Preventing unnecessary invasive cancer-diagnostic tests: changing the cut-off points

To determine a cut-off point of tPSA and PSAD to prevent unnecessary invasive cancer-diagnosing tests in the community. This study was performed on 688 consecutive patients referred to our center due to prostatism, suspicious lesions on digital rectal examination and/or elevated serum PSA levels. All patients underwent transrectal ultrasound guided biopsies and obtained PSAD. Serum levels of tPSA and fPSA were measured by chemiluminescence. Comparisons were done using tests of accuracy [AUC-ROC]. Prostate cancer was detected in 334 patients, whereas the other 354 patients were suffering from benign prostate diseases. The mean tPSA in case and control groups were 28.32 +/- 63.62 ng/ml and 7.14 +/- 10.04 ng/ml; the mean f/tPSA ratios were 0.13 +/- 0.21 and 0.26 +/- 0.24 in PCa and benign prostate disease groups; the mean PSAD rates were 0.69 +/- 2.24, 0.12 +/- 0.11, respectively. Statistically significant differences were found [P<0.05]. Using ROC curve analysis, it was revealed that AUC was 0.78 for tPSA and 0.80 for f/tPSA. Sensitivity was 71% for the cut-off value of 7.85ng/ml. For f/tPSA ratio, the optimal cut-off value was 0.13 which produced the sensitivity of 81.4% and for PSAD, it was15%. As this trial is different from the European and American values, we should be more cautious in dealing with the prostate cancer upon the obtained sensitivity and specificity for PCa diagnosis [7.85ng/mL fortPSA, 15% for PSAD and 0.13 for f/tPSA ratio]

[ effect of honey bee venom and all-trans retinoic acid on proliferation and differentiation of HL-60 cell line]

Differentiation therapy is one of the methods for treatment of cancer. In this method, some drugs such as All-trans Retinoic Acid, are used which can inhibit proliferation of cancerous cells and induce cell differentiation. Previus experiments showed some anti-proliferation materials, anti-inflammation materials or anti-oxidant materials could influence the effects of such drugs and decrease resulting side effects. In this research the effects of the Honey Bee Venom, on All-trans Retinoic Acid functions was measured. In this experimental project we used HL-60 cell line belonging to acute promyelocyte leukemia. The HL-60 cell line was obtained from Pasteur Institute of Iran, and were grown in RPMI 1640 medium [Roswell Park Memorial Institute] containing 10% FBS [Fetal Bovine Serum] and 1% streptomycin-penicilin. We used MTT [3-[4, 5-Dimethylthiazol-2-yl]-2, 5-Diphenyltetrazolium bromide] and NBT [Nitro-Blue tetrazolium chloride] tests for these purposes. All experiments were done three times and data were analyzed using SPSS. Both Honey Bee Venom and All-trans Retinoic Acid could inhibit proliferation of HL-60 cells, also All-trans Retinoic Acid could induce differentiation in this cells. The amount of differentiation was increased significantly [p<0.01] in the presence of Honey Bee Venom. On the base of our findings, it seemed that Honey Bee Venom could increase anti cancer effects of All-trans Retinoic Acid

[Designing of human cartilage tissue, by differentiation of adipose-derived stem cells with BMP-6 in alginate scaffold]

In the present study the effect of BMP-6 was investigated on chondrogenesis of adipose-derived stem cells. Mesenchymal stem cells derived from subcutaneous adipose tissue were cultured on alginate scaffold to induce chondrogenesis in experimental group, with chondrogenic medium having BMP-6 growth factor for 3weeks. In control group medium without BMP-6 was applied. The harvested constructs were examined with immunohistochemical and RT-PCR methods for assessment of cartilage-specific characteristics. The results of immunohistochemical method revealed the presence of typical cartilage extracellular matrix components such as type II collagen and aggrecan in constructs induced by BMP-6 growth factor on alginate scaffold. In addition evaluation of the results of RT-PCR analysis confirmed the expression of cartilage- specific genes, such as type II collagen and aggrecan, in the differentiated cells under the influence of growth factor BMP-6. It can be concluded that BMP-6 promotes chondrogenesis of ADSC in 3-D and adipose-derived stem cells could be used for cartilage tissue engineering

Tolerance induction by CD40 silenced dendritic cells through antisense

One of the valuable tools for inhibiting the specific gene expression is antisense technique. To determine T cell responses, co-stimulatory molecule expression on the antigen presenting cells is important. In the present study, the effects of high affinity antisense against CD40 mRNA on the function and phenotype of DCs [dendritic cells] were investigated. The DCs were separated from the mice spleens and then cultured in vitro. By means of lipofectamine 2000, the antisense was delivered into the cells and the efficacy of transfection was estimated by flow cytometry. Also, the mRNA expression and protein synthesis were assessed by real time PCR and flow cytometry, respectively. The DCs were transfected with 6 M antisense and 2 l lipofectamine 2000. The percentage of CD40 expression in DCs was 38%. The results showed that CD40 expression is reduced in DCs to 22% and 24%. By annexine V and propidium iodine staining, we could evaluate the viability of the transfected cells. The inhibition of CD40 gene expression was associated with the increase in IL-4 secretion. This shifted the DCs to stimulate Th2 cytokine production from the allogenic T cells. In addition, in the MLR, the DCs without CD40 expression showed poor allostimulatory effects. This finding is valuable in the study of the costimulatory molecules of DCs. These data demonstrate that direct interference of the cell surface expression of CD40 at transcriptional level by antisense confers tolerogenecity potential of DCs. This approach is a useful tool through which DCs become tolerogenic and can be studied as a potential therapeutic option for the autoimmune diseases and allograft rejection

Isolation of mesenchymal stem cells from dental pulp of exfoliated human deciduous teeth

The exfoliated human deciduous tooth [SHED] contain multipotent stem cells that identified to be a population of highly proliferative and clonogenic .These cells are capable of differentiating into a variety of cell types including neural cells, adipocytes, and odontoblasts. Normal exfoliated human deciduous incisors collected from six- to nine-years-old children. The pulp was separated from the crown and digested with collagenase .Single cell solutions were cultivated in alpha-MEM supplemented with ES-FCS. After two to three days, the cells reached confluency and were trypsinized and cultured for further passages. The passage-4 cells were analyzed with CD34, CD45, CD105, CD166, CD31, CD90 and CD146 markers that indicated these cells had a mesenchymal stem cell [MSC] identity. We examined the cells for Alkaline Phosphatase activity to investigate the mesenchymal [stromal] nature.Finally, the cells were differentiated into the osteoblastic and adipocytic lineages in different subcultures and analysed by RT-PCR and different staining protocols. Viable cells growing out of the explants showed elongated shapes in clusters. These cells showed alkaline phosphatase activity. Flow cytometry results revealed high expression of pluripotent stem cell markers .In some area of the osteoinductive cultures nodule-like structures were observed that showed red mineralizing area upon staining with Alizarin Red.In adipogenic cultures lipid vesicles appeared after five weeks of induction with Oil Red. This study show that pulp contains cells with high plasticity and proliferation capacity and can be easily isolated without any serious intervention

[Cross-resistance to anticancer drugs in acute myeloblastic leukemia Pgp expressing K562 cell line]

Drug resistance remains one of the most important clinical obstacles in the treatment of some cancers. This drug resistance referred to as Multidrug Resistance [MDR] induces cross-resistance to many chemotherapy agents such as anthracyclines, vinca alkaloides, epipodophyllotoxins and Taxol. MDR is most likely due to the reduction of drug accumulation with an energy-dependent drug efflux pump. This drug pump is a 170 kDa transmembrane glycoprotein [Pgp]. We developed a resistance subline of K562 by stepwise increase in concentration of Doxorubicin, and Pgp expression was verified by flowcytometry and RT-PCR methods. Cross resistance of the resistant cell line to Etoposide, Vincristine and Taxol was analyzed by MTT assay. IC[50] [the level of drug concentration inhibiting 50% of cell growth] of Doxorubicin, Etoposide and Taxol of parental K562 came out to be 100ng/ml and it was 50 ng/ml for vincristine. IC[50] levels of these drugs on resistant K562 were 500, 500, 450 and 450ng/ml. These drugs also displayed 5-, 5-, 4.5-, and 9- fold resistance respectively. According to the results, expression of Pgp confers MDR phenotype to the K562 cell line and makes it resistant to most of anticancer drugs including anthracyclines, vinca alkaloides, epipodophyllotoxins and taxans. This MDR phenotype is a major obstacle of cancer treatment and in recent years investigators are trying to reverse it by gene therapy

value of thyroid fine needle aspiration in evaluation of follicular neoplasms

The presence of single or multiple nodules with in the thyroid glandis a common clinical problem. It is immposible to differentiate benign and malignant follicular neoplasms in cytologic examinations. Estimation of risk of malignancy in thyroid nodules who are cytologically assumped to be follicular neoplasm has a critical importance. The aim of this research is evaluation of diagnostic value of thyroid FNA in follicular neoplasm. This survey was done on 476 patients with thyroid nodule whome were biopsied in a 7 years period [in 5th Azar hospital, Gorgan]. Standard aspirations and biopsies techniques performed and necessary smears prepared. Wet-fixed smear glass slides are placed immediately in alcohol [70%] 7-10 slides from the aspiration. The patients with cytologic report whome considered follicular neoplasm followed total thyroidectomy or lobotomy. From 476 patients for thyroid nodules 412 cases [86.5%] are suitable for cytologic evaluation which 312 cases [77.9%] of them were benign and 14 cases [3.3%] were malignant. The rest [77 cases] reported to have follicular neoplsm, this patients also undergone thyroid surgery. The result of histological findings appeared in 32 cases [57.1%] follicular adenoma, 15 cases[26.7%] non-euplastic nodules, 5 cases[8.9%] follicular carcinoma and 4 cases [7.1%]follicular type of papillary carcinoma. 9 cases [16%] found malignant among cases which cytologic reports considered as "follicular neoplasms" and subsequently undergone thyroid surgery [consist of follicular carcinoma and follicular type of papillary carcinoma], confirme the importance of histopathologic study of follicular lesions for differentiation between malignant and benign lesions