Effects of Human Umbilical Cord Mesenchymal Stem Cells on Infected State of Human Alveolar type Ⅱ Epithelial Cells / 中山大学学报(医学科学版)

[Objective]To investigate the effect of human umbilical cord mesenchymal stem cells on infected state of human alve?olar type Ⅱ epithelial cells.[Methods]Human alveolar type Ⅱ epithelial cells A549(1×105/mL)2 mL and PA(3×104 CFU/mL)2 mL has grown after 6 hours,add hUCMSC(1 × 106/mL)2 mL as the experimental group,add equal amounts of phosphate buffer (PBS)for infection,A549 and PBS and the medium has grown as the control group. A549 cells morphological changes between the compared groups(Transmission electron microscope,TEM),A549 cell viability(new CCK-8 cell proliferation assay Kit),A549 cells apoptosis(Annexin V-FITC/PI double staining flow cytometry)and the expression of A549 pulmonary surfactant A(SP-A) (Western blot).[Results]Transmission electron microscope cell morphology observation displayed ,infection group A549 cell dam?aged obviously,cell quality appeared empty bubble degeneration,chromatin height agglutination,visible apoptosis bodies;experi?ment group cell package film structure full,nuclear film full,nucleolus obviously,nuclear chromatin electronic density low,chroma? tin uniform,no apoptotic bodies;control group A549 cell structure full,membrane surface micro-fluff rich,nuclear film full,nucle?ar week clearance structure normal,chromatin uniform;infection group and control group compared,Infection group A549 cell sur?vival significantly reduced[(70.35±2.89)% and(97.37±2.07)%,n=3,P<0.01],apoptosis rate significantly increased[(8.63%± 0.16)%and(2.55±0.11)%,n=3,P<0.01],In the infected group,PA could damage A549 cells and decrease the amount of SP-A ex?pression(n=5,P<0.05). In the experiment group,the protective effect of hUCMSC on the A549 cells after infection may increase the expression of SP-A(n=5,P<0.05);[Conclusions]HUCMSC inhibits the infection of A549 cells apoptosis and protection of A549 cells secrete SP-A.

Therapeutic effect of Tat-LK15-mediated the interference of siRNA with nNOS expression in the spinal dorsal horn on neuropathic pain in rats / 解放军医学杂志

Objective To investigate the therapeutic effect of cell penetrating peptide Tat-LK15 mediating small interfering RNA (siRNA) interference with the expression of neuronal nitric oxide synthase (nNOS) in rat spinal dorsal horn on neuropathic pain. Methods The transfection reagent, Tat-LK15, was used to mediate the transfection of rat spinal dorsal horn (SDH) neuronal cells with carboxyfluorescein (FAM), and then the transfection effect was observed under inverted fluorescence microscope. Fifty healthy male SD rats were randomly divided into 5 groups (n=10): control group, sham operation group (sham group), neuropathic pain group (SNL group), Tat-LK15-nNOS siRNA group (TS group) and Tat-LK15-NC siRNA group (TN group). Neuropathic pain was induced by spinal nerve ligation (SNL), rats in control group did not receive operation and only the spinal nerve was exposed in sham group. Groups SNL, TS and TN were made into the models by SNL and implanted intrathecal catheter, intrathecal administration was performed from the 7th day after model establishment, and 10μl normal saline, 10μl TS complex (including 5μg siRNA) and 10μl TN (including 5μg siRNA) were injected intrathecally each day for 7 days. Paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) were measured at 1 day before (baseline) and 3, 7, 10 and 14 days after model establishment. Then animals were sacrificed on the 14th day after the operation and the lumbar segment (L4-6) of the spinal cord was removed to detect the expressions of nNOS mRNA and protein using q-PCR and Western blotting analysis. Results Tat-LK15 effectively mediated FAM-siRNA into SDH neuronal cells. Compared with sham group, SNL significantly decreased PWMT and PWTL and increased expressions of nNOS mRNA and protein from the 3rd day (P<0.01), but there was no significant difference between the sham and control group. Tat-LK15-nNOS siRNA complex significantly increased PWMT and PWTL and down-regulated nNOS mRNA and protein expressions in TS group compared with SNL group on the days 10 and 14. There was no significant difference between TN and SNL group. Conclusion Tat-LK15 not only can mediate successful nNOS siRNA transfection and inhibit the expression of nNOS, but also effectively relieve SNL-induced neuropathic pain in rats.

Analysis of 35 Cases of ADR Induced by Low-molecular Dextran and Amino Acid Injection in Our Hospital / 中国药房

OBJECTIVE:To investigate the characteristics and the regularity of ADR induced by Low-molecular dextran and amino acid injection,and to provide reference for safe use of drugs in clinic.METHODS:Among 35 cases of ADR induced by Low-molecular dextran and amino acid injection were collected from our hospital during Jan.2010-Jan.2017.Those ADR cases were analyzed statistically in respects of patients'gender and age,type and occurrence time of ADR cases,primary disease,aller gic history,organs/systems involved in ADR,clinical manifestations,outcomes,etc.RESULTS:Among 35 cases of ADR induced by Low-molecular dextran and amino acid injection,there were 18 male (51.43％) and 17 female (48.57％).The youngest patient was 11 years old,the oldest patient was 80 years old,the average age was 48.2 years.There were 4 cases of severe ADR (11.43 ％) and 31 cases of common ADR (88.57 ％).ADR mostly occurred 10 min-1 h after medication (71.43 ％).Bone fractures (14 cases,40.00％) and knee inflammation (8 cases,22.86％) were main original disease involved in ADR.Main organs or systems involved in ADR were digestive system(34.29％),circulatory system(22.86％),skin and its appendants (20.00％),etc.All cases were recovered or cured after drug withdrawal and systematic treatment.CONCLUSIONS:Low-molecular dextran and amino acid injection induce low incidence of ADR but can induce severe ADR.The nursing staff should strengthen ADR monitoring to reduce the occurrence of complications and avoid medical statf-patient dispute.

Effect and mechanism of miR-206/miR-613 on expression of OATP1B1 / 药学学报

This study was designed to explore the effect and mechanism of miR-206/miR-613 on the expression of OATP1B1 gene. Bioinformatic analysis was used to predict the potential miRNAs target sites in 3'-untranslated region (3'-UTR) of OATP1B1 mRNA. The expression level of miR-206/miR-613 and OATP1B1 mRNA and protein was determined with RT-qPCR and Western blot, respectively. Luciferase assay was used to explore the exact mechanism of the effect of miR-206/miR-613 on the expression of OATP1B1 mRNA and protein. The results showed that the seed sequences of miR-206/miR-613 has perfect complementary with 3'-UTR of OATP1B1 mRNA in terms of sequence specificity. The secondary structure between miR-206/miR-613 and 3'-UTR of OATP1B1 mRNA was rather stable. The OATP1B1 protein level was down-regulated by 24.7%, 38.8% by overexpression of miR-206/miR-613. The expression was up-regulated by 25%, 38.2% by inhibition of miR-206/miR-613. However, overexpression or inhibition of miR-206/miR-613 had no effect on the expression of OATP1B1 mRNA. The luciferase activity of pMIR/OATP1B1-WT luciferase reporter gene was decreased by 35% and 30% through overexpression of miR-206/miR-613. The expression was increased by 33.1% and 32.5% through inhibition of miR-206/miR-613. When the binding sites in the 3'-UTR of OATP1B1 mRNA complementary with miR-206/miR-613 was mutated, overexpression or inhibition of miR-206/miR-613 had no effect on the luciferase activity. Collectively, miR-206/miR-613 post-transcriptionally regulates the expression of OATP1B1 protein by directly targeting the 3'-UTR of OATP1B1 mRNA.

Tat-LK15-mediated delivery of nNOS siRNA in rats with chronic inflammatory pain / 实用医学杂志

Objective To investigate the potential application of a non-viral gene carrier , TAT-LK15 , for delivering nNOSsiRNAin vivo and to study whether TAT-LK15/siRNA can be a new treatment method for chronic inflammatory pain. Method TAT-LK15 was complexed with nNOSsiRNA or scrambled control siRNA. The expression of nNOS was determined in SCDH of chronic inflammatory pain rats by western-blot assay. Pain control efficacy was evaluated by mechanical withdrawal threshold (MWT) and thermal withdrawal duration (TWD) assays. Results nNOS protein expression was efficiently inhibited by intrathecal injection of TAT-LK15/siRNA complexes , with the reduction of nNOS protein by 52%. Moreover , injection of TAT-LK15/siRNA com-plexes significantly could decrease MWT , but increase TWD in rats with chronic inflammatory pain. Conclusions TAT-LK15 can efficiently deliver nNOSsiRNAin vivo and nNOSsiRNA can relieve chronic inflammatory pain in rats.

Delivery of therapeutic siRNA by Tat-LK15 targeting nNOS fusion gene in RGC-5 cells / 中国药理学通报

Aim To investigate the potential applica-tion of a non-viral gene carrier Tat-LK15 for delivering siRNA targeting nNOS in vitro, which provides evi-dence of Tat-LK15 mediating siRNA targeting nNOS in vivo for treatment of neuropathic pain. Methods 1. Tat-LK15 was mixed with siRNA, then the mixture was analyzed the best ratio by Gel retardation. The trans-fection efficiency of FAM-siRNA mediated by Tat-LK15 on RGC-5 cells was examined by Flow Cytome-try. The apoptosis ratio of RGC-5 was identified by Flow Cytometry 24 h after treated with the different do-ses of Tat-LK15 (1, 2. 5, 5, 10 and 20 μg). 2. The model of RGC-5 cell overexpression of nNOS protein was prepared. 3. RGC-5 cells were randomly divided into 5 groups:control group,model group, Tat-S group ( Tat-LK15 mediate nNOS/siRNA transfection model cell) , Lipo-S group ( LipofectamineTM RNAiMAX me-diate nNOS/siRNA transfection model cell) and Tat-N group ( Tat-LK15 mediate NCsiRNA transfection model cell) . Real-time Quantitative polymerase chain reac-tion( Q-PCR) and Western blot were used to evaluate nNOS expression level assay. Results It indicated that the Tat-LK15/siRNA complex completely formed at the weight ratio of 2∶ 1 (μg/μg) , and the transfec-tion efficiency was (84. 4 ± 3. 9)%. It caused cotytox-icity when Tat-LK15 dose was 20 μg ( 6. 1 μmol · L-1 ) , and the apoptosis rate more than control group [(10. 3 ± 1. 1)% vs (7. 4 ± 0. 9)%, P<0. 05]. The nNOS protein level of RGC-5 cells was significantly el-evated after modeling. Compared with that of model group, Tat-LK15/siRNA efficiently inhibited the ex-pression of nNOS at transcriptional level or protein leve1 of Tat-S group ( P <0. 05 ) , and there was no significant difference of the efficiency inhibited between Tat-S group and Lipo-S group. Conclusions Tat-LK15’ advantage is with high efficiency, low cytotox-icity. The Tat-LK15 can deliver siRNA targeting nNOS in vitro efficiently and safely.

Effects of CO2 pneumoperitoneum on cell cycle and cell cycle protein of a gastric cancer cell line / 中华普通外科杂志

Objective To investigate the effects of different CO2 pneumoperitoneum on cell cycle and cell cycle protein of a gastric cancer cell line. Methods Human gastric cancer cell line MNK-45 were exposed to 0,10,12 and 15 mm Hg CO2 pneumoperitoneum in vitro for 4 hrs. Cell cycle was measured by flowcytometry, the expression of CDK4 ,Cyclin D1、Rb and pRb was studied by Western-blot, and the binding ability of CDK4 and Cyclin D1 was evaluated by immunoprecipitation. Results The cell proliferation index in 15 mm Hg CO2 pneumoperitoneum group dropped significantly (27.4% ± 3. 7%) vs. (36. 4% ±3. 3%) ,P <0. 05, while that in other groups did not change significantly. The protein of CDK4、Cyclin D1and binding ability of Cyclin D1 and CDK4 dropped dramatically in the 15 mm Hg CO2 pneumoperitoneum group (0.71%±0.12%),(0.93% ±0.21%),(0.54%±0.11%),(0.18% ±0.02%) vs. (1.05% ±0.16%),(1.40% ±0.24%),(0.75% ±0.14%),(0.31% ±0.02%), all P<0.05. There were no changes of Rb in protein levels, while the phosphorylated Rb dropped obviously. Conclusion There was no obvious effects of clinical CO2 pneumoperitoneum on gastric cancer cells growth and proliferation.

Effects of CO2 pneumoperitoneum pressures on growth and proliferation of gastric cancer cells in nude mice transplanted tumor / 中华普通外科杂志

Objective To investigate the effects of different CO2 pneumoperitoneum pressures on gastric cancer cells' growth and proliferation in nude mouse model of implanted tumor. Methods Human gastric cancer cell lines MNK-45 were exposed under 0、10、12 and 15 mm Hg CO2 pneumoperitoneum for 4 hrs respectively. 2 × 106 processed cells were inplanted into nude mice subcutaneously. Three weeks later, mice were sacrificed and the weight and bulk of the tumor measured. Then we observed the transplantation tumor by HE stain and Ki-67 stain. Results There was no significant difference in tumor's growing time, bulk and weight between 0, 10, 12 mm Hg CO2 pneumoperitoneum groups (7. 8 d, 7. 2 d, 7. 8 d; 1. 2 cm3, 1. 3 cm3, 1. 3 cm3; 1.5 g, 1. 9 g, 1. 6 g)and the control group (7. 3 d, 1. 2 cm3, 1.4 g) (P > 0. 05 ). The growing time of tumor in 15 mm Hg CO2 pneumoperitoneum (12. 5 d) was obviously longer than the control group ( P < 0.05 ) , the bulk and weight of tumor in 15 mm Hg CO2 pneumoperitoneum (0. 5 cm3, 0. 5 g) group significantly decreased compared with the control group (P <0.05). The positive rate of Ki-67 in 15 mm Hg CO2 pneumoperitoneum (27. 5% ) group was obviously lower than the control group (59.6%) (P<0.01). However, there were no significant differences between 0, 10, 12 mm Hg CO2 pneumoperitoneum groups (61.2%, 60.5%, 63.4%) and the control group (P > 0.05). Conclusion Clinically adopted CO2 pneumoperitoneum pressures have no significant effect on gastric cancer cells growth and proliferation.

Effects of hypoxia-inducible factor-1α on human gastric cancer cell apoptosis under in vitro CO(2) pneumoperitoneum environment / 中华外科杂志

<p><b>OBJECTIVE</b>To study the effect of hypoxia-inducible factor-1α (HIF-1α) on human gastric cancer cells apoptosis in simulated CO2 pneumoperitoneum environment.</p><p><b>METHODS</b>Applied closed box to simulated CO2 pneumoperitoneum environment under the pressure of 0, 5, 10 and 15 mm Hg (1 mm Hg = 0.133 kPa). Compared HIF-1α mRNA and protein expression of MKN-45 cells before and after silencing HIF-1α by RT-PCR and Western blot. Study the changes of bcl-2/bax expression in MKN-45 cells before and after silencing HIF-1α by immunohistochemistry. The apoptosis ratio of MKN-45 was measured by using Annexin V-FITC/PI double labelled staining.</p><p><b>RESULTS</b>In 15 mm Hg group, HIF-1α mRNA and protein expression of MKN-45 cells (1.48 ± 0.22, 1.34 ± 0.09) and HIF-1α protein expression in 10 mm Hg group (1.25 ± 0.10) were significantly higher than those in control group (0.55 ± 0.17, 0.83 ± 0.04) (P < 0.05). But there was no significant differences among 0, 5, 10 mm Hg group and control group in HIF-1α mRNA (P > 0.05); and no obvious difference was found among 0, 5 mm Hg group and the control group in HIF-1α protein expression (P > 0.05). In 15 mm Hg CO2 pressure, bcl-2/bax expression (0.78 ± 0.05) was obviously lower than that in the control group (1.43 ± 0.15) (P < 0.05) and the apoptosis ratio (11.70 ± 0.12) was significantly higher than the control group (0.22 ± 0.07) (P < 0.01) before silencing HIF-1α. But once HIF-1α was silenced, HIF-1α mRNA (0.52 ± 0.11), HIF-1α protein expression (0.92 ± 0.02), bcl-2/bax ratio (1.57 ± 0.04) and apoptosis ratio (0.45 ± 0.11) in MKN-45 were not significantly different between 15 mm Hg group and the control group (P > 0.05).</p><p><b>CONCLUSIONS</b>The apoptosis ratios of MKN-45 under 0, 5, 10 mm Hg CO2 pneumoperitoneum are comparable with that in the control group before the silencing of HIF-1α. The apoptosis ratio of MKN-45 is increased under 15 mm Hg CO2 pneumoperitoneum environment and HIF-1α may be one of the important factor to improve the apoptosis of human gastric cancer cell.</p>

Role of connexin 43 gene in peritoneal metastasis of gastric cancer / 中华消化外科杂志

Objective To investigate the relationship between the expression of eonnexin 43 (Cx43) and clinicopathologieal characteristics of gastric cancer, and to study the role of Cx43 in peritoneal metastasis of gastric cancer. Methods Thirty-two patients who had gastric cancer and with peritoneal metastasis had been admitted to Southwest Hospital from January 2000 to December 2008. Gastric cancer tissues, adjacent tissues and metastatic peritoneal tissues were obtained postoperatively, and the expression of Cx43 was detected by immunohistochemistry. The relationship between Cx43 expression and clinicopathological characteristics of gastric cancer was analyzed. All data were analyzed via Spearman rank correlation coefficient, Fisher exact probability and chi-square test. Results The expression of Cx43 was mainly detected in the cell membrane and cytoplasm. The positive expres-sion rates of Cx43 in gastric cancer tissues, adjacent tissues and metastatic peritoneal tissues were 34% (11/32), 100% (32/32) and 94% (30/32), respectively. There were significant differences in the Cx43 expression between gastric cancer tissues and adjacent tissues (X~2=28.350, P < 0.01), and between gastric cancer tissues and metastatic peritoneal tissues (X~2 = 21.989, P < 0.01). The expression of Cx43 did not correlate with age and sex of patients (r = -0.030, - 0.169, P > 0.05), but with tumor differentiation, histological type and lymph node metastasis (r = 0.750, 0.642, - 0.357, P < 0.05). Conclusions There is a decreased expression of Cx43 in gastric cancer tissues and a up-regulated expression of Cx43 in metastatic peritoneal tissues. Cx43 may play a positive role in the peritoneal metastasis.

Effects of HIF-1α on human gastric cancer cell's apoptosis in CO2 pneumoperitoneum environment / 中华普通外科杂志

Objective To study the effect and mechanism of human gastric cancer cell's apoptosis in simulated CO2 pneumoperitoneum environment by HIF-1α. Methods We used a closed box to simulate CO2 pneumoperitoneum environment, a standard surgical insufflator maintained pressure of the box at 0,5,10 and 15 mm Hg respectively. MKN-45 cell's HIF-1α mRNA and protein expression were observed before and after silencing cell's HIF-1α by real time RT-PCR and Western blot. Cell's bcl-2/bax was studied by immunohistochemistry before and after silencing HIF-1α. Cell's apoptosis ratio was measured using Annexin V-FITC/PI double labelled staining. Results In 15 mm Hg group, MKN-45 cell's HIF-1α mRNA and protein expression were significantly higher than control group (P<0.01 ), while the difference was not significant among 0,5,10 mm Hg group and control group (P>0.05). In 15 mm Hg CO2 pressure, cell's bcl-2/bax was obviously lower than in control group (P<0.05) and apoptosis ratio was more than control group(P<0.01). When HIF-1α was silenced, cell's bel-2/bax and apoptosis ratio weren't significantly different between 15 mm Hg group and control group (P>0.05 ). Conclusions MKN-45 cell's apoptosis did not experience any alterations under 0,5,10 mm Hg.15 mm Hg CO2 pneumoperitoneum environment enhanced cell's apoptosis probably by way of HIF-1α.

Effects of different CO2 pressure pneumoperitoneum on the migration and cytoskeleton in gastric cancer cells / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the influence of CO(2) and He insufflation administered at different pressures on the migration and cytoskeleton of cultured human gastric cancer cells.</p><p><b>METHODS</b>The cultured gastric cancer cells MKN-45 were exposed to a CO(2) or He environment maintained at different pressures (12, 15 mm Hg). After 0, 2, 4, 6, 8 hours exposure to CO(2) or He environment, pH of the MKN-45 cells culture media was measured with blood gas analysis. The cell migration was detected with Transwell technology. The cell cytoskeleton was observed with laser confocal microscope.</p><p><b>RESULTS</b>The media pH was acid after exposure to CO(2) environment, while it was basic in the He group. The number of cells passing millipore in 12 mm Hg CO(2) or He insufflation pressure were not significantly different with control group (P>0.05), however in 15 mm Hg pressure CO(2) group, it was significantly decreased as compared to control group (P<0.01). The microfilament and microtubule in gastric cancer cell were ambiguous in 15 mm Hg pressure CO(2) group.</p><p><b>CONCLUSIONS</b>There are no obvious effects on the migration and cytoskeleton of MKN-45 cells under 12 mm Hg CO(2) insufflation pressure. The migration and cytoskeleton of MKN-45 cells can be inhibited in 15 mm Hg CO(2) pneumoperitoneum environment.</p>

Inhibitive effects of gastric cancer cell-dendritic cell fusion vaccine on tumor cell proliferation cycle / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To study the inhibitive effects of gastric cancer cell-dendritic cell fusion vaccine on tumor cells of proliferation cycle.</p><p><b>METHODS</b>Peripheral blood mononuclear cells were separated from gastric cancer patients and co-cultured with granulocyte macrophage colony stimulating factors(GM-CSF), interleukin-4(IL-4) and tumor necrosis factor-alpha(TNF-alpha) to generate mature dendritic cells. The dendritic cells and SGC7901 cells were fused by polyethylene glycol, and the pure fusion cells were screened out by selective culture systems. The inhibitive effects of gastric cancer cell-dendritic cell fusion vaccines on tumor cell proliferation cycle in vivo and in vitro were detected by flow cytometry.</p><p><b>RESULTS</b>Treated with the fusion vaccine in vitro, the percentages of G(0)/G(1), S and G(2)/M cells of tumor cells were (76.77+/- 4.38)%, (16.50+/- 2.90)% and (6.73+/- 1.59)% respectively. There were significant differences in the percentages of different cell cycle tumor cells between the tumor cells treated with the fusion vaccine and those co-cultured with dendritic cell or T cells alone(P< 0.01). The proliferative index of the tumor cells treated with the fusion vaccine was 23.34+/- 3.51, significantly lower than those co-cultured with dendritic cell and controls (P< 0.05).</p><p><b>CONCLUSIONS</b>Fusion vaccines can affect cell cycle of the tumor cells, thus inhibit tumor cell proliferation and growth.</p>

Expressions and clinical significance of CD83 and S100 on gastric cancer infiltrating dendritic cells / 中国普通外科杂志

Objective To investigate the expression and clinical significance of tumor infiltrating dendritic cells(TIDCs) within gastric tumor tissues.Methods Immunohistochemistry(IHC),in-situ hybridization(ISH) and flow cytometry were applied to detect the expression of S100,CD83 mRNA and CD83 on DCs in 45 gastric adenocarcinoma tissues.The co-relationship of the S100 and CD83 expression with clinical(pathological) features was analyzed.Results IHC showed that S100 expression was unevenly distributed(within) 42 cancer tissue and CD83 was only expressed in tumor-adjacent tissue and normal tissue.ISH showed that CD83 mRNA was limitedly expressed within 7 samples.S100 expression had no significant(correlation) with clinical pathological features,while CD83 reversely correlated with TNM stages.Detected by flow cytometry,CD83 was expressed in low level in all 45 gastric cancer tissues and negative correlations were found with lymph node metastasis and TNM stages of gastric cancer(r=-0.879,P

Experimental study of dendritic cells inducing immunoeffects on anti-gastric cancer / 中国普通外科杂志

Objective To investigate the anti tumor effects of cytotoxic T lymphocytes induced by tumor antigen specific dendritic cells. Methods Soluble tumor antigen was prepared by four freeze thaw cycles of gastric tumor cell line SCG7901. Anti gastric tumor vaccine was acquired by co incubation of tumor antigen and mouse bone marrow derived dendritic cells and cultured with granulocyte/macrophage colony stimulating factor and interleukin 4. Antigen specific cytotoxic T lymphocytes were induced by this tumor vaccine from the spleen and were applied to tumor bearing nude mice. Results Tumor growth was significantly inhibited and the apoptosis of tumor cells was promoted extensively. Conclusions Antigen specific dendritic cell tumor vaccine may play an important role in future immunotherapy of gastric cancer.

A study on the biological characters of SGC7901 gastric cancer cell-dendritic cell fusion vaccine / 中国普通外科杂志

Objective To provide experimental basis for the study of SGC7901 gastric cancer cell-dendritic cell fusion vaccine. Methods The fusion of SGC7901 gastric cancer cells and dendritic cells was induced by PEG. We obtained pure fusion cells by selective culture with HAT/HT culture systems. The growth curve characters, clone abilities, abilities to irritate T lymphocytes and the ability to grow were detected. Results The fusion cells had weaker proliferative abilities and clone abilities with increased abilities to irritate T lymphocytes. The CPM value of T lymphocytes were 15083?231、 9608?83、 4214?135、 3020?28 respectively when they were co-cultured with the fusion cells on different proportions. Conclusion SGC7901 gastric cancer cell-dendritic cell fusion cells keep strong abilities to irritate T lymphocytes while inable to grow into new planted tumors in immunodeficiency animals.

Study on the activation effects to T lymphocytes by gastric cancer cell-dendritic cell fusion vaccines / 中国普通外科杂志

Objective To study the activation effects to cytotoxic T lymphocytes by gastric cancer cell-dendritic cell(DC) fusion vaccines and to provide theoretical data for biological therapy of gastric cancer based on(anti-tumor) fusion vaccines.Methods(1)Peripheral blood mononuclear cells from gastric cancer patients were separated and co-cultured with granulocyte macrophage colony stimulating factors,interleukin-4 and tumor necrosis factor-? to generate mature dendritic cells.(2)The dendritic cells and SGC7901 cells were fusioned by use of polyethylene glycol,and the pure fusion cells were screened out and cultured with HAT and HT(selective) culture systems.(3)The ability of fusion cells to activate the cytotoxic T lymphocytes was(investigated) by using MLA method.Results(1)Mature dendritic cells were gained from gastric cancer(patients)′ peripheral blood mononuclear cells by co-culturing with granulocyte-macrophage colony stimulating factors,interleukin-4 and tumor necrosis factor-?.(2)Dendritic cells and SGC7901 cells could be fusioned by PEG and could get pure fusion cells by culture with HAT/HT selective culture systems.(3)The fusion vaccines could strongly activate cytotoxic T lymphocytes,and there were significent differences between the(fusion) vaccine group and the control group.(P

Study on the induction culture and phenotypes of dendritic cells originating from peripheral blood of gastric cancer patients / 中国普通外科杂志

Objective To analyse the morphological and phenotypic characteristics of dendritic cells obtained by induction of peripheral blood monocytes of gastric cancer patients and provide an experimental basis of (dendritic) cells for the biological treatment of gastric cancer. Methods Peripheral blood monocytes of gastric cancer patients were co-cultured with GM-CSF, IL-4 and TNF-? to obtain the dendritic cells. The (morphological) characteristics of the dendritic cells were observed by light and electron microscopes and the (phenotypes) were studied by flow cytometry. Results Mature dendritic cells were oblained seven days after (induction) culture. These cells showed numerous typical dendritic protuberances on their surface.The (phenotypes) of the cells increased gradually with their maturation and reached stable levels seven days later. Conclusions Mature dendritic cells could be obtained from peripheral blood monocytes of gastric cancer (patients) after induction culture with GM-CSF,IL-4 and TNF-?. The obtained cells had typical morphological and phenotypic characteristics. These cells could be used for further research in the biological treatment of (gastric) cancer.