Objective To prepare an antiserum of human putative magnesium transporter protein (HPT protein) to study its biological effects on the drug resistance of gastric cancer cell by observing its expression level. Methods The cDNA encoding HPT protein was cloned and sequenced. Prokaryotic expression vector was constructed and HPT protein fused with histidine tag was expressed. Mice were immunized with the polyacrylamide gel particles containing the fusion protein, and the antisera of them were detected for antibody activity by Western blot. Using the antiserum, HPT protein expression level was assessed in different gastric cancer cell line. Results The sequence of cloned cDNA fragment was consistent with that of HPT. After BL21/pRSETB bacterium was induced with IPTG, a new protein band with a relative molecular weight of 55kD was showed on SDS PAGE profile, and further proved to be fusion protein with histidine tag by Western blot. The antiserum of immunized mice only recognized a unique protein band with a relative molecular weight of 50kD. HPT protein expression level in SGC7901/ADR was higher than in SGC7901. Conclusion The antiserum of HPT protein was successfully prepared, and HPT was found to probably take part in the gastric cancer cell MDR

Adolescent ; Alkanes ; poisoning ; Alkenes ; poisoning ; Daphne ; chemistry ; Humans ; Male

Administration, Oral ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Peracetic Acid ; administration & dosage ; poisoning ; Poisoning ; diagnosis ; therapy

Objective To explore the dysfunction of dendritic cells (DC) related to TGFβ reversed after blocking the TGFβ signal pathway by recombinant adenovirus vector encoding for Smad7.Methods Smad7 by recombinant adenovirus vector was transfected into dendritic cells.Expression of immunologic phenotypes was detected by FCM,and CTL activity induced by DC was compared.Results The DC modified with Smad7 still expressed high adhesiveness factor related to maturation even if existing exogenous TGFβ1,which was significant statistically compared with DC transfected with control adenoviral vector (P ＜0.01).Even if existing exogenous TGFβ1,the DC modified with Smad7 pulsed with soluble antigen associated with Lewis pulmonary carcinoma could still induce potent CTL activity against Lewis pulmonary carcinoma,which showed significant difference with DC-Ad-c (P ＜0.01).Conclusion The inhibitory effects on function of DC of TGFβ may be reversed by blocking the Smad signal of TGFβ pathway.

OBJECTIVE To investigate the prevalence of nasal colonization among health care workers(HCWs).METHODS Nasal swabs from 93 ICU workers and 98 other clinical workers were cultured and isolated and the tests of antibiotic susceptibility were performed by using paper diffusion method.RESULTS In total,214 isolates of 8 species from 191 health care workers were recovered,of which 187 isolates were coagulase-negative Staphylococcus(CNS)(the carriage rate of 93.71%) and 8 isolates were Staphylococcus aureus(the carriage rate of 4.19%).While the total Gram-negative bacteria carriage rate was 14.14%(27 isolates).The most frequent CNS species were S.epidermidis and S.haemolyticus.The antibiotic susceptibility profiles of S.aureus and CNS differed sharply: all 8 S.aureus strains were resistant to penicillin but were fully susceptible to oxacillin,in contrast,most of CNS were resistant to both penicillin and oxacillin.The carriage rate of CNS(60.2%)and Gram-negative bacteria(26.9%)in HCWs of ICU were higher than other HCWs(P

Objective To detect the changes of hemodynamics in patients with portal vein embolization (PVE) before surgery for hilar cholangiocarcinoma, and analyze the relationship between hemodynamics and liver hypertrophy. Methods The clinical data of 21 patients with hilar cholangiocarcinoma who were admitted to the Eastern Hepatobiliary Surgery Hospital from April 2008 to December 2009 were retrospectively analyzed.Relevant hemodynamic variables were detected and analyzed before and 3, 7, 14 days after PVE. Data were processed using Student t test or linear correlation analysis. Results The main portal vein pressure after PVE was (25.9 ± 4.1 ) cm H2O ( 1 cm H2O = 0.098 kPa), which was ( 3.5 ± 2.5 ) cm H2O higher than that before PVE [( 22.4 ± 4.1 ) cm H2O] ( t = - 6. 504, P ＜ 0.05 ). The blood flow velocity in the non-embolized branch of portal vein increased after PVE, and reached peak [(26 ±9)cm/s] at the seventh day after PVE. A positive correlation was found between the hypertrophic rate of the non-embolized lobes and the ratio of embolized lobes to total liver volume ( r = 0. 593, P ＜ 0. 05 ). Conclusion Greater scope of the embolized vascular bed of portal vein induces higher hypertrophic rate of non-embolized liver.

BACKGROUND:Mesenchymal stem cells (MSCs) exist in human tissues.Presently,cell source is single;culture method has great differences;obtained results are not consistent.Thus,it cannot verfy that isolated and cultured cells are identical calls,which is difficult to compare.OBJECTIVE:To compare the biological features of MSCs derived form bone marrow (BM),perpheral blood (PB) and cord blood (CB) under in vitro culture conditions.DESIGN,TIME AND SETTING:The cytological in vitro controlled study was performed at the Department of Hematology,Navy General Hospital of Chinese PLA from June 2007 to December 2008.MATERIALS:A total of 10 donors of hemopoietic stem cell transplantation at the Department of Hematology,Navy General Hospital of Chinese PLA were selected.MB and PB cells were obtained from the same donor,and cell volumes were respectively 20 mL and 2 mL.CB cells (30 mL) were obtained from healthy primipara at the Department of Obstetrics,Navy General Hospital of Chinese PLA.METHODS:MSCs were obtained from BM,PB and CB by Percoll density gradient + adherence method,and then incubated in DMEM/F12 medium containing 10% fetal bovine serum.When 80%-90% confluency,cells were digested in trypsin-EDTA and made into 5×10~8/L cell suspension as P_0.Above-described operation was performed as P_1,and the rest may be deduced by analogy as P_2-P_5.MAIN OUTCOME MEASURES:The following parameters were measured:cell growth morphology;results of Wright-Giemsa staining;results of cytochemistry;cell proliferation amount;cell surface markers using flow cytometry.RESULTS:Time of adherence,time to 50% confluency and time to 80% confluency of BMSCs were earlier comarped with the PBMSCs and UCMSCs.Adherent cells from BM grew in whirpool-like type,while CB and PB did not at 5-7 days.Majority of aderent cells from BM were fibroblast-like cells,and small parts were endothelioid cells.Aderent cells from PB and CB at the fifth generation contained more endothelioid cells and mononuclear and macrophage-like cells besides fibroblast-like cells.PAS stain,Sudan black B stein,alkaline phosphatase (AKP) staining of adherent cells from BM,PB and CB were negative from P_1 to P_5.Compared with P0 cells,number of BMMSCs till P5 was significantly more in PBMSCs and UCMSCs (P ＜ 0.05).Positive rates of CD29,CD44,CD90,CD71,CD105,CD166 and HLA-ABC were 55.9% 92.8% at P0 to P5,but ≤6% following BMMSCs were incubated;19.7%-33.4% at P0 to P5,but ≤10% following PBMSCs were incubated;35.4%-93.2% at P_0 to P_5,but ≤20% following CBMSCs were incubated.Positive rates of CD34,CD45 and HLA-DR were low in BM-,PB-and CB-MSCs.Positive rates of CD14 and CD31 were low in BMMSCs;12.1%-28.3% in PBMSCs,and 8.1%-21.3% in CBMSCs.CONCLUSION:MSCs can be attained from BM,PB and CB.Quantities of MSCs form BM are the highest,with single component,followed by CBMSCs and PBMSCs,with multiple components.

Obesity is an established risk factor for endometrial cancer. Leptin, a secreted protein of the ob gene by white adipose tissue, plays an important role in the regulation of food intake and energy consumption in the brain and acts as a potential growth stimulator in normal and neoplastic cancer cells. However, a direct role for leptin in endometrial cancer has not been demonstrated. In the present study, the effect of leptin on the proliferation of Ishikawa endometrial cancer cells was investigated as well as the possible mechanism(s) underlying this action in endometrial cancers which express both short and long isoforms of leptin receptors. The expression of leptin receptor (ObRb) in Ishikawa cells was detected by RT-PCR and Western blotting. The cells after serum starvation, were treated by leptin with various concentrations (0, 10, 50, 100, 150 ng/mL) for different durations (6, 12, 24 h). The effect of leptin treatment on cell proliferation was examined by MTT assay. Meanwhile, inhibitory effect of Janus tyrosine kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) inhibitor AG490 or extracellular signal-regulated kinase 1/2 (ERK1/2) inhibitor PD98059 on the proliferation of Ishikawa cells induced by leptin was also studied. Ishikawa cells were treated with 100 ng/mL leptin for various periods (0, 20, 40, 60 min), and the levels of STAT3 phosphorylation and ERK1/2 phosphorylation were examined by Western blotting. The results showed that leptin induced the phosphorylation of STAT3 and the activation of ERK1/2 in a time- and dose-dependent manner in the Ishikawa endometrial cancer cells. Blocking STAT3 phosphorylation with the inhibitor AG490, or blocking ERK1/2 activation by the specific ERK1/2 kinase inhibitor, PD98059, abolished leptin-induced proliferation of Ishikawa cells. In addition, leptin was found to potently induce the invasion of endometrial cancer cells in a Matrigel invasion assay. Leptin-stimulated invasion was effectively blocked by pharmacological inhibitors of STAT3 (AG490) and ERK1/2 kinase (PD98059). These results suggested that leptin promotes endometrial cancer growth and invasiveness by activating STAT3 and ERK1/2 signaling pathways and therefore blocking its action at the receptor level can be a rational therapeutic strategy.

Adolescent ; Brain Diseases ; diagnostic imaging ; Brain Neoplasms ; diagnostic imaging ; surgery ; Diagnosis, Differential ; Hematoma, Epidural, Cranial ; diagnostic imaging ; Hernia ; diagnostic imaging ; Humans ; Male ; Plasmacytoma ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed

OBJECTIVETo observe the impact of tonifying Qi traditional Chinese medicines contained in Yiqi Qingwen Jiedu mixture on mRNA expression of lung inflammatory cytokines and pulmonary pathological injury of mice infected by influenza virus, in order to discuss the mechanism of tonifying Qi traditional Chinese medicines against pulmonary immune inflammatory injury of infected mice.

METHODIn different time phases after mice were infected with influenza virus FM1, the RT-PCR method was adopted to observe the impact of tonifying Qi traditional Chinese medicines contained in Yiqi Qingwen Jiedu mixture on five inflammatory cytokines TNF-α, IL-1, IL-6, IL-10 and IFN-γ, and the changes in pulmonary pathological injury of mice with viral pneumonia after intervention with tonifying qi traditional Chinese medicines.

RESULT(1) Tonifying Qi traditional Chinese medicines significantly reduced the mRNA expression of TNF-α at 1-5 d and IL-1 mRNA expression at 7 d, may increase IL-1 mRNA expression in mouse lung at 3 d, significantly reduced IL-6 mRNA expression in mouse lung and increased IL-10 mRNA expression at 3-7 d, and significantly increased IFN-γ mRNA expression at 1 d. (2) Tonifying Qi traditional Chinese medicines could significantly inhibited and repaired pulmonary immune inflammatory injury of mice infected by FM1, which was most remarkable at 3-7 d after the infection with influenza virus FM1.

CONCLUSIONTonifying Qi traditional Chinese medicines contained in Yiqi Qingwen Jiedu mixture could resist pulmonary immune inflammatory injury and repair inflammatory injury by regulating the mRNA expression of imbalance inflammatory cytokines of organisms infected with influenza virus.

Animals ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Influenza A virus ; drug effects ; immunology ; Influenza, Human ; drug therapy ; genetics ; immunology ; Interferon-gamma ; genetics ; immunology ; Interleukin-1 ; genetics ; immunology ; Interleukin-10 ; genetics ; immunology ; Interleukin-6 ; genetics ; immunology ; Lung ; immunology ; virology ; Male ; Mice ; Mice, Inbred BALB C ; Tumor Necrosis Factor-alpha ; genetics ; immunology