased (113.18±58.59 vs 364.54±88.67 vs 524.35±162.51)(F＝68.369,P＜0.01) and the HO-1 protein expression also increased significantly (22.84±9.98 vs 36.72±15.85 vs 58.1±15.93)(F＝31.302,P＜0.01). There was a positive correlation among the HO-1 mRNA, protein expression and MDA level(r＝0.407,0.429,P＜0.05). Conclusions There existed a severer oxidative stress condition in patients with diabetic nephropathy compared with the patients without diabetic nephropathy. HO-1 could be a potential pathway to ameliorate oxidative stress in diabetic kidney disease patients.
ABSTRACT
Objective To explore the hierarchical chain management model of type 2 diabetes and determine its evaluation.Method Based on the hierarchical chain management of the three community health service institutions and Dahua hospital in Shanghai Xuhui district,215 cases of type 2 diabetes had been involved in the study.Results Compared with the baseline before management,lasting blood glucose (FBG),2 h postprandial glucose (2hPBG),glycosylated hemoglobin (HbA1c),low density lipoprotein cholesterol (LDL-C),systolic blood pressure (SBP) and diastolic blood pressure (DBP) of the diabetes after 12 months' management declined [(8.50 ±2.81) mmol/L,(11.09 ±4.01) mmol/L,(8.56 ±2.41)% ,(3.31 ± 1.06) mmol/L,(139.06 ±20.68) mm Hg (1 mm Hg = 0.133 kPa),(78.20 ± 12.11) mm Hg vs.(7.41 ±2.04) mmol/L,(9.03 ±2.46) mmol/L,(7.34 ± 1.59)% ,(3.00 ± 1.06) mmol/L,(135.48 ± 17.82) mm Hg,(77.27 ±11.83) mm Hg],and the differences were statistically significant(P＜0.01 );control rate of FBG,2hPBG,HbA1c,LDLC,SBP,DBP had improved significantly [19.5% (42/215),20.9% (45/215),24.7%(53/215),20.0%(43/215),27.4%(59/215),30.2%(65/215) vs.50.7%(109/215),53.0% (114/215),54.0%(ll6/215),42.3%(91/215),47.0%(101/215),45.6%(98/215)](P＜0.01).Conclusion Primary and secondary-care hospital based hierarchical chain management model is valid and can be implemented for type 2 diabetes.
ABSTRACT
Rhesus monkeys (Macaca mulatta) are human's closest evolutionary relatives next to Chimpanzees, and they are widely used in biomedical researches. Analyses of the rhesus monkey trasnscriptome and the sequence divergence between monkey and human are of importantce to the development of scientific analyses and to the application and interpretation of the results from animal experiments. In this study, we analyzed the genetic and transcriptional information. Four hundred and one clones were randomly selected from a liver tissue cDNA library of rhesus monkey, and the expressed sequence tags (ESTs) were sequenced. We acquired 393 effective ESTs that were assembled into 221 Unigenes with Phrap software. Alignments of the sequences showed that 188 Unigenes matched with known proteins in Swiss_prot database, of which 16 Unigenes matched the known rhesus proteins, and 171 Unigenes had high homology with human proteins. Then the result of BLASTN comparison showed that 26 of another 33 Unigenes matched the known rhesus genes. Finally, the remaining Unigenes were aligned in dbEST and rhesus genome database, and the results suggested 3 Unigenes be newly discovered ESTs of rhesus.
Subject(s)
Animals , Expressed Sequence Tags , Chemistry , Gene Library , Liver , Chemistry , Macaca mulatta , Genetics , Sequence Analysis, DNAABSTRACT
In the conventional treatments of type I diabetes, there are various problems. As a new adequate treatment of diabetes, cell replacement therapy of diabetes has been applied and given research priority. We have investigated the applications of cell transplantation in the treatment of diabetes and have retrieved the relevant articles on cells transplantation for the treatment of diabetes. In this paper, we review the history, development, merits and demerits of cell transplantation and the recent advances in pancreatic islet transplantation research. The latest progress in the induction of stem cell to differentiate into the insulin-producing cells was also introduced.
Subject(s)
Animals , Humans , Diabetes Mellitus, Type 1 , General Surgery , Therapeutics , Insulin-Secreting Cells , Cell Biology , Islets of Langerhans Transplantation , Methods , Stem Cell TransplantationABSTRACT
This study sought to clone Chinese Banna minipig inbred-line (BMI) alpha1,3-galactosyltransferase (alpha1,3-GT) gene and construct its recombinant eukaryotic expression vector. Total RNA was isolated from BMI liver. Full length cDNA of alpha1,3-GT gene was amplified by RT-PCR and cloned into pMD18-T vector to sequence. Subsequently, alpha1,3-GT gene was inserted into pEGFP-N1 to construct eukaryotic expression vector pEGFP-N1-GT. Then the reconstructed plasmid pEGFP-N1-GT was transiently transfected into human lung cancer cell line A549. The expression of alpha1,3-GT mRNA in transfected cells was detected by RT-PCR. FITC-BS-IB4 lectin was used in the direct immunofluorescence method, which was performed to observe the alpha-Gal synthesis function of BMI alpha1,3-GT in transfected cells. The results showed that full length of BMI alpha1,3-GT cDNA was 1116 bp. BMI alpha1,3-GT cDNA sequence was highly homogenous with those of mouse and bovine, and was exactly the same as the complete sequence of those of swine, pEGFP-N1-GT was confirmed by enzyme digestion and PCR. The expression of alpha1,3-GT mRNA was detected in A549 cells transfected by pEGFP-N1-GT. The expression of alpha-Gal was observed on the membrane of A549 cells transfected by pEGFP-N1-GT. Successful cloning of BMI alpha1,3-GT cDNA and construction of its eukaryotic expression vector have established a foundation for further research and application of BMI alpha1,3-GT in the fields of xenotransplantation and immunological therapy of cancer.
Subject(s)
Animals , Animals, Inbred Strains , Base Sequence , China , Cloning, Molecular , Galactosyltransferases , Genetics , Metabolism , Genetic Vectors , Genetics , Molecular Sequence Data , Recombinant Proteins , Genetics , Metabolism , Sequence Analysis, DNA , Swine , Swine, Miniature , Genetics , TransfectionABSTRACT
Mesenchymal stem cells from bone marrow of Rhesus monkey (RhBMSCs) were isolated by density gradient centrifugation, purified by adherence separation, and further identified by phenotype and karyotype analysis. Growth characteristics of RhBMSCs were investigated by observation of cell proliferation and detection of apoptosis. Density gradient centrifugation and adherence separation revealed a simple method to obtain fairly pure RhBMSCs. Fusiform was the most common form of cell under observation, and cell karyotype was normal. Phenotyping assay revealed that the RhBMSCs are highly positive (99.2%) for CD29 when compared with the low positive rates (less than 3.2%) for CD34, CD45 and HLA-DR, which indicated a successful isolation of high purity RhBMSCs and a vigorous activity of cells to proliferate. But the proliferation activity of RhBMSCs gradually decreased following the increasing of cell generations. The methods and results of isolation, expansion, identification and growth characteristics of RhBMSCs were discussed in this paper, which may be helpful to understanding the bone marrow mesenchymal stem cells of human, and may serve as the groundwork for orientated differentiation of RhBMSCs and tissue repairment on experimental animal model of rhesus monkey.
Subject(s)
Animals , Bone Marrow Cells , Cell Biology , Cell Culture Techniques , Cell Separation , Cells, Cultured , Integrin beta1 , Blood , Macaca mulatta , Mesenchymal Stem Cells , Cell Biology , PhenotypeABSTRACT
Gene chip has been an important way to detect gene changes of organism in different condition. It is a new tendency to use gene chip to research graft rejection at gene level. Ischemia reperfusion injury (IRI) accurs early in organ transplantation. Studying IRI with gene chip is beneficial to understand the mechanism of graft rejection and will provide guidance for surveying and treating graft rejection after organ transplantation.
Subject(s)
Animals , Mice , Rats , Disease Models, Animal , Graft Rejection , Genetics , Oligonucleotide Array Sequence Analysis , Reperfusion Injury , Genetics , TransplantsABSTRACT
The specific anti-tumor immune response induced by mouse bone marrow dendritic cells (DCs) transfected with recombinant adenovirus carrying mutant k-ras genes was investigated. DCs were generated from mouse bone marrow in the presence of rmGM-CSF (3.3 ng/mL) and rmIL-4 (1.3 ng/mL) and detected by FACS, and then transfected with the recombinant adenovirus encoding mutant k-ras gene. The efficacy of transfection and T cell stimulating activity of DCs were detected. CTL activity of the mice vaccinated with DCs was observed. The results showed that DCs had dendritic veiled morphology. BmDCs highly expressed B7-1 (80 %), B7-2 (77 %), MHC Ⅱ (70 %), CD11c (65 %), CD40 (70 %) and CD54 (96 %) with FACS, and no significant difference in the expression was observed before and after the transfection (P>0.05). The DCs transfected by mutant k-ras gene could significantly stimulate lymphocytes proliferation as compared with those transfected by Ad-c or non-modified DCs (P<0.05). DC vaccine transfected by mutant k-ras gene could induce CTL activity against Lewis lung cancer, but not against B16. The specific cytotoxicity against Lewis lung cancer in Ad-k-ras/12-transduced DC group was significantly higher than those in the control, vector and non-transfected DCs groups (P<0.05). It was concluded that special antitumor response could be induced by DCs transfected with recombinant adenovirus carrying mutant k-ras genes.
ABSTRACT
<p><b>BACKGROUND</b>To study the relationship between the serum levels of endostatin,VEGF and clinical and pathophysiological characteristics in the non-small cell lung cancer (NSCLC) patients.</p><p><b>METHODS</b>The serum levels of endostatin and VEGF were detected in 46 patients with NSCLC and 14 patients with benign pulmonary diseases as control by ELISA mothod.</p><p><b>RESULTS</b>(1)The preoperative serum level of endostatin [( 20.85 ±4.56) μg/L] and VEGF [(1.75±0.37) μg/L] in lung cancer patients was significantly higher than those in patients with benign pulmonary diseases [(15.68±2.78) μg/L and (1.05±0.32) μg/L). (2)The preoperative serum level of endostatin and VEGF in lung cancer patients was closely related to P-TNM stages, distant metastasis, grade of cell differentiation and the size of the primary tumors ( P < 0.05), but not to the histological classification, type of the tumor, lymph node status, age, sex of the patients and smoking or not ( P > 0.05). (3)The serum level of endostatin in lung cancer patients on the 7th postoperative day [(23.41± 5.12 ) μg/L] was significantly higher than that before operation [(20.85±4.56) μg/L] ( P < 0.05) and on the 1st postoperative day [(18.89±4.67) μg/L] ( P < 0.001). The serum level of VEGF in lung cancer patients on the 7th postoperative day [(3.75±0.71) μg/L] was also significantly higher than that before operation [(1.72±0.46) μg/L] and on the 1st postoperative day [(2.22±0.58) μg/L] ( P < 0.001). (4)The preoperative serum level of endostatin was highly negatively correlated to serum VEGF level in lung cancer patients ( r=-0.380, P < 0.01).</p><p><b>CONCLUSIONS</b>Elevation of serum endostatin and VEGF exists in patients with NSCLC. The serum levels of endostatin and VEGF in patients with NSCLC might be helpful to evaluate the biological behavior of lung cancer.</p>
ABSTRACT
<p><b>BACKGROUND</b>To construct a DC-Ad-Ki-ras(V12) vaccine and investigate the anti-tumor activities of lung cancer dendritic cell vaccine modified by mutant Ki-ras gene in vitro.</p><p><b>METHODS</b>Ki-ras(V12) cDNA was transfected into cultured bone marrow-derived DC with the recombinant adenovirus [(Ad-Ki-ras(V12)] containing human mutant Ki-ras gene. Anti-tumor activity of the vaccine was studied in vitro by flow cytometry, PCR, MLR and cytotoxicity assay.</p><p><b>RESULTS</b>(1) The DC vaccine was confirmed not only to express Ki-ras(V12) gene, but also to remarkably stimulate lymphocyte proliferation and improve CTL activity. (2) The DC vaccine modified by mutant Ki-ras gene could induce specifical CTL activity of immunized mice against Lewis lung carcinoma that could express Ki-ras(V12) gene, but not to B16.</p><p><b>CONCLUSIONS</b>The DC vaccine modified by mutant Ki-ras gene can induce obvious anti-tumor activities against Lewis lung carcinoma that can express Ki-ras(V12) gene.</p>
ABSTRACT
In order to compare the effects of GM-CSF + IL-4 with the effects of IL-3 + IL-4 on the differentiation and development of dendritic cells (DCs) from mouse bone marrow, we cultured bone marrow hematopoietic progenitor cell in vitro and examined whether DCs induced by IL-3 + IL-4 (IL-3 DCs) differed from the DCs induced by GM-CSF + IL-4 (GM-CSF DCs) in morphology, yields, phenotype and endocytic activity. Scanning electron microscope, laser scanning confocal microscope (LSCM) and flow cytometer (FCM) were used. The results showed that the DCs induced by IL-3 + IL-4 and the DCs induced by GM-CSF + IL-4 had similar morphology, and the DCs induced by IL-3 + IL-4 expressed high level of major histocompability complex (MHC) class II. Besides, IL-3 DCs had the characteristics of higher yields and better consistency. But CD80 and CD86 were expressed at low levels or absent on IL-3-treated DCs. The capability of uptaking antigen of IL-3 DCs was more powerful than that of GM-CSF DCs. IL-3 could substitute for GM-CSF in culturing DCs from mouse bone marrow cells and could obtain the tolerance DCs that lack costimulatory molecules.
Subject(s)
Animals , Male , Mice , Antigen-Presenting Cells , Cell Biology , Bone Marrow Cells , Cell Biology , Cell Differentiation , Cells, Cultured , Dendritic Cells , Cell Biology , Granulocyte-Macrophage Colony-Stimulating Factor , Pharmacology , Interleukin-3 , Pharmacology , Interleukin-4 , Pharmacology , Mice, Inbred BALB CABSTRACT
<p><b>BACKGROUND</b>To study the cell immunity induced by lung cancer B7 vaccine, FLB2C cells.</p><p><b>METHODS</b>Compared withparental lung cancer cell LA795 line, proliferation of mouse T lymphocytes stimulated by FLB2C was observed through mixed lymphocyte culture. CTLL cell MTT test was used to detect whether FLB2C stimulated T lymphocyte to secrete IL-2. After immunized with the FLB2C and LA795 cells, the CTL activity of mouse was observed in vivo.</p><p><b>RESULTS</b>The spleen lymphocyte proliferation stimulated by FLB2C cells was remarkably stronger than that by LA795 . FLB2C might stimulated the T lymphocyte to secrete IL-2 in vitro, but LA795 didn't. FLB2C cell could induce CTL activity in vivo and the induced CTL killed FLB2C cells and LA795 in vitro, with the killing rate of 34% and 25.3% respectively; while LA795 induced CTL killing rate being 10.5% and 12.25% respectively (P < 0.01).</p><p><b>CONCLUSIONS</b>FLB2C can stimulate T cell immunity in vivo or in vitro. The effect of FLB2C is significantly stronger than that of LA795 . The results suggest that FLB2C may be used to treat lung cancer through improving immunity. This provides immunological basis for applying FLB2C as a vaccine to clinical use.</p>
ABSTRACT
<p><b>BACKGROUND</b>To establish an ADM resistant Lewis lung cancer cell line and to investigate its biological characteristics.</p><p><b>METHODS</b>The multidrug-resistant cell line was gradually induced by ADM from Lewis lung cancer cell line (L3-8), its growth characteristics and cancinogenicity were observed. The fluorescent density of ADM and Rh-B in the cells were analyzed by IPP image analysis system. The ADM concentration in cells was assayed with fluorescent meter. The IC50 was evaluated by MTT assay and the drug resistant index was counted.</p><p><b>RESULTS</b>The drug resistant Lewis lung cancer cell line, L3-8/ADM, which can grow in the medium containing 0.4mg/l ADM was acquired after 14 months selective culture. Its tumor generatic rate was 10/10. When the L3-8/ADM was cultured in 0.15mg/l ADM and general 1640 medium, the doubling time was 22.0h and 21.8h separately. After culturing in 4mg/l ADM and Rh-B medium, the fluorescent density ratio of L3-8/ADM and L3-8 were 2.82:1 and 2.65:1 separately. The ADM concentration of L3-8/ADM was only 64.7% of its parental, but there was no significant difference in their ADM release rate. Except for ADM, L3-8/ADM was resistant to DNR, VCR, MIT and CDDP in different degrees.</p><p><b>CONCLUSIONS</b>L3-8/ADM cell is a typical MDR cell line. The cell membrane obstruction of drug infiltration might be the cause of drug resistance. This study will provide a basis for the establishment of lung cancer MDR animal model.</p>