Objective:To investigate the radiosensitization and the cell-cycle of mild hyperthermia(≤42℃)on human pulmonary adenocarcinoma cell line SPC-A-1 in vitro. Methods: The human pulmonary adenocarcinoma cell line SPC-A were treated with radiation and the combination of radiation with mild hyperthermia. Radiosensitivity was determined by clonogenic assay and quantified by calculating the thermal enhancement ratio (TER). Flow cytometry was used to observe the cell-cycle. Results: Do, Dq calculated from the dose-response curve for radiation combined with 41.5℃ were 1.390 Gy, 1.426 Gy, whereas 1.693 Gy, 2.453 Gy for radiation alone, respectively. TER was 1.218. The proportion of cells in S phase was found to be 14.81% in the radiation group. The values, after 48 hours and 72 hours, with 6Gy radiation combined immediate 41.5℃ one hour mild hyperthermia, were 5.89% and 9.08%, respectively, versus 18.8% and 31.91% with 6 Gy radiation alone. Conclusion:Radiosensitization of mild hyperthermia in SPC-A-1 cells associated with the hyper-radiosensitization of the cells in S phase.

Objective: To understand the relationship between chloroquine resistance and the virulence of Plasmodium berghei. Met hods: Dynamic changes of histopathologic features of livers, spleens, brains, hearts, lungs and kidneys of mice infected with the chloroquine-sensitive (N) and the chloroquine-resistant (RC) strains of P. berghei were compared. Results: In mice infected with the N strain, deposition of heavy hemoz oin in livers and spleens, congestive edema in lungs, and congestion and embolis m in the brain capillaries were observed. The histopathologic features revealed ac ute inflammatory reaction. In mice infected with the RC strain, histopathologic variations of livers and spleens were associated with changes of parasitemia. In terstitial pneumonia was displayed in lungs. There were chronic histopathologic changes of the organs in the mice infected with RC strain. Conclusion: The mice infected by the N strain with potent virulence die due to adher ence of the erythrocytes to microvascular endothelia and embolism of the microva scula, especially in their brain. Immune responses of the mice infected by the R C strain with poor virulence may be a delayed-type hypersensitive inflammation a ssociated with CD4+Th1 at an early stage of the infection, but may become anti body-dependent immune response assisted with CD4+Th2, which play a key role in elimination of the malaria parasites at later stage of the infection.

Objective To investigate the structural and functional changes of lower motor neuron distal to the site of spinal cord injury in rats. Methods Seventies Sprague-Dawley rats were divided randomly into 6 groups: sham-operation group (controls, n=10) and 3 day group (n=10), 1 week group (n=10), 2 week group (n=10), 4 week group (n=15) and 8 week group (n=15) after spinal cord transaction at T10. Neuronal apoptosis and acetylcholinesterase (AChE) activity of spinal cord at L4- 6 were observed by using the terminal deoxynucleotidal transferase- mediated DUTP-biotin nick end labeling (TUNEL) method and the semiquantitative enzyme cytochemistry, respectively. Results The assessment of apoptosis by TUNEL labeling showed that fluorescent markers were observed occasionally in anterior horn distal to the site of injury. The optical density (OD) value of AchE positive motor neurons (area > 300 μm2) initially decreased about 3 days after transaction and then overshot 1 week or so. However, after that, the OD value decreased again, the lowest about 4 weeks. Then the OD value increased again, though at 8 weeks was still lower than that of controls (P<0.05). Conclusion The findings on indistinctive apoptosis provided the proof of no significant changes of lower motor neuron distal to the site of transection. Semiquantitative histochemical results about AChE reflected marked metabolic changes of motoneurons caudal to the transaction, which represented as part of functional reorganization.

OBJECTIVETo evaluate the expression of CD4+, CD8+ T cells and cell apoptosis in oral lichen planus (OLP) and investigate the role and the relationship of immunological reaction and cell apoptosis in the pathogenesis of OLP2.

METHODSImmunohistochemical technique was used to study the expression of CD4+, CD8+ T cells in 27 OLP cases. TUNEL was used for detecting the cell apoptotic index (AI) in 17 OLP2 cases.

RESULTSThe expression of CD4+, CD8+ T cells were obviously elevated in lamina propria of OLP group compared with control group (P<0.05). There was a strong significance when compared the ration of CD4/CD8 in both group. AI was remarkably increased in epithelia cells and significantly decreased in lymphocytes in lamina propria in OLP cases compared with its expression in the control group respectively.

CONCLUSIONThe increased amount of CD4+, CD8+ T cells in lamina propria of OLP and the change ration of CD4/CD8 suggest that immune response is involved in the pathogenesis of OLP. The abnormal cell apoptosis plays an important role in the pathogenesis of OLP.

To develop a safe and broad-spectrum effective hepatitis C virus (HCV) T cell vaccine,we constructed the recombinant adenovirus-based vaccine that carried the hepatitis C virus truncated NS3 and core fusion proteins. The expression of the fusion antigen was confirmed by in vitro immunofluorescence and western blotting assays. Our results indicated that this vaccine not only stimulated antigen-specific antibody responses,but also activated strong NS3-specific T cell immune responses. NS3-specific IFN-γ+ and TNF-α+ CD4+ T cell subsets were also detected by a intracellular cytokine secretion assay. In a surrogate challenge assay based on a recombinant heterologous HCV (JFH1,2a) vaccinia virus,the recombinant adenovirus-based vaccine was capable of eliciting effective levels of cross-protection. These findings have im- portant implications for the study of HCV immune protection and the future development of a novel vaccine.
Adenoviridae ; genetics ; metabolism ; Animals ; CD4-Positive T-Lymphocytes ; immunology ; Cross Protection ; Female ; Genetic Vectors ; biosynthesis ; genetics ; Hepacivirus ; genetics ; immunology ; Hepatitis C ; immunology ; prevention & control ; virology ; Humans ; Interferon-gamma ; immunology ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; administration & dosage ; genetics ; immunology ; Viral Core Proteins ; administration & dosage ; genetics ; immunology ; Viral Hepatitis Vaccines ; administration & dosage ; genetics ; immunology ; Viral Nonstructural Proteins ; administration & dosage ; genetics ; immunology

Objective To investigate the immunogenicity and cross protective effects of two novel HCV DNA vaccines in a mice model.Methods Two self-replicating alphavirus vector-based HCV DNA vaccines, pSCK CE1E2Y and pSCK H155, were constructed based on the genes encoding the structural pro-teins (Core, E1 and E2) and structural and NS3 fusion proteins (Core, E1 , E2 and NS3) of a HCV strain isolated from a Chinese patient (genotype 1b, Hebei strain), respectively.Western blot analysis was per-formed to detect the expression of fusion antigens.The BALB/c mice were intradermally immunized with the recombinant DNA vaccines by using electroporation.The immune responses induced in mice and the cross protective effects of the recombinant DNA vaccines were evaluated.Results The DNA vaccines effectively expressed the target antigens in vitro.The antigen-specific antibody responses and specific T cell immune re-sponses were induced in mice by the immunization of replicative DNA vaccines.However, no effective cross protection was provided by either of the DNA vaccines in the surrogate challenge model based on a recombi-nant heterologous HCV (JFH1, 2a) vaccinia virus strain.Conclusion Although no effective cross protec-tion was observed, both of the two replicative DNA vaccines could induce strong humoral and cellular im-mune responses against multi-target antigens of HCV strains.This study has paved the way for further inves-tigation on the development of novel HCV vaccines.

BACKGROUND: Tumor-adopted immunity and gene transduction technique are used to introduce tumor necrosis factor-α vector into carrier cells, which are then re-infused into the body so that cancer cells can be killed by tumor necrosis factor-α more directly and effectively with fewer side effects on the other tissues due to high local expression.OBJECTIVE: To study the bioactivity of in vitro cultured tumor necrosis factor-α transduced tumor-infiltrating lymphocytes as well as the inhibitory effects on cancer cells of cancer-loaded rats infused in different ways.DESIGN: A randomized controlled study based on experimental animals.SEETING: Cancer Research Institute of China Medical University.MATERIALS: This study was carried out at the Cancer Research Institute and the Experimental Animal Department, China Medical University,between January 2000 and December 2001. TJ8510 cell line (human brain glioblastoma cell line) was provided by the Neurological Research Institute of Tianjin Medical University Affiliated Hospital. The experimental animals were 36 BALB/C nude mice congenitally having no thymius.METHODS: Based on the establishment of tumor necrosis factor-α retroviral transduction system and the preparation of cartier cells tumor-infil-trating lymphocytes, the monoclonal virus cell line PLC-2 and PLJC-5available were used to introduce marked gene NeoR and targeted gene tumor necrosis factor-α into tumor-infiltrating lymphocytes, respectively.Then cell proliferation, tumor necrosis factor expression and in vitro antitumor activity were examined. After cancer cell inoculation, the 36 nude mice were randomly divided into 6 groups: local infusion control group, local tumor-infiltrating lymphocytes infusion group, local tumor necrosis factor-tumor-infiltrating lymphocytes infusion group, venous infusion control group, venous tumor-infiltrating lymphocytes infusion group and venous tumor necrosis factor-tumor-infiltrating lymphocytes infusion group, and the therapeutic effects on the cancer-loaded mice were observed.proliferation and tumor necrosis factor-α expression in tumor-infiltrating oR-tumor-infiltrating lymphocytes and tumor necrosis factor-tumor-infiltrating lymphocytes was not significantly different from each other (P ＞ 0.05).NeoR-tumor-infiltrating lymphocytes, though not significantly different (P ＞0.05), significantly differ from that of tumor necrosis factor-tumor-infiltrating lymphocytes (P ＜ 0.01); moreover, tumor necrosis factor-tumor-infiltrating lymphocytes were found to express higher tumor necrosis factor-α conactivity did not significantly differ between tumor-infiltrating lymphocytes and NeoR-tumor-infiltrating lymphocytes (P ＞ 0.05), but obviously increased come of the animal experiment: 40 days after tumor necrosis factor-tumorinfiltrating lymphocytes infusion, cancer size in local tumor necrosis factortumor-infiltrating lymphocytes infusion group was found smaller than that in local infusion control group [(307±42) and (2 048±278) mm3, P ＜ 0.01],and it was also smaller in venous tumor necrosis factor-tumor-infiltrating lymphocytes infusion group than that in venous control group [(954±195)and (1 989±305) mm3 , P ＜ 0.05].CONCLUSION: Tumor necrosis factor-α gene transduced tumor-infiltrating lymphocytes could effectively express tumor necrosis factor, exerting higher and in vivo anti-tumor effects than tumor-infiltrating lymphocytes in cancer-loaded nude mice. No obvious inhibitory effects on the growth of subcutaneous solid carcinoma could be observed in nude mice after venous infusion of human brain glioblastoma tumor-infiltrating lymphocytes, but the inhibitory effects became obvious due to venous infusion of tumor necrosis factor-tumor-infiltrating lymphocytes and significant due to local tumor necrosis factor-tumor-infiltrating lymphocytes infusion, indicating that local infusion is the preferable way in the treatment of glioblastoma by immuno-gene therapy.

Objective To investigate the development strategy of novel T cell based vaccine against HCV infection.Methods BALB/c mice were primed with pSCK-based DNA vaccine and boosted with type 5 adenoviral vector-based vaccine, which expressed the structural proteins ( Core, E1 and E2) de-rived from a Chinese HCV patient (genotype 1b, Hebei strain).Enzyme linked immunospot assay (ELIS-POT) and intracellular cytokine staining ( ICS) were used to analyze the elicited antigen-specific immune re-sponses and the efficacy of cross-protection.Results Immunization of mice with the prime-boost vaccination strategy elicited stronger T cell immune responses against multiple HCV antigens than using the DNA vac-cines alone, especially the IFN-γ-secreting T cell responses against E1 protein as indicated by ELISPOT as-say.ICS data indicated that the prime-boost regimen elicited more TNF-α-producing CD4+and IFN-γ-produ-cing CD8+T cells against E1 protein and high levels of IFN-γ-producing CD4+and CD8+T cells against E2 protein in comparison with immunization with DNA vaccines.Moreover, the prime-boost vaccination was ca-pable of eliciting effective cross-protection in a surrogate challenge model based on a recombinant heterolo-gous HCV (JFH1, 2a) vaccinia virus.Conclusion The prime-boost vaccination using DNA and rAd5-based vaccine expressing HCV structural antigens induced significant cellular immune response and cross-protection in mice, suggesting the possibility of using it as a promising T cell based vaccine against HCV in-fection.

A comparable study were carried out by determination of trace elements on five marine-derived shell traditional Chinese medicine (TCM) (Ostreae Concha, Haliotidis Concha, Margaritifera Concha, Meretricis Concha, and Arcae Concha), which were recorded in the Chinese Pharmacopoeia (2010 version). Seven trace elements in 51 batches of this type of shell TCM were analyzed by Inductively Coupled Plasma Mass Spectrometry (ICP-MS), combined with principal component analysis (PCA) methods. The content of element Se, which exhibited significant differences among different drugs, could be used as a key element to distinguish this type of drugs. Meanwhile, the contents of elements Co, Cu, Mo, and Ba in Haliotidis Concha, Co and As in Margaritifera Concha, Mo and As in Meretricis Concha, Mo, As, and Ba in Arcae Concha, and Zn in Meretricis Concha were relatively stable. In the PCA plot, Arcae Concha and Meretricis Concha could be efficiently distinguished from Ostreae Concha together with Haliotidis Concha, and Margaritifera Concha. The results also showed a correlation with their medicinal function. In conclusion, trace elements in marine-derived shell TCM could not be neglected for their quality control.
Animal Shells ; chemistry ; Animals ; Aquatic Organisms ; chemistry ; Bivalvia ; chemistry ; Mass Spectrometry ; Medicine, Chinese Traditional ; Trace Elements ; analysis

Objective To analyze the characteristics of laboratory test resuits of dengue fever(DF)patients in Guangzhou area.Methods Routine tests were performed in the patients admission to hospital. Serology examination was performed in the patients in acute phase or recovery phase.The clinieal symptoms and teatures were analyzed and positive numbers and positivity ratios were calculated.Results The clinical symptoms of the dengue fever were typieal,with the features of fever,headache,myalgia and rash.The leukopenia rate was 76.0%,and the thromboeytopenia rate was 62.6%.The levels of ALT increased in 56.7%patients,and the levels of AST increased in 84.0%patients.Hypopotassemia was found in 46.1%patients.Dengue virus antibody IgM(DF-IgM)was detected positive from the first day to the 16th day of the onset,and the positive rate was 85.9% on the 8th day.Virus loads were positive by fluorescence real-time PCR in seven acute serum samples(within 3 days of the onset)of 51 cases whose DE-IgM were negative all the time, and the results was 105 -106 copies/ml(＜103 copies/ml means negative).Conclusions Clinical manifestations of this DF epidemic were typical including fever,headache,myalgia and skin rash.Most of the patients had decreased leukocyte and thrombocyte obviously.Liver damage was common but kidney damage was seldom.Halt of the patients got hypopotassemia.DF-IgM appeared in very early and persisted for a long time.The detection of DF-IgM within 7 days of the onset was helpful for diagnosis as early as possible.Viral load detected by real-time PCR could be another indicator of early pathogen diagnosis which provides complementation for antibody detection.