Objective To observe the expression of STEAP4 gene(a novel obesity-related gene) during the period of human preadipocyte differentiation and to explore the relationship between the STEAP4 gene expression and adipocytes differentiation,adipogenesis.Methods Human preadipocytes were cultured and differentiated into the matured adipocytes in vitro.Adipocytes morphology and lipid accumulation were observed during this process.Total RNA was extracted from adipocytes at various time points (preadipocyte,Day 0,Day 4,Day 6,Day 8,Day 11,Day 14,and Day 17) and the level of STEAP4 mRNA expression was measured by fluorescent real-time quantitative reverse transcriptase-polyme-rase chain reaction(RT-PCR).Results The level of STEAP4 mRNA expression remained high in preadipocytes.In the presence of differentiation medium (Day 4),there was a transient upregulation in the expression of STEAP4 gene.After that,with the human preadipocytes being differentiated into matured adipocytes,the expression of STEAP4 mRNA was downregulated and reached the lowest level in fully differentiated adipocytes.There was a significant difference between any 2 detected phases in the level of STEAP4 mRNA expression (Pa

OBJECTIVETo observe the effects of Dioscornin Tablet (DT) containing serum on nuclear factor of kappa B (NF-kappaB) p65, signal transducer and activator of transcription 3 (STAT3), and vascular endothelial growth factor (VEGF) mRNA expressions in rats' synovial cell strain 364 (RSC-364) induced by interleukin-17 (IL-17) and tumor necrosis factor-alpha (TNF-alpha), and to investigate the underlying mechanisms for DT to inhibit angiogenesis of rheumatoid arthritis (RA).

METHODSIn this experiment, the vehicle control group, the cell model group, the DT containing serum group, and the positive control group (Tripterygium containing serum) were set up. The DT containing serum and the Tripterygium containing serum were prepared. The RA cell model was established by IL-17 combined TNF-alpha induced injury in RSC-364. The RA cells were intervened by DT containing serum and Tripterygium containing serum respectively. The DNA binding activity of NF-kappaB p65 was detected using TransAM NF-kappaB p65. The expression of STAT3 was observed using Western blot. The VEGF mRNA expressions were detected by real-time quantitative PCR.

RESULTSCompared with the vehicle control group, the NF-kappaB p65 activity, the expressions of STAT3 and VEGF mRNA increased significantly in RSC-364 induced by IL-17 +TNF-alpha (P < 0.01, P < 0.05). Compared with the model group, the NF-kappaB p65 activity, the expressions of STAT3 and VEGF mRNA decreased significantly in the DT containing serum group and the positive control group (P < 0.01, P < 0.05). There was no statistical difference between the two groups (P > 0.05).

CONCLUSIONDT inhibited the VEGF mRNA expression through inhibiting the NF-kappaB p65 activity and the STAT3 protein expression in the Janus kinase (JAK)-signal transducer and activating transcription factor pathway, thus inhibiting the angiogenesis of RA.

Animals ; Arthritis, Rheumatoid ; pathology ; Cells, Cultured ; Diosgenin ; analogs & derivatives ; pharmacology ; Interleukin-17 ; adverse effects ; Male ; Neovascularization, Pathologic ; pathology ; RNA, Messenger ; pharmacology ; Rats ; Rats, Wistar ; STAT3 Transcription Factor ; metabolism ; Serum ; Signal Transduction ; Synovial Membrane ; cytology ; drug effects ; metabolism ; Transcription Factor RelA ; metabolism ; Tumor Necrosis Factor-alpha ; adverse effects ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo observe the primary prevention role of Wuling Capsule (WC) on poststroke depression (PSD) patients.

METHODSAcute stroke patients were recruited and randomized into 2 groups by stratification, 55 in each group. All patients received same routine treatment of cardiovascular diseases. Patients in the experimental group additionally took WC (0.33 g each pill), 3 pills per day, three times per day; while those in the control group additionally took placebos, 3 pills per day, three times per day. Two weeks consisted of one therapeutic course. The diagnosis of PSD was performed once every other week. Those in accordance with PSD diagnosis discontinued any drug therapy. Those not in accordance with PSD diagnosis continued the drug therapy for 1-12 therapeutic course(s) (in total of 6 months). If they were still not in accordance with PSD diagnosis, then they discontinued the drug therapy. The morbidity of PSD, the average time of depression occurrence, Hamilton depression rating scale (HAMD) score, and adverse reactions were observed.

RESULTSThe 1-, 3-, and 6-month morbidity of PSD was 8%, 16%, and 34% in the experimental group, while they were 19.6%, 29.4%, and 54.9% in the control group. The occurrence rate was lower in the experimental group than in the control group. Besides, there was statistical difference in the 6-month occurrence rate between the two groups (chi2 = 4.465, P < 0.05). The average time of PSD occurrence was longer in the experimental group than in the control group (14.96 +/- 8.31 weeks vs. 9.36 +/- 6.06 weeks; t=6.762, P < 0.05). The HAMD score at the PSD occurrence was 11.96 +/- 2.14 in the experimental group, lower than that of the control group (14.57 +/- 4.24), showing statistical difference (t=5.641, P < 0.05).

CONCLUSIONWC was superior to the placebos in lowering the incidence of PSD, delaying the occurrence time of PSD, attenuating the depression degree of PSD, and had certain preventive effect on the incidence of PSD.

Aged ; Capsules ; Depression ; etiology ; prevention & control ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Primary Prevention ; Stroke ; complications

Developing AIDS education course in college and university is urgent and important,especially for medical college students on account of the characteristic of their specialty.This paper is on the basis of practice and consideration for AIDS education course among medical college students.

Objective To approach the clinical presentation,treatment and diagnosis of primary seminal vesicle carcinoma.Methods The records of 4 patients who diagnosed seminal vesicle carcinoma were retrospectively reviewed,including the symptoms signs and examination results as well as operation program,postoperative therapy.Considered to the literature reports.Bilateral seminal vesicles,bladder, prostate and rectum were totally removed in one case.Seminal vosiculectomy and partial cystoprostotectomy were performed in 2 cases,and the another one,bilateral lower ureterectomy and ileum substitute bladder was be done.Results Followed up for 3 months to 5 years,and no recurrence had been observed so far, one died of colon cancer after 2 years.Conclusions Early symptoms of primary seminal vesicle carcinoma are unobvious,so that early diagnosis of seminal vesicle carcinoma is difficult and the misdiagnosis is so usually.Thus,accurate recognition is important for early diagnosis.Radical surgery appears to offer the best chance and different approaches can be selected according to tumor stage and invasive condition of the circumambient organ.Comprehensive treatment like hormonal therapy,along with the 5-fluorouracil,paclitaxel,and oxaliplatin chemotherapy regimen appears to be effective against adenocarcinoma.

OBJECTIVETo investigate the inhibitory effect and mechanism against the growth of human gastric carcinoma cell line HS-746T by the combination of the survivin antisense oligonucleotide (ASODN) and P53 gene and its mechanism.

METHODSGastric carcinoma cell line HS-746T was treated by P53 gene and survivin antisense oligonucleotide was designed. There were four regimen groups treated by different agents:ASODN alone, P53 gene alone and the combination of ASODN and P53 gene, blank control. Cell proliferative ability and cell growth were determined by cells counting and MTT. The expression of survivin mRNA and protein was detected by RT-PCR and Western blot respectively. Cell apoptotic index was detected by TUNEL.

RESULTSASODN alone, P53 alone and the combination of ASODN and P53 could inhibit not only the growth of gastric carcinoma cell, but also down-regulate the survivin mRNA and protein expression. The inhibitory effect was stronger, and the apoptosis index was higher in the combined transfection group than those in the other two single transfection groups.

CONCLUSIONThe combination of survivin ASODN and P53 gene is more efficient to inhibit cell growth and induce apoptosis than that of agent alone.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; genetics ; Oligonucleotides, Antisense ; genetics ; pharmacology ; Stomach Neoplasms ; genetics ; pathology ; Transfection ; Tumor Suppressor Protein p53 ; genetics

This study examined the effect of Notch-1 signaling on malignant behaviors of breast cancer cells by regulating breast cancer stem cells (BCSCs). BCSCs were enriched by using serum-free medium and knocked out of Notch-1 by using a lentiviral vector. Real-time polymerase chain reaction (RT-PCR) and Western blotting were used to detect the Notch-1 expression levels in breast cancer cell lines and BCSCs, and flow cytometry to detect the proportion of BCSCs in BCSC spheres. The BCSC self-renewal, migration, invasion, and tumorigenicity were examined by the tumor microsphere-forming assay and transwell assay and after xenotransplantation. The results showed that the Notch-1 silencing reduced the number of BCSC spheres, the proportion of BCSCs, and the number of cells penetrating through the transwell membrane. It also decreased the size of tumors that were implanted in the nude mice. These results suggest that Notch-1 signaling is intimately linked to the behaviors of BCSCs. Blocking Notch-1 signaling can inhibit the malignant behaviors of BCSCs, which may provide a promising therapeutical approach for breast cancer.

OBJECTIVEHuman STEAP4, a novel obesity-related gene, is associated with insulin sensitivity regulation in human adipocytes. This study aimed to explore the regulative role of TNFalpha on STEAP4 gene in matured human adipocytes.

METHODSHuman preadipocytes were cultured and differentiated into matured adipocytes in vitro. Fully differentiated adipocytes (Day 17) were treated with different concentrations of TNFalpha (0, 5, 10, 25 and 50 ng/mL) for 24 hrs. Total RNA and protein were extracted from the adipocytes. Levels of STEAP4 mRNA and protein expression were determined by real-time quantitative RT-PCR and Western blot respectively.

RESULTSDifferent concentrations (5, 10, 25 and 50 ng/mL) of TNFalpha treatment for 24 hrs resulted in a significant increase in the STEAP4 mRNA expression of human matured adipocytes.The maximal effect was seen in the 50 ng/mL of TNFalpha treatment group. In parallel, STEAP4 protein synthesis in matured adipocytes increased in response to TNFalpha treatment of different concentrations (5, 10, 25 and 50 ng/mL) for 24 hrs. The maximal up-regulated effect was seen in the 25 ng/mL of TNFalpha treatment group.

CONCLUSIONSTNFalpha can up-regulate STEAP4 mRNA expression in human matured adipocytes.

Adipocytes ; metabolism ; Cells, Cultured ; Dose-Response Relationship, Drug ; Gene Expression Regulation ; drug effects ; Humans ; Membrane Proteins ; genetics ; Oxidoreductases ; genetics ; Recombinant Proteins ; pharmacology ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo screen the differentially expressed genes in human renal clear-cell carcinoma (RCC) cells using suppression subtractive hybridization (SSH), and to explore their biological function and underlying mechanism in RCC cells.

METHODSTotal RNAs were extracted from human renal clear-cell carcinoma cell line RLC-310 and human normal renal cell line HK-2 cells, and SSH technology was used to construct a RCC cell library of differential expression genes and to screen the most differentially expressed genes. RNA interference vector was constructed to silence the expression of the differentially expressed gene SIPL1 in human renal cell lines RLC-310 and GRC-1. Proliferation index was estimated by cell counting, MTT and tumor xenograft assay. Cell cycle analysis was performed using fluorescence activated cell sorting. Drug resistance potential to adriamycin was assessed by MTT.

RESULTSA subtractive cDNA library of highly expressed genes in the RCC cells was constructed and 12 differentially expressed genes were screened from the subtractive library, in which SIPL1 was the most differently expressed gene in the RCC cell line. SIPL1 overexpression in the RCC cells and clinical samples was confirmed by RT-PCR and Western blot analyses. The shRNA expression plasmid targeting to SIPL1 gene was constructed and transfected into RLC-310 and GRC-1 cells, resulting in downregulation of SIPL1. SIPL1 knockdown inhibited the cell proliferation (P < 0.05) and tumorgenesis. The tumor weights formed by RLC-310 cells transfected with SIPL1 shRNA was (0.22 ± 0.07)g and that of negative control vector was (0.85 ± 0.06)g. The tumor weight formed by GRC-1 cells was (0.32 ± 0.07)g and that of control vectors was (1.21 ± 0.11)g (P < 0.05). SIPL1 shRNA-transfected RLC-310 cells showed that more cells were arrested at G0/G1 phase [(71.13 ± 4.58)%] than that in the negative control RLC-310 cells [(53.27 ± 3.34)%, P < 0.05]. The proportion of G0/G1 phase in the SIPL1 shRNA transfected GRC-1 cells was (73.83 ± 3.97)%, significantly higher than that of (59.33 ± 3.03)% in the negative control GRC-1 cells (P < 0.05), and enhanced their sensitivity to adriamycin (P < 0.05). Silence of SIPL1 caused inactivation of AKT signaling and up-regulated expression of P27(Kip1) and P21(Cip1) proteins.

CONCLUSIONSA differentially expressed gene SIPL1 in the renal clear-cell carcinoma is successfully screened using SSH technology. SIPL1 functions as an oncogene in RCC, and may become a novel molecular target for RCC diagnosis and therapy.

Adenocarcinoma ; metabolism ; pathology ; Adult ; Aged ; Animals ; Antibiotics, Antineoplastic ; pharmacology ; Carcinoma, Renal Cell ; metabolism ; pathology ; Carrier Proteins ; genetics ; metabolism ; Cell Cycle ; Cell Line ; Cell Line, Tumor ; Cell Proliferation ; Cyclin-Dependent Kinase Inhibitor p21 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; Female ; Humans ; Kidney ; cytology ; Kidney Neoplasms ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Middle Aged ; Neoplasm Transplantation ; Nucleic Acid Hybridization ; Proto-Oncogene Proteins c-akt ; metabolism ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Transfection ; Tumor Burden

OBJECTIVETo explore the association between the genetic polymorphisms of mannose-binding protein (MBP) alleles and susceptibility to pulmonary tuberculosis.

METHODS125 pulmonary tuberculosis cases and 198 healthy controls were collected. A case-control study was conducted. Three structural gene mutations in exon 1 of MBP gene (codon 52, codon 54 and codon 57) were studied. Polymerase chain reaction with sequence-specific primers (PCR-SSP) was carried out in the polymorphism in MBP alleles. Information on related risk factors of tuberculosis was collected, using a pre-tested questionnaire. Univariate and multivariate logistic analyses were conducted with SPSS software package.

RESULTSThe frequencies of mutant heterozygote or homozygote of MBP-52, 54, 57 were 8.0%, 7.2% and 0.4% for cases and 5.3%, 4.3%, 0.5% for controls, respectively. The distribution of mutant genotypes of MBP did not show significant difference between tuberculosis patients and control by Mantel-Haenszel chi2 on sex. The univariate analysis demonstrated that body mass index, marital status, vaccinal vestige, bacillus of Calmette-Guerin vaccine immunization, contacted with pulmonary tuberculosis patients, familial traits were the risk factors of pulmonary tuberculosis. After adjusting those related environmental factors in the multivariate logistic analyses, the total MBP (MBP-52, MBP-54 and MBP-57) and MBP-52 heterozygote genotypes were significantly overrepresented in cases, with adjusted OR (95% CI) being 2.182 (1.058-4.499) and 2.574 (1.028-6.446).

CONCLUSIONTotal MBP and MBP-52 mutant genotypes might be associated with the susceptibility to pulmonary tuberculosis.

Case-Control Studies ; Genetic Predisposition to Disease ; Humans ; Mannose-Binding Lectin ; genetics ; Polymorphism, Genetic ; Tuberculosis, Pulmonary ; genetics