Objective To probe the effects of nuclear factor kappa B (NF-?B) on cell apoptosis and left ventricular segmental function in acute ischemia repeffusion in dogs.Method Twenty-four dogs were randomly divided into three groups:without left anterior artery (lAD) ligation group (C group),LAD was occluded 30 min following reperfusion 120 minutes in isehemical reperfusion group (IR group),and dogs were administered with PDTC before LAD ligation in ischemical reperfusion plus pyorrole dithitocarbamate group (PDTC group).The left ventricular segmental function was detected by echo cardiography using strain rate anlysis software.EF measured by Simpson's method.Cardiac myocyte apoptosis numbers were determined by terminal deoxynudeotidy transferease-mediated biotinylated deoxyuridine triphosphate nick end labeling (TUNEL).lmmunohistochemistry and western-blot anylysis of NF-?B protein expression.Results NF-?B was obviously expression on injury myocardium of IR group,and increased significantly in contrast to control group (P0.05)Conclusions NF-?B might play an important role in acute myocardial ischemia reperfusion.PDTC reduces myocardial iscbemia/repeffasion injury by preventing expression of factor NF-?B.

BACKGROUND: Isoflavone isolated from Trifolium pratense L. has been found to be able to effectively inhibit bone resorption, reduce bone turnover rate, improve osteocyte activity and bone mineral density by enhancing the effect of estrogen, which is helpful for the prevention and treatment of osteoporosis. OBJECTIVE: To investigate the effect of Trifolium pratense L. extracts on the bone resorption and differentiation of osteoclasts.METHODS: Rat bone marrow cells were extracted, isolated by lymphocyte separation and cultured for 5 hours; then, the non-adherent cells were selected followed by induced by 30 μg/L macrophage colony stimulating factor and 75 μg/L RANKL (control groups), or different concentrations of Trifolium pratense L. extracts (0.3, 0.6 and 1.2 g/L) to observe their effect on the osteoclast differentiation and bone resorption. The levels of osteoclast differentiation-associated proteins c-fos and NFATcl were determined by western blot assay. RESULTS AND CONCLUSION: Compared with the control group, different concentrations of Trifolium pratense L. extracts could suppress osteoclast differentiation and bone resorption to different degrees. Tartrate-resistant acid phosphatase staining showed that Trifolium pratense L. extracts could significantly reduce the number of osteoclasts. Western blot assay results suggest that Trifolium pratense L. extracts significantly inhibited the expression levels of c-fos and NFATcl. These results reveal that Trifolium pratense L. extracts can inhibit osteoclast differentiation and bone resorption.

Objective To explore the clinical characteristics of nosocomial infection in children with acute leukemia and the strategy of prevention and treatment.Methods One hundred and thirty-three cases of nosocomial infection in children with acute leukemia were analyzed by retrospective study.The relationship between nosocomial infection and stage of leukemia,hospitalization duration,and the rate of infection were investigated.Results Nosocomial infection rate was 53.4%(71/133 cases),significant difference of infection rate between acute lymphoblastic leukemia and nonlymphoblastic leukemia group was found(P0.05).The main pathogens of septicaemia were gram negative bacilli,and they were generally sensitive to Amicacin and Pi-peracillin/tazobactam.Conclusions Children with acute leukemia have high nosocomial infection rate.The occurrence of nosocomial infection was related to the type and stage of leukemia and hospitalization duration but not to the prognosis.The main pathogens of septicaemia were gram negative bacilli.

Objective To investigate the status of the disease of the fluorosis and arsenism caused by coal-burning in Ankang city of Shaanxi. Methods Nine survey spots were chosen to carry out the epidemiological investigation of adult skeletal fluorosis and arsenism in the coal-polluted areas of Ankang, respectively using Determination of Fluorine in Coal (GB/T 4633-1997) to determine the coal fluorine and using hydride generation atomic fluorescence spectrophotometry(HCAFS) to determine coal arsenic. The diagnose of the adult skeletal fluorosis followed the Diagnosis of Clinical Classification for Endemic Skeletal Fluorosis Standard(GB 16396-1996), that of arsenism using Standard of Diagnosis for Endemic Arsensim (WS/T 211-2001). Results Totally 569 adults were investigated over the age of 16, among which 121 cases were skeletal fluorosis, with a total detection rate of 21.27%. Four cases of II degree and higher skeletal fluorosis patients were identified, accounting for 0.70% of the number of subjects. One hundred and thirty-two cases of arsenic poisonin were detected, in a rate of 23.20%. Ninety-five patients were identified with moderate or severe arsenic poisoning, accounting for 16.69% of subjects. A positive correlation was found between the detection rates of the skeletal fluorosis and the arsenism(r = 0.816, P < 0.01), as well as between the detection rate of skeletal fluorosis and fluoride content of coal(r = 0.775, P < 0.05). The detection rate of arsenism and arsenic content of coal also had close relationship (r = 0.761, P < 0.05). The detection rate of skeletal fluorosis in the group aged 40 - ,50 - , and 60 - [27.20%(34/125) ,29.27%(36/123), 28.13%(36/128)] was increased, compared the group of less than 40 years age[7.77%( 15/193), X~2 = 21.969,25.648,23.856,P<0.01].For the detection rate of arsenism,male[33.67%(99/294)]was obviously higher than female[12.00%(33/275),)(X~2=37.162,P<0.01].Conclusions A high detection rate of fhorosis is correlated with arsenic poisoning,but the probability of the two diseases simultaneously occurred in a person is not high.In this polluted area.when fluoride accumulates to a certain level as in aduh,the detection rates no longer varies obviously;however,that of arsenism increases along with the age.

AIMTo investigate the effects of NPPB, a chloride channel blocker, on proliferation of mesangial cells.

METHODSCell proliferation was determined by measuring cell number and 3H-thymidine incorporation. The LDH activity released from these cells was measured as evaluation of cell viability. The phase of cell cycle was detected by flow cytometry.

RESULTSCell proliferation assays showed that treatment with both NPPB (50 and 25 micromol x L(-1)) and in hypertonic media (100% increased osmolarity with D-mannitol ) significantly reduced the number of human MC and 3H-thymidine incorporation in a dose-dependent manner. But the LDH activity was not significantly altered in the treatment with 50 micromol x L(-1) NPPB. Flow cytometry experiments showed that 50 and 25 micromol x L(-1) NPPB arrested (84.2 +/- 2.4) % and (80.8 +/- 2.9) % of cells at G0/G1 stage, versus (70.5 +/- 1.4) % of control cells. Conclusion NPPB suppresses cell proliferation and produces growth arrest at G0/G1 phase in human MC by a mechanism probably associated with changes in cell volume.

Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Chloride Channels ; antagonists & inhibitors ; Dose-Response Relationship, Drug ; Humans ; L-Lactate Dehydrogenase ; metabolism ; Mesangial Cells ; cytology ; metabolism ; Nitrobenzoates ; administration & dosage ; pharmacology

Complete mannanase gene with two introns was cloned from Trichoderrna reesei by PCR. The two introns were then removed by overlap extension PCR. The gene encoding the mature mannanase protein was inserted into the expression vector pPIC9K, downstream of a alpha-factor signal peptide sequence. The resultant recombinant vector was named pM242. After linearized with Sac I , pM242 was transformed to Pichia pastoris GS115 by electroporation. After screening, the recombinant strain Gpmf25 that expresses the secretory protein at high level was obtained. The activity of the recombinant mannanase reached 12.5 IU/mL. Optimum pH and temperature for the recombinant enzyme were 5.0 and 80 degrees C, respectively. The enzyme was stable at pH 5.0-6.0 and maintained over 50% of original activity after incubation at 70 degrees C for 30 min.
Fungal Proteins ; biosynthesis ; genetics ; Hydrogen-Ion Concentration ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; Temperature ; Trichoderma ; enzymology ; genetics ; beta-Mannosidase ; biosynthesis ; genetics

OBJECTIVETo study the effect of all-trans retinoic acid (ATRA) on U937 cell growth and its mechanism.

METHODSCell cycle was detected by flow cytometry (FCM), expressions of cell cycle associated protein and the p27 related protein were detected by Western blot. The binding of P27 and Skp2 was detected by immunoprecipitation.

RESULTSFCM displayed that ATRA could inhibit the proliferation of U937 cells. At 72 h on 1 micromol/L ATRA treatment, 72% of the cells were arrested at G0/G1 phase. Western blot displayed that ATRA could decrease the expression of cyclin A, up-regulate the expression of p21 and p27, and down-regulate the expression of p27 related proteins Skp2. p27 could bind with Skp2 in U937 cells as detected by immunoprecipitation.

CONCLUSIONATRA may arrest the proliferation of U937 cells through the reduction of Skp2 expression, and finally the induction of the accumulation of p27.

Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Humans ; S-Phase Kinase-Associated Proteins ; metabolism ; Tretinoin ; pharmacology ; U937 Cells

OBJECTIVETo investigate the expression and relationship of p27(kip1) and its nuclear export factor Jab1 during proliferation process of lymphoma cell.

METHODSJurkat and Raji cells were treated with serum starvation and then serum release. The protein and mRNA expression of p27(kip1), Jab1 in the cells were detected by Western blot and RT-PCR respectively. LMB were used for stimulating Jurkat cells during their proliferation process, and then the expression changes of p27(kip1) and Jab1 were detected. An eukaryotic expression plasmid(pcDNA3. 1-myc) containing Jab1 was constructed. Jurkat cell were transfected in vitro with or without pcDNA3. 1-myc-Jab1. Double immunolabelling was used to identify the localization of p27(kip1). Immunoprecipitation was used to detect the combination of p27(kip1) and Jab1.

RESULTSThe growth of Jurkat and Raji cells were blocked by serum starvation. The total protein amount of p27(kip1) increased while that of Jab1 decreased. The reverse changes were happened after serum release, but the mRNA expression of p27(kip1) has no significant change. LMB could inhibit the cell proliferation caused by serum release. The expression of p27(kip1) was up-regulated and Jab1 down-regulated when Jurkat cells were treated with LMB. After pcDNA3. 1-myc-Jab1 infected Jurkat cells for 48 h, the distribution of p27(kip1) was translocated from nucleus into cytoplasma. p27(kip1) and Jab1 could form compound in Jurkat and Raji cells detected by Immunoprecipitation.

CONCLUSIONJab1 may influence the location and expression of p27(kip1) through integrating with p27(kip1), and then participates in regulating the growth of NHL cell through interfering with the function of p27(kip1).

COP9 Signalosome Complex ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Humans ; Intracellular Signaling Peptides and Proteins ; metabolism ; Jurkat Cells ; Peptide Hydrolases ; metabolism ; Plasmids ; RNA, Messenger ; metabolism ; Transfection

Soxhlet extraction is a common method of sample preparation. However, there has been no discussion about the efficiency of Soxhlet extraction from different batches and the factors that cause content fluctuation. In this study, Panax ginseng was selected as a model sample. Soxhlet extraction by means of a water bath, which has always been neglected, was identified as a novel key factor in the poor repeat-ability in different batches of Soxhlet extraction, as it can affect the siphon times and reflux time, which have been positively correlated with the ginsenoside contents. By substituting round bottom flasks in the same column, the relative standard deviation of the most fluctuated compound, ginsenoside Rb1, was decreased from 24.6% to 5.02%. Scanning electron microscopy analysis confirmed that the breakdown of the surface of the ginseng powder in the Soxhlet extraction led to a better dissolution of ginsenosides, indicating that chloroform may promote the extraction of ginsenosides by disrupting the cell structure. Moreover, 70% methanol was regarded as the better solvent for extracting the ginsenosides. Overall, this work offers a practical and effective protocol for improving the accuracy and repeatability of Soxhlet extraction methodology for ginsenosides and other analytes.