Effect of 25-hydroxyvitamin D3 and 1 α,25 dihydroxyvitamin D3 on differentiation and apoptosis of human osteosarcoma cell lines

Osteosarcoma (OS) is a malignant bone tumor predominantly affecting children and adolescents. OS has a 60% survival rate with current treatments; hence, there is a need to identify novel adjuncts to chemotherapeutic regimens. In this pilot study, we investigated the dose‐response to 1α,25‐dihdroxyvitamin D₃ (1,α 25(OH)₂D₃) and 25‐hydroxyvitamin D₃ (25(OH)D₃) by human OS cell lines, SaOS‐2, and 143B. We hypothesized that 1,α 25(OH)₂D₃ and 25(OH)D₃ would stimulate differentiation and induce apoptosis in OS cells in a dose‐dependent manner. Human OS cell lines, SaOS‐2, and 143B, were treated with 1,α 25(OH)₂D₃ or 25(OH)D₃ or an ethanol control, respectively, at concentrations ranging from 1 to 1,000 nM. Ki67 (a marker of cellular proliferation) immunocytochemistry revealed no significant changes in the expression of Ki‐67 or MIB‐1 in 1α,25(OH)₂D₃ or 25(OH)D₃ treated SaOS‐2 or 143B cells. Both control and 1α,25(OH)₂D₃ treated SaOS‐2 and 143B cells expressed vitamin D receptor (VDR). Markers of osteoblastic differentiation in 143B cells and SaOS‐2 cells were induced by both 25(OH)D₃ and 1α,25(OH)₂D, and evident by increases in alkaline phosphatase (ALP) activity, osteocalcin (OCN) mRNA expression, and mineralization of extra‐cellular matrix (ECM) by alizarin red staining. An increasing trend in apoptosis in response to 25(OH)D₃, in both SaOS‐2 and 143B cells was detected by terminal deoxynucleotidyl transferase (TdT)‐mediated dUTP nick end labeling (TUNEL) staining. With 1α,25(OH)₂D₃ treatment, apoptosis was evident at higher concentrations only. These preliminary findings suggest that OS cells express VDR and respond to 25(OH)D₃ and 1α,25(OH)₂D₃ by undergoing differentiation and apoptosis. © 2011 Orthopaedic Research Society Published by Wiley Periodicals, Inc. J Orthop Res 30:831–844, 2012     

Boldine induces cell cycle arrest and apoptosis in T24 human bladder cancer cell line via regulation of ERK,AKT, and GSK-3β

ObjectiveBladder cancer is one of the most prevalent genitourinary malignancies. Despite active chemotherapy regimens, patients with bladder cancer suffer from a high rate of tumor recurrence. Thus, new approaches and agents to improve quality of life and survival still need to be developed. The objective of the present study was to evaluate the effect and underlying mechanisms of boldine, an aporphine alkaloid of Peumus boldus, on bladder cancer proliferation and cell death.MethodsSulforhodamine B assay, Tetrazolium reduction assay, Flow Cytometry Analysis, Ecto-5’-nucleotidase activity and Western blot assay were performed.ResultsThe results showed that boldine was able to reduce cell viability and cell proliferation in T24 cells. In addition, boldine arrests the cell cycle at G2/M-phase and cause cell death by apoptosis. Boldine-induced inhibition of cell growth and cell cycle arrest appears to be linked to inactivation of extracellular signal–regulated kinase protein (ERK). Additionally, the efficacy of boldine in apoptosis-induced in T24 cells is correlated with modulation of AKT (inactivation) and glycogen synthase kinase-3β (GSK-3β) (activation) proteins.ConclusionsThe present findings may, in part, explain the therapeutic effects of boldine for treatment of urinary bladder cancer.     

Effects of chitosan on cell proliferation and collagen production of tendon sheath fibroblasts, epitenon tenocytes, and endotenon tenocytes

Flexor tendon repair in zone II is often complicatedby adhesions to the surrounding fibro-osseoustendon sheath. Adhesion between the tendon andtendon sheath will impair the gliding function oftendons and result in marked impairment of handfunction. Experimental strategies to decrease adhesionformation, with the long-term goal of improving clinicaloutcome after tendon injury and repair, have been thesubject of numerous studies over the past 20 years.Biochemical agents such as antihistamines, s…     

Survival and prognostic factors of germ cell tumors of the testis in the departments of Bas-Rhin, Isère, and Somme]

The authors report a study of the prognosis of germ cell tumours of the testis in populations of the Bas-Rhin, Isère and Some departments. This study was performed on French Eurocare data (European testicular cancer register study). 247 patients diagnosed between 1987 and 1993 were studied. Estèv's method and model were used to estimate the relative survival and for prognosis analysis with reference to the French age-matched male population. 84.6% of patients were between the ages of 20 and 49 years. The 5-years relative survival was 90.1%; 95 CI% [85.5-94.8]. The 5-year relative survival was lower (63.9%) in advanced forms (stage III). Univariate analysis in 223 patients demonstrated the following significant prognostic factors: stage, metastatic sites, treatment, retroperitoneal lymph node dissection and testicular vascular invasion. Multivariate analysis identified: stage, age, CNS metastases, retroperitoneal lymph node dissection and treatment. This study confirms the generally good prognosis of this cancer. As reported in the literature, factors negatively influencing relative survival are those related to advanced disease. Age, a controversial factor in the literature, was correlated with relative survival in this study.     

Photodetection and photodynamic therapy of ‘early’ squamous cell carcinomas of the pharynx,oesophagus and tracheo-bronchial tree

The efficacy of photodynamic therapy (PDT) alone was evaluated on 41 early squamous cell carcinomas of the pharynx (10), oesophagus (15) and tracheo-bronchial tree (16). All lesions but two were synchronous second primaries in ENT-patients suffering from a more extensive cancer, governing the overall oncological prognosis.Photofrin I (3 mg/kg) or Photofrin II (2 mg/kg) were injected 72 h prior to the red light irradiation, supplied by an argon pumped dye laser. A diffusing cylinder was used to obtain a homogeneous light distribution at the tumour site (60 J to 150 J/cm²). In the oesophagus and bronchi, the results are good for cancers staged in situ or microinvasive at endoscopy (two recurrencies for 23 lesions treated). For more advanced cancers (submucosal in the oesophagus or invading the bronchial cartilage), the results are less satisfactory (three recurrencies for eight lesions treated). In the pharynx where light dosimetry is more difficult, the rate of recurrencies is higher (3/10 lesions treated). In the bronchi (one case) and oesophagus (one case), the longest disease-free survival is now 5 years.The irradiation of a non-cancerous zone of normal buccal mucosa on 25 patients having received HPD showed necrosis in all cases with light doses as low as 50mW/cm² for 20 min (60 J cm^–2), even with Photofrin II.We encountered six complications (three cicatricial stenosis, two fistulae, one severe sunburn), most of them resulting from the lack of selectivity of HPD. According to these experiments, PDT is efficient at destroying early squamous cell carcinomas in the pharynx, oesophagus and bronchi, but the tumour selectivity of HPD is poor in the digestive tract lined with squamous cell epithelium. The only hope for the future lies in the synthesis of a more selective and more stable photosensitizer. This discussion reviews possible directions of research for the development of new dyes (cationic dyes, dyes attached to monoclonal antibodies, etc), for PDT and hyperthermia, for photodetection of early cancers using a fluoro-endoscope, and finally, for tumour depth profiling in hollow organs using lasers of different wavelengths.     

Metachronous gonadal and extragonadal primaries, or late relapse of germ cell tumor?

We report five distinct cases of apparently metachronous extragonadal and gonadal germ cell tumors (GCT) occurring in the same patient. Two patients had metachronous GCT of the central nervous system and the testis. One of these patients had been successfully treated for a germinoma of the pineal gland and developed a nonseminomatous GCT of the testis 10 years later. The second patient had a primary seminoma of the testis treated by orchiectomy followed by radiotherapy and developed a germinoma of the sphenoidal sinus 17 months later. Three other patients had an apparently metachronous retroperitoneal and testicular GCT with 22, 44, and 66 months elapsing, respectively, between the first and second neoplasms. These cases suggest that the remaining testis is not only at risk for a second primary GCT, but that a second GCT may emerge at an extragonadal site. A genetic predisposition may account for some of these cases and the occurrence of bilateral testicular GCT. In none of these cases, however, could we ascertain whether testicular GCT was truly a second primary or a relapse of a "primary" retroperitoneal GCT in our cases.     

Low utility of serum 25–hydroxyvitamin D3 and 1, 25-dihydroxyvitamin D3 in predicting peripheral Treg and Th17 cell counts in ESRD and renal transplant patients

BackgroundVitamin D has shown an immune-modulatory effect in different studies. Vitamin D stimulates Tregs and inhibits Th17 cells. The immune-modulatory role of vitamin D in chronic kidney disease (CKD) and renal transplant patients is unclear. We measured whether different serum levels of vitamin D were associated with an increased or decreased presence of lymphocyte subsets including Treg and Th17 cells in end-stage renal disease (ESRD) and renal transplant recipients.MethodsEighty-seven renal transplant recipients and 53 end-stage renal disease (ESRD) patients were enrolled in this study. The absolute counts of CD4 + and CD8 + T, CD16 + CD56 + NK, CD19 + B, CD4 + CD25 + CD127- Foxp3 + (Tregs), Helios + Tregs, CD38 + Tregs, and CD4 + CD17 + (Th17) cells were analyzed in peripheral blood in both patient groups. In addition, serum 25 (OH) D₃, 1, 25 (OH)₂ D₃, IL-6, IL-17, IL-23, and TGF-β₁ were measured. The association between lymphocyte subset counts and 1, 25 (OH)₂ D₃ or 25 (OH) D₃ was studied, as was the association between serum IL-6, IL-17, IL-23, or TGF-β₁ and 1,25 (OH)₂ D₃ or 25 (OH) D₃.ResultsSerum 25 (OH) D₃ and 1,25 (OH)₂ D₃ levels were not independently associated with peripheral CD4 + T, CD19 + B, CD16 + CD56 + NK, Treg, or Th17 cell counts. In contrast to serum 25 (OH) D₃, serum1, 25 (OH)₂ D₃ was positively associated with CD8 + T cells counts in renal transplant recipients.ConclusionOur findings indicate low utility of serum 25 (OH) D₃ and 1, 25 (OH)₂ D₃ levels in predicting a change in lymphocyte subset counts in ESRD and renal transplant patients.     

Langerhans’ cell density and degree of keratinization in foreskins of Chinese preschool boys and adults

Background  

Recent epidemiological studies have proposed that male circumcision reduces the relative risk of acquiring HIV-1. Here, we evaluated the density of Langerhans’ cell and degree of keratinization in the foreskins of Chinese preschool boys and adults.     

Increased expression of collagens,transforming growth factor-β1, and -β3 in gluteal muscle contracture

Backgroud  

Gluteal muscle contracture (GMC) is a multi-factor human chronic fibrotic disease of the gluteal muscle. Fibrotic tissue is characterized by excessive accumulation of collagen in the muscle's extracellular matrix. Transforming growth factor (TGF)-β1 and -β2 are thought to play an important role in fibrogenesis, while TGF-β3 is believed to have an anti-fibrotic function. We hypothesize that the expression of collagen and TGF-βs would be up-regulated in GMC patients.     

Testicular torsion-induced germ cell apoptosis and expression of gene iNOS

To investigate the relationship between germ cell apoptosis and expres-sion of iNOS in experimentally torsed testes in rats. Adult male rats weresubjected to unilateral testicular torsion of 720 g for two hours. Six days lat-er, testes were collected for the detection of iNOS expression with immuno-histochemical method and germ cell apoptosis with TdT-mediated dUTP-X     

Detection of vital germ cell tumor cells in short-term cell cultures of primary tumors and of retroperitoneal metastasis —Clinical implications

By establishing short-term cell cultures derived from retroperitoneal metastasis after neoadjuvant chemotherapy, our aim was to improve the diagnosis and prognosis in patients with advanced testicular germ cell tumors. The histological evaluation of surgically removed metastatic tissue by retroperitoneal lymphadenectomy (RLA) is extremely complicated after previous chemotherapy, but knowledge of persistence of vital tumor cells in residual lesions is of great prognostic value and therapeutic consequence in patients with testicular germ cell tumors. We therefore investigated whether vital tumor tissue could be detected in short-term cell cultures derived from such metastatic lesions by measuring the concentration of the tumor markers beta human chorionic gonadotropin (HCG) and alpha-1 fetoprotein (AFP) in cell culture supernatants. We initially demonstrated the specificity of the determination in cell cultures of human transitional-cell carcinoma cell lines, human foreskin fibroblasts and normal testicular tissue. In a group of 20 patients with untreated primary testicular germ cell tumors, detection of HCG and AFP was increased about threefold in cell culture supernatants in comparison to the serum concentration. Finally, we prepared primary cell cultures from surgically removed retroperitoneal metastasis of 12 patients with testicular germ cell tumors after chemotherapy. The serum concentrations of HCG and AFP of all patients were at normal values when RLA was performed. However, pathologically increased concentrations of HCG (3/3) and AFP (2/3) in cell culture supernatants were found in 3 of 12 cell cultures. Interestingly, these three patients with a pathological increase in HCG and AFP as determined in the supernatant of the short-term cell cultures had tumor progression within a mean follow-up of 3 ± 1 months (P < 0,01), whereas 9 of 12 patients who had no pathological increase in HCG and AFP as determined in the supernatant of the short-term cell culture were in complete remission (CR) after a mean follow-up of 40 ± 11.6 months.     

Loss of phosphoinositide 3-kinase γ decreases migration and activation of phagocytes but not T cell activation in antigen-induced arthritis

Background  

Phosphoinositide 3-kinase γ (PI3Kγ) has been depicted as a major regulator of inflammatory processes, including leukocyte activation and migration towards several chemokines. This study aims to explore the role of PI3Kγ in the murine model of antigen-induced arthritis (AIA).     

The effects of sodium hyaluronate on mRNA expressions of matrix metalloproteinase-1,-3 and tissue inhibitor of metalloproteinase-1 in cartilage and synovium of traumatic osteoarthritis model

teoarthritis (OA)isadegenerativeprocessofjointscharacterizedbyprogressivedeteriorationanderosionofcartilage .TraumaticOAcausedbyjointsinjuryandoperationiscommonlyobserved .Intra articularinjectionofsodiumhyaluronate (HA )isnowwidelyusedintreatmentofOA .It…     

The Prognostic Value of E-Cadherin, α-, β-, and γ-Catenin in Urothelial Cancer of the Upper Urinary Tract

Objectives

To determine the value of loss of the expression of E-cadherin and cadherin-associated molecules as useful markers for both prognosis and bladder recurrence in patients with upper urinary tract cancer.

Materials and methods

In 61 paraffin-embedded nephroureterectomy specimens, the expression of E-cadherin and α-, β-, and γ-catenin was examined by immunohistochemical staining. To evaluate the prognostic significance, Kaplan-Meier survival curves were calculated and compared by the log-rank test. A multivariate test was performed to detect prognostic markers.

Results

Normal expression was found in 32 cases (52.5%) for E-cadherin, 41 cases (67.2%) for α-catenin, 42 cases (68.9%) for β-catenin, and 31 cases (50.8%) for γ-catenin. The expression patterns of E-cadherin and α-, β- and γ-catenin were significantly correlated with each other. Survival analysis showed a significant difference between normal and aberrant expression in each staining. Multivariate analysis revealed that tumor stage and the expression of E-cadherin were independent prognostic factors for cause-specific survival. In contrast, there was no significant correlation between the expression of E-cadherin and α-, β-, and γ-catenin and bladder recurrence.

Conclusion

Our data suggest E-cadherin may be a good prognostic marker for patients with upper urinary tract cancer.     

Construction and identification of immortalized rat astrocyte cell line expressing enkephalin

Objective ： To provide a sound cell source for further ex-vivo gene therapy for chronic pain, we attempt to develop an immortalized rat astrocyte cell line that expresses enkephalin regulated by doxycycline. Methods. Retrovirus infection method was employed to develop an immortalized rat astrocyte cell line that could express enkephalin regulated by doxycycline. The hPPE gene expression level of immoralized astroyte cells （IAC）/ hPPE was detected by RT-PCR, indirect immunofluorescence staining and radioimmunoassay. Results. IAC carrying Tet-on system transfected with preproenkephalin gene could secrete enkephalin that was regulated by doxycycline in a dose-dependent manner and hPPE gene activation could be repeated in on-off-on cycles through administration or removal of doxycycline. Conclusion： An immortalized rat astrocyte cell line that secrete enkephalin under the control of doxycycline is established successfully, which provides a research basis for transgenic cell transplantation for analgesia.