Abnormal increase in CD8(low) T lymphocyte in patients with occupational chronic lead poisoning / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To analyze the changes in CD8(low) T lymphocyte subsets in patients with occupational chronic lead poisoning.</p><p><b>METHODS</b>Flow cytometric analysis was used to count the numbers of CD8+ cells. 23 patients with occupational chronic lead poisoning and 20 controls were examined.</p><p><b>RESULTS</b>Compared with control group (8.21% ± 3.02%), the CD8(low) T lymphocyte (12.98% ± 5.62%) were significantly increased in patients with occupational chronic lead poisoning.</p><p><b>CONCLUSION</b>Although the ratio of CD+ T lymphocyte is normal, the CD8 level is significantly decreased. The increase of CD8(low) T lymphocyte may be an important phenomenon of immuno-injury induced by lead. CD8(low) T lymphocyte could be an new direction for research of lead immuno-toxicity.</p>

Anti-tumor effects induced by fusion of interleukin-18 gene transfected NCI-H460 lung cancer cell line with dendritic cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the anti-tumor effects induced by fusion of interleukin (IL)-18 gene transfected lung cancer cell line NCI-H460 cells with dendritic cells (DC).</p><p><b>METHODS</b>(1) DC were induced from human monocytes and fused with IL-18 transfected NCI-H460 cells. Fusion was selected using MACS microbeads. (2) Four groups (group GT, group PT, group NT and group BC) were set up. T cells activated by IL-18 gene transfected fusion or pcDNA3. 1 + vector transfected fusion and non-transfected fusion were taken as effetor cells. No effector cells was in group BC. Lactic dehydrogenase ( LDH) method was used to evaluate the antitumor effect in vitro. (3) Tumor-bearing nude mice were inoculated with effector cells mentioned above. The tumor size and weight in the 4 groups were compared.</p><p><b>RESULTS</b>The killing rate in vitro of 3 groups were 53. 14% ,30. 10% and 31.49% , respectively. The tumor size and weight in the 3 groups were lower than group BC, among which group GT was the lowest.</p><p><b>CONCLUSION</b>Fusion of IL-18 gene transfected NCI-H460 lung cancer cells with dendritic cells can effectively induce anti-tumor immunity in the host.</p>

Expression of sialylated carbohydrate antigens and nm23-H1 gene in prognosis of breast cancer / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To assess the significance of expression of sialylated carbohydrate antigens and nm23-H1 gene in metastasis and prognosis of breast cancer.</p><p><b>METHODS</b>Tissue specimens from 102 cases of primary breast cancer were stained with antibodies against sialyl Lewis A (SleA) and salyl Lewis X (SleX), and nm23-H1 proteins by immunohistochemical methods.</p><p><b>RESULT</b>Of the 102 cases, the positive cases of SleA and SleX were 24.5% (25/102) and 59.89% (61/102),respectively; the reduced expression of nm23-H1 was showed in 37.3% (38/102) of the cases. The positive expression of SleX and the reduced expression of nm23-H1 gene were significantly associated with lymph node involvement. Among the 100 patients who underwent curative surgery, the disease-free survival rate was significantly correlated with nm23-H1 and SleX expression, respectively,but not with SleA expression. In multivariate analysis using Cox regression model, combination assay of nm23 H1 and SleX expression emerged as independent prognostic factors.</p><p><b>CONCLUSION</b>These results suggest that nm23-H1 gene and SleX may be involved in the metastatic process in human breast cancer, and immunohistochemical detection of SleX and nm23-H1 may be used as a biologic marker of prognosis.</p>

Construction of human MAGE-3 DNA vaccine and its immune effects observed in vivo / 生物工程学报

To investigate the antitumor immune responses induced by MAGE-3 DNA vaccine, the recombinant mammalian expression plasmid pcDNA3.1/MAGE-3 was constructed by ligating MAGE-3 gene, which was amplified by RT-PCR, and the pcD-NA3.1 + vector. The recombinant plasmids were transfected into B16 cells by liposome, the expression of MAGE-3 was checked by RT-PCR, immunocytochemistry and Western blot. Then, 100 ug recombinant plasmids were injected intramuscularly per C57BL/6 mouse on 0, 10 and 20 days, with pcDNA3.1 + plasmid and PBS as controls. Splenocytes CTLs, the level of antibodies against MAGE-3 the changes of the T lymphocyte subsets and the levels of cytokines were checked after 3 times immunization. As a result, the mice immunized with pcDNA3.1/MAGE-3 plasmid can produce MAGE-3 specific immune response. The CTLs kill activities against B16/MAGE-3 cells was 51.08 +/- 7.41%, and had significant difference (P < 0.01) compared with that of pcDNA3.1 + group (8.44 +/- 1.89%) and PBS group (5.76 +/- 1.75%). The titre of antibody against MAGE-3 was 1:15, while controls were negtive. The number of CD4 + CD8 + and the levels of IFN-gamma IL-2 increased significantly after immunization with pcDNA3.1/MAGE-3 plasmid as compared with those of control groups (P < 0.01). It is concluded that the pcDNA3.1-MAGE-3 DNA vaccine are able to induce both cellular and humoral immune responses in vivo.

Construction of MAGE-3 prokaryotic expression plasmid and its expression in Escherichia coli / 生物工程学报

To express the GST-MAGE-3 protein in E. coli, and investigate the antitumor immune responses induced by Dendritic cells (DCs) pulsed with GST-MAGE-3 protein, the recombinant expression plasmid pGEX-MAGE-3 was constructed by ligating MAGE-3 gene, which was amplified by RT-PCR and confirmed by sequencing, and the pGEX-4T-2 vector. The recombinant plasmid was transformed into BL-21 E. coli. The expression of GST-MAGE-3 was induced with IPTG. The GST-MAGE-3 protein expressed as high as 32% of the total cellular protein. After purification with Glutathione Sepharose 4B, the purity of the protein was more than 90%, and 3mg GST-MAGE-3 was obtained from 100 mL BL-21 lysate. Dendritic cells from gastric carcinoma patients were pulsed with GST-MAGE-3 protein, and these DCs were used to stimulate the autologous T. lymphocytes. After 7 days, the T. lymphocytes cocultured with DCs pulsed with GST-MAGE-3 antigen exhibited specific cytotoxicity against MAGE-3 positive SGC-7901 cells. It is concluded that the GST-MAGE-3 protein are able to present antigen to T. lymphocytes, activate antigen-specific CTLs and induce special antitumor immune responses in vitro. Our results lay the groundwork for further research of the MAGE-3 vaccine.

Measurement of the mRNA level of Polo-like kinase 1 in bronchoscopic bioptic specimens by realtime quantitative PCR / 中华检验医学杂志

0.05)and the TNM staging (P=0.55).A mild elevated compared other pathological classification was found in small cell lung cancer (0.191?0.275).Conclusions The results showed that RFQ-PCR was suitable for measurement of the mRNA level of PLKI in bronchoscopic bioptic specimens.This study suggest elevated expression of PLK1 might play a important role in development of lung cancer,so that PLK1 might be a potential tumor marker for Lung cancers.Advanced studies will be needed to clarify that PLKI mRNA level do not relate to TNM staging and pathological classification.

A fMRI study of item memory and source memory / 中华神经科杂志

Objective To examine the neural basis of item memory and source memory with fMRI approach.Methods Eight male and eight female healthy fight-handed native Chinese speakers were involved in this study.The item memory and source memory task were conducted with 504 highly frequent Chinese double-character words in the Block-designed experiment.Participants underwent such a double- round procedure as fMRI scanning following study.The fMRI data collected from a GE 1.5T MRI system were analyzed to generate corresponding activation maps for females and males respectively(P20)using statistical parametric mapping software(SPM).Results For females,item memory task activated the bilateral dorsolateral prefrontal cortex(BA6,the number of activated voxel clusters was 62 or 11 in the left and the right,respectively),source memory more activated the left dorsolateral prefrontal cortex(BA6/46,the number was 59).For males,item memory activated the right dorsolateral prefrontal cortex(BA6/46,the number was 64),source memory activated the bilateral dorsolateral prefrontal cortex(BA6,9 and 40 in the left and the right).Conclusion On the neural basis of item or source memory,there exists dissociation,which is that right dorsolateral prefrontal areas are more activated by item memory while left dorsolateral prefrontal areas by source memory.For the difference of gender,it is suggested that left dorsolateral prefrontal areas(BA6/46)are more activated in females while right dorsolateral prefrontal cortex(BA6/46)more in males.

Ultrasmall superparamagnetic iron oxide-enhanced MRI in a rabbit model of antigen-induced arthritis:a preliminary study / 中华放射学杂志

Objective To investigate the feasibility of ultrasmall superparamagnetic iron oxide- enhanced(USPIO)-enhanced MR imaging for monitoring synovitis of antigen-induced arthritis in rabbit model and explore the optimal MR imaging sequences.Methods Nine female white rabbits with antigen(0.5 ml mBSA,2 mg/ml)induced arthritis of the right knees were used in the study.The left knees of these rabbits and both knees of another 3 rabbits served as the control.Nine to 28 days(mean 21.3 d)after successful model induction,all knees were imaged before and 24 h after intravenously injection of USPIO (0.3 ml/kg),among which 2 rabbits were also imaged at 48 and 72 h after administration of USPIO respectively.The MR protocol included spin-echo(SE) T_1WI,fast spin-echo(FSE)T_2WI,gradient echo (GRE)T_2~* WI and short tau inversion recovery(STIR).Images were analyzed quantitatively and qualitatively based on signal characteristics and patterns of the synovium.Paired t-test was used for the analysis of the signal intensity of inflammatory synovial membrane before and 24 h after injection of USPIO. MR findings were correlated with histopathology.Results Arthritis was successfully induced in all 9 right knees with intraarticular injection of mBSA.Pathological examination revealed hyperplasia of synovium with infiltration of USPIO-loaded-macrophages.MR depicted synovial thickening(thickness 2.07?0.97 mm) and joint effusion.Synovium and joint fluid appeared as slightly hypo- or iso-intense on T_1 WI and hyper- intense on T_2 WI or T_2~* WI.Twenty four hours after USPIO injection,significant T_1 enhancement(ASNR 41.91%?27.94%),negative T_2 and T_2~* enhancement(△SNR -34.92%?11.77% and -57.24%? 16.05%)were demonstrated in the region of synovial inflammation respectively.The signal at 48 h and 72 h changed less than that at hour 24.No signs of arthritis occurred in all left knees and in all knees of the artificial model group.Conclusion Iron oxide phagocytized into macrophages can be a root cause resulted in signal change on USPIO-enhanced MR images.The gradient echo sequence should be the optimal sequence to be used in USPIO-enhanced MR imaging in antigen-induced arthritis.