Construction of lung cancer cell model overexpressing human MutS homologue 2(hMSH2) / 基础医学与临床

Objective To construct human lung cancer cell model with human MutS homologous protein 2 (hMSH2) overexpression for exploring the effect of hMSH2 molecule in the cytotoxicity of γδ T cell against lung cancer cells.Methods hMSH2 coding sequence was cloned by PCR for construction of recombinant vector which over expressed hMSH2-EGFP fusion protein using homologous recombination.The recombinant vector was transfected to lung cancer cell line NCI-H520 to construct human lung cancer cell model overexpressing hMSH2 molecule.The expression of hMSH2 molecule in NCI-H520 was detected by Western blot.The expression of hMSH2 on the cell membrane was measured by flow cytometry.Cytotoxicity of expanded γδ T cells from peripheral blood mononuclear cells against NCI-H520 cells was detected by LDH release assay in vitro.Results hMSH2 coding sequence (2805 bp) was cloned and the result of restriction endonuclease digestion of Fugw-hMSH2 recombinant vector was accordance with the anticipated objective strip size.Exogenous hMSH2-EGFP fusion protein was expressed in NCIH520 cells.The level of hMSH2 molecule on the surface of NCI-H520 cells with overexpression of hMSH2 was significantly increased (P＜0.001).Cytotoxicity of γδ T cells against NCI-H520 cells with overexpression of hMSH2 was significantly increased compared to the wild type NCI-H520 cells (P＜0.05).Conclusions Lung cancer cell model that overexpresses hMSH2 molecule is successfully constructed,hMSH2 molecule on the cell membrane is increased and the cytotoxicity of γδ T cells against lung cancer cells is enhanced.

Composition and clinical significance of γδ T lymphocytes in peripheral blood of Gestational Diabetes Mellitus patients / 国际检验医学杂志

Objective To compare the composition of γδ T cells in the peripheral blood of gestational diabetes mellitus(GDM) patients with those of normal pregnant women,so as to explore the association of γδ T cells with the onset and progression of GDM.Methods The peripheral blood mononuclear cells(PBMC) of GDM patients(case group) and normal pregnant women(con-trol group) were separated by Ficoll-Hypaque density gradient centrifugation.γδ T cells were labeled with FITC conjugated anti TCRγδ monoclonal antibody(McAb) and PE conjugated anti-CD3 McAb and analyzed by flow cytometry.Results γδ T cells in GDM patients' peripheral blood were about(6.89 ± 3.22) ％,which was higher than normal control group(4.26 ± 1.64) ％.The percentage of γδ T cells in non-treated GDM group was significantly increased to(8.79 ± 2.33)％,while in treated GDM group,the percentage of γδ T cells was drop to normal(3.76±1.62)％.Conclusion γδ T cells in GDM patients' peripheral blood and CD3+ T Cells were slightly increased.The alteration may be related to the onset of GDM.

Composition and clinical significance of γδ T lymphocytes in peripheral blood of Gestational Diabetes Mellitus patients / 国际检验医学杂志

Objective To compare the composition of γδ T cells in the peripheral blood of gestational diabetes mellitus(GDM) patients with those of normal pregnant women,so as to explore the association of γδ T cells with the onset and progression of GDM.Methods The peripheral blood mononuclear cells(PBMC) of GDM patients(case group) and normal pregnant women(con-trol group) were separated by Ficoll-Hypaque density gradient centrifugation.γδ T cells were labeled with FITC conjugated anti TCRγδ monoclonal antibody(McAb) and PE conjugated anti-CD3 McAb and analyzed by flow cytometry.Results γδ T cells in GDM patients' peripheral blood were about(6.89 ± 3.22) ％,which was higher than normal control group(4.26 ± 1.64) ％.The percentage of γδ T cells in non-treated GDM group was significantly increased to(8.79 ± 2.33)％,while in treated GDM group,the percentage of γδ T cells was drop to normal(3.76±1.62)％.Conclusion γδ T cells in GDM patients' peripheral blood and CD3+ T Cells were slightly increased.The alteration may be related to the onset of GDM.

NaF· EDTA-K2 negatively bias blood lipids and lipoproteins measurement in pregnant women / 中华检验医学杂志

Objective To explore the impact of NaF · EDTA-K2 on blood lipids measurement in pregnant women , determine whetherthe empty-stomach plasma for OGTT could be used as substitute for serumin blood lipids analyses.Methods Fastingplasma with NaF · EDTA-K2 and serumfrom 100 pregnant women werecollected,andconcentration of CHO,TG, HDL,LDL,apoA1 and apoB were tested with Hitachi 7 600 automatic biochemical analyzer.Data were analyzed by SPSS 19.0 software.Results The concentrations oflipids and lipoproteins with NaF · EDTA-K2 were slightly lower than those of serum.There were statistical significances between the means of CHO , HDL, LDL, apoA1 and apoBof two groups ( P<0.05) except TG.Correlation coefficients of the six analyteswere CHO 0.968, TG 0.995, HDL 0.979, LDL 0.991, apoA1 0.692, apoB 0.846respectively.Standard Error of Estimate for plasma were: CHO 0.281, TG 0.094, HDL 0.077, LDL 0.112, apoA1 0.230, apoB 0.111 respectively.The modified coefficients were 1.08 for CHO, HDL and LDL, 1.14 for apoA1 and 1.07 for apoB.Conclusion NaF· EDTA-K2 negatively biased the blood lipidsmeasurement of CHO , HDL, LDL, apoA1 and apoB in pregnant women,yet lipids concentrations in plasma with NaF · EDTA-K2 were closely related to those in serum , which may be used to predict the bloodlipidslevel of pregnant women afterajusted.

MutS homolog 2 plays a role in Vγ9δ2 T cell-mediated anti-gastric carcinoma immunity / 中华微生物学和免疫学杂志

Objective To study the role of human MutS homolog 2 (hMSH2), a newly identified protein ligand that was recognized by Vγ9δ2 T cells , in innate anti-gastric carcinoma immunity .Methods Flow cytometry and laser confocal microscopy were used to identify hMSH 2 that ectopically expressed on gas-tric carcinoma cell line 803.An anti-hMSH2 antibody was used to block hMSH2 to evaluate its effects on the cytotoxicity of Vγ9δ2 T cells and their cytokines secretion .Subcellular distribution of hMSH 2 in gastric car-cinoma tissues was examined by tissue microarray immunohistochemistry analysis .Results Ectopic mem-brane expression of hMSH 2 was observed on 803 cells at a relatively high level .Vγ9δ2 T cells blocked with specific anti-hMSH2 antibody showed a decreased cytotoxicity and a reduced IFN-γbut an increased TNF-αsecretion.Ectopic expression of hMSH2 was found in various types of gastric carcinoma tissues at different stages.Enhanced expression of hMSH2 was detected in specimens collected from patients with chronic super-ficial gastritis.Conclusion Ectopically expressed hMSH2 served as a stress-induced endogenous ligand which could promote the cytotoxicity of Vγ9δ2 T cells against gastric carcinoma cells and enhance their IFN-γsecretion.hMSH2 played an essential role in innate anti-gastric carcinoma immunity .

Realization of dose verification tool for IMRT plan based on DPM / 中华放射医学与防护杂志

Objective To build a Monte Carlo dose verification tool for IMRT Plan by implanting an irradiation source model into DPM code and to extend the ability of DPM to calculate any incident angles and irregular-inhomogeneous fields.Methods The virtual source and the energy spectrum unfolded from the accelerator measurement data were used,in combination with optimized intensity maps,to calculate the dose distribution of the irradiation irregular-inhomogeneous field.The irradiation source model of accelerator was substituted by a grid-based surface source.The contour and the intensity distribution of the surface source were optimized by IMRT.The dose calculation was realized by combining the position of the emitter with the fluence map from the IMRT plan.The weight of the emitter was decided by the grid intensity.The direction of the emitter was decided by the combination of the virtual source and the emitting position.The weighted fraction of the emitter was also combined with the flux grid intensity based on the particle transport model of DPM code.Results The accuracy of calculation was verified by comparing with the measured data.It was illustrated that the differences were acceptable (＜ 2％ inside the field,2-3 mm in the penumbra).The dose calculation of irregular field by DPM simulation was also compared with that of FSPB (Finite Size Pencil Beam).The passing rate of gamma analysis was 95.1％ for peripheral lung cancer.The regular field and the irregular rotational field were all within permissible range of error.The calculation time of regular fields were less than 2 h,and that of the test of peripheral lung cancer was 160 min.Conclusions The adapted DPM code with its simple irradiation source model is faster than that with classical Monte Carlo procedure.Its computational accuracy and speed satisfy the clinical requiremcnt,and it can be useful as a Monte Carlo dose verification tool for IMRT Plan.

A research of correlation between renal damage of patients with gestational diabetes mellitus and level of serum cystatin C / 国际检验医学杂志

Objective To explore the correlation between renal damage of patients with gestational diabetes mellitus (GDM ) and level of serum cystatin C(Cys C) .Methods Eutocia puerperas with single birth in this center from January 2011 to December 2012 were retrospectively analyzed .400 puerperas ,who were confirmed as GDM by oral glucose tolerance test (OGTT) detection ,were collected .Among them ,200 puerperas with 24 h urinary protein content more than 0 .15 g during their 30 to 40 weeks of pregnancy were served as GDM group ,while another 200 puerperas with those not more than 0 .15 g as GDM control group .200 healthy puer-peras without GDM were served as the normal group .Serum uric acid ,creatinine ,urea ,Cys C and 24 h urinary protein content of them during their 30 to 40 weeks of pregnancy were detected .Results Serum uric acid and Cys C of patients in GDM group were obviously higher than those in the GDM control group(P0 .05) .24 h urinary protein content was positively correlated with the serum uric acid and Cys C(r=0 .715 ,0 .863 ,P0 .05) .Conclusion Serum Cys C may be used as indicators for early diagnosis and monitoring of renal damage in patients with GDM .

The role of a novel protein ligand hMSH 2 in Vγ9δ2 T cell-mediated anti-cervix cancer immunity / 中华微生物学和免疫学杂志

Objective To explore the role of hMSH2, a novel endogenous tumor-associated pro-tein ligand recognized by Vγ9δ2 T cells, in innate anti-cervix cancer immunity .Methods hMSH2 that ex-pressed on the surface of cervical cancer cell line HeLa cells was blocked by specific antibody .Then the differences in their effects on Vγ9δ2 T cells before and after antibody blockage were evaluated by cytotoxicity of Vγ9δ2 T cells and cytokines secretion .The serum levels of hMSH2 in patients with cervical cancer were detected by ELISA .Distribution of hMSH2 in cervical cancer tissues was analyzed by TMA immunohisto-chemistry .Results Decreased cytotoxicity and IFN-γsecretion were observed in Vγ9δ2 T cells against He-La cells blocked with specific anti-hMSH2 antibody .Serum concentration of hMSH 2 in patients with cervical cancer was slightly higher than that of healthy control .Altered distribution pattern of hMSH 2 was observed in cervical cancer tissues .Conclusion hMSH2 is involved in Vγ9δ2 T cells-mediated innate anti-cervical cancer immunity by enhancing their cytotoxicity and IFN-γsecretion.