The levels of c-fos mRNA in U937 cell line were determined with in situ hybridization.Theresults showed that TPA induced the expression of c-fos gene with a relationship of dosage-ef-fect.It was also proved that GM-CSF,TNF,IL-9,IL-3 and IL-6 increased the expression of c-fos gene.

Animals ; Antineoplastic Agents ; therapeutic use ; Gene Silencing ; Genetic Therapy ; methods ; Humans ; MicroRNAs ; antagonists & inhibitors ; therapeutic use ; Neoplasms ; genetics ; therapy ; RNA Interference ; physiology ; RNA, Small Interfering ; antagonists & inhibitors ; therapeutic use ; RNA-Induced Silencing Complex ; metabolism

Objective:To construct and express the fusion protein between ZZ affinity peptide and alkaline phosphatase and examine its biological activities.Methods:The alkaline phosphatase gene was cloned into pEZZ 18 vector containing ZZ peptide gene resulted in the pEZZ-AP recombinant vector.Then the vector was transformed and expressed in E.coli DH5α.And HisTrap affinity chromatography was employed to separate and purify the target protein.After analyzed by Western blot , ZZ-AP fusion protein was applied to immunocytochemistry as an alterative second antibody.Results:The result of SDS-PAGE showed that the fusion protein with a relative molecular weight was consistent with the theoretical values (62 kD).The fusion protein has rabbit IgG-binding ability and en-zymatic activity of alkaline phosphatase ,those were validated in Western blot;and it produced a good signal that was comparable in its staining pattern to that generated with goat anti-rabbit IgG-AP in immunocytochemistry.Conclusion: The ZZ-AP fusion protein was constructed successfully ,it has rabbit IgG-binding ability and enzymatic activity of alkaline phosphatase.We anticipate that the ZZ-AP fusion protein has a potential application in immunoassay field.

OBJECTIVE Autosomal dominant polycystic kidney disease (ADPKD) is a common-monogenetic disease characterized by progressive development of renal cysts. Thereis still further need for effective therapy.Based on our precious study that Ganoderma triterpenes(GT),which is the major secondary metabolites of Ganoderma lucidum,is able to attenuate renal cyst development.The aim of this study was to investigate the effect of a monomer,Ganoderic acid A(GA-A)that was purified from the GT,which has been reported to exhibit antinociceptive,antioxidative,hepatoproctive and anti-cancer activities,to have a potent anti-cyst effect in ADPKD. METHODS We first evaluated the potential cytotoxicity of GA-A on MDCK cells using a CCK-8 assay.Then we used MDCK cyst model,cultivated MDCK cells in vitro to form fluid-filled cysts surrounded by monolayer cells.MDCK cells were co-incu-bated with 10 μmol·L-1FSK with or without GA-A(25 μg·mL-1)and equal concentration GT as positive control from day 0 to day 6 to investigate the inhibitory effect of GA-A on cyst formation.And to further investigate the inhibitory effect of GA-A on cyst enlargement, MDCK cysts were treated with different concentration of GA-A(6.25,25 and 100 μg·mL-1)from day 5 to day 12.Next we used an embryonic kidney cyst model, wile-type mice kidneys were taken out on embryonic day 13.5 to form renal cysts stimulated with 8-Br-cAMP to prove the renal cyst inhibition at organ level.Meanwhile,we explored the possible mechanisms underlying GA-A inhibition on renal cyst development using MDCK cells treated with 10 μmol·L-1FSK co-incubated with GA-A(25 μg·mL-1)and equal concentration GT.Several key components of Ras/MAPK pathway was evaluated by Western blot,the protein expression of H-ras,B-raf, p-ERK, Egr-1 and c-fos was evaluated. RESULTS MDCK cell viability was not affected by GA-A that were used ofincreasing concentrations up to 200 μg·mL-1. GA-A had no significant influence on cyst formation,but inhibited cyst enlargement dose-dependently and the inhibitory effect is significantly better than that of the same concentration of GT which proved that GA-A may be an effective monomer from GT.And after washing out GA-A on day 8,MDCK cysts regrew to a large size,suggesting that the inhibitory effect of GA-A on MDCK cyst enlargement was reversible. GA-A inhibited embryonic kidney cyst development significantly in a dose-dependent and reversible manner proving GA-A cyst inhibition at organ level,which is also more effective than equal concentration GT.Treatment of MDCK cells with FSK caused a significant elevation of H-ras,B-raf,p-ERK,Egr-1 and c-fos signaling molecules,while treatment with GA-A reduced the level of H-ras,B-raf,p-ERK,Egr-1 and c-fos expression.GA-A down-regulated Ras/MAPK signaling pathway could contribute to its inhibitory effect on cyst development. CONCLUSION Ganoderic acid A from Ganoderma lucidum retard ADPKD renal cyst development via down-regulating Ras/MAPK signaling pathway.

OBJECTIVEThis study aimed to evaluate the effort of applying frontal and occipital bones in extensive cranioplasty and preserving multiple cranial bone flaps adhered to the dura mater in the treatment of sagittal synostosis.

METHODSFrom April 2008 to June 2013, sixty-three children with sagittal synostosis, aged 5 months to 3 years, were included in the study. The frontal bone flap was removed using an air drill. The occipital and bilateral temporal bone flaps were cut open but not detached from the dura mater or fixed to produce floating bone flaps. The skull bone was cut into palisade-like structures. Brain compression from both sides and the base of the skull was released and the brain expanded bilaterally through the enlarged space. Only a long strip-shaped bone bridge remained in the central parietal bone. Subsequently, the frontal bone flaps and occipital bone flap were pushed towards the midline and fixed with the parietal bone bridge to shorten the anteroposterior diameter of the cranial cavity and allow the brain to expand bilaterally to correct scaphocephaly. The CT images showed that both sides of the parietal bone of artificial sagittal groove gradually merged postoperative 1 year, and skull almost completely normal healing after operation 2 or 3 years, without deformity recurrence within 5 years. Among them all, 61 children's intelligence is normal and 2 children's lagged behind normal level, no further improvement.

RESULTSPatients were followed up 1 - 5 years (an average of 43 months). Skull growth was excellent in all patients, the anteroposterior diameter was shortened by 14.6 mm averagely, the transverse diameter was increased by 12.3 mm averagely, the prominent forehead was corrected, and scaphocephaly improved significantly. There were no complications such as death and skull necrosis.

CONCLUSIONSThe application of frontal and occipital bones in extensive cranioplasty and preserving multiple cranial bone flaps adhered to the dura mater can be used in the treatment of sagittal synostosis. Surgery without removing bone flaps is less traumatic and results in no massive bleeding. It can effectively relieve brain compression and promotes transversal expansion of the brain during surgery and subsequent normal brain development.

Bone and Bones ; Brain ; growth & development ; Child, Preschool ; Craniosynostoses ; surgery ; Dura Mater ; Frontal Bone ; surgery ; Humans ; Infant ; Parietal Bone ; surgery ; Recurrence ; Surgical Flaps ; Temporal Bone ; surgery

Objective To detect the value of autoantibedy profile in primary biliary cirrhosis (PBC). Methods 96 serum samples of patients with primiary biliary cirrhosis, 100 serum samples of other antoimmune disease and 49 serum samples of healthy were tested for anti- M2, anti-3E(BPO), anti-Sp100,anti-PML,anti-gp210,anti-LKM-1 ,anti-LC-1 ,anti-SLA/LP by EUROLine. Results The positive of the anti-M2,anti-BPO, anti-Sp100, anti-PML and anti-gp210 for PBC was 76. 0%, 84.4%, 32. 3%, 28. 1% and 35.4% ,respectively. The positive of the anti-M2, anti-BPO, anti-Sp100, anti-PML and anti-gp210 for other autoimmune disease was 13.0% ,9. 0% ,3.0% ,2.0% and 1. 0%, respectively. The sensitivity of the anti-M2 for PBC was 76. 0% ,with specificity of 87. 0%. The sensitivity of the anti-BPO for PBC was 84. 4%, with specificity of 91.0% ;the sensitivity of the anti-Sp100 for PBC was 32. 3%, with specificity of 97.0%. The sensitivity of the anti-PML for PBC was 28. 1% ,with specificity of 98.0%. The sensitivity of the anti-gp210 for PBC was 35.4%, with specificity of 99. 0% . Anti-LKM-1, anti-LC-1, anti-SLA/LP positive patients with PBC were not detected;the incidence rate of liver function failure in anti-gp210 positive serum higher than anti-gp210 negative serum (χ2 = 11.17, P < 0. 01). Conclusions Multiple autoantibedies can be detected in the sera of PBC patients. The detection of autoantibody profile is useful for the diagnosis and differential diagnosis of PBC, and may he helpful for therapy and prognosis of PBC.

OBJECTIVETo observe the clinical therapeutic effect of Shuizhitong capsule (SZT) in treating senile vascular dementia (VD) and on changes of related physio-chemical criteria.

METHODSEighty-five patients with VD were randomly divided into the treated group (n = 51) and the control group (n = 34) in the ratio of 3:2, treated with SZT and Piracetam respectively. Before and after treatment, the Zhang's Dementia Scoring (HDS) and Function of Social Activity Questionnaire (FAQ) scoring, cerebral blood flow and hemorrheologic properties were determined.

RESULTSSZT could significantly improve the patients' clinical symptoms, intracranial hemodynamic condition and the hyperviscosity, hypercoagulation and hyperaggregation status, lower the whole blood and plasma specific viscosity, raise the living standard of partial patients, with significant difference between before and after treatment (P < 0.05).

CONCLUSIONThe therapeutic effect of SZT in treating senile VD is definite.

Aged ; Blood Viscosity ; drug effects ; Capsules ; Cerebrovascular Circulation ; drug effects ; Dementia, Vascular ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Hemorheology ; drug effects ; Humans ; Male ; Middle Aged ; Nootropic Agents ; therapeutic use ; Phytotherapy

OBJECTIVE: To study the effects of iron oxide nanoparticles (IONPs) and prussian blue nanoparticles (PBNPs) on the stemness of epithelial ovarian cancer (EOC) cell lines, which may provide experimental basis and reference significance for the application of nanoparticles in the treatment of EOC. METHODS: SKOV3 and HO8910 cell lines were treated with IONPs and PBNPs respectively, then the drug-resistant genes in mRNA and protein levels were detected by RT-PCR and Western blot. The change of cell proliferation ability were detected by cell proliferation and clone formation experiments, and the expression of surface stemness-related molecules were detected by flow cytometer. RESULTS: The expression of ABCG2 and ALDH1A1 were both down regulated in SKOV3 and HO8910 cell lines treated with IONPs and PBNPs, respectively, while IONPs decreased the CD44+CD117+ subpopulation, and PBNPs increased the CD44+CD117+ subpopulation. Compared with the control cells, the proliferation of SKOV3 cells treated with IONPs and PBNPs were significantly reduced, however, there were no effects on HO8910 cell's proliferation after the cells treated with PBNPs. CONCLUSION: Drug-resistance genes and stem cell subpopulation of EOC SKOV3/HO8910 cell lines treated with IONPs were markedly reduced. However, PBNPs can improve the stem cell subpopulation and promote the cancer stem cell's characteristic of EOC cells but the drug-resistance genes were decreased.

Accidents, Occupational ; Adolescent ; Adult ; Arsenic Poisoning ; Child ; Female ; Humans ; Male ; Middle Aged ; Water Pollution, Chemical

The purpose of this study is to investigate the reversal effect and its mechanism of arsenic trioxide (As2O3) on multidrug resistance of gastric carcinoma cells. The concentration of vincristine (VCR) increased gradually to induce the drug resistance of gastric carcinoma cell SGC7901. MTT assay was used to determine the lethal effect of anticarcinogens on tumor cells and Western blotting assay was applied to determine the expression of P-glucoprotein (P-gp) and glutathione S-transferase (GST-s) in tumor cells. As a result, the resistance of SGC7901/VCR cells to VCR, fluorouracil and epirubicin was 16.56, 2.69 and 13.05 times, respectively, more than that of SGC7901 cells. After 24 h precondition with As2O3, RI of vincristine, fluorouracil and epirubicin decreased significantly (P < 0.05). Expression of P-gp and GST-s in resting SGC7901/VCR cells was significantly higher than that in carcinogen-sensitive SGC7901 cells. As2O3 decreased the expression of P-gp and GST-s in SGC7901/VCR cells significantly, while it showed no significant effect on carcinogen-sensitive SGC7901 cells. The result suggested that As2O3 could partly reverse drug resistance of SGC7901/VCR cells by probably the mechanism of decreasing the expression of P-gp and GST-s.
ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Adenocarcinoma ; metabolism ; pathology ; Antineoplastic Agents ; pharmacology ; Arsenicals ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Epirubicin ; pharmacology ; Fluorouracil ; pharmacology ; Glutathione Transferase ; metabolism ; Humans ; Oxides ; pharmacology ; Stomach Neoplasms ; metabolism ; pathology ; Vincristine ; pharmacology