Objective To investigate the role of PKCθ signal pathway on regulation of L-selectin (CD62L) expression in human activated γδT cells. Methods Human peripheral blood mononuclear cells (PBMC) were stimulated with Mycobacterium tuberculosis antigen (Mtb-Ag) and cultured for 6-8 d to generate Mtb-Ag activated T cells(MtbAT) as γδT cells enrichment T cell line. The MtbAT were stimulated with PMA or PMA + IMN for 3, 6, 12 and 24 h, respectively, or MtbAT cultured for 8 d were stimulated with Mtb-Ag, or PMA, with or without PKC0 inhibitor Rottlerin for 4 h. After the treated cells stained with fluo-rescent labeled monoclonal antibodies, the expression of CD62L on γδT cells were measured by flow cytome-try (FCM ). Results The expression of CD62L on γδT cells cultured for 6-8 d were 75.0%-87.0%. Decrease of CD62L from the surface of γδT cells by 3 h to 12 h after exposure to PMA (42.3% to 23.5% ), but CD62L expression increased to 53.2% when γδT cells were exposed to PMA for 24 h. The expression of CD62L of γδT cells decreased to 52.1% and 39.3% respectively when γδT cells were exposed to PMA + IMN for 3 h and 6 h. After treated with PMA + IMN for 12 h and 24 h, the expression of CD62L were 52.9% and 35. 3% respectively. The CD62L expression of γδT cells treated with PMA and Rottlerin (47.9%) were higher than that treated with PMA alone (31.8%). After Mtb-Ag restimulated MtbAT for 4 h, the CD62L level of γδT cells decreased from 70.0% to 54.8%, Rottlerin could inhibite Mtb-Ag down regulation CD62L level of γδT cells (63.1%). Conclusion The CD62L expression of γδT cells could be ingibited partly by the inhibitor of PKCθ signal pathway may regulate L-selectin (CD62L) expression of activated human γδT cells.

AIM: To investigate the expression and effects of sialyl Lewis X (SLeX) on the invasion and migration of human hepatocellular carcinoma HepG2 cells.METHODS: The expression of α1,3-fucosyltransferase VII (FUT7) in HepG2 cells and L-02 cells was detected by RT-qPCR and Western blot.The SLeX expression in HepG2 cells and L-02 cells was determined by Western blot and immunocytochemical staining.The invasion and migration abilities of the treated cells were evaluated by Transwell assay.RESULTS: The expression of FUT7 and SLeX in the HepG2 cells, but not in the L-02 cells, was observed.The invasion rates of the HepG2 cells treated with SLeX monoclonal antibody at 0.05, 0.5 and 5 mg/L were significantly decreased as compared with control group (P<0.05).The migration ability of the HepG2 cells treated with SLeX monoclonal antibody at 0.05, 0.5 and 5 mg/L was also significantly reduced as compared with control group (P<0.05).The invasion rate and migratory cell number were significantly different between any 2 groups in the HepG2 cells treated with SLeX monoclonal antibody at 0.05, 0.5 and 5 mg/L (P<0.05).CONCLUSION: HepG2 cells express SLeX.SLeX is closely related to the migration and invasion abilities of the HepG2 cells.

Objective:To investigate the effect of Artesunate (Art) on the expression of transferrin receptor (TtR)in K562/ADM cells.Methods:The drug-resistant K562/ADM cells were cultured with 1000 ng/mL doxorubicin for two weeks followed by Artesunate treatment with different concentrations (12.5 μg/mL,25μg/mL and 50 μg/mL) or different time (12 h,24 h,36 h,and 48 h).The content of transferrin receptor in K562/ADM cells was determined by flow cytometry.The effect of Artesunate on the expression of transferrin receptor protein in K562/ADM cells was measured by Westem blot.Cell counting kit-8 (CCK-8) assay was used to evaluate inhibitory effect of Art combined with doxorubicin (ADM) in K562/ADM cells.The reversal index was defined as the IC50 of the experimental group divided by the IC50 of the control group in K562/ADM cells.Results:Art effectively decreased the content of transferrin receptor and the expression of transferrin receptor protein in K562/ADM cells in a dose-dependent manner.Moreover,Art also inhibited transferrin receptor protein expression in K562/ADM cells in a time-dependent manner.The different concentrations of Art(12.5 μg/mL,25μg/mL and 50 μg/mL) could induce reversal of drug-resistance with the reversal index being 1.38,2.12 and 2.95 times (P＜0.05).Art inhibited cell proliferation of K562/ADM cells,and the IC50 werel9.7 μmol/L.Conclusions:Art effectively down-regulated the transferrin receptor content as well as transferrin receptor protein expression in K562/ADM cells,which resulted in reversal of drug resistance of K562/ADM cells.Art also inhibited K562/ADM cells proliferation,which has great value in clinical treatment of leukemia.

Background Cornea astigmatism can be effectively corrected by implanting Toric intraocular lens (IOL) during cataract surgery and therefore improve visual acuity of patients.However,the decentration and rotation position errors were inevitable sometime.What's the difference of effect of position errors on quality of image between spherical IOL and Toric IOL needs further research.Objective This study was to evaluate the optical performance and wavefront with rotation and decentration of Toric IOL.Methods Different decentration for SN60AT IOL(spherical IOL) and Toric IOL in Hwey-Lan Lion model eyes was set with the role as follows:decentration 0.25 mm to 0.75 mm in a 5°-interval from 0° to 90°.Furthermore,Toric IOL was rotated at 5° and 10°,respectively.Then the image performances of SN60AT IOL and Toric IOL at different decentration distances and rotated degrees were evaluated with modulation transfer function (MTF) and value of wavefront aberration under all conditions.Results At the centration,the MTF curves of spherical IOL and Toric IOL were similar under 3,4 and 5 mm pupil diameter at each spatial frequency.Under the condition of 4 mm pupil diameter,when the decentration was 0.25 mm,the MTF values of SN60AT IOL at 6 c/d and 12 c/d were 0.581 087 and 0.411 960,respectively.T3 IOL were 0.454 259 and 0.382 313,T4 IOL were 0.426 020 and 0.360 490,T5 IOL were 0.425 606 and 0.359 877.When the decentration was 0.50 mm,the MTF values of SN60AT at 6 c/d and 12 c/d were 0.573 073 and 0.412 787,respectively.T3 IOL were 0.450943 and 0.379481,T4 IOL were 0.423 153 and 0.356 664,T5 IOL were 0.422 881 and 0.356 230.When the decentration was 0.75 mm,the MTF values of SN60AT at 6 c/d and 12 c/d were 0.560 038 and 0.413 624,respectively.T3 IOL were 0.445 597 and 0.374 322,T4 IOL were 0.418 522 and 0.350 087,T5 IOL were 0.418 468 and 0.349 976.When the IOL decentralized along 0°,5°,10°,90°meridian line,the MTF values were almostly same.The root mean square (RMS) of spherical IOL and Toric IOL was increased when the IOL decentralized from 0 mm to 0.75 mm,with the most increasing level in coma aberration and slight increase in trefoil aberration.When the T4 IOL decentralized from centre to 0.75 mm,the coma increased from 0 to C(3,-1)-0.049 79 μm,C (3,1)-0.037 59 μm and the trefoil aberration increased from 0 to C (3,3) 0.005 72 μm,C (3,-3) 0.004 64 μm.With the increase of rotation degrees (from 5°to 10°) of Toric IOL,the MTF was worse at high spatial frequency.Toric IOL rotation caused the increase of astigmatism and residual astigmatism and spherical error,but not high order aberration.Conclusions The tolerance of Toric IOL to decentration is very close to the spherical IOL,and optical performance is only associated with the amount of decentration but not direction.The aberration caused by Toric IOL decentration is mainly coma.The rotation of Toric IOL causes astigmatism error but not high order aberrations.

OBJECTIVETo investigate the effect of occupational exposure to toluene diisocyanate (TDI) on the workers' health.

METHODSA total of 76 workers exposed to TDI (exposure group) and 64 management staff members (control group) were selected from a factory as the study subjects. Area sampling was performed for the place with exposure to TDI according to the method in GBZ 159-2004 Specifications of air sampling for hazardous substances monitoring in the workplace, and gas chromatography was applied to measure the concentration of TDI in workplace air. The workers' personal information was collected with questionnaire, pulmonary ventilation function was determined with a portable spirometer, hematological parameters were analyzed by automatic blood analyzer and blood chemistry analyzer, and the indicators of oxidative damage and energy metabolism were measured by the reagent kit provided by Nanjing Jiancheng Bioengineering Institute. SPSS 17 software was applied for statistical analysis.

RESULTSThe exposure group had significantly lower forced vital capacity (FVC), forced expiratory volume in 1 second(FEV1.0), and FEV1.0/FVC ratio than the control group (P <0.05). Compared with the control group, the exposure group had significantly higher red blood cell count, platelet distribution width, mean platelet volume, lymphocyte count, and neutrophil count(P<0.01), and significantly lower activities of lactate dehydrogenase(LDH), superoxide dismutase, and succinodehydrogenase (SDH)(P <0.01). In the exposure group, the length of exposure was negatively correlated with the activities of SDH and LDH in the serum (r=-0.319, P <0.05; r=-0.239, P <0.05), and the length of exposure was not found to be correlated with the activity of SOD and pulmonary function indices.

CONCLUSIONTDI can induce inflammatory response and lung ventilation function impairment in workers exposed to TDI, as well as oxidative stress and imbalance of energy metabolism. Therefore, it can cause damage to workers' health, and protective measures should be enhanced.

Case-Control Studies ; Erythrocyte Count ; Forced Expiratory Volume ; Humans ; Inflammation ; physiopathology ; L-Lactate Dehydrogenase ; blood ; metabolism ; Leukocyte Count ; Lung ; physiopathology ; Occupational Exposure ; adverse effects ; Pulmonary Ventilation ; Succinate Dehydrogenase ; blood ; metabolism ; Superoxide Dismutase ; metabolism ; Toluene 2,4-Diisocyanate ; adverse effects ; Vital Capacity

OBJECTIVE: To explore the effects of low molecular weight heparin (LMWH) on cisplatin (DDP)- induced apoptosis in human hepatocellular carcinoma cell and the underlying mechanisms.
 METHODS: Hepatocellular carcinoma SMMC-7721 cells were divided into a control group, a LMWH group, a DDP group and a LMWH plus DDP group. The effect of the drugs on proliferation of hepatocellular carcinoma SMMC-7721 cells were evaluated by 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di- phenytetrazoliumromide (MTT) assay, and the apoptosis were detected by acridine orange/ethidium bromide (AO/EB) double fluorescence method and flow cytometry method. The expression levels of apoptosis-related protein Fas, Bcl-2 and Bax were detected by quantitative real-time PCR and Western blot.
 RESULTS: Compared with the control group, the proliferation of SMMC-7721 cells was inhibited in the DDP group and the LMWH plus DDP group (both P<0.05). The mRNA and protein levels of Fas, Bcl-2, Bax in the SMMC-7721 cells were not obviously changed in the LMWH group (all P>0.05). The Fas level was increased obviously (P<0.05), while the Bcl-2 level moderately reduced (P<0.05), Bax were not obviously changed in the DDP group (P>0.05). Compared with the control group and the DDP group, the Bcl-2 level was reduced significantly in the LMWH plus DDP group (both P<0.05), while the level of Bax was increased obviously (both P<0.05). Compared with control group, the Fas level was increased significantly in the LMWH plus DDP group (P<0.05), but the increase was not significant compared with the DDP group (P>0.05).
 CONCLUSION LMWH can enhance the cisplatin-induced apoptosis in SMMC-7721 cells, which might be related to activation of mitochondrial pathway.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; drug effects ; Cisplatin ; pharmacology ; Flow Cytometry ; Heparin, Low-Molecular-Weight ; pharmacology ; Humans ; Liver Neoplasms ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Real-Time Polymerase Chain Reaction ; bcl-2-Associated X Protein ; metabolism ; fas Receptor ; metabolism

OBJECTIVE: To explore the rate and distribution of Runt- related transcription factor 1 (RUNX1) gene mutations in patients with acute myeloid leukemia (AML) and the correlation of these mutations with the clinical characteristics and survival outcomes of the patients. METHODS: The genomic DNA extracted from the bone marrow of 158 patients with newly diagnosed AML for PCR amplification of RUNX1 gene and sequence analysis to identify the mutations. The mutations of ASXL1, DNMT3A, TET2, FLT3, CEBPA, NPM1, IDH2, NRAS and c-KIT genes were also examined to analyze their association with RUNX1 gene mutations. RESULTS: Among the 158 AML patients, 19 (12.0%) were found to have RUNX1 mutations in A166G (9 cases), A142T (6 cases) and A162L (4 cases). RUNX1 mutations were more frequent in elderly patients ( CONCLUSIONS RUNX1 gene mutations are associated with an adverse prognosis of patients with AML.
Core Binding Factor Alpha 2 Subunit/genetics* ; Humans ; Leukemia, Myeloid, Acute/genetics* ; Mutation ; Prognosis ; Remission Induction