OBJECTIVETo investigate the changes of gene expression file in transitional cell carcinoma of bladder after hepatocyte cell adhesion molecule(hepaCAM) overexpression.

METHODSAffymetrix Human Genome U133 Plus 2.0 Array was used to investigate the changes of gene expression profile between adenovirus-green fluorescent protein(GFP) -hepaCAM group and GFP group in transitional cell carcinoma of bladder EJ cells.Significant Analysis of Microarray(SAM) was used to screen the differentially expressed genes, DAVID software was used to conduct gene ontology analysis and wikiPathway analysis based on the differentially expressed genes. Reverse transcription-polymerase chain reaction and Western blot were applied to verify microarray data.

RESULTSCompared with the GFP group, a total of 2469 genes were up-regulated or down-regulated by more than 2 times in the GFP-hepaCAM group. Among these genes, 1602 genes were up-regulated and 867 were down-regulated.Most of the differentially expressed genes were involved in the function of cell proliferation and cell cycle regulation. The mRNA expressions of nibrin, liver kinase B1, and cyclin D1 detected by reverse transcription-polymerase chain reaction in three different bladder cancer cell lines were consistent with the microarray data.The protein expressions of nibrin and liver kinase B1 in these three cell lines measured by Western blot were consistent with the mRNA expression.

CONCLUSIONSHepaCAM can alter the gene expression profile of bladder cancer EJ cells. The well-known anti-tumor effect of hepaCAM may be mediated by regulating the gene expression via multiple pathways.

Carcinoma, Transitional Cell ; genetics ; pathology ; Cell Cycle Proteins ; metabolism ; Cell Line, Tumor ; Cyclin D1 ; metabolism ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; physiology ; Humans ; Nuclear Proteins ; metabolism ; Protein-Serine-Threonine Kinases ; metabolism ; Proteins ; genetics ; physiology ; Urinary Bladder Neoplasms ; genetics ; pathology

The aim of this study was to expand hematopoietic progenitor cells at large scale by magnet stirred culture system. Mononuclear cell from umbilical cord blood were cultured in serum-free medium with stem cell factor, FIT-3 ligand and thrombopoietin. Firstly, the role of magnet on cell growth and colony-forming was studied by static culture on 0, 25 and 50 mT. Then the expansion multiple of cells, colony-forming and expression of surface markers were studied in magnet stirred culture by cell counting, colony-forming assay and flow cytometry. The results indicated that there was no difference in multiple of total cell expansion and numbers of hematopoietic colonies between 0, 25 and 50 mT groups and spinner groups (all p > 0.05). After 7 day cultures, the multiple of total cell expansion in magnet stirred culture was higher than that in static culture (p < 0.01). The numbers of CFU-GM (colony-forming unit-granulocyte/macrophage) and CFU-E (erythroid colony forming unit) in magnet stirred culture were higher than those in static culture, (p < 0.05). The primitive cells (CD34(+), CD34(+)/CD38(-) or CD133(+)) of the expanded cells in magnet stirred culture were less than those in static culture (p < 0.05). However, the CD184(+) or CD62L(+) expanded cells were more than that in static culture (p < 0.05). It is concluded that magnet stirred culture favors the expansion of hematopoietic progenitor cells. The results will be finally confirmed in further in vivo experiments and clinical applications.
Cell Culture Techniques ; methods ; Cell Differentiation ; physiology ; radiation effects ; Cells, Cultured ; Electromagnetic Fields ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans ; Leukocytes, Mononuclear ; cytology

This study was purposed to investigate the relationship between the levels of soluble receptor activator of nuclear factor kappa B ligand (sRANKL) and osteoprotegerin (OPG) in serum of the patients with multiple myeloma (MM) and multiple myeloma bone disease (MBD). The serum levels of sRANKL, OPG, tartrate-resistant acid phosphatase-5b (TRAP-5b) and C-terminal telopeptide of collagen I (CTP-I) which both are indexes for metabolism of osteoclast (OC) in newly diagnosed MM patients (n=42, experimental group) and healthy persons (n=25, control group) were detected by enzyme-linked immunosorbent assay. The roentgenography was used to determine bone damage in MM patients at the same time. According to these results acquired, the correlation of sRANKL/OPG ratio with levels of TRAP-5b/CTP-I, the incidence and degree of bone destruction were analyzed. The results indicated that the level of sRANKL (median value 9.33 microg/L) increased and level of OPG (median value 4.93 microg/L) decreased and the sRANKL/OPG ratio (2.65) increased significantly in experimental group. Compared with control group, the differences in all the corresponding indicators were statistically significant (p<0.05). The sRANKL/OPG ratio was closely related to levels of TRAP-5b (r=0.512, p<0.05) and CTP-I (r=0.481, p<0.05) in MM patients. After all patients in experimental groups were divided into group with bone destruction (n=29) and without bone destruction (n=13), the sRANKL/OPG ratio in the group with bone destruction was 5.13 and much higher than that in group without bone destruction (1.12) (p<0.05). A close correlation between the sRANKL/OPG ratio and degree of bone destruction (r=0.445, p<0.05) was acquired when all MM patients were divided into three groups according to degree of bone destruction, but no difference between the ratio and clinical classification and International Staging System (ISS) in MM patients was found. It is concluded that the sRANKL/OPG ratio in serum of MM patients is significantly elevated, which may be closely related to increase metabolism of OC along with the incidence and degree of bone destruction. In short, the sRANKL/OPG ratio can be used as a reference index for the diagnosis of MBD.
Adult ; Aged ; Bone Diseases ; blood ; diagnosis ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; blood ; diagnosis ; Osteoprotegerin ; blood ; RANK Ligand ; blood

OBJECTIVETo investigate the expression profiles of bone morphogenetic protein and activin membrane-bound inhibitor (BAMBI) during the development of mouse adipose tissue.

METHODSThe total RNA was extracted for real-time PCR for amplification of BAMBI mRNA from the suprascapular brown adipose tissue (BAT) and subcutaneous (inguinal) and visceral (gonadal) white adipose tissue (sWAT and vWAT, respectively) of mice at various embryonic and postnatal stages, as well as from isolated primary preadipocytes during differentiation.

RESULTSIn BAT, BAMBI mRNA levels exhibited a transient increase, peaking at day 0 (D0) and declined thereafter. sWAT and vWAT could be isolated from mice from postnatal D21 onwards, in which BAMBI mRNA levels were the highest and decreased at 8 weeks and 6 months. BAMBI mRNA levels were also significantly reduced in primary preadipocytes isolated from vWAT after induced differentiation. BAMBI mRNA expression level was higher in vWAT than in sWAT and BAT at the same developmental stages.

CONCLUSIONBAMBI is differentially expressed in different adipose tissues and developmental stages, which supports the hypothesis that BAMBI plays a pivotal role in the development of adipose tissues.

Adipocytes ; metabolism ; Adipose Tissue ; metabolism ; Animals ; Bone Morphogenetic Proteins ; metabolism ; Cell Differentiation ; Membrane Proteins ; metabolism ; Mice ; Phosphorylation ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Signal Transduction

This study was purposed to investigate the expression and clonal proliferation of receptor (TCR) Vbeta subfamilies of the T-cells in acute leukemic patients at different disease status (onset, complete remission or relapse) and to analyze the influence of the leukemic cell load on anti-leukemic effect of peripheral T-lymphocytes of the patients. Gene sequences of peripheral TCR Vbeta 24 families from 11 leukemic patients and 3 normal donors were expanded by RT-PCR. Genescan technique was applied to evaluate clonal expression of the TCRVbeta subfamilies, clonal characteristics of the CDR3 from peripheral blood of AML patients at different disease status. The application, clonal proliferation, cellular complexity of T-cells, and the variation of immunotypes of T-cells were compared. The results indicated that the lower and partial distribution of TCR Vbeta subfamily was found in all 11 patients when firstly diagnosed; the expression of TCR Vbeta subfamilies after induction in vitro increased; obvious elevation of TCR Vbeta subfamilies was observed in patients at complete remission although expression level was still lower than normal, whereas the significant descent of TCR Vbeta subfamilies was detected in 4 relapsed patients. Only 1 - 2 clonal proliferation of TCR Vbeta subfamilies existed in 9 out of 11 patients at initial diagnosis which increased at remission. The status of clonal proliferation of Vbeta subfamily T-cells continued regardless of any different disease status in most patients. There was an obvious decrease of CDR3 complexity at initial diagnosis or relapse, while CDR3 complexity would be partially improved at remission. It is concluded that the restrict distribution and expression of TCR Vbeta subfamilies were found in AML patients. Clonal proliferation of T-cells Vbeta subfamily continuously exists regardless of any different disease status in most patients. Some Vbeta subfamilies sustain clonal proliferation at different disease status. Some clonal proliferations of Vbeta subfamilies are associated with the effects of leukemic cells, CDR3 complexity obviously decreases under disease status which can be partially improved at remission.
Adult ; Aged ; Clone Cells ; Complementarity Determining Regions ; genetics ; Female ; Humans ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Receptors, Antigen, T-Cell ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Young Adult

Objective To explore the responses and mechanisms of peripheral primary afferent neurons to adenosine 5'-triphosphate (ATP) and bradykinin (BK) applied separately or in combination by electrophysiological recording and behavioral observation. Methods The experiments were done on samples of acutely isolated rat dorsal root ganglion (DRG) neurons by the whole-cell patch clamp recording technique, to record ATP-activated current (IATP) and the regulating effect of BK on IATP and to observe the global behavior with pain behavioral experiment. Results ATP added after the pretreatment of BK in the majority of detected cells, IATP would be reinforced significantly, the degree of increment depending on the concentration of BK (BK 10-6 -10-4 mol/L), while the EC50 values of the concentration-response curve with and without pretreatment of BK were very close to each other (1.65×10-5 mol/L vs 2.0×10-5 mol/L). In the behavioral experiment, subcutaneously intraplantar injection of BK and ATP separately in hind limbs of rats both induced concentration-dependent pain behavioral (paw lifting) responses, while the duration of hindpaw lifting was prolonged dramatically with the increase in the ATP concentration, when BK (10-6 mol/L) was injected in combination with ATP (10-5, 10-4 and 10-3 mol/L). Conclusion Inflammatory mediators like BK and ATP etc play an important role in the production, transmission and modulation of pain information in peripheral sensory nerve endings. Both electrophysiological and behavioral experiments demonstrate that there is a synergic effect between ATP and BK, which is thought to be non-competitive. BK may reinforce IATP remarkably, and the pain responses induced by the increment in ATP concentration increase with the existence of BK.

Objective To investieate the reproductive health of surviving women whose children had died or were disabled in the carthage in Beiehuan Qiang-autonomous County in China,and to provide the theoretical foundation in the establishment and implementation of nursing intervention.Methods Questionnaires of General Scale and the Reproductive Status Scale were designed according to those women.And survey and analysis was carried out.Results The proportion of these women whose productivity is impaired and lost is 33.2%.The actual difficulties confronted consist of advanced age(>35 years old)with a ratio of 55.14%,menstrual disorrier which is affected greatly by earthquake with a ratio of 23.89%,and complication such as gynecological diseases with a ratio of 34.76%,etc.Conclusions The possibility of reproduction of these surviving women is relatively low.Positive medical supports should be provided to make sure both the physical and mental status of these women are at their best,meanwhile,a high level of long-term health edueation on the premenopausal women and their spouses has a great positive significance on the reconstruction and the stableness in the disaster-hit area.

Objective:To establish a graded method to avoid mean fluorescence intensity(MFI)threshold of HLA Class I antibodies corresponding antigen,and the HLAMatchmaker program has been used to select the minimum mismatch value of donor-patient epitopes.Evaluate the application value of combining both methods in selecting HLA compatible platelets(PTL)for patients with immune platelet transfusion failure(IPTR)in improving platelet the corrected count increment(CCI).Methods:A total 7 807 PLT cross-matching compatible were performed by the solid-phase red cell adherence(SPRCA)method for 51 IPTR patients.The Luminex single antigen flow cytometry was used to detect HLA Class I antibodies in patients,and detected the MFI value for different specificity antigens of HLA Class I antibodies,was graded into strong positive group(MFI＞4 000,level 1),medium positive group(1 000＜MFI 4 000,2),weak positive group(500＜MFI≤1 000,3),and one negative control group(MFI≤500).The results of 7 807 SPRCA their negative/positive reaction wells were enrolled and statistically analyzed in different grades and the four groups,the statistical differences between the four groups were compared.Multiple applications for the select HLA Class I compatible donor events were made for patients in two cases,and HLAMatchmaker program was used to calculate the number of HLA Class I epitopes mismatches between the donors and patients.The donor with the minimum number of epitopes mismatches was selected,while avoiding the corresponding antigens of HLA Class I antibodies in levels 1 and 2,the provision of HLA compatible platelets for IPTR.After the transfusions,the CCI value of the platelet transfusion efficacy evaluation index was calculated,and the clinical evaluation of the transfusion effect was obtained through statistical analysis.Results:There were statistically significant differences in the positive results of SPRCA immunoassay among the strong positive group,medium positive group,and weak positive group of 51 IPTR patients with different specific of HLA-I class antibodies and corresponding antigens(all P＜0.001).The positive results showed a range from high to low,with strong positive group＞medium positive group＞weak positive group.There were a statistical difference among between the strongly positive or moderately positive groups and the negative control group(P＜0.001).There was no statistical difference between the weakly positive group and the negative control group(P＞0.05).The strong positive group was set as the corresponding specific HLA Class I site corresponding antigen grade 1 avoidance threshold,the medium positive group as the grade 2 avoidance thresholds,and the weak positive group as the grade 3 avoidance threshold.In the case of donor platelet shortage,it is not necessary to avoid the weak positive group.Avoiding the strategy of donor antigens and HLAMatchmaker program scores≤7 corresponding to HLA Class I antibodies of levels 1 and 2,with CCI values＞4.5 × 109/L within 24 hours,can obtain effective clinical platelet transfusion conclusions.Conclusion:When selecting HLA Class I compatible donors for IPTR patients,the grading avoids HLA Class I antibodies corresponding to donor antigens,and the donor selection strategy with the minimum scores of HLAMatchmaker program is comprehensively selected.The negative result confirmed by platelet cross-matching experiments has certain practical application value for improving platelet count in IPTR patients.

Objective To evaluate the performance of the main indicators in the blood analysis of an occupational health exam ination agency by Sigma verification of performance chart and guide quality improvement.Methods Collected blood testing laboratory internal quality control (IQC) data in October 2016 and 2016 second national external quality assessment (EQA) results of the blood analysis laboratory in the agency.Regarded the accumulated coefficient of variability as the estimation value of imprecision.Used the percentage difference in EQA results as the estimation value of bias (％) of the laboratory.The total allowable error in health industry standard WS/T406-2012 was adopted as the quality specification (％).Sigma verification of performance chart of the main eight projects in the blood analysis were drawn with professional software.Results The sigma value of WBC reached σ≥6 level,as the first-class performance,and the sigma values of HGB in the 4≤σ ＜5 level,test performance was good.The sigma value of Hct,HCV and MCHC all were in the 3≤σ＜4 level,as the critical level,and the sigma values of RBC,platelet and MCH in the 2≤σ＜3 level,as poor performance.Conclusion Sigma verifica tion of performance chart could reflect the level of the laboratory testing performance on different test items of blood count,promote the quality improvement.

OBJECTIVETo explore epidemiological features and risk factors of severe acute respiratory syndrome (SARS) in Guangdong Province of China, so as to work out effective strategies for its better control.

METHODSA total of 1 511 clinically confirmed SARS cases in Guangdong Province of China from November 16, 2002 to Jun 15, 2003 were retrospectively analyzed.

RESULTSThe first SARS case was identified in Foshan municipality on November 16, 2002, followed by 1 511 clinically confirmed cases (including 58 deaths) up to May 15, 2003. Of all cases, health care workers and community family cluster cases accounted for 19.38% and 12.04%. 65.86% SARS patients aged 20 - 49 years, and increased incidence was positively related to their ages. 95.97% cases lived in the following five cities around Pearl Delta Area: Foshan, Guangzhou, Shenzhen, Zhongshan, and Jiangmen. Eleven early reported cases in the communities took animal-related positions. Face-to-face contacts with infected droplets were the main transmission route. An epidemic peak occurred during January 28 to February 26, and those cases accounted for 50.69% of total. Incidence, mortality, and case fatality of SARS were 1.77/100,000, 0.07/100,000, and 3.84% respectively. The mean incubation period was 4.5 days.

CONCLUSIONThe most effective way to control SARS is to break the chain of transmission from infected to healthy persons-early identification, prompt and effective isolation, and vigorous close contact tracing. Hospital infections among health care workers is critical. Several observations support the hypothesis of an animal origin for the disease.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; epidemiology ; Disease Outbreaks ; Female ; Follow-Up Studies ; Humans ; Incidence ; Infant ; Infectious Disease Transmission, Patient-to-Professional ; Male ; Middle Aged ; Retrospective Studies ; Severe Acute Respiratory Syndrome ; epidemiology ; prevention & control ; transmission