OBJECTIVETo investigate the expression of B lymphocyte stimulator (BlyS) and its receptors in multiple myeloma (MM) cells, and to explore the relationship between BLyS and the development of human multiple myeloma.

METHODSFlow cytometry, RT-PCR and Western blot were used to examine the expression of BLyS and its receptors in MM (KM3 and CZ1) cells. Fluorescence immunocytochemical method and confocal laser scanning technique were applied to the localization of BLyS in KM3 cell. WST proliferation assay was used to examine the effect of BLyS on MM cells growth and survival. Linear correlation analysis was used to detect LDH and beta 2-microglobulin (beta2M) levels with BLyS protein and mRNA expressions in MM patients.

RESULTS(1) BLyS and its receptors were expressed in MM cells. (2) BLyS protein was localized on the KM3 plasma membrane. (3) BLyS promoted survival and proliferation of MM cells. (4) MM patients had significantly higher expression levels of BLyS [77.42% (24/31)] BLyS mRNA [93.55% (29/31)], which were significantly correlated with the levels of LDH and beta 2-microglobulin (beta2M).

CONCLUSIONBLyS and its receptors in MM cell lines and MM patient bone marrow might have a potential role in the growth and survival of malignant plasma cells.

Aged ; B-Cell Activating Factor ; genetics ; metabolism ; B-Cell Activation Factor Receptor ; genetics ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; metabolism ; pathology ; RNA, Messenger ; genetics ; beta 2-Microglobulin ; metabolism

OBJECTIVETo investigate the relationship of time-concentration of bisphenol A (BPA) in male Sprague-Dawley (SD) rats after single oral BPA administration.

METHODSA total of 66 specific pathogen free (SPF) SD male rats were divided into 10 experimental groups and control group (n = 6). The experimental group rats were treated with BPA of 300 mg/kg by oral gavage and blood samples were taken from one group at 0.5, 1, 2, 4, 6, 12, 24, 36, 60, 84 h time point after oral administration, respectively. The serum BPA concentration was determined by fluorescence-high performance liquid chromatography (FL-HPLC) analysis.

RESULTSAfter oral administration of 300 mg/kg, the total serum BPA concentration of 17.13 microg/ml was the highest in rats at 1 h, then decreased, but it increased to 15.18 microg/ml again at 24 h, then gradually decreased to 0.51 microg/ml at 84 h. The level of serum free BPA was lower than that of total serum BPA after oral administration, the serum free BPA was 0.57 microg/ml at 0.5 h after oral administration. The serum free BPA level decreased to 0.06 microg/ml at 1 h, 0.03 microg/ml at 4 h, 0.01 microg/ml at 36 h after oral administration. The free BPA was only 4.15% (0.57/13.73) in total BPA in serum at 0.5 h after oral administration of 300 mg/kg BPA.

CONCLUSIONThese results suggested that conjugated BPA was the main metabolite of BPA in rat serum after single oral administration. Enterohepatic circulation of BPA glucuronide in rats may results in two peak levels of total BPA in serum.

Animals ; Benzhydryl Compounds ; Male ; Phenols ; blood ; pharmacokinetics ; toxicity ; Rats ; Rats, Sprague-Dawley ; Serum ; metabolism ; Time Factors

OBJECTIVEB cell multiplication plays a key role in infections mononucleosis. The present study was designed to detect the expression of B-lymphocyte stimulator (BLyS) mRNA in peripheral blood using real-time fluorescence quantitative polymerase chain reaction (RFQ-PCR) in children with infectious mononucleosis in order to explore the role of BLys in this disorder.

METHODSSpecific primers and TaqMan probes of BLyS were designed, and fluorescence of the PCR products were detected continuously during amplification. According to the standard curves created by plasmid DNA, the expression level of target genes in clinical samples were calculated using Stata Software version 8.0, and the results were presented as the ratio of copies of target gene mRNA to beta2 microglobulin (beta2M) mRNA copies. BLyS mRNA expression in peripheral blood was measured by RFQ-PCR in 18 children with infectious mononucleosis and the results were compared with those measured in 15 healthy controls.

RESULTSThe range of target gene mRNA detected by REQ-PCR was from 109 ng/L to 101 ng/L. The coefficient of variation for intra-experimental and inter-experimental reproducibility ranged from 1.88% to 5.89% and 6.32% to 12.34%, respectively. BLyS mRNA expression in peripheral blood in children with infectious mononucleosis were significantly higher than that in controls (1.65+/-0.10 vs 0.56+/-0.08; P < 0.01).

CONCLUSIONSRFQ-PCR has a high sensitivity and reproducibility for the measurement of BLyS mRNA expression. BLyS may be involved in the development of infectious mononucleosis.

B-Cell Activating Factor ; genetics ; Child ; Child, Preschool ; Female ; Fluorescence ; Humans ; Infectious Mononucleosis ; etiology ; metabolism ; Male ; Polymerase Chain Reaction ; methods ; RNA, Messenger ; analysis

OBJECTIVETo explore the regulating effect of miR-202 on B cell-activating factor, and check whether the regulation influences the growth of multiple myeloma cells.

METHODSThe potential binding sites of BAFF for miR-202 were predicted using bioinformatics software. Luciferase reporter gene analysis was used to evaluate the regulatory effect of miR-202 on BAFF. Human multiple myeloma U266 cells were transfected with has-miR-202-mimics, has-miR-202-inhibitor, siBAFF and their negative controls, respectively. After above treatments, BAFF mRNA and protein levels were detected by real-time PCR and Western blot analysis, and the proliferation and apoptosis in the multiple myeloma (MM) cells were examined by WST-1 and annexin V-FLUOS assay, respectively.

RESULTSThe BAFF mRNA expression levels in the untransfected group, has-miR-202-3P-mimics transfected group, has-miR-202-3P-inhibitor transfected group and siBAFF transfected group were 1.040 ± 0.057, 0.573 ± 0.073, 1.205 ± 0.097 and 0.368 ± 0.052, respectively. BAFF mRNA expressions in U266 cells transfected with has-miR-202-3P-mimics and siBAFF were significantly decreased compared with that in the untransfected group (P < 0.05). The BAFF protein expression level of each group was consistent with the mRNA assay result. The absorbance value in 450 nm of the untransfected group, has-miR-202-3P-mimics transfected group, has-miR-202-3P-inhibitor transfected group and siBAFF transfected group were 1.063 ± 0.052, 0.714 ± 0.045, 0.936 ± 0.066 and 0.764 ± 0.053, respectively. In comparison with the untransfected group, the absorbance value at 450 nm of has-miR-202-3P-mimics and siBAFF transfected groups was significantly reduced (P < 0.05). The cell apoptosis rates of untransfected group, has-miR-202-3P-mimics transfected group, has-miR-202-3P-inhibitor transfected group and siBAFF transfected group were 26.2%, 49.6%, 21.1% and 30.7%, respectively. Therefore, the cell apoptosis rate of has-miR-202-3P-mimics transfected group was significantly increased than that of the untransfected group (P < 0.05). p-JNK protein expression level was decreased in the has-miR-202-3P-mimics transfected cells.

CONCLUSIONSMiR-202 can inhibit the proliferation and induce apoptosis in MM cells via regulating BAFF. JNK/SAPK signaling pathway is involved in the regulation of BAFF by miR-202.

Apoptosis ; B-Cell Activating Factor ; genetics ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; HEK293 Cells ; Humans ; Luciferases ; metabolism ; MAP Kinase Signaling System ; MicroRNAs ; genetics ; metabolism ; Multiple Myeloma ; metabolism ; pathology ; Plasmids ; RNA, Messenger ; metabolism ; Transfection

OBJECTIVETo investigate the effects of a proliferation-inducing ligand (APRlL) on colorectal cancer (CRC) cell growth and migration, and to observe the role of APRIL in CRC biological behavior.

METHODSThe siRNA plasmid vector targeting APRIL gene (APRIL-siRNA) was transfected into human colorectal cancer SW480 cells and recombinant human APRIL (rhAPRIL) was used to stimulate human colorectal cancer HCT-116 cells. Cell proliferation activity was analyzed using cell counting kit-8 (CCK-8), cell cycle was detected by flow cytometry, and the protein expression of cyclin D1, p21 and Bcl-2 was detected by Western blot analysis. Tumor cell migration and invasion were measured by Transwell chambers. RT-PCR was applied to examine the mRNA expression level of MMP-2 and MMP-9. APRIL-siRNA was used to transfect directly SW480 cells, which were injected subcutaneously into nude mice, then the tumor growth and metastasis were observed.

RESULTSCell proliferation ability of APRIL-siRNA-transfected SW480 cells was drastically repressed, and the percentage of G0/G1 phase cells was significantly increased (t = 4.12, P < 0.05), accompanied with depressed cyclin D1, Bcl-2 expression and elevated p21 expression. Cell proliferation ability of rhAPRIL-stimulated HCT-116 cells was promoted with a decreased G0/G1 phase ratio (t = 3.31, P < 0.05). cyclin D1 and Bcl-2 protein expression was up-regulated while p21 was down-regulated by rhAPRIL stimulation. Metastatic and invasive capacities of APRIL-siRNA-transfected SW480 cells were significantly inhibited compared with their respective controls (both P < 0.05), accompanied with the deregulated MMP-2 and MMP-9 mRNA expression. Metastatic and invasive capacities of rhAPRIL-stimulated HCT-116 cells were promoted with up-regulated MMP-2 and MMP-9 mRNA expression(both P < 0.05). Tumor growth in the group transfected with APRIL-siRNA appeared to be slower than that in the control groups and the expression of MMP-2, MMP-9 in tumor tissues was depressed in the APRIL-siRNA group.

CONCLUSIONSAPRIL facilitates tumor growth and metastasis, and is associated with carcinogenesis and prognosis. Our findings suggest that APRIL might be used as a novel target for the intervention and therapy of colorectal cancer.

Animals ; Cell Cycle ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Colorectal Neoplasms ; metabolism ; pathology ; Cyclin D1 ; metabolism ; Female ; Genetic Vectors ; HCT116 Cells ; HT29 Cells ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Mice, Nude ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasm Transplantation ; Plasmids ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Proto-Oncogene Proteins p21(ras) ; metabolism ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Transfection ; Tumor Burden ; Tumor Necrosis Factor Ligand Superfamily Member 13 ; genetics ; metabolism

OBJECTIVETo evaluate the efficacy of Bufei Yishen Granule BFYSG) combined with Shufei Tie acupoint sticking therapy on quality of life of patients with stable chronic obstructive pulmonary disease (COPD).

METHODSA multi-center, double-blinded, double-dummy and randomized controlled method was adopted in this trial. A total of 244 patients were randomly assigned to a trial group and a control group according to the random number, each with 122 patients; treatment allocation occurred when the participants met the inclusion criteria and signed the informed consent form. In the trial group, patients were treated with BFYSG combined with "Shufei Tie" acupoint sticking therapy and sustained-release theophylline dummy, and in the control group patients were treated with oral sustained-release theophylline and BFYSG dummy combined with "Shufei Tie" acupoint sticking therapy dummy. The therapeutic course for two groups was 4 months and the follow-up was 6 months. The frequency and duration of acute exacerbation calculated by adding up each frequency and duration of acute exacerbation in treatment and follow-up time respectively, the quality of life measured by the World Health Organization Quality of Life (WHOQOL)-BREF scale and adult COPD quality of life (COPD-QOL) scale were observed.

RESULTSAmong the 244 enrolled patients, 234 were screened for full analysis set (FAS); 221 were screened for per-protocol analysis set (PPS). After 4-month treatment and 6-month follow-up there were differences between the trial group and the control group in frequency of acute exacerbation (FAS: P=0.013; PPS: P=0.046); duration of acute exacerbation (FAS: P=0.005; PPS: P=0.006); scores of physiological, psychological and environment aspects of the WHOQOL-BREF questionnaire (FAS: P=0.002, P=0.006, P=0.000; PPS: P=0.00, P=0.001, P=0.000); scores of daily living ability, social activity, depression symptoms aspects of the COPD-QOL questionnaire (FAS: P=0.000, P=0.000, P=0.006; PPS: P=0.002, P=0.001, P=0.001).

CONCLUSIONBFYSG combined with acupoint sticking therapy could improve the quality of life of patients with stable COPD.

Acupuncture Points ; Adult ; Aged ; Anxiety ; complications ; psychology ; Combined Modality Therapy ; Depression ; complications ; psychology ; Disease Progression ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Pulmonary Disease, Chronic Obstructive ; complications ; drug therapy ; psychology ; Quality of Life ; Surveys and Questionnaires ; Treatment Outcome

AIMTo determine the mechanisms of glucocorticoids in inhibiting advanced prostate cancer growth.

METHODSThe cell proliferation and cell cycle of prostate cancer DU145 cells following dexamethasone treatment were determined by proliferation assay and fluorescence-activated cell sorter. Western blot analysis was carried out to evaluate the effects of dexamethasone on phosphorylation of extracellular signal-regulated kinase (ERK)1/2 and expression of cyclin D1 in DU145 cells with or without glucocorticoid receptor (GR) antagonist RU486. Reverse transcription-polymerase chain reaction verified the expression of GR mRNA in DU145 cells.

RESULTSDexamethasone significantly inhibited DU145 cell proliferation at the G(0)/G(1) phase. Western blot analysis showed a dramatic reduction of ERK1/2 activity and cyclin D1 expression in dexamethasone-treated cells. The decreased phosphorylation of ERK1/2 in dexamethasone-treated cells was attenuated by GR blockade. Additionally, the effects of dexamethasone in inhibiting cyclin D1 expression were altered by GR blockade.

CONCLUSIONDexamethasone suppresses DU145 cell proliferation and cell cycle, and the underlying mechanisms are through the inhibition of phosphorylation of ERK1/2 and cyclin D1 expression. The inhibition of ERK1/2 phosphorylation and cyclin D1 expression is attenuated by GR blockade, suggesting that GR regulates ERK1/2 and cyclin D1 pathways. These observations suggest that dexamethasone has a potential clinical application in prostate cancer therapy.

Antineoplastic Agents, Hormonal ; pharmacology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin D1 ; drug effects ; metabolism ; Dexamethasone ; pharmacology ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Male ; Mitogen-Activated Protein Kinase 1 ; drug effects ; metabolism ; Mitogen-Activated Protein Kinase 3 ; drug effects ; metabolism ; Prostatic Neoplasms ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Receptors, Glucocorticoid ; metabolism ; Signal Transduction ; drug effects

OBJECTIVETo evaluate the feasibility and clinical effect of transperitoneal laparoscopic enucleation of renal angiomyolipoma (RAML) without obstruction of renal pedicle.

METHODSTen patients with renal angioleiomyoma (tumor diameter < 4 cm) were operated by transperitoneal laparoscopy without obstruction of renal pedicle. The operating time, blood loss, hospital stay after operation, intraoperative and postoperative complications and the operative effect were observed.

RESULTSAll the 10 patients underwent the operation successfully. The average operating time was 90 min, average blood loss was 80 ml, the average hospital stay after operation was 7 d. No intraoperative or postoperative complications occurred. Follow-up period was 3-19 months and no tumor metastasized or occurred again.

CONCLUSIONThis mininvasive procedure is a more precise and complete method than before, which can minimize the blood loss and make patients recover quickly, so it is well worth clinical applying.

Adult ; Angiomyolipoma ; surgery ; Female ; Follow-Up Studies ; Humans ; Kidney Neoplasms ; surgery ; Laparoscopy ; Male ; Middle Aged ; Nephrectomy ; methods ; Treatment Outcome

This study was purposed to investigate the effects of compound Zhe-Bei granule (CZBG) combined with doxorubicin on expressions of P-gp, MRP, LRP in K562/A02 cell xenografts. Tumor xenograft model were established by injecting the multidrug resistant cell line K562/A02 in the axillary flank of BALB/c-nu-nu mice. CZBG-intragastric administration and doxorubicin-intraperitoneal injection in combination were given to the BALB/c-nu nude mice. The tumor xenografts were made into slice after the dissection, and the expression of P-gp, MRP, LRP in K562/A02 tumor xenografts in mice were investigated by immunohistochemical technique. The integral optical density (IOD) of P-gp, MRP, LRP in K562/A02 tumor xenografts were measured by Image Pro Plus 6.0. The results showed that as compared with the doxorubicin alone, the combination of the doxorubicin and CZBG with high, middle and low dosage could decrease IOD of P-gp, MRP in K562/A02 tumor xenografts with statistical significance (p < 0.05). The LRP expression in K562/A02 tumor xenografts was not observed in five groups. It is concluded that the combination of CZBG with doxorubicin decreases the expressions of P-gp, MRP in K562/A02 tumor xenografts of mice.
Animals ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gene Expression Regulation, Leukemic ; drug effects ; Humans ; K562 Cells ; Membrane Transport Proteins ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Xenograft Model Antitumor Assays

OBJECTIVE: To noninvasively assess the neurodegenerative changes in the brain of patients with Niemann-Pick type C (NPC) disease by measuring the lesion tissue with the iterative decomposition of water and fat with echo asymmetry and least square estimation-iron quantification (IDEAL-IQ). MATERIALS AND METHODS: Routine brain MRI, IDEAL-IQ and 1H-proton magnetic resonance spectroscopy (1H-MRS, served as control) were performed on 12 patients with type C Niemann-Pick disease (4 males and 8 females; age range, 15–61 years; mean age, 36 years) and 20 healthy subjects (10 males and 10 females; age range, 20–65 years; mean age, 38 years). The regions with lesion and the normal appearing regions (NARs) of patients were measured and analyzed based on the fat/water signal intensity on IDEAL-IQ and the lipid peak on 1H-MRS. RESULTS: Niemann-Pick type C patients showed a higher fat/water signal intensity ratio with IDEAL-IQ on T2 hyperintensity lesions and NARs (3.7–4.9%, p < 0.05 and 1.8–3.0%, p < 0.05, respectively), as compared to healthy controls (HCs) (1.2–2.3%). After treatment, the fat/water signal intensity ratio decreased (2.2–3.4%), but remained higher than in the HCs (p < 0.05). The results of the 1H-MRS measurements showed increased lipid peaks in the same lesion regions, and the micro-lipid storage disorder of NARs in NPC patients was detectable by IDEAL-IQ instead of 1H-MRS. CONCLUSION: The findings of this study suggested that IDEAL-IQ may be useful as a noninvasive and objective method in the evaluation of patients with NPC; additionally, IDEAL-IQ can be used to quantitatively measure the brain parenchymal adipose content and monitor patient follow-up after treatment of NPC.
Brain ; Female ; Follow-Up Studies ; Healthy Volunteers ; Humans ; Magnetic Resonance Imaging ; Magnetic Resonance Spectroscopy ; Male ; Methods ; Niemann-Pick Diseases ; Proton Magnetic Resonance Spectroscopy ; Water