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 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

OBJECTIVETo investigate quinalizarin-induced apoptosis in gastric cancer cells in vitro and explore the molecular mechanisms.

METHODSMTT assay was used to determine the cytotoxic effects of quinalizarin on human gastric cancer AGS, MKN-28 and MKN-45 cells. Annexin V-FITC/PI staining and flow cytometry were used to assess quinalizarin-induced apoptosis in AGS cells and its effect on intracellular ROS levels; the expression levels of apoptotic proteins in the cells were determined with Western blotting.

RESULTSQuinalizarin dose-dependently reduced the cell viabilities of the 3 gastric cancer cells (P<0.05). The ICvalues of quinalizarin in AGS, MKN-28 and MKN-45 cells were 7.07 µmol/L, 22.55 µmol/L and 14.18 µmol/L, respectively. Quinalizarin time-dependently induced apoptosis of AGS cells and potentiated the generation of intracellular reactive oxygen species (ROS) levels. Pretreatment with NAC, a scavenger of ROS, inhibited quinalizarin-induced apoptosis (P<0.001). Western blotting results showed that quinalizarin also up-regulated the expression levels of the apoptotic proteins including p-p38, p-JNK, Bad, cleaved caspase-3, and cleaved PARP-1 (P<0.05), and down-regulated the expression of the anti-apoptotic proteins p-Akt, p-ERK, and Bcl-2 (P<0.05).

CONCLUSIONQuinalizarin inhibits the proliferation and induces apoptosis in gastric cancer cells in vitro through regulating intracellular ROS levels via the MAPK and Akt signaling pathways.

BACKGROUNDA multi-center large scale study is needed to confirm the efficacy and safety of domestic peritoneal dialysis (PD) solutions. Some researchers believe that 6 L/d is enough for adequate dialysis, but there is no multi-center prospective study on Chinese population to confirm this. In this study, we evaluated the efficacy and safety of domestic PD solution (Changfu) and its difference between 6 L and 8 L dosage.

METHODSAdult PD patients who had taken PD therapy for at least one month were selected and divided into four groups according to two dialysis solution brands and two dialysis dosages, i.e., 6 L dose with Changfu dialysis solution, 6 L dose with Baxter dialysis solution, 8 L dose with Changfu dialysis solution, and 8 L dose with Baxter dialysis solution. After 48 weeks, the changes of primary and secondary efficacy indices were compared between different types and different dosages. We also analyzed the changes of safety indices.

RESULTSChanges of Kt/V from baseline to 48 weeks between Changfu and Baxter showed no statistical differences; so did those of creatinine clearance rate (Ccr). Normalized protein catabolic rate (nPCR) from baseline to 48 weeks between Changfu and Baxter showed no statistical differences; so did those of net ultrafiltration volume (nUF) and estimated glomerular filtration rate (eGFR). Changes of nPCR from baseline to 48 weeks between 6 L and 8 L showed no statistical differences; so did those of nUF and eGFR. The decline of Kt/V from baseline to 48 weeks in 6 L group was more than that in 8 L group. Change of Ccr was similar. During the 48-week period, the mean Kt/V was above 1.7/w, and mean Ccr was above 50 L×1.73 m(-2)×w(-1). More adverse events were found in Changfu group before Changfu Corporation commenced technology optimization, and the statistical differences disappeared after that.

CONCLUSIONSThe domestic PD solution (Changfu) was proven to be as effective as Baxter dialysis solution. During 48-week period, a dosage of 6 L/d was enough for these patients to reach adequate PD. Clinical study promotes technological optimization, further helps to improve the safety indices of the medical products.

Adolescent ; Adult ; Aged ; Dialysis Solutions ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Peritoneal Dialysis ; methods ; Young Adult