OBJECTIVETo study the effect of epigallocatechin gallate (EGCG) against lactacystin induced PC12 cell injury.

METHODSThe inoculated rat PC12 cells were cultured for 24 h, followed by intervention. The cells were divided into 5 groups, i.e., the normal control group, 10 micromol/L lactacystin injury group, and the EGCG pretreated groups (at the final concentration of 5, 10, and 50 micromol/L, respectively). The cytoactive was detected by MTT colorimetry. Morphological changes of the cell nucleus were observed by Hoechst 33,258 staining, and the apoptosis ratio was detected by flow cytometry (FCM).

RESULTSEGCG at different doses showed protective effect on lactacystin-induced PC12 cell injury. Compared with the lactacystin injury group [(61.22 +/- 1.02)%], the cytoactive in EGCG pretreated groups at the final concentration of 5, 10, and 50 micromol/L, respectively increased obviously to (66.99 +/- 1.30)%, (66.67 +/- 0.65)%, and (73.4 +/- 0.67)%, respectively. Hoechst 33 258 staining found that more nuclear pyknosis and aggregation occurred in the lactacystin injury group, but less occurred in EGCG pretreated groups. FCM indicated that the apoptosis ratio was reduced by EGCG pretreatment. It was 3.0%, 60.4%, 59.8%, 57.5%, and 38.6%, respectively in the normal control group, the lactacystin injury group, and EGCG pretreated groups (at the final concentration of 5, 10, and 50 micromol/L, respectively).

CONCLUSIONEGCG could attenuate lactacystin induced PC 12 cell injury.

Acetylcysteine ; adverse effects ; analogs & derivatives ; Animals ; Apoptosis ; drug effects ; Catechin ; analogs & derivatives ; pharmacology ; Flow Cytometry ; PC12 Cells ; Rats

AIM: To investigate the early changes of retinal function in diabetic patients detected by multifocal electroretinogram (mfERG).METHODS: The first-order kernel responses of mfERG were recorded from eyes of 33 normal control subjects,63 diabetic patients without retinopathy and 43 diabetic patients with background retinopathy. The response densities and implicit times of N1 and P1 were compared among the control, diabetic patients without retinopathy and diabetic patients with retinopathy.RESULTS: The response densities of N1 and P1 in central 3 rings were reduced significantly in diabetic eyes with and without retinopathy. And the implicit times of N1 and P1 were delayed significantly only in diabetic eyes with retinopathy.CONCLUSION: mfERG can detect the early changes of retinal function quantitatively in diabetic patients. Analysis of response densities and implicit times of N1 and P1 can reflect the progress of local retinal dysfunction in diabetes.

Objective To investigate new type cytokine induced killer cells expansion using advanced breast cancer′s peripheral blood .Methods peripheral blood mononuclear cells were isolated from 8 advanced breast cancer volunteers and co -cultured with Cytokine induced killer cells .These cells were placed in plastic flasks containing CIK-MediumTM supplemented with 10% auto-plasma in the presence of IL -2 ( 1 000 IU/mL) .The cultures were fed with CIK-MediumTM supplemented with IL -2 following the proliferation capacity . Cell proliferation was measured by cell counting during the cultivation .Fourteen days after cultivation ,cell mark-ers CD3/CD16/CD56 were examined by flow cytometry .51Cr and MTT assays were employed in cytotoxicity as-says.Cytokines were assayed by ELISA method .Results CD16+,CD16+CD56+,CD56+CIK cells were 5.8~11.6%in 2 ×107 fresh PBMCs and 95.2~97.6%in co-cultured cells after 18 days cultivation .The in vitro ex-pansion rate of new type cytokine induced killer cells was up to more than 8.2 ×108 in total,the cytotoxicity are ef-fective killing cells against MCF 7 and BT20 breast cancer cell lines .New type cytokine induced killer cells expand-ed from all PBMCs and secreted cytokines IFN -and TNF-.Conclusion The present culture could be useful to clarify the mechanisms of CIK cells expansion in vitro and feasible for breast cancer immmuno cell therapy .

Animals ; Antioxidants ; pharmacology ; Apoptosis ; drug effects ; Atherosclerosis ; prevention & control ; Ceramides ; analysis ; Dietary Fats ; administration & dosage ; Emodin ; pharmacology ; Lipids ; blood ; Male ; Rabbits ; Signal Transduction ; Sphingomyelin Phosphodiesterase ; metabolism ; Sphingomyelins ; metabolism

OBJECTIVETo evaluate the efficacy and safety of adefovir dipivoxil in treatment of decompensated liver cirrhosis patients with YMDD motif mutation during lamivudine therapy.

METHODSThe disease relapsed in 14 hepatitis B patients with decompensated liver cirrhosis during lamivudine treatment due to the YMDD motif mutation. All 14 patients had positive HVBDNA and active hepatitis, and were evaluated as Child-Pugh Score C (CPS-C). The patients were treated with lamivudine 50 mg/d and adefovir dipivoxil 10 mg/d for 6 months.

RESULTSOne patient signed off due to non-hypoxemic hyperlactacidemia; other 13 patients showed decreased serum HBVDNA. All patients had serum HBVDNA < or =10(5) copies/ml and 7 patients had HBVDNA < or =10(4) copies/ml. Six patients regained normal serum ALT level and Child-Pugh scores decreased in all patients.

CONCLUSIONAdefovir dipivoxil has satisfied efficacy and safety in treatment of decompensated liver cirrhosis patients with YMDD motif mutation during lamivudine treatment.

Adenine ; adverse effects ; analogs & derivatives ; therapeutic use ; Adult ; Aged ; Amino Acid Motifs ; genetics ; Antiviral Agents ; therapeutic use ; DNA-Directed DNA Polymerase ; genetics ; Female ; Hepatic Encephalopathy ; drug therapy ; genetics ; virology ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; complications ; drug therapy ; Humans ; Lamivudine ; therapeutic use ; Liver Cirrhosis ; drug therapy ; genetics ; virology ; Male ; Middle Aged ; Mutation ; Organophosphonates ; adverse effects ; therapeutic use ; Protein Structure, Tertiary ; genetics ; Recurrence ; Reverse Transcription ; genetics

Objective To develop appropriate evaluation methods of local basic public health services which are suitable to county level and above.Methods Data on basic public health services of 1 1 cities in Zhejiang province in 201 2 was evaluated by different evaluation methods including weighted synthetic scored method,weighted synthetic index method, Weighted Technique for Order Preference by Similarity to Ideal Solution (Topsis ) and Weighted Rank -Sum Ratio (RSR).The consistency of evaluation results were tested by Kendall's coefficient of concordance W test.Combination evaluation was conducted to evaluate four single synthetic evaluation results through average method,weighted average combination evaluation method and hierarchical clustering analysis.Results Different synthetic evaluation methods had different evaluation results.However,in the order,the top two were all Hangzhou and Ningbo.Kendall's W test showed good consistence of four evaluation results.Rank of 1 1 cities were Hangzhou,Ningbo,Shaoxing,Jiaxing,Huzhou, Taizhou,Jinhua,Zhoushan,Lishui,Wenzhou and Quzhou based on combination evaluation value by average method, which was the same to the rank based on weighted average combination evaluation result.Eleven cities could be classified into four categories by hierarchical clustering analysis with statistical significance (P <0.01 ):Excellent (Hangzhou, Ningbo),Good (Huzhou,Jiaxing,Shaoxing),Middle (Zhoushan,Jinhua,Taizhou)and Poor (Wenzhou,Quzhou, Lishui).Conclusion These four synthetic evaluation methods used in this study are all suitable to county level and above in basic public health services evaluation.Various synthetic evaluation methods could be used in practice with combination evaluation of various evaluation results.Average method which is convenient and accurate is preferred when consistency of various synthetic evaluation results was testified.Hierarchical clustering analysis could be used for combination evaluation when no precise rank is needed.

Objective To investigate the effect of glutathione peroxidase-1 (GPX1) on cispatin resistance in endometrial carcinoma cells and its related mechanism.Methods The HEC-1-A cisplatin resistance (HEC-1-A CR)cell was constructed by intermittent concentration gradient method.The control HEC-1-A cell was constructed with equal amount of normal saline.Lentivirus transfection was used to up-regulate the expression of GPX1 in the HEC-1-A CR cell as HEC-1-A CR GPX1 cell,and blank vector transfected was regarded as HEC-1-A CR control cell.The half maximal inhibitory concentration (IC50) value of cisplatin in cells was detected by thiazolyl blue (MTS) assay.The expression of mRNA and protein of GPX1 and HIF-1 α were detected by fluorescent quantitative chain reaction (qPCR) and Western blotting.Results The IC50 of HEC-1-A cells and HEC-1-A CR cells were (2.13 ±0.24),(7.89 ± 1.17)μg · mL-1.The IC50of HEC-1-A CR control cells and HEC-1-A CR GPX1 cells were (7.91 ±0.98),(3.87 ±0.52)μg · mL-1.The expression level of GPX1 mRNA in HEC-1-A cells and HEC-1-A CR cells were 0.14 ±0.01,0.07 ±0.01.The expression level of HIF-1α mRNA in HEC-1-A cells and HEC-1-A CR cells were 0.32 ±0.03,0.72 ±0.08.The expression level of GPX1 mRNA in HEC-1-A CR control cells and HEC-1-A CR cells were 0.09 ± 0.02,0.24 ± 0.03.The expression level of HIF-1 α mRNA in HEC-1-A CR control cells and HEC-1-A CR cells were 0.76 ±0.08,0.54 ±0.05.The above indexes were compared between groups,the difference was statistically significant (all P ＜ 0.05).The results of the changes of GPX1 and HIF-1 α protein levels were in accordance with the changes of mRNA levels.Conclusion GPX1 can enhance the sensitivity of cisplatin in endometrial cancer cells via down-regulating the expression of HIF-1 α.

AIMTo explore the effects of tRNA on the growth of mammalian cells.

METHODSL929, NIH3T3, MCF-7 and PC12 cells were seeded in 96 well culture plate individually, and incubated at 37 degrees C in 5% CO2 for 4 h, the tRNAs from different species were added to the culture media individually. After certain time of incubation, the viability of the cells was evaluated by the MTT methods. Sub-confluent L929 cells were incubated with 200 microg/ml ytRNA for different times, then the cells were pooled and analyzed with flow cytometry assay.

RESULTStRNA specifically inhibited the growth of L929 cells in a dose-dependent manner. The sizes of tRNA-treated cells showed larger sizes and longer processes than those of untreated cells. Flow cytometric analysis further showed that most of tRNA-treated cells were arrested in S phase of the cell cycle.

CONCLUSIONThe cell growth inhibitory effects of tRNAs were caused mainly by their degraded fragments. The results suggested that tRNA or its degraded fragments might play important roles in regulation of cell proliferation.

Animals ; Cell Cycle Checkpoints ; physiology ; Cell Line ; Cell Proliferation ; Fibroblasts ; cytology ; Flow Cytometry ; Mice ; RNA, Transfer ; physiology

OBJECTIVETo explore the relationship of clinic features, lab findings, the origin of tumor cell as well as prognosis in Chinese patients with diffuse large B-cell lymphoma( DLBCL).

METHODSSeventy four cases of primarily diagnosed DLBCL were analyzed. Immunohistochemistry stain was used to check the expressions of Bcl-6,CD10 and MUM1.

RESULTSAmong the 74 patients, the average age was 58.5 years, the ratio of male to female was 1.64:1. 23.2% (16/69) cases developed in lymph node, 15.9% (11/ 69) in the extra node area. Among 55 follow-up cases, 13 (23.6%) died, and 12 (92.3%) died in the first year after diagnosis. The prognosis analysis showed that diagnosed at age > 65 years (P = 0.036), and the international prognostic index (IPI) (P = 0.009) were independent prognostic factors; origin of tumor cell had a trend to be a prognostic factor, but no statistic difference (P = 0.086). beta2-MG and Bcl-6 expression had no relation with the prognosis.

CONCLUSIONThe middle and old-aged male patients are the most common in DLBCL and the first-year mortality rate is higher. The age at diagnosis and IPI can predict the clinical outcome. The origin of tumor cell might suggest the prognosis.

Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Child ; Female ; Follow-Up Studies ; Humans ; Immunohistochemistry ; Lymphoma, Large B-Cell, Diffuse ; diagnosis ; metabolism ; pathology ; therapy ; Male ; Middle Aged ; Prognosis ; Survival Analysis

OBJECTIVETo investigate the frequencies of HLA-Alu repeat polymorphisms (AluMICB, AluTF, AluHJ, AluHG and AluHF) in Chinese Lisu and Nu ethnic populations.

METHODSThe frequencies of HLA-Alu repeat polymorphisms in above populations were determined with polymerase chain reaction (PCR). The associations between HLA-Alu repeat polymorphisms and HLA-A, HLA-B and HLA-C alleles were also analyzed. Phylogenetic trees were constructed with genetic distance calculated from the frequencies of HLA-Alu repeat polymorphisms.

RESULTSFrequencies of AluTF*2 and AluHF*2 were different between the two populations (P< 0.05), while those of other three insertions were similar. The strength of association between HLA-Alus and HLA alleles were different (P< 0.05) in the two populations. Although AluMICB*2 were associated with HLA-B*56:01 in both populations, the association was stronger in Lisu population (74.0%) but moderate in Nu population (30.7%). HLA-Alus were associated with particular HLA subtypes, e.g., AluHG*2 with certain HLA-A*02 subtypes. By phylogenetic analysis, Lisu and Nu were clustered together with southern Chinese and Thai populations.

CONCLUSIONThe distribution of HLA-Alus and the strength of associations between HLA-Alus and HLA class I alleles have varied between the two populations. Study of this association may facilitate identification of origins, evolution, progenitor haplotypes and recombination within the HLA class I region.

Adolescent ; Adult ; Aged ; Alleles ; Alu Elements ; Asian Continental Ancestry Group ; genetics ; Child ; Female ; Genes, MHC Class I ; Humans ; Male ; Middle Aged ; Phylogeny ; Polymorphism, Genetic ; Young Adult