Objective To compare the gene expression difference between 0.1 and 5 Gy X-ray irradiated cells,and to explore its possible mechanism.Methods A cDNA microarray corresponding to 45033 human genes was used to analyze the transcriptional profiles of normal human lymphoblastoid AHH-1 cells at 4 h after 0.1 or 5 Gy irradiation.The genes with a fold change ≥ 2.0 were identified as the differentially expressed genes.real-lime PCR and Western blot were used to confirm the expression of PERP.Results The microarray assay showed that there were 760 up-regulated genes and 1222 down-regulated genes in the cells at 0.1 Gy,while there were 744 genes down-regulated and 457 genes up-regulated in the cells at 5 Gy.In addition,55 genes were commonly up-regulated and 339 genes commonly down-regulated at 0.1 and 5 Gy.The predominant biological processes of the differential genes responding to low-dose radiation include cell-cell signaling transduction and DNA damage response,and the altered genes after 5 Gy irradiation were related to cell proliferation,differentiation,and apoptosis.Moreover,the expression of PERP gene was down regulated,which was consistent with the data of microarrey assay.Conclusions The quantitative and qualitative differences in the gene expressions may contribute to the diversed biological effects induced by low or high doses of ionizina radiation.

Objective To observe the radiosensitivity by targeting HIF-lα in human lung cancer and the effects on tumor growth in nude mice.Methods Radiosensitivity of A549 and A549/HIF-1α ( － ) cells were tested by clonogenic forming assay.A549/HIF-1 α( － ) cells and A549 cells were injected into the male BALB/C nude mice.Tumor growth was observed.The expression of HIF-1α and microvessel density were detected by immunohistochemistry method.Results SERs of HIF-1α gene silencing were 1.03 in normoxia and 1.65 in hypoxia.The sizes of tumor xenografts derived from A549/HIF-1α( － ) cells were significantly reduced compared to those of the xenografts derived from A549 cells.HIF-1 α protein staining result showed a dramatic decrease in tumors from A549/HIF-1α ( － ) mice.The microvessel densities (MVD) were 19.83 ± 4.09 in A549 group and 11.61 ±3.04 in A549/HIF-lα (-)group(F=15.57,P ＜0.05 ).Conclusions Hypoxia-induced radio-resistance in lung cancer A549 cells could be reversed by silencing the HIF-1α.It also retards the growth of tumor xenografts,decreases HIF-1α expression and reduces the vascularity.

Objective To investigate the expression and distribution of Diacylglycerol Kinase α (DGKα) mRNA in human hepatocellular carcinoma (HCC),and to explore the function of DGKα in the metastasis of hepatocellular carcinoma.Methods Tissues from 30 cases of HCC and 5 normal liver tissues were collected immediately after surgical resection.Semi-quantitative RT-PCR and in situ hybridization were used to detect the expression levels and distribution of DGKα mRNA,respectively.Results Semi-quantitative RT-PCR showed that the expression level of DGKα mRNA in HCC (0.798±0.317) and normal tissues (0.908±0.425) was significantly higher than those in carcinoma adjacent tissue with cirrhosis (0.205±0.102,P<0.05).In situ hybridization demonstrated that the number of DGKα mRNA positive hepatocytes in HCC [(57.6±6.3)/mm2] and normal tissues [69.8±8.7)/mm2] was significantly higher than those in carcinoma adjacent tissue with cirrhosis [(26.3±4.9)/mm2,P<0.05]; DGKα mRNA was expressed in the cytoplasm of hepatocytes in HCC and carcinoma adjacent tissue with cirrhosis,and in the nuclear of hepatocytes in normal tissues.Conclusion The present study suggests that DGKa may play important roles in carcinogenesis and progressing of HCC.

Objective To investigate the chromosome damage in peripheral lymphocytes of underground gold miners.Methods Conventional method and cytokinesis-block micronuclens assay were used to analyze frequency of chromosome aberrations and micronucleus in peripheral lymphocytes in 58 gold miners,respectively.Results Frequencies of chromosome-type aberrations,ehromatid-type aberrations and total aberrations were higher in the miners than those in the control group(0.72%,0.41%,1.16% vs 0.14%,0.18%,0.33,X2=44.322,9.501,50.476,P＜0.01).Both micronucleated cell rate and micronucleus rate were higher in the miners group than those in the control group(10.8‰ and 11.6‰ vs 8.7‰ and 9.0‰,X2=8.672,12.546,P＜0.01).Frequencies of chromosomal aberrations and micronucleus proportionally increased with underground working years.Compared with those miners who had worked underground 6 years or shorter,both frequencies were statistically higher among the miners who had worked underground for more than 21 years(P＜0.05).No difference was found among other groups of working years(P＞0.05).Compared with the control group,both frequencies increased in the miner group,and the differences were statistically significant(X2=2.395,P＜0.05 for chromosomal aberrations and X2=2.319,P＜0.05,respecfvely).The common types of chromosome aberrations were acentrie fragments,while chromatid break and dicenrics were subordinate.Conclusions Chromosomal damages were observed in the gold workers who exposed high radon in the underground mining.

objective To study the TCR gene mutation in peripheral blood lymphocytes induced by X-ray exposure using cultured lymphocytes cloning method.Methods Freshly isolated peripheral lymphocytes from healthy aduh donors were irradiated with X-ray in doses ranging from 0 to 8 Gy and cultured with interleukin2 and phytohemagglutinin for 7 days.The mutant frequencies of TCR gene(TCR MF)were detected by flow cytonletry and the dose response curves were fitted.Results TCR MF increased with the dose going up.An aquadratic polynomial dose response model was fitted.Conclusions TCR gene mutation could which serve as a potential biological dosimeter.It might be applied for the estimation of biological dose in emergency exposure.

Objective To observe the consistency of T-cell receptor (TCR) genes mutation in lymphocytes in rats after irradiation in vivo and in vitro.Methods A total of 48 female rats were randomly divided into 6 equal groups.Peripheral blood samples from them were collected to separate the lymphocytes and then irradiated to X-ray irradiation with the dose rate of 200 cGy/min at the doses of 0,0.5,0.75,1.0,2.0,and 3.0 Gy,respectively.Then all the lymphocyte samples were cultured for 7 days.Flow cytometry with direct immunofluorescence was used to detect the TCR gene mutation.The levels of TCR gene mutant frequency (TCRMF) of different groups were calculated.Results The TCRMF levels of different groups after irradiation in vivo and in vitro all displayed a dose-dependent manner and there were no significant differences in the TCRMF between different dose irradiation groups(t = -1.1-0.3 ,P ＞0.05).Conclusions A consistency of TCRMF after irradiation in vivo and in vitro is proven.The results of TCRMF of peripheral blood lymphocytes irradiated in vitro by flow cytometry can precisely reflect the TCR genes mutation after whole-body irradiation.

OBJECTIVETo assess the therapeutic effect of Qufengtongluo (QFTL) recipe against proteinuria and glomerular filtration membrane damage in rats with adriamycin-induced nephropathy (AN).

METHODSFifty-six SD rats were randomized into normal control (A) and AN model groups. In the AN model group, the rat AN models established by a single intravenous injection of adriamycin via the tail vein were subdivided into model (B), QFTL recipe (C), prednisone (D), and benazepril (E) groups 3 weeks after adriamycin injection. The 24-h urinary protein level was measured and the expression of anionic sites on the filtration membrane was evaluated using electron microscope with PEI staining. Nephrin expression on the glomerular filtration membrane was detected with indirect immunofluorescence assay.

RESULTSCompared with group A, the model group showed significantly increased level of 24-h urinary protein (P<0.01), suggesting successful establishment of the AN model. Treatment with QFTL recipe obviously lowered the 24-h urinary protein (P<0.01), and increased the expression of anionic sites and nephrin on the glomerular filtration membrane in the AN rats (P<0.01).

CONCLUSIONQFTL recipe can effectively decrease 24-h urinary protein, improve the symptoms, and up-regulate the expressions of anionic sites and nephrin on the glomerular filtration membrane in rats with AN.

Animals ; Doxorubicin ; Drugs, Chinese Herbal ; therapeutic use ; Glomerular Basement Membrane ; drug effects ; Male ; Nephrosis ; chemically induced ; drug therapy ; Phytotherapy ; Proteinuria ; drug therapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo observe the effects of Qufengtongluo Recipe (QFTLR) on the expression of podocin mRNA and podocyte morphology in rats with adriamycin-induced nephropathy (AN), and explore the possible mechanism mediating the therapeutic effect of QFTLR on nephropathic proteinuria.

METHODSSD rats were randomized into normal control group, AN model group (established by a single injection of adriamycin via the tail vein), and 3 intervention groups with QFTLR, prednisone, or benazepril treatment. After the corresponding treatments, the expression of podocin mRNA in the renal tissues was detected by RT-PCR methods, and the morphological changes of the podocytes were examined by electron microscope.

RESULTSCompared with the normal control group, the AN model group showed significantly lowered expressions of podocin mRNA (P<0.01) with reduced podocytes and widening, fusion or even absence of the foot processes (FP). Intervention with QFTLR significantly increased the expression of podocin mRNA (P<0.01) and the number of podocytes, and obviously lessened the structural changes of the FP.

CONCLUSIONQFTLR can produce therapeutic effect against nephropathic proteinuria possibly by up-regulating the expression of podocin mRNA and improving the morphological changes of the podocytes.

Animals ; Doxorubicin ; Drugs, Chinese Herbal ; pharmacology ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Male ; Membrane Proteins ; genetics ; metabolism ; Nephrosis ; chemically induced ; metabolism ; pathology ; Podocytes ; pathology ; Proteinuria ; etiology ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo establish the dose-effect curve between TCR MF and ionizing radiation.

METHODSPeripheral lymphocytes were collected from 8 healthy adults (4 males and 4 females) and cultured in vitro with 12 well culture plates. They were stimulated by PHA-P and IL-2 after exposed to different doses of irradiation (0.00 - 8.00 Gy) and cultured for 7 d. The dose-effect curve was established after measuring TCR MF using flow cytometry. Also, using the same method, we separated and cultured the peripheral lymphocytes collected from 16 radiotherapy cancer patients, whose radiation styles and doses were different, and then measured TCR MF to estimate the whole equivalent dose of radiotherapy patients through the dose-effect curve. Peripheral blood was collected and cultured, chromosome aberration (dicentric and ring) was determined under microscope to estimate irradiation dose.

RESULTSThe relationship of dose-effect between the TCR MF and ionizing radiation (0.00 - 8.00 Gy) was well, the curve of large dose group (2.00 - 8.00 Gy), low dose group (0.00 - 1.00 Gy) and 0.00 - 8.00 Gy dose group were met with the quadratic polynomial model, the equation was TCR MF = -32.8579 + 20.5436D + 0.6341D(2), TCR MF = 1.796 + 0.017D + 5.155D(2) and TCR MF = -0.6229 + 6.305D + 0.6919D(2), respectively. D was the radiation dose (Gy). Using the established curve and the chromosome aberration method to estimate the systemic exposure dosage, the average relative deviation was 16.8%.

CONCLUSIONThe curve established by the TCR gene mutation analysis technology can be applied to exposure dose estimation of victims in ionization radiation accidents.

Case-Control Studies ; Dose-Response Relationship, Radiation ; Female ; Genes, T-Cell Receptor ; genetics ; Humans ; Male ; Mutation Rate ; Radiation, Ionizing ; Receptors, Antigen, T-Cell ; genetics ; radiation effects

OBJECTIVETo observe the effect of yishen capsule (YSC) on preventing the recurrence of chronic glomerulonephritis (CGN) and to explore its mechanism preliminarily.

METHODSCGN patients were assigned to the treated group (61 cases) and the control group (48 cases) and all of them were orally administered with 4 mg of Perindopril twice a day, but 3 capsules of YSC, thrice a day, were given additionally to patients in the treated group. The therapeutic course for both groups was 18 months. The recurrence rate of CGN at the 6th, 12th, and 18th month in the two groups was observed and compared, and the changes of 24-h urinary protein quantity and T-lymphocyte subsets before and after treatment were observed as well.

RESULTS(1) Comparison of recurrence rate between the two groups showed insignificant difference at the 6th month, but it did show significant difference at the 12th and the 18th month, which was significantly decreased in the treated group than in the control group (P<0.05, P<0.01); (2) The 24-h urinary protein quantity at the 18th month decreased significantly in both groups (P<0.05, P<0.01), but in the treated group was more significant (P<0.01); (3) T-lymphocyte subsets showed no obvious change in the control group after treatment (P>0.05), while in the treated group, it showed significant increase in CD3, CD4 and CD4/CD8 (P<0.05 or P<0.01) and significant decrease in CD8 (P<0.05), and also the difference after treatment in T-lymphocyte subsets between the two groups was significant (P<0.05 or P<0.01).

CONCLUSIONYSC has marked effects in reducing the recurrence of CGN and in decreasing urinary protein, and its mechanism might be related with its function in regulating the ratio of T-lymphocyte subsets to enhance the immunity of patients.

Adolescent ; Adult ; Capsules ; Case-Control Studies ; Chronic Disease ; prevention & control ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Glomerulonephritis ; drug therapy ; prevention & control ; Humans ; Lymphocyte Subsets ; cytology ; drug effects ; Male ; Patient Dropouts ; Proteinuria ; Secondary Prevention ; T-Lymphocytes ; cytology ; drug effects