Objective To develop a convenient method efficiently expands the frequency of specific CTLS.Methods We used different concentrations of CMV-speeific epitope peptides pp65 to stimulate PBMCs for expansion of CMV-specific CTLs.CMV-specifie CTLs were doubly labeled by tetramers-PE and CD_8-FITC for FACS analysis.Results The method expands CMV-speeific CTLs efficiently.CMV-specific CTLs were expanded from 1% to 20% of PBMCs quickly(namely 40% of CD_8~+ T cells).The method provided a large number of cells with tetramer staining of CD_8~+ T cells for FACS analysis from a single blood sampling.Conclusions Peptides stimulation methods are convenient,easy to operate and expanded CMV- specific CTLs efficiently.The increased frequencies of CMV-specific CTLs allowed the data of different individuals to be easily compared and sequentially evaluated.The methods lay the base for adoptive immunotherapy to prevent CMV disease.

BACKGROUND:Umbilical cord as childbirth waste has wide variety of sources and can be easily obtained, without any ethical and legal restrictions. Therefore, human umbilical cord mesenchymal stem cells break al the restrictions originated from other sources of mesenchymal stem cells. OBJECTIVE:To review the application of human umbilical cord mesenchymal stem cells in cartilage diseases, neuroglioma, ischemic brain injury, lung disease, liver disease and models of myocardial infarction. METHODS:The PubMed and Wanfang databases were searched by the first author using the keywords of“human umbilical cord mesenchymal stem cells, disease models, celltherapy”in English and Chinese, respectively. Seventy-three articles were searched and final y, 35 articles were included in result analysis. RESULTS AND CONCLUSION:Human umbilical cord mesenchymal stem cells have multilineage differentiation capacity similar to bone marrow mesenchymal stem cells. Compared with bone marrow mesenchymal stem cells, human umbilical cord mesenchymal stem cells have lower immunogenicity. Human umbilical cord mesenchymal stem cells show certain curative effects on cartilage disease, neuroglioma, ischemic brain injury, lung disease, liver disease and myocardial infarction, indicating that human umbilical cord mesenchymal stem cells can be used for celltransplantation to treat various diseases.

BACKGROUND:Reprogramming somatic cells to generate pluripotent stem cells has a wide application in biomedical research. OBJECTIVE:To analyze the effect of different molecular weight proteins in chicken egg-white extract to elevate expression of pluripotent genes Oct-3/4 and Nanog in 293T cells. METHODS:The extracts of chicken egg-white were separated into more than 3 ku and less than 3 ku ingredients to be used for co-culture with 293T cells. There were four groups, 1×105 293T cells per wel , total 500μL. In the control group, 500μL culture medium was added;in the other three groups, 500μL chicken egg-white extract, more than 3 ku and less than 3 ku ingredients were respectively added. Quantitative PCR was used to determine the relative expression levels of pluripotent genes Nanog and Oct-3/4 in 293T cells. RESULTS AND CONCLUSION:By using co-culture method, more than 3 ku ingredients have a role to increase the expression of pluripotent genes Oct-3/4 and Nanog, but less than 3 ku ingredients cannot elevate the expression of pluripotent genes. This indicates that the ingredient of chicken egg-white extract to elevate the expression of pluripotent genes is more than 3 ku proteins.

Animals ; Brain Edema ; Brain Ischemia ; Cerebral Infarction ; Cerebrovascular Circulation ; Rats ; Reperfusion ; Reperfusion Injury

Objective: To explore the relationship between dietary patterns and non-alcoholic fatty liver disease(NAFLD) among college freshmen of Hangzhou area, and provide scientific evidence for the prevention of NAFLD in universities. Methods: The 1 752 freshmen students from six universities were selected to perform a questionnaire survey, including general information, and dietary survey in the last year, by using a random sampling method. Factor analysis was used to achieve the major dietary patterns. Results: Three dietary patterns were obtained from the analysis, including the animal food, Western fast food, and traditional Chinese patterns. After adjusting for confounding factors, participants in the highest tertile of the animal food and Western fast food pattern had significantly higher BMI (P<0.01). Besides, participants in the highest tertile of the Western fast-food pattern had higher ALT and AST than those in the lowest tertile(P<0.05). Conversely, participants in the highest tertile of the traditional Chinese pattern had lower BMI, ALT and AST than those in the lowest tertile(P<0.05). Logistic regression analysis showed that Western pattern might increase the risk of NAFLD (OR=1.26, 95%CI=1.09-1.72, P<0.05), whereas the traditional Chinese pattern might decrease the risk of NAFLD (OR=0.51, 95%CI=0.38-0.74, P<0.05). Conclusion Different dietary patterns are closely related to NAFLD in college freshmen. Chinese traditional food should be kept their diet, red meat and fried food should be reduced to decrease. the risk of NAFLD.

Purpose To investigate the difference of expression of autophagy-related gene (Beclin1,LC3,mTOR) in the development of esophageal squamous cell cancer.Methods Immunohistochemical EnVision method was adopted to detect the expression of autophagy-related gene Beclinl,LC3 and mTOR in 30 cases of normal esophageal mucosa,32 cases of low-grade intraepithelial neoplasia (LGIN),34 cases of highgrade intraepithelial neoplasia (HGIN),35 cases of early carcinoma and 126 cases of advanced esophageal carcinoma,respectively.The correlation between their expression with clinicopathologic factors was also analysed.Results The expression of Beclin1 in advanced esophageal carcinoma was obviously higher than that in another four groups (P ＜ 0.005).LC3 expression in advanced esophageal carcinoma was significantly higher than that in normal esophageal mucosa,LGIN and early carcinoma (P ＜ 0.005).The expression of mTOR in advanced esophageal carcinoma was significantly higher than that in normal esophageal mucosa,LGIN and HGIN (P ＜ 0.005).In advanced esophageal carcinoma group,the expression of Beclin1,LC3 and mTOR was related to tumor TNM stage and lymph node metastasis (P ＜ 0.05).Beclin1 expression was positively associated with LC3 and mTOR expression in advanced squamous cell carcinoma (P ＜ 0.05).Positive correlation was also observed between the expression of mTOR and LC3 in advanced esophageal carcinoma and HGIN (P ＜ 0.05).Conclusion In the carcinogenesis and development of esophageal cancer,Beclin1,as a tumor suppressor gene,activates autophagy and leads to excessive self consumption and death of tumor cells.mTOR promotes tumor growth by inhibiting autophagy and promoting angiogenesis.The combined detection of Beclinl,LC3 and mTOR may be beneficial to evaluate the progression and prognosis of esophageal squamous cell cancer.

OBJECTIVEThe wetland at Dalian Bay in the Northeast of China has been polluted by oil severely. The effect of various microbes and operation parameters on the bioremediation of oil-polluted wetlands at Dalian Bay was investigated and reported previously. In the study, other operation conditions related to the status of medium were investigated via statistical experimental design and analysis and a necessary information is involved to use micro-technology in the application.

METHODSThe method used involved the direct inoculation of selected bacteria, which were capable of degrading oil. The operation conditions were further optimized and evaluated by gravimetrical assay.

RESULTSThe optimal pH and temperature for the studied bacteria to degrade the existing oil pollutants were established as pH 8.0 and 27 degrees C. The mixed of various bacteria showed better results in terms of oil degradation than any single one. Among the selected four factors, disturbance, oxidant, nutrients, and biosurfactant, the former two contributed more impacts on the oil degradation in the early stage of process, while the latter two became the limiting factors in the late stage. Three sets of optimal conditions were obtained for each individual stage, but no one was suitable for the overall process.

CONCLUSIONThe study demonstrated the technical feasibility of using direct inoculation into the contaminated soil samples to remove oil pollutants. It suggested that the operation conditions should be monitored and adjusted during the different stages of bio-reactions in the process to achieve the best result of oil degradation.

Bacteria ; metabolism ; Biodegradation, Environmental ; China ; Ecosystem ; Hydrogen-Ion Concentration ; Oceans and Seas ; Petroleum ; metabolism ; Soil ; analysis ; Soil Pollutants ; chemistry ; metabolism ; Temperature

Objective To investigate selcnium(Se) levels of environment and human body in Gaomi City and Zichuan District of Shandong. Methods Lijiaying Township in Gaomi City of Weifang City, Zhaili Township and Longquan Township in Zichuan District of Zibo City were selected. Two farming soil samples at different spot, local wheat and corn, residents nail samples from 3 to 4 families were collected in each natural village in the investigated towns. The contents of Se were detected by 2,3-diamino naphthalene fluorescence method. Results Se level of the soil, wheat, corn, and nails in Lijiaying [(0.054 ± 0.019), (0.022 ± 0.009), (0.018 ± 0.007), (0.365 ± 0.108)mg/kg] was significantly lower than that in Zhaili [(0.425 ± 0.080), (0.130 ± 0.043), (0.098 ± 0.026), (0.751 ± 0.134)mg/kg] and Longquan[(0.487 ± 0.153), (0.112 ± 0.030), (0.097 ± 0.029), (0.735 ± 0.145)mg/kg;P < 0.01]. In Lijiaying, Se was deficient in soil, wheat, corn(< 0.200, < 0.025 mg/kg), above Se deficiency diagnosis and below Se-adequate level in the nail, while in Zhaili and Longquan, the Se level in the soil (0.425, 0.487 mg/kg), wheat(0.130, 0.112 mg/kg), corn (0.098, 0.097 mg/kg), nails (0.751, 0.735 mg/kg) was adequate (≥0.400 mg/kg). Conclusions The external environment is Se-deficient in Lijiaying, Se-adequate in Longquan and Zhaili. The selenium level in human body is consistent with the external environment.

OBJECTIVEThe aim of this investigation was to study the effects of fat-soluble extracts from vegetable powder (FEFVP) and beta-carotene on the proliferation and apoptosis of cultured YTMLC-90 lung cancer cells.

METHODSThe lung cancer cells were continuously exposed to a broad range of concentration of FEFVP and beta-carotene. The proliferation was evaluated in MTT test. The induction of apoptosis was evaluated by morphological change, DNA fragmentation analysis, and DNA content analysis combined with flow cytometric analysis.

RESULTSBoth FEFVP and beta-carotene were found to inhibit cell proliferation and to induce morphologic changes consistent with apoptosis in YTMLC-90 cancer cells, including cellular shrinkage, chromatin condensation and nuclear fragmentation. DNA agarose gel electrophoresis showed DNA fragmentation 'ladder'. Flow cytometric analysis revealed decreased DNA content and the presence of a sub-G1 apoptotic peak.

CONCLUSIONThese findings are consistent with the induction of apoptosis. Moreover, the effects of FEFVP are stronger than those of beta-carotene. FEFVP inhibits the growth of YTMLC-90 probably via the induction of apoptosis cancer cells.

Antioxidants ; pharmacology ; Apoptosis ; Cell Division ; DNA Damage ; Flow Cytometry ; Humans ; Lung Neoplasms ; pathology ; Plant Extracts ; pharmacology ; Powders ; Tumor Cells, Cultured ; Vegetables ; chemistry ; beta Carotene ; pharmacology

Objective To introduce the technique of interictal 18F-fluorodeoxyglucose positron emission tomography (18F-FDG-PET) examination and explore the value of 18F-FDG-PET in the localization of epileptogenic focus of temporal lobe epilepsy (TLE) confirmed by surgical result.Methods Clinical data were retrospectively analyzed in 82 TLE patients having received interictal 18F-FDG-PET preoperative evaluation and got EngleⅠ grade epileptic surgical outcome, and the sensitivity and specialty of interictal 18F-FDG-PET were compared with those of MRI and scalp video-EEG. Results Epileptogenic foci showed hypometabolism on 18F-FDG-PET, and the hypometabolism zones were localized in ipsilateral temporal lobe in 68 cases,beyond ipsilateral temporal lobe in 9 cases; the other 5 had no hypometabolism zone. Accuracy rate of localization of epileptogenic foci by interictal 18F-FDG-PET was 82.9% (68/82), significantly higher than that by MRI or EEG(P＜0.05).77.4%(41/53)epileptogenic foci where MRI showed negative and 75%(15/20)where EEG with imbedded electrode was applied were precisely localized by 18F-FDG-PET. The accuracy was higher in the cases with positive pathological result than in the ones with negative result. Conclusions Interictal 18F-FDG-PET possesses excellent sensitivity and specialty in preopemtive assessment for TLE, and is of good value in the localization of epileptogenic focus where MRI shows negative or invasive electrophysiologic monitoring is needed.Rational application may raise the accuracy rate.