Objective To investigate the histogenesis and clinicopathologic features of carcinosarcoma of the urinary bladder. Methods Two cases of carcinosarcoma (one man and one woman)were included.They were admitted to hospital with discontinuous,painless,macroscopic hematuria.Clinicopathalogic features of all findings of the 2 cases were reviewed. Results Cystoscopy of both cases showed that the tumors displayed polypous or cauliflower-like shape,and grew invassively.CT examination of the bladder wall confirmed the presence of parenchymatous tumor.Both of the 2 cases underwent partial cystectomy.Intraoperative examination showed the tumors were similar to carcinoma of bladder.Histopathology showed biphasic neoplasms with distinct high grade epithelial and mesenchymal components.Morphologic characteristic of the tumor was abnormally proliferative and the mitotic figures could be seen.One of the patients died within 11 months and the other,16 months following the operation. Conclusions Carcinosarcoma of the urinary bladder is a kind of rare malignant tumor that is most often in an advanced stage at presentation and has an unfavorable prognosis. It should be identified promptly and treated appropriately.Those who have discontinuous,painless,macroscopic hematuria should be warned of the risk of the disease.

Strict regulation of HIV-1 PR function is critical for efficient production of mature viral particles. During viral protein expression and viral assembly, HIV-1 PR located within Gag-Pol precursor must be inactive to prevent premature cytoplasmic processing of the viral Gag and Gag-Pol precursors. Premature activation of HIV-1 precursors leads to major defects in viral assembly and production of viral particles. A cell-level premature activation of HIV-1 precursors assay using bioluminescence resonance energy transfer (BRET) was established. Three thousand compounds were screened to evaluate this assay. The results showed that the assay is sensitive, specific and stable (Z' factor is 0.905).

The HIV-1 Rev protein facilitates nuclear export of unspliced and singly spliced viral transcripts containing RRE RNA through the CRM1 export pathway. Inhibition of Rev-mediated RNA nuclear export can arrest HIV-1 transcriptional process, which clearly, reveals a target for anti-HIV drug development. In this work, a cell-based assay has been established for screening anti-HIV compounds targeting the Rev-mediated RNA nuclear export. This assay utilized a codon-optimized green fluorescent protein (GFP) as reporter gene, which expression is in a Rev-dependent manner. Any compound that inhibits the Rev-mediated RNA nuclear export is identified by reducing emission of GFP. The Z' score of this model is 0.8220. Three thousands compounds were screened and the positive rate was 9.3% with a cutoff at 50% inhibition. IMB7C7, one of the positive compounds, efficiently inhibits viral production from HIV-1 infected cells.

AIM: To solve the two difficulties of bone resorption and inflammation in rheumatoid arthritis, clone the recombinant human osteoprotegerin (OPG) and mycobacteria heat shock protein 70 (HSP70) functional gene,and study the expression and activity of OPG-HSP70 fusion protein in E.coli. METHODS: Experiments were performed at the Laboratory of Department of Immunology, Capital Medical University from May 2006 to September 2007. Complementary DNA encoding full length OPG protein was amplified by reverse transcription-polymerase chain reaction (RT-PCR) from human osteosarcoma cell line MG63 and cloned into pGEMT-Easy vectors. Then, using the recombinant plasmid as the template,the DNA encoding the fusion protein OPG-HSP 70 was amplified by PCR,and was inserted into prokaryotic expression vector pET-28a. Construct pET-28a-OPG-HSP 70 was used to transform into competent E.coli. BL21(DE3) which were induced by isopropyl B-D-thiogalactopyranoside (IPTG),and the fusion protein from above E.coli was collected. Sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) and Western-blotting were performed to identify OPG-HSP 70 fusion protein. The experiments of osteoclast inhibition and restraining inflammation were used to detect the bioactivity of fusion protein. RESULTS: ①The complementary DNA encoding full length OPG protein was obtained. OPG-HSP 70 fusion gene obtained in this experiment was successfully inserted into pET-28a vector. OPG-HSP70 fusion protein was expressed when transformed into E.coli.BL21(DE3). ②SDS-PAGE indicated that the fusion protein was large expressed at the molecular weight of Mr22 000, but there were no band in the total lysate of bacteria harboring pET-28a-OPG-HSP70 without IPTG induction group. ③Western-blotting indicated that the OPG-HSP70 fusion protein could specifically react with anti-human OPG monoclonal antibodies. ④The osteoclast inhibition test demonstrated that the fusion protein could reduce the number of osteoclast, and had the ability to inhibit bone absorptions in vitro. ⑤The experiment of restraining inflammation showed that the fusion protein could significantly reduce the inflammation of delayed type hypersensitivity (DTH) mice, which explained that HSP70 in fusion protein had the inflammation inhibitory bioactivity. CONCLUSION: OPG-HSP70 fusion protein is expressed in E.coli.BL21(DE3),and function study in vitro illustrates the bioactivity of fusion protein.

Objective:To establish an experimental model for intracellular antibacteria and endotoxin neutralization in vitro to detect the antibacterial and endotoxin neutralization activity of the muBPI_(25) protein.Methods: RAW264.7 cells were transfected with pcDNA3.1(+)muBPI_(36-259), and then were infected with intracellular bacterial of either G ~+/G~-to establish the experimental model of intracellalar antibacteria.The RAW264.7 cells were co-transfected with the pSecTag2B-muBPI_(36-259) and dual-luciferase reporter gene plasmids for establishment of the experimental model of endotoxin neutralization.Results:The experimental model of intracellular antibacteria confirmed that the muBPI_(25) protein could inhibit/kill Salmonella typhi.The experimental model of endotoxin neutralization indicated that the muBPI_(25) protein could neutralize endotoxin.Conelusion: We firstly demonstrate that murine BPI N-terminal functional fragment(muBPI_(25) protein)can inhibit/kill Salmonella typhi,and can neutralize, its lysating product, endotoxin.

The volume change of tumor during radiotherapy processes indirectly reflects the short-term efficacy and the quality of radiotherapy planning. We analyzed clinical data of radiotherapy using a mathematical model in our study. First, we selected eight esophageal carcinoma patients with only using 3DRT and conventional dose fractionation schemes. And then we observed and measured the change of tumor volume during the radiotherapy. Based on the LQ model, repopulation and re-oxygenation in 4Rs, and the kinetics of doomed tumor disintegration, we established the mathematical model of tumor evolution in radiotherapy. And then we used the model to analyze the clinical trial data about esophageal carcinoma with radiotherapy. It was proved that the results of the model almost coincided with the clinical trial data. According to the analysis results, we could get the related radiobiology parameters to estimate biological effective dose and repopulation of patients. The mathematical model could provide reference for assessment of prognosis and further scheme of treatment.
Algorithms ; Esophageal Neoplasms ; pathology ; radiotherapy ; Humans ; Models, Theoretical ; Radiotherapy Planning, Computer-Assisted ; methods ; Tumor Burden

Objective:To summarize the CT imaging features of obturator hernia.Methods:From July 2009 to May 2019, the clinical and CT imaging features of 26 cases with obturator hernia diagnosed by multi-slice spiral CT(MSCT) were retrospectively analyzed.The locations of hernia sac, contents of hernia and intestinal obstruction were observed by multi-planer reconstructions(MPR).Results:There were 5 cases of bilateral obturator hernia, 9 cases of right obturator hernia and 12 cases of left obturator hernia.The contents of hernia were small intestine in 19 cases, mesentery in 9 cases and bladder in 5 cases.There were 3 cases of bilateral small intestine obturator hernia, 7 cases of right and 9 cases of left small intestine obturator hernia, presented with abruptly collapse and narrowing of small intestinal cavity at the entrance of obturator canal.The small intestine was herniated between the external obturator muscle and the pubic muscle and adductor brevis, between the upper and lower bundles of external obturator muscle and the internal obturator muscle and superior pubic sulcus.There were 15 cases of small intestinal obstruction, including 9 cases of incarcerated small intestinal obstruction and 6 cases of strangulated small intestinal obstruction, of which 15 cases presented with small intestinal effusion, 3 cases with a little accumulation of gas, 14 cases with small intestinal wall edema and 2 cases with intestinal wall defect.The dilatation effusion and pneumatosis could be found in the proximal small intestine, of which 14 cases with gas-liquid level, 12 cases with small intestinal wall edema and mesenteric edema.There were 4 cases of bilateral mesenteric obturator hernia, 2 cases of right mesenteric obturator hernia and 3 cases of left mesenteric obturator hernia.There were 2 cases of right mild bladder obturator hernia and 3 cases of left mild bladder obturator hernia, presented with cystic water-like density at the entrance of obturator canal.Conclusion:MSCT reconstruction can intuitively display imaging features of obturator hernia, which has a rather high diagnostic value for small intestinal ischemic, necrosis and perforation.

Objective: To investigate the signal transduction mechanisms of time-dependent effects of 50 Hz 1.8 mT sinusoidal electromagnetic fields(SEMFs)on osteoblastic activity. Methods: Newborn rat calvarial osteoblasts(ROBs)were treated with 50 Hz 1.8mT SEMFs for 30, 60, 90, 120, and 150 min, respectively.Intracellular alkaline phosphatase(ALP)activity was assayed, and protein expression of Runt-related transcription factor 2(Runx2), Smad1/5/8 and mitogen-activated protein kinases(MAPKs)were examined by Western blot.The Dual-Luciferase Reporter Assay System was used to measure the activity of the Wnt pathway.Immunofluorescence staining and laser confocal microscopy were applied to examine the nuclear translocation of Smad1/5/8 and β-catenin.Changes in ALP activity were determined after inhibiting p38 MAPK using a specific inhibitor. Results: ALP activity of ROBs increased after 30, 60, 90, 120 and 150 min of SEMFs treatment, compared with the control group(51.41±5.21, 59.47±4.02, 67.56±4.68, 63.69±3.92, and 58.16±3.61 vs.45.53±3.24), and reached the highest value at 90 min and then started to decline.Protein expression of Runx-2, p-Smad1/5/8, p-p38 and β-catenin increased after SEMFs treatment, and reached the highest value at 90 min and then gradually returned to baseline levels.The values for Wnt pathway activity for the control group and with SEMFs treatment at 30, 60, 90, 120 and 150 min were 0.49±0.06, 0.52±0.09, 0.75±0.05, 0.77±0.42, 0.58±0.08 and 0.42±0.09, respectively.Wnt pathway activity, the nuclear translocation of β-catenin and Smad1/5/8 reached the highest level at 90 min.Values of ALP activity in the control group and the SEMFs group were 44.60±3.84 and 71.54±7.34, respectively, before specifically inhibiting p38 MAPK, and were 52.08±0.83 and 52.15±10.77, respectively, after p38 MAPK inhibition, indicating that ALP activity could not be increased with inhibition. Conclusions 50 Hz 1.8 mT SEMFs increase osteoblastic activity by activating the BMP-2/Smad1/5/8, Wnt/β-catenin and p38 MAPK signal pathways.The optimal duration of treatment is 90 min per day.

Deregulation of endothelial nitric oxide synthase (eNOS) plays an important role in the development of multiple cardiovascular diseases. Our recent study demonstrated that genistein supplementation attenuates pulmonary arterial hypertension in broilers by restoration of endothelial function. In this study, we investigated the molecular mechanism by using broiler pulmonary arterial endothelial cells (PAECs). Our results showed that genistein stimulated a rapid phosphorylation of eNOS at Ser(1179) which was associated with activation of eNOS/NO axis. Further study indicated that the activation of eNOS was not mediated through estrogen receptors or tyrosine kinase inhibition, but via a phosphatidylinositol 3-kinase (PI3K)/Akt-dependent signaling pathway, as the eNOS activity and related NO release were largely abolished by pharmacological inhibitors of PI3K or Akt. Thus, our findings revealed a critical function of Akt in mediating genistein-stimulated eNOS activity in PAECs, partially accounting for the beneficial effects of genistein on the development of cardiovascular diseases observed in animal models.
Animals ; Cell Line ; Chickens ; Disease Models, Animal ; Endothelium, Vascular/drug effects/*metabolism/pathology ; Enzyme Activation/drug effects ; Female ; Genistein/*pharmacology ; Humans ; Hypertension, Pulmonary/drug therapy/*metabolism/pathology ; Nitric Oxide Synthase Type III/genetics/*metabolism ; Oncogene Protein v-akt/*metabolism ; Phosphorylation ; Signal Transduction/drug effects

Objective To determine the effect of isopsoralen(ISO)on the healing of tibia fracture in mice and explore its underlying mechanism.Methods Fifty male C57BL/6 mice(2 month old,20±2 g)were randomly divided into model group and ISO treatment group,with 25 animals in each group.From the 3rd day after modeling,the mice from the ISO group were given an intragastric gavage of 40 mg/kg ISO,once per day for 28 consecutive days,while those of the model group was given same volume of normal saline in same way.On the 7th,14th,21st,and 28th day after gavage,the tibia on the surgical side was taken,and the fracture area was quantified by bone volume/total volume(BV/TV)after micro-CT scanning.The healing and shaping of the fracture end were observed through HE staining.ELISA was used to detect the serum contents of bone alkaline phosphatase(BALP)and procollagen type I N-terminal peptide(PINP)on the 14th day of gavage.Western blotting was employed to determine the expression levels of Collagen Ⅰ,Runx2,BMP2,OSX,and VEGF in the tibial callus tissue in 7 and 14 d after gavage.Vascular perfusion was applied to observe the callus microvessels in 28 d to quantitatively analyze the vascular volume fraction and vessel diameter.Immunohistochemical staining was conducted to observe the expression of VEGF in the callus in 14 d after gavage.Results HE staining displayed that the ISO group had faster healing process than the model group.Micro-CT quantification results showed that the ISO group had higher BV/TV ratio in 7 d after gavage though no statistical difference,significantly higher ratio in 14 d(P＜0.05),but obviously lower ratio in 21 and 28 d after gavage(both P＜0.05)when compared with the model group.The serum contents of BALP and PINP were also remarkably higher in the ISO group than the model group(P＜0.05).Western blotting results indicated that the expression levels of Collagen Ⅰ,Runx2,BMP2,OSX and VEGF in the ISO group were higher than those in the model group(P＜0.05).The results of angiography revealed that the vascular volume fraction and vessel diameter were notably increased in the ISO group than the model group(both P＜0.05).Immunohistochemical assay showed that the expression of VEGF was higher in the ISO group than the model group(P＜0.05).Conclusion ISO can improve the activity of osteoblasts,increase the expression of osteogenesis-related proteins,and accelerate the angiogenesis to promote fracture healing.