Objective To establish a stable cell line expressing transmembrane form of human blood group A antigen mimotope vaccine by transfecting malignant melanoma cell line B16, and to detect the cytotoxicity of the vaccine against melanoma cells. Methods Cultured B16 cells were classified into 4 groups, i.e.,P/F-M-pIRES group [transfected with the recombinant plasmid mimotope peptide/Fas-macrophage inflammatory protein (Mip)-pIRES], P/F-pIRES group (transfected with the recombinant plasmid mimotope peptide/FaspIRES), M-pIRES group (transfected with the recombinant plasmid Mip-pIRES), and pIRES group (transfected with the empty plasmid pIRES). B 16 cells were transfected through Lipofectamine 2000. Subsequently, RT-PCR and Western blotting were performed to detect the mRNA and protein expressions of the mimotope peptide/Fas fusion gene and Mip3β in transfected B16 cells. Cell counting kit-8 (CCK-8) was used to evaluate the vaccinemediated complement dependent cytotoxicity (CDC) and antibody-dependent cell-mediated cytotoxicity (ADCC)against B16 cells. Results RT-PCR yielded specific DNA fragments with expected size. Western blotting revealed the anti-A antibody-binding activity of the recombinant mimotope peptide/Fas fusion protein. Factor analysis indicated significant differences in CDC (F = 244.522, P ＜ 0.01 ) and ADCC (F = 71.593, P ＜ 0.01 )against B16 cells between the 4 groups. Group comparisons demonstrated more intense CDC and ADCC in P/FM-pIRES and P/F-pIRES groups compared with M-plRES and pIRES groups, stronger ADCC in P/F-M-pIRES group in comparison with P/F-pIRES group (F = 15.42, P ＜ 0.05), but no significant difference in CDC was observed between M-pIRES and pIRES group. Conclusions The transmembrane form of human blood group A antigen mimotope vaccine could be stably expressed in B16 cells, and mediate ADCC and CDC against B16 cells in vitro.

OBJECTIVEIn order to develop plans for effective intervention measures, prevalence and health-seeking behavior related to reproductive tract infection among floating married women of childbearing age in Fengtai district in Beijing were studied.

METHODSCross-sectional study was carried out. Two thousand and sixty-nine eligible women were randomly selected from strata based on their home provinces. From June to July 2001, the subjects were given face-to-face interview at the Fengtai family planning clinic in Beijing using standard questionnaire followed by gynecologic examination and laboratory tests.

RESULTSThirty point three percent of the subjects were found to have reproductive tract infections (RTI) by laboratory tests. Prevalence rates of bacterial vaginosis, candida and trichomonas vaginitis were 22.2%, 4.9% and 2.1% respectively. Prevalence rates of chlamydia, gonorrhea, condyloma acuminatum and syphilis were 2.2%, 1.6%, 0.5% and 0.2% respectively. Of these infected women, only 43.1% (270/626) were symptomatic, and 61.5% (166/270) of these women with symptoms had sought treatment.

CONCLUSIONCompared to other results in the literature, we found a relatively high prevalence of RTI in our study population. Only a small proportion of these infected women were symptomatic but only few of them sought treatment. We suggested that the provision of more family planning service and promotion of RTI knowledge to the floating women of childbearing age.

Adolescent ; Adult ; China ; epidemiology ; Cross-Sectional Studies ; Female ; Humans ; Infection ; epidemiology ; Middle Aged ; Prevalence ; Surveys and Questionnaires ; Travel ; Trichomonas Vaginitis ; epidemiology ; Urban Health ; Vaginitis ; epidemiology ; microbiology ; Vaginosis, Bacterial ; epidemiology ; Women's Health Services

The purpose of this study was to investigate the effect of simvastatin (SIM) on proliferation and apoptosis of acute monocytic leukemia cell line SHI-1 and its mechanism. Experiments were divided into control and test groups (5 µmol/L, 10 µmol/L, 20 µmol/L SIM groups). The growth inhibitory rate of SHI-1 cells was detected using methyl thiazolyl tetrazolium (MTT) method. The cell cycle distribution and apoptotic rate were measured by using flow cytometry. The expression of BCL-2, caspase-3 mRNA were determined by reverse transcription polymerase chain reaction (RT-PCR). The expression of BCL-2, caspase-3 protein levels were analyzed by Western blot. The results demonstrated that SIM inhibited the growth of SHI-1 cells in time- and does-dependent manners. Cell cycle analysis showed that SHI-1 cells significantly arrested in S phase (p < 0.05) after treating with SIM for 48 hours, as compared with control group. 5 µmol/L SIM in test group significantly blocked cell cycle progression, but can not induce apoptosis. The expressions of BCL-2 mRNA and protein were down-regulated and caspase-3 mRNA and protein were up-regulated along with the increase of SIM concentration (p < 0.05). It is concluded that SIM is able to inhibit proliferation and induce apoptosis of SHI-1 cells, the mechanism may be associated with downregulating the expression of apoptosis-related gene BCL-2, upregulating the expression of caspase-3.
Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Gene Expression Regulation, Leukemic ; Humans ; Leukemia, Monocytic, Acute ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Simvastatin ; pharmacology

Most patients with differentiated thyroid cancer have a good prognosis after radioiodine-131 therapy,but a small number of patients are insensitive to radioiodine-131 therapy and even continue to develop disease.At present,some targeted drugs can improve progression-free survival in patients with radioactive iodine-refractory differentiated thyroid cancer(RAIR-DTC),such as sorafenib and levatinib,have been approved for the treatment of RAIR-DTC.However,due to the presence of primary and acquired drug resistance,drug efficacy in these patients is unsatisfactory.This review introduces the acquired drug resistance mechanism of sorafenib in the regulation of mitogen-activated protein kinase(MAPK)and phosphatidylinositol-3-kinase(PI3K)pathways and proposes related treatment strategies,in order to provide a reference for similar drug resistance mechanism of sorafenib and effective treatment of RAIR-DTC.

BACKGROUNDThe hairpin cell-penetrating peptides (hCPPs) demonstrate an interesting characteristic of conditioned activation by molecules. We hypothesized that hCPPs have the potential to selectively deliver a paramagnetic gadolinium probe into the matrix metalloproteinase 2 (MMP-2) positive human ovary adenocarcinoma cell lines, SKOV-3.

METHODShCPPs were synthesized and labeled with 1,4,7,10-tetraazacyclododecane-N,N',N'',N''' tetraacetic acid gadolinium (III) (Gd-DOTA) and fluorescein isothiocyanate (FITC) by f-moc strategy using a standard solid phase peptide synthesis protocol. MMP-2 expression and activity were demonstrated by reverse transcriptase polymerase chain reaction (RT-PCR) and zymography. Internalization and location of hCPPs in SKOV-3 cells were observed by fluorescein imaging and flow cytometery. Selective delivery of Gd-DOTA in SKOV-3 cells was observed by magnetic resonance imaging (MRI) and transmission electron microscopy (TEM).

RESULTSThe uptake of hCPPs by SKOV-3 cells depended on the activity of MMP-2. T1WI signals of SKOV-3 cells treated with Gd-DOTA-hCPPs suggested the uptake of Gd-DOTA-hCPPs increased in a time- (r = 0.990, P < 0.01) and concentration-dependent manner (r = 0.964, P < 0.001), but was inhibited by a MMP-2 inhibitor. Electron-dense particles observed in the cytoplasm and nucleus by transmission electron microscopy proved the intracellular penetration of gadolinium.

CONCLUSIONShCPPs can be used as an effective vector for an MRI molecular probe to assess the activity of MMP-2.

Cell Line, Tumor ; Cell-Penetrating Peptides ; adverse effects ; chemical synthesis ; chemistry ; metabolism ; Flow Cytometry ; Heterocyclic Compounds ; adverse effects ; chemical synthesis ; chemistry ; metabolism ; Humans ; Magnetic Resonance Imaging ; Matrix Metalloproteinase 2 ; chemistry ; metabolism ; Microscopy, Electron, Transmission ; Organometallic Compounds ; adverse effects ; chemical synthesis ; chemistry ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo explore the morphology, immunophenotype, cytogenetics and clinical features of TCF3-PBX1 fusion gene positive adult acute lymphoblastic leukemia (ALL).

METHODSR banding was used to analyze conventional cytogenetics (CC), interphase fluorescence in situ hybridization (iFISH) and RT-PCR to detect the TCF3-PBX1 fusion gene, and flow cytometry to immunophenotype. The clinical and laboratory features and long-term follow-up of the patients were analyzed.

RESULTSThe incidence of 19 TCF3-PBX1-positive adult ALL was 3.13% of total ALL patients. Of them, 12 and 7 cases were diagnosed as L(1) and L(2) morphology respectively; 7 cases with balanced translocation of chromosome 1 and 19; 10 with der(19) t(1;19) formed from unbalanced translocation and 2 with normal karyotypes. TCF3-PBX1 fusion gene was detected by RT-PCR in 9 cases, and by iFISH in 17. 16 cases were B-phenotype and the other 2 T-phenotype; 17 cases had lymph node, spleen or liver infiltration. Of 18 patients received chemotherapy, 17 (94.7%) achieved complete remission (CR); the median relapse-free survival (RFS) and median overall survival was 3.2 months and 7.2 months, respectively.

CONCLUSIONSTCF3-PBX1-positive adult ALL had unique clinical and pathological features with high remission rate, high relapse rate and short survival time and should be considered to receive intensified treatment strategies. iFISH combined with CC and RT-PCR can increase the detection rate of t(1;19)/TCF3-PBX1 fusion gene.

Adult ; Chromosomes, Human, Pair 1 ; Humans ; In Situ Hybridization, Fluorescence ; Oncogene Proteins, Fusion ; genetics ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; Translocation, Genetic

Objective To investigate the clinical characteristics and therapeutic methods of bac-terial colonization and infection of Acinetobacter baumannii on the wound of earthquake induced patients. Methods A retrospective study was done on 42 Wenehuan earthquake induced patients with positive wound germiculture of Acinetobacter baumannii. There were 24 males and 18 females, at mean age of 37 years (12-96 years). Open injury was located at the upper arm in one patient, at the forearm in four, at the thigh in 12, at the calf in 23 and at the trunk in two. The time between injury and treatment varied from 3 to 7 days. The clinical characteristics including the bacteria identification and drug sensitivity test were studied to compare drug resistance to 15 antibiotics. Results Bacterial colonization of Acineto-bacter baumannii was found in 31 patients (8.2%) and infection of Acinetobacter baumannii in 11 (2.9%). After debridement, pruphylactic antibiotics and nutrition support, 15 patients with bacterial colonization were managed with Ⅱ stage suture or skin grafting. The other 16 patients were transferred to hospitals of other provinces after germiculture turned negative. Through debridement and drainage, antibi-otic therapy and nutrition support, the infection was controlled and the wound eliminated in six patients through Ⅱ stage suture but four were concomitant with pulmonary infection and one with septicemia. Drug sensitivity test showed that sensitive rate to imipenem, amikacin, levofloxacin, ticarcillin-clavulanic acid, tobramycin were 59.5%, 21.4%, 21.4%, 19.5% and 19.0% respectively. Conclusions The risk factors of infection of Acinetobacter baumannii include severe tissue trauma, severe wound contamination, delayed treatment and weak body resistance. During treatment, the bacterial colonization and infection of Acinetobacter baumannii should be distinguished and treated respectively. Correct wound treatment, suit-able antibiotic therapy and increased body resistance are key to improvement of clinical curative effect.

The objective was to observe the influence of previously activated psoralens on the proliferation of K562 cells, and to provide laboratory data for its clinical usage. K562 cells were treated separately with previously and late activated psoralens, then their trypan blue exclusion inhibited rates (TBIR), cell proliferation inhibited rates (CPIR) and colony forming inhibited rates (CFIR) after culture were compared. The results showed that previously activated psoralens displayed an inhibiting effect on the proliferation of K562 cells with a dose-effect relationship. There was no obvious difference between previously and late activated psoralens on TBIR, CPIR and CFIR. In order to exert the inhibitive effect of previously activated psoralens, the time of ultraviolet ray exposure should be 10 minutes at least, and longer than 12 hours for inhibiting K562. The inhibitive effect of previously activated psoratens decreased as the time interval from activation to its use was prolonged. The inhibiting effect of previously activated psoralens was strongest within 6 hours after activation. In conclusion, both previously and late activated psoralens show inhibiting effects on the proliferation of K562, which may be able to use an antineoplastic drug in clinic.
Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Furocoumarins ; pharmacology ; Humans ; K562 Cells ; Time Factors

OBJECTIVETo study the expression of annexin II (AnnII) and the fibrinolytic activity in NB4 cells and their alterations in the presence of arsenic trioxide (ATO) and daunorubicin (DNR).

METHODSLeukemia cell line NB4 was treated with ATO or DNR for 24 ∼ 72 h. Cell surface expression of AnnII and its mRNA were analysed by flow cytometry and real time PCR, respectively, the fibrinolytic activity by chromogenic assay.

RESULTSCompared with other acute leukemia cell lines, the expression of AnnII on untreated NB4 cells was relatively higher. The AnnII positive cell rates on NB4, HL-60, K562, and A3 cells were (94.5 ± 1.6)%, (40.1 ± 2.1)%, (36.3 ± 1.5)% and (11.8 ± 2.5)%, respectively. The fibrinolytic activity of NB4 cells was the greatest with a A value of 0.68 ± 0.02. The fibrinolytic activity of NB4 cells was obviously decreased by ATO, DNR or monoclonal antibody against AnnII, being decreased by 60.4%, 35.8% and 26.0% of the pretreatment level, respectively. The expressions of AnnII and its mRNA in NB4 cells were decreased dramatically after ATO and DNR treated for 48 h. Annexin II positive cells rate were (55.46 ± 4.72)% and (27.00 ± 6.18)%, respectively.

CONCLUSIONNB4 cells have strong ability to enhance the catalytic efficiency of the t-PA-dependent plasminogen activation and AnnII on the cell membrane contributes to this activity. Its high fibrinolytic activity can be corrected by ATO and DNR through down-regulating AnnII.

Annexin A2 ; Apoptosis ; Daunorubicin ; HL-60 Cells ; Humans ; Leukemia ; metabolism

Objective:To explore the effect of the X-BL mixed teaching mode on Pharmacology course. Methods:In Pharmacology course of the 2017 pharmacy major of our university, 3 teaching units were randomly selected as the control group while the rest 3 teaching units were selected as the experimental group. Traditional teaching mode was carried out in control group. In the experimental group, we designed a X-BL mixed teaching mode composed of web-based learning (WBL), case-based learning (CBL), and team-based learning (TBL). Teaching effects of the two groups were compared using online unit tests and questionnaires. Test scores were analyzed by SPSS 20.0, and differences between groups were analyzed by t test. Results:The test scores of each unit of the experimental group were significantly higher than those of the control group ( P<0.01), and the low scores were all zeroed. The questionnaires showed that the two groups showed similar learning willingness, but the experimental group students were more satisfied with teaching method, teaching quality, classroom atmosphere, teacher guidance and learning effect than the control group. Conclusion:The X-BL mixed teaching mode, which focused on case teaching and group learning, integrated online and offline teaching, and information teaching, has showed a better teaching effect than traditional teaching in the Pharmacology courses. This teaching mode may have certain promotion value in the future teaching applications.