Objective To evaluate the prognostic significance of MMP-2 , MMP-9 and Cathepsin-D expression in lymph node-negative human breast carcinoma. Methods Specimens from the primary breast tumors of 270 axillary lymph node negative breast cancer patients were studied for MMP-2 , MMP-9 and Cathepsin-D protein expression by immunohistochemical staining. Results The positive expression of MMP-2, MMP-9 and Cathepsin-D was 56.7%(153/270),59.6%(161/270) and 55.6%(150/270), respectively. In univariate analysis, MMP-2, MMP-9 and the co-expression of MMP-2 and MMP-9 were significantly associated with tumor free survival. In multivariate analysis, co-expression of MMP-2 and MMP-9 was unfavorablely related with tumor free survival(RR=2.487; P

Objective To analyze the local control rate and the desimetric patterns of local recurrence in nasopharyngeal carcinoma(NPC)patients having been treated with standardized conventional radiotherapy and to evaluate the delineation of rational target volume.Methods From Jan.2000 to Dec.2000,476 patients with untreated NPC were treated by standardized conventional radiotherapy alone at the Sun Yat-sen University Cancer Center.The radiation ports were designed on a X-ray simulator.The nasopharyngeal lesion demonstrated by CT scan and the subclinical spread regions adjacent to the nasopharynx were defined as the target volume.Kaplan- Meier method was used to calculate the cumulative local recurrence rate.For patients with locad recurrence,the primary and recurrent local tumor volumes(V_(nx),V_(recur))were delineated with three-dimensional treatment planning system(3DTPS),and the dataset of radiation ports and delivered prescription dose to the 3DTPS were transferred according to the first treatment.The dose of radiation received by V_(recur)was calculated and analyzed with dose- volume histogram(DVH).Local recurrence was classified as:1.“in-port”with 95% or mere of the recurrence volume((recur)_V_(95))was within the 95% isedase;2.“marginal”with 20% to95% of _(recur)V_(95)within the 95% isedese; 3.“outside”with only less than 20% of _(recur)_V_(95)within the 95% isodose curve.Results With the median follow- up of 42.5 months(range 8～54 months),52 patients developed local recurrence.The 1-,2-,3 and 4-year cumulative local failure rate was 0.6%,3.9%,8.7% and 11.5%,respectively.Among the 42 local recurrent patients who could be analyzed by 3DTPS,52% were in-port,40% were marginal and 7% were outside.For most of the marginal recurrence and all the outside recurrence patients,the main reason of recurrence were related to the unreasonable design of the radiation port and inaccuracy in the interpretation image findings.Conclusions The outcome of better local control rate and the dosimetric pattern of local recurrence show that the target volume is reasonable for NPC in Sun Yat-sen University Cancer Center.Enhancing the capability of correct interpretation of images,accurate design of the radiation pouts and making most useful molecular or functional imaging techniques to escalate the local radiation dose are promising ways to improve the local control further and better.

Objective To investigate blood plasma melatonin level in night-shift nurses and explore the relationship of blood plasma melatonin level with nervous system symptom (insomnia、anxiety、depression). Methods ELISA were used for detection of blood plasma melatonin level in 80 night-shift nurses of different age group. Results Blood plasma melatonin level of shift work nurses (36to40、41to45 yearold) were significant lower than the corresponding age group of the control group, the nervous system symptom of these age group night-shift nurses correlated to melatonin level of melatonin. Conclusions Blood plasma level of melatonin have a close relation to nervous system symptom(insomnia、anxietydepression).

BACKGROUNDTreatment for malignant glioma generally consists of cytoreductive surgery followed by radiotherapy and chemotherapy. In this study, we intended to investigate the effects of 2-propylpentanoic acid (VPA), a histone deacetylase inhibitor, on chemosensitivity and radiosensitivity in human glioma cell lines.

METHODSHuman glioma cell lines, T98-G, and SF295, were treated with temozolomide (TMZ) or irradiation (IR), with or without VPA (1.0 mmol/L). Then, cytotoxicity and clonogenic survival assay was performed. Cell cycle stage, apoptosis, and autophagy were also detected using flow cytometry and dansyl monocadaverin (MDC) incorporation assay. One-way analysis of variance (ANOVA) and t-test were used to analyze the differences among variant groups.

RESULTSMild cytotoxicity of VPA was revealed in both cell lines, T98-G and SF295, with the 50% inhibiting concentration (IC50) value of (3.85 ± 0.58) mmol/L and (2.15 ± 0.38) mmol/L, respectively; while the IC50 value of TMZ was (0.20 ± 0.09) mmol/L for T98-G and (0.08 ± 0.02) mmol/L for SF295. Moreover, if combined with VPA (1.0 mmol/L) for 96 hours, the sensitivity of glioma cells to TMZ was significant increased (P < 0.05). The surviving fractions at 2 Gy (SF2) of T98-G and SF295 cells exposed to IR alone were 0.52 and 0.58. However, when VPA was combined with IR, the SF2 of T98-G and SF295 dropped to 0.39 (P = 0.047) and 0.49 (P = 0.049), respectively. Treatment with VPA plus TMZ or IR also resulted in a significant decrease in the proportion of cells in the G2 phase and increased apoptotic rates as well as autophagy in T98-G and SF295 cell lines (P < 0.01).

CONCLUSIONVPA may enhance the activities of TMZ and IR on glioma cells possibly through cell cycle block and promote autophagy, and thus could be a potential sensitizer of glioma treatment.

Apoptosis ; drug effects ; radiation effects ; Blotting, Western ; Cell Line, Tumor ; Cell Survival ; drug effects ; radiation effects ; Dacarbazine ; analogs & derivatives ; pharmacology ; Flow Cytometry ; Glioma ; metabolism ; Histone Deacetylase Inhibitors ; pharmacology ; Humans ; Valproic Acid ; pharmacology

OBJECTIVETo establish a method of acutely isolating dorsal root ganglion (DRG) neurons for patch clamp study of single-channel.

METHODSDRG neurons of rats were acutely isolated by enzymatic digestion and mechanical blowing.

RESULTSThe acutely isolated DRG cells were easy to form the higher sealing resistance (> 5G Omega), which lowered noise level, so that pA-level single channel currents could be recorded.

CONCLUSIONThe acutely isolated DRG neurons in this study are an ideal for patch-clamp study of single-channel.

Animals ; Cell Separation ; methods ; Female ; Ganglia, Spinal ; cytology ; Ion Channel Gating ; physiology ; Ion Channels ; Male ; Neurons ; cytology ; physiology ; Patch-Clamp Techniques ; Rats ; Rats, Sprague-Dawley

Objective To investigate the current status and influencing factors associated with the health literacy of the elderly. Methods 24 communities were randomly selected from Jinan City, and 1 201 elderly people were surveyed by the eHealth literacy scale. Univariate analysis was performed using the Chi-square test and multivariate analysis was performed using binary Logistic regression. Results The qualification rate of eHealth literacy among 1 201 older adults was 11.1%. Multivariate analysis showed that primary school education and below (OR=4.50, 95% CI:1.924-10.530, P=0.001), family pension (OR=3.08, 95% CI:1.326-7.165, P=0.009), poor self-rated health (OR=2.12, 95% CI:1.022-4.406, P=0.044), great self-rated life pressure (OR=4.09, 95% CI:1.686-9.938, P=0.002) were risk factors for eHealth literacy in the elderly; urban household registration (OR=0.52, 95% CI:0.337-0.815, P=0.004), the main person to taking care of grandchildren (OR=0.43, 95% CI:0.273-0.682, P<0.001 ), urban basic medical insurance or NCMS medical insurance (OR=0.22, 95% CI:0.047-0.998, P=0.05), commercial medical insurance (OR=0.10, 95% CI:0.019-0.552, P=0.008) and the parents being alive (OR=0.44, 95% CI:0.264-0.719, P=0.001) were protective factors for the elderly eHealth literacy . Conclusion The type of household registration, the level of education, the type of medical insurance, the way of caring for grandchildren, the way of providing for the elderly, the self-rated of health status, with the parents being alive, and the self-rated life pressure are the influencing factors of the eHealth literacy of the elderly.

Antibiotic-associated diarrhea(AAD) is a frequent adverse effect of antibiotic in children.AAD is associated with longer hospitalization, higher healthcare cost and even lead to death.Pediatricians usually do not pay enough attention to AAD.Domestic experts from pulmonary medicine, infection and gastroenterology are organized to develop the consensus, to improve the diagnosis, treatment and prevention of AAD, and contribute the children health in future.

OBJECTIVEIn order to clarify the exact molecular weight of tumor metastasis suppressor gene-1 (TMSG-1) protein and its cellular localization, a monoclonal antibody against TMSG-1 was prepared, characterized and applied to evaluate the metastatic potential of human tumors.

METHODSA dominant epitope-TMSG-1(15)-derived from TMSG-1 was synthesized based on Fmoc method, and the hapten was conjugated to Imject Maleimide activated mcKLH as a carrier protein. The antigen preparation was used to immunize BAL B/C mice. Hybridomas were generated and screened by ELISA for specific monoclonal antibodies, which were further characterized by western blotting and immunohistochemical staining.

RESULTSOne hybridoma cell line secreting anti-TMSG-1 antibody, designated as C8, was eventually established after primary ELISA screening, followed by rapid limited dilution procedure. It was confirmed that C8 was of IgM isotype. Result of competitive inhibition assay showed that the antibody was TMSG-1 specific. Using this antibody, an expected protein band of about 45,000 (relative molecular mass) was detected in the non-metastatic variants PC(3)-2B4 and PG-LH7 cells by Western blotting, but not in the isogenetic metastatic variants of PC3-1E8 and PG-BE1 cells. Immunohistochemistry using C8 showed a positive staining of cell membrane and cytoplasm of 2B4 and LH7 cells, whereas 1E8 and BE1 cells were non-reactive. Immunostaining using C8 of paraffin sections of 52 breast carcinomas and 41 colon cancers demonstrated a strong positivity in non-metastatic tumors, but none to weakly reactive in metastatic tumors (P < 0.05).

CONCLUSIONC8 monoclonal antibody against the synthetic peptide is TMSG-1 specific and is effective for Western blot and immunohistochemistry assays to detect TMSG-1 expression in cancer cells. TMSG-1 protein is about 45 000 (relative molecular mass) at cell membrane and cytoplasm of tumor cells. Expression of TMSG-1 protein correlates well, inversely with the tumor metastatic potential.

Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Membrane ; metabolism ; Colonic Neoplasms ; metabolism ; pathology ; Cytoplasm ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Hybridomas ; immunology ; secretion ; Male ; Membrane Proteins ; immunology ; metabolism ; Mice ; Mice, Inbred BALB C ; Neoplasm Metastasis ; Sphingosine N-Acyltransferase ; Tumor Suppressor Proteins ; immunology ; metabolism

OBJECTIVETo detect the quantity, proportion and function of producing cytokines of Th1 and Th2 cells in aplastic anemia (AA) patients and their contribution to the hematopoietic failure.

METHODS(1) Eleven patients with severe aplastic anemia (SAA) at diagnosis were observed by Marsh's method for the CFU-E, BFU-E and CFU-GM before and after depletion of CD(4)(+) T lymphocytes from bone marrow mononuclear cells (BMMNC); (2) Th1 (CD(4)(+) IFN-gamma(+)) and Th2 (CD(4)(+) IL-4(+)) cells in peripheral blood mononuclear cells (PBMNC) of 21 SAA patients and 17 normal controls were counted by FACS. (3) mRNA expression of IFN-gamma and IL-4 gene in unstimulated BMMNC from 16 SAA patients, 11 chronic aplastic anemia (CAA) patients, 26 other hematological diseases patients and 11 normal controls were measured by reverse transcriptase polymerase chain reaction (RT-PCR).

RESULT(1) CFU-E, CFU-GM and BFU-E increased significantly after depletion of CD(4)(+) T lymphocytes from BMMNC of SAA patients. (2) The percentage of IFN-gamma producing CD(4)(+) T cell (Th1) of SAA patients was significantly higher than that of controls, the percentages of IL-4 producing CD(4)(+) T cells (Th2) had no difference between SAA patients and normal controls. (3) IFN-gamma mRNA was detected in unstimulated BMMNC in 13 of 16 SAA patients, 6 of 11 CAA patients and one of 6 paroxysmal nocturnal hemoglobinuria (PNH) patients. The IFN-gamma mRNA was not detected in unstimulated BMMNC of 11 normal controls and other hematological diseases patients.

CONCLUSIONSDisbalance of CD(4)(+) T lymphocytes subsets and increases in quantity and IFN-gamma producing function of Th1 cells might be important for the development of bone marrow failure in AA and in distinguishing AA from other kinds of pancytopenic diseases.

Adolescent ; Adult ; Anemia, Aplastic ; blood ; etiology ; Colony-Forming Units Assay ; Erythroid Precursor Cells ; cytology ; Female ; Granulocytes ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans ; Interferon-gamma ; genetics ; Interleukin-4 ; genetics ; Macrophages ; cytology ; Male ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Th1 Cells ; cytology ; metabolism ; physiology ; Th2 Cells ; cytology ; metabolism

OBJECTIVETo investigate the relationship between DNA-dependent protein kinase (DNA-PK) activity and anti-cancer drug sensitivity in human glioma tissues.

METHODSHuman glioma specimens were primarily cultured and its sensitivity to several anti-cancer drugs were evaluated by MTT assay. Nuclear protein was extracted from the glioma sample of the same patient and its DNA-PK activity was determined by a biotinylated DNA-PK assay with p53-derived peptide as a specific substrate.

RESULTSDNA-PK activity varied widely among these glioma samples. Of all 36 samples, 16 showed higher DNA-PK activity (relative activity > or = 0.40) and 20 samples with lower DNA-PK activity (relative activity < 0.40). The gliomas sensitive to DDP and VCR as evaluated by inhibition rate (IR > or = 50%) under plasma peak concentration (PPC) showed lower DNA-PK activity than the resistant ones (IR < 50%) (t = -3.445, P < 0.01). Furthermore, the gliomas with higher DNA-PK activity showed lower inhibition rate (IR < 50%) than those with lower DNA-PK activity ones (t = -2.145, P < 0.05).

CONCLUSIONDNA-PK activity is significantly associated with anti-cancer drug sensitivity to DDP and VCR in human gliomas. DNA-PK activity could be used as a new biomarker for the chemotherapy sensitivity of human gliomas.

Antineoplastic Agents ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Cisplatin ; pharmacology ; DNA-Activated Protein Kinase ; metabolism ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Glioma ; enzymology ; pathology ; Humans ; Nuclear Proteins ; metabolism ; Vincristine ; pharmacology