Objective To investigate the fuction of tumor necrosis factor alpha(TNF-?),interleukin-1 beta(IL-1?) and interleukin-6(IL-6) in children with intracranial infection.Methods TNF-?,IL-1? and IL-6 levels of serum and cerebrospinal fluid(CSF) were determined in the purulent meningitis group(25 cases),tuberculous meningitis group(17 cases),viral meningitis group(30 cases)and control group(20 cases)by enzyme-linked immunosorbent assay(ELISA).Results The levels of TNF-?,IL-1? and IL-6 obviously increased in CSF compared with that in the serum (Pa

OBJECTIVETo study the effect of ursolic acid (UA), apentacyclic triterpene acid, on MCF-7 cell apoptosis, and probable mechanism involved by detecting the expressions of caspase-3 and poly ADP-ribose polymerase(PARP) at protein level.

METHODMCF-7 cells were cultured with different concentrations of UA. Growth inhibition of UA on MCF-7 cells was evaluated by MTT assay. Cell cycle and sub-G1 peak were performed by FCM. Morphologic changes of UA-treated cells were observed by light microscope. Apoptotic cells with condensed or fragmented nuclei were visualized by Ho 33258 staining by a fluorescence microscope (EX: U. V.). The protein expression of caspase-3 and PARP was analyzed by immunofluorescence cell staining (SABC-Cy3).

RESULT24 hours after UA treatment, inhibition of MCF-7 cell growth was concentration-dependent. The IC50 value for UA was (22.6 +/- 3.0) micromo x L(-1). Cell cycle anaysis by FCM showed that 50 micromol x L(-1) of UA arrested MCF-7 cell cycle at G0 - G1 phase. Morphological changes of MCF-7 Cells exhibited many of the hallmark features of apoptosis, including chromatin clumps and aggregation and DNA fragmentation. UA increased caspase-3 protein expression.

CONCLUSIONThe results suggest that UA evokes MCF-7 cell apoptosis is correlation with the up-regulation of caspase-3. Our study indicated that UA might be a potential Chinese medical component for breast neoplasm.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Breast Neoplasms ; enzymology ; pathology ; Caspase 3 ; Caspases ; metabolism ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Female ; Humans ; Poly(ADP-ribose) Polymerases ; metabolism ; Triterpenes ; pharmacology

OBJECTIVETo study the changes in expression and activity of protein phosphatases type 2A (PP2A ) during differentiation of NB4 and NB4-MR2 cells induced by all-trans retinoic acid (ATRA), and evaluate the role of PP2A in MR2 resistance to ATRA.

METHODSATRA, okadaic acid (OKA) and ATRA + OKA at the same dosage were incubated with NB4 and MR2 cells respectively. Wright's staining and NBT reduction test were employed to evaluate the change in the cells. The CD11b expression was measured by flow cytometry. The activity of PP2A was evaluated by serine/threonine phosphatase assay system, and the level of PP2A subunits was detected by Western blot.

RESULTS1) Wright's staining, NBT reduction test and flow cytometry results showed OKA could augment the differentiation of NB4 induced by ATRA, and OKA + ATRA induced slight differentiation of MR2 cells. 2) Phosphatase assay showed a decrease in PP2A phosphatase activity [(534 +/- 43) pmol x min(-1) x microg protein(-1)] in NB4 after ATRA treatment, accompanied with that activity [(959 +/- 83) pmol x min(-1) x microg protein(-1)] in untreated NB4 cells. OKA enhanced the inhibitory effect of ATRA on the activity in NB4. When OKA + ATRA was incubated with MR2, PP2A in the cells was significantly decreased [(229 +/- 23) pmol x min(-1) x microg protein(-1)]. 3) Western blot analysis showed that the level of PP2A catalytic subunit (PP2A/C) was decreased during the course of ATRA-induced NB4 cell differentiation, whereas expressions of every subunits of PP2A in MR2 cells were somewhat unaltered.

CONCLUSIONExpression of PP2A/C and activity of PP2A is decreased during differentiation of NB4 induced by ATRA, and no repression of the PP2 activity maybe related to MR2 resistance to ATRA.

Cell Differentiation ; drug effects ; Cell Line, Tumor ; Humans ; Leukemia, Promyelocytic, Acute ; metabolism ; pathology ; Okadaic Acid ; pharmacology ; Phosphoprotein Phosphatases ; metabolism ; Protein Phosphatase 2 ; antagonists & inhibitors ; metabolism ; Tretinoin ; pharmacology

Objective To study the effects of verbascoside on the proliferation and osteogenic differentiation of rat bone marrow stromal cells (BMSCs) and its mechanism.Methods BMSCs were obtained and identified from rat bone marrow by density gradient centrifugation.BMSCs were randomly divided into high,medium,low dose test groups and control group,and treated with 100,50,25,0 μmol · L-1verbascoside.The cell counting kit-8 (CCK-8) method was used to evaluate the BMSCs proliferation at 24,48,72 h after administration the medication.The alkaline phosphatase(ALP) activities were detected at 24,48,96 h after medication.The ALP staining was performed after 72 h of medication.Western blot and real time-polymerase chain reaction (PCR) were used to detect the osteogenic related proteins and genes expression.Results After 72 h,the proliferation of BMSCs in high dose test group was 1.22 ± 0.05,lower than that in control group,which was 1.50 ± 0.09 (P ＜ 0.05).The ALP expression levels in high,medium and low dose test groups were higher than control group at 24,48,96 h.After 72 h,ALP staining showed that the color of culture dishes in high,medium and low dose test groups were deeper than that in control group.Western blot showed that osteogenic related proteins of bone morphogenetic protein-2 (BMP2),Smad1,Smad5,Smad8,Runx2,Osterix in high,medium and low dose test groups were higher than those in control group (P ＜0.05),but there was no significant difference among those three test groups (P ＞ 0.05).Real time-PCR showed that the osteogenic related genes of BMP2,Smad1,Smad5,Smad8,Runx2,Osterix expression levels in high,medium and low dose test groups were higher than those in control group (P ＜ 0.05).However,there was no significant difference among three test groups(P ＞ 0.05).Conclusion The verbascoside at different concentrations has no obvious effect on BMSCs proliferation and promote osteogenic differentiation of BMSCs through BMP2-Smads -Runx2-Osterix pathway.

OBJECTIVETo investigate the role of bacteria in the etiology of chronic prostatitis.

METHODSA total of 162 complete prostate specimens were obtained at autopsy from organ donors (aged 20 -38 yr) who died of non-prostatic diseases. Each of the samples from the peripheral zone of the prostate was divided into two parts, one for routine pathological examination and immunohistochemical studies of interleukin (IL)-1beta, tumor necrosis factor-alpha (TNF-alpha) and the nerve growth factor (NGF), and the other for PCR assay to detect the bacterial 16S rRNA gene (16S rDNA).

RESULTSFifty-one (31.5%) of the total specimens presented pathological changes of chronic prostatitis, of which 44 had mild focal stromal, 5 mild focal stromal and periglandular and 2 mild focal periglandular inflammation. The positive rate of 16S rDNA was 19.1% (31/162), 51.0% (26/51) in the chronic prostatitis and 4.5% (5/111) in the non-prostatitis specimens (chi2 = 29.783, P < 0.01). In the specimens with chronic prostatitis, the expressions of IL-1beta, TNF-alpha and NGF were significantly higher in the 16S rDNA positive than in the 16S rDNA negative group (P < 0.01).

CONCLUSIONBacterial inflammation may play an important role in the etiology of chronic prostatitis.

Adult ; Chronic Disease ; Genes, rRNA ; Humans ; Interleukin-1beta ; metabolism ; Male ; Nerve Growth Factor ; metabolism ; Prostate ; metabolism ; microbiology ; pathology ; Prostatitis ; metabolism ; microbiology ; pathology ; RNA, Bacterial ; genetics ; RNA, Ribosomal ; RNA, Ribosomal, 16S ; genetics ; Tumor Necrosis Factor-alpha ; metabolism ; Young Adult

OBJECTIVETo determine the prevalence of sleep disordered breathing (SDB) in elderly patients with chronic congestive heart failure (CHF) and explore the relations between SDB and left ventricular function.

METHODSBy means of polysomnography, 56 elderly patients with CHF were divided into non-SDB, mild SDB, moderate SDB, and severe SDB groups, and the left ventricular ejection fraction (LVEF) was measure by (99)Tc equilibrium radionuclide angiography.

RESULTSIn the 56 elderly patients with CHF, 38 (67.9%) had SDB, including 12 (21.4%) mild SDB, 14 (25.0%) moderate SDB, and 12 (21.4%) severe SDB patients. Thirty (53.6%) of the 56 patients with CHF had obstructive sleep apnea (OSA), 4 (7.1%) had central sleep apnea and 22 (39.2%) had mixed sleep apnea. The moderate and severe SDB groups had lower minimum arterial oxyhemoglobin saturation during sleep than the non-SDB groups, and the apnea-hyponea index was closely related to LVEF (r=-0.74, P<0.01).

CONCLUSIONThe prevalence of SDB, predominantly OSA, is high in elderly patients with CHF. Moderate and severe SDB might affect the left ventricular function in these patients, who require polysomnography monitoring.

Aged ; Aged, 80 and over ; China ; epidemiology ; Chronic Disease ; Female ; Heart Failure ; complications ; physiopathology ; Humans ; Male ; Middle Aged ; Polysomnography ; Sleep Apnea Syndromes ; complications ; epidemiology ; Ventricular Dysfunction, Left ; physiopathology

OBJECTIVE: To investigate the expressions of Survivin protein and nm23 protein and the relationship among the expressions and axillary lymph node metastasis in breast cancer. METHODS: The expression of Survivin and nm23 in 80 cases of breast cancer tissues were detected by immunohistochemistry SP method, and their correlation with axillary lymph node metastasis and 5-year disease free survival (DFS) were analysed. RESULTS: Survivin protein positive expression rate was 68.75% (55/80) in breast cancer tissues, which had positive correlation with the axillary lymph nodes metastasis but negative correlation with 5 years FS (P < 0.05); nm23 protein expression had negative correlation with the axillary lymph nodes metastasis but positive to 5 years FS (P < 0.05). Survivin and nm23 proteins expression had no obvious correlation with the breast cancer pathology type, patient age and clinical stage (P > 0.05). CONCLUSION The anti-apoptosis effect of Survivin protein and the anti-metastasis effect of nm23 protein may be important in the occurrence and advancement of breast cancer, suggesting that it may be a new indicator of prognostic and judgement in breast cancer.
Adult ; Aged ; Axilla ; Biomarkers, Tumor ; Breast Neoplasms ; metabolism ; pathology ; surgery ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Mastectomy ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; Middle Aged ; NM23 Nucleoside Diphosphate Kinases ; Neoplasm Proteins ; biosynthesis ; genetics ; Nucleoside-Diphosphate Kinase ; biosynthesis ; genetics ; Prognosis ; Retrospective Studies ; Survivin

This study was aimed to explore the effects of peptidoglycan (PGN) on proliferation and cell cycle of human bone marrow mesenchymal stem cells (MSCs). MSCs were isolated from human bone marrow by density gradient centrifugation. The purity of MSCs with the spindle fibroblastic morphology was identified by microphotography and the phenotypes were detected by flow cytometry (FCM). MSCs incubated with different doses of PGN (1, 10, 20 μg/ml) were used as test groups, and those incubated without PGN were regarded as control group. The isolated and cultured MSCs were inoculated into 96-well plates according to a certain concentration. Cell cycle was measured by flow cytometry after incubated with PGN for 72 hours. The results showed that the cell proliferation index was significantly increased in dose and time dependent manners after MSCs was incubated with PGN. Its effects on the proliferation of MSCs were highest in 10 μg/ml group. Compared with the control group, PGN could significantly decrease proportion of MSCs in G₀/G₁ phase and increase them in S and G₂/M phases (p < 0.05). It is concluded that PGN can promote more MSCs to enter the DNA synthesis phase and proliferate many much MSCs in dose and time dependent manners.
Bone Marrow Cells ; cytology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Flow Cytometry ; Humans ; Mesenchymal Stromal Cells ; cytology ; Peptidoglycan ; pharmacology ; Toll-Like Receptor 2

OBJECTIVETo detect the expression and variation of p53 gene during tree shrews' hepatocarcinogenesis induced by hepatitis B virus (HBV) and aflatoxin B1 (AFB1).

METHODSTree shrews were divided into four groups: the tree shrews were infected with HBV and fed with AFB1 in group A, only infected with HBV in group B, fed with AFB1 alone in group C, and normal control in group D. All the tree shrews were performed liver biopsy every 15 weeks. The tissues of liver and tumor were detected by immunohistochemistry and molecular biotechnologies.

RESULTS(1) The incidence of hepatocellular carcinoma (HCC) in group A (66.7%) was higher than that in Group B and C (30%). HCC appearance in group A was earlier than that in group C (120.0 weeks +/-16.6 weeks vs 153.3 weeks +/-5.8 weeks, t = 3.336, P<0.01). (2) Mutated p53 protein was not found before the 75th week of the experiment in each group. (3) At the 105th week, the expression rates of mutated p53 protein were 78.6%, 60% and 71.4% in group A, B and C respectively, which were much higher than that (10%) in group D (x2 > or = 5.03, P<0.05). An abnormal band of p53 gene was detected in both group A and C. (4) The mutation points of p53 gene in liver cancer of tree shrew were at codon 275, 78 and 13. The nucleotide sequence and amino acids sequence of tree shrew's wild-type p53 showed 91.7% and 93.4% homology with those of human p53 respectively.

CONCLUSIONSThere is a remarkable synergistic effect between HBV and AFB1 on HCC. Mutated p53 protein is expressed before HCC occurrence, which promotes the development and progress of HCC. HBV and AFB1 may synergistically induce p53 gene mutation.

Aflatoxin B1 ; toxicity ; Animals ; Carcinoma, Hepatocellular ; genetics ; Cocarcinogenesis ; Gene Expression Regulation, Neoplastic ; Genetic Variation ; Hepatitis B ; virology ; Hepatitis B virus ; Liver Neoplasms, Experimental ; genetics ; Point Mutation ; RNA, Neoplasm ; analysis ; Tumor Suppressor Protein p53 ; genetics ; Tupaiidae

Carcinoma, Hepatocellular ; metabolism ; Glypicans ; biosynthesis ; genetics ; Humans ; Liver Neoplasms ; metabolism ; RNA, Messenger ; biosynthesis ; genetics