Silibinin-Induced Apoptosis and Downregulation of MicroRNA-21 and MicroRNA-155 in MCF-7 Human Breast Cancer Cells / 한국유방암학회지

PURPOSE: MicroRNAs (miRNAs) have received much attention owing to their aberrant expression in various stages of cancer. In many biological processes, miRNAs negatively regulate gene expression, and may be useful in therapeutic strategies. The present study evaluated the effects of silibinin (silybin), a natural flavonoid, on miRNA expression and attempted to elucidate therapeutic targets in MCF-7 breast cancer cells. METHODS: The rates of cell proliferation and apoptosis were determined in silibinin-treated and untreated MCF-7 cells. Furthermore, the expression levels of miR-21 and miR-155 were measured in MCF-7 cells after incubation with silibinin (100 µg/mL), and the putative targets of the miRNAs within the apoptotic pathways were predicted using bioinformatic approaches. The expression levels of some of these targets were evaluated by quantitative reverse transcription polymerase chain reaction (qRT-PCR). RESULTS: Silibinin induced apoptosis in MCF-7 cells in a dose- and time-dependent manner. qRT-PCR analysis revealed a decrease in miR-21 and miR-155 expression levels in silibinin-treated cells relative to the levels in the untreated cells. Potential miR-21 and miR-155 targets within the apoptotic pathways, such as CASP-9, BID, APAF-1, CASP-3, CASP-8, and PDCD4, were predicted by in silico analysis. qRT-PCR analysis showed upregulation of some of these potential targets including caspase-9 (CASP-9) and BID after silibinin treatment for 48 hours. CONCLUSION: Our results suggest a correlation between the expression of miR-21 and miR-155, and MCF-7 cell proliferation. The antiproliferative activity of silibinin may partly be attributable to the downregulation of miR-21 and miR-155, and the upregulation of their apoptotic targets. Furthermore, the upregulation of CASP-9 and BID indicates that silibinin induces apoptosis through both the extrinsic and intrinsic pathways.

Comparison of Mesenchymal Stem Cell Markers in Multiple Human Adult Stem Cells

OBJECTIVES: Mesenchymal stem cells (MSCs) are adult stem cells which identified by adherence to plastic, expression of cell surface markers including CD44, CD90, CD105, CD106, CD166, and Stro-1, lack of the expression of hematopoietic markers, no immunogenic effect and replacement of damaged tissues. These properties led to development of progressive methods to isolation and characterization of MSCs from various sources for therapeutic applications in regenerative medicine. METHODS: We isolated MSC-like cells from testis biopsies, ovary, hair follicle and umbilical cord Wharton's jelly and investigated the expression of specific cell surface antigens using flow cytometry in order to verify stemness properties of these cells. RESULTS: All four cell types adhered to plastic culture flask a few days after primary culture. All our cells positively expressed common MSC-specific cell surface markers. Moreover, our results revealed the expression of CD19and CD45 antigens in these cells. CONCLUSION: According to our results, high expression of CD44 in spermatogonial stem cells (SSCs), hair follicle stem cells (HFSCs),granulosa cells (GCs)and Wharton's jelly-MSCs (WJ-MSCs)may help them to maintain stemness properties. Furthermore, we suggest that CD105+SSCs, HFSCs and WJ-MSCs revealed the osteogenic potential of these cells. Moreover, high expression of CD90 in SSCs and HFSCs may associate to higher growth and differentiation potential of these cells. Further, the presence of CD19 on SSCs and GCs may help them to efficiency in response to transmembrane signals. Thus, these four types of MSCs may be useful in clinical applications and cell therapy.

Stem cell isolation from human Wharton's jelly: a study of their differentiation ability into lens fiber cells

Recently, the use of stem cells has expanded into numerous areas including cell therapy. In this study, we investigated the differentiation capacity of human Wharton's jelly stem cells [hWJSCs] into lens fiber cells. Morphological changes and expressions of four crystallin genes [Alpha A, Alpha B, Beta B1 and Beta B3] were studied. The bovine vitreous body has been shown to induce expression of crystallin genes in hWJSCs. By using the vitreous as a lens fiber cell inducer, we showed that Alpha B-, Beta B1- and Beta B3-crystallin genes expressed in hWJSCs

[Efficacy and safety of topical or oral methoxsalen plus ultraviolet A in the treatment of vitiligo]

Vitiligo is a common skin disorder that presents as depigmented patchs and associated with psychosocial morbidity. Photochemotherapy with methoxsalen plus ultraviolet A [PUVA] is an effective and commonly used modality in treatment of vitiligo and is available in Khorasan Province since more than ten years ago. This uncontrolled clinical trial included patients with vitiligo referred to phototherapy center in Emam Reza Hospital in Mashhad during 1998-1999. Treatment was done topically on patients with involvement less than 2% of the body area and systemic in cases with involvement more than that. Data including improvement rate and side effects were analyzed by PSS software version 11.5. One-hundred and thirty-five patients, 79 female and 56 male, were enrolled. One-hundred and twenty-six patients were treated systemically and 9 patients underwent topical therapy. Complete [100%] repigmentation was observed in one patient. Repigmentation of 80-99%, 60-79%, 40-59%, 20-39% and less than 20% were observed in 53, 43, 12, 6 and 20 patients; respectively. Results were significantly better in generalized vitiligo, face involvement, and cases whom received more than 200 treatment sessions, but there was not significant relationship between improvement and patients' age, family history, the first involved area, beginning time of repigmentation, the observed interval between initiation of treatment and disorder appearance, and skin type. Common side effects were pruritus [69.6%], erythema [52.6%], nausea and vomiting [28.9%], and headache [17.8%]. PUVA therapy is an effective and safe treatment in patients with vitiligo, and has good results in nonsegmental vitiligo and facial involvement

Accuracy rate for clinical diagnosis of dermatofibroma in patients referred to dermatology and pathology department of Emam Reza hospital

Considering its typical clinical manifestation, Dermatofibroma [DF] can be clinically diagnosed in most cases. It has been nevertheless, common practice to rely on pathologic diagnoses. We decided to estimate the rate of correct clinical diagnosis and also the relative importance of other lesions in differential diagnosis of DF. We studied archived biopsies in Dermatology and Pathology Department of Emam Reza hospitals from 1984 until 1 2004 and Dermatofibroma cases were selected, then slides were reviewed. Demographic and other relevant data such as age, sex, etc were gathered and analysed. Eighty two of 127 clinically diagnosed dermatofibroma cases were confirmed by histologic examination [positive predictive value 64.5%]. The most common histological diagnosis in the remaining 45 cases were nodular hidradenoma [7 cases] and comedon and epidermal cyst [11 cases collectively]. In addition, 32 cases of histologically diagnosed dermatofibroma were found with different clinical diagnosis. According to our data, the sensitivity of clinical diagnosis was 72%. It was also found that the lesion is more common in females [57%] and in extremities [58%]. It often appears as a nodular [72%] lesion measuring 5 to 15 millimeters. Histopathologic examination is important for diagnosis of Dermatofibroma. According to our data, the probabilities of false positive and false negatives were 35.5% and 28%, respectively, which is equivalent to 64.5% positive predictive value [PPV]

Evaluation of subtypes of peripheral blood lymphocytes in patients with vitiligo

Vitiligo is characterized with white patches on the skin and alteration of melanocytes in dermoepidermal junction. Autoimmune mechanisms with an underlying genetic predisposition are the most likely causes of vitiligo. This study was performed to evaluate immune disturbance in vitiligo and clarify its more details. A total of 29 vitiligo patients and 21 healthy controls were included in this case control study. Complete blood count was measured and peripheral blood samples were evaluated floweytometrically for surface antigenic markers including CD3, CD4, CD8, CD19, CD16, CD56 and CD25 for determining the percentage and total number of various lymphocyte subgroups. Patients with different clinical subtypes were compared with each other and controls in terms of the flowcytometry results. Obtained information was assessed by SPSS statistical software. Total numbers of CD3+, CD8+ T cells, B cells and CD25+ cells were significantly increased in generalized type vitiligo patients in comparison with localized type. CD25+ cells were also increased significantly in generalized and stable types of vitiligo compared with healthy controls and finally the total number of lymphocytes was significantly decreased in localized type vitiligo patients in comparison with healthy controls.. Our data indicate cellular immune disturbance in vitiligo. Disorders of immune regulatory system may play a major role in this context. Significant CD25+ or regulatory T cells increment in different clinical subtypes of the disease is in favor of the above hypothesis. Later and larger studies may result in new and effective routs of treatment for vitiligo acting through regulating immune system

evaluation of [sun protection factor] SPF accuracy of some sunscreens

The prevalence of photo-aging and skin cancers has been increasing in many parts of the world. To reduce cutaneous photodamage and skin carcinogenesis, primary prevention that an integral component of it is the use of sunscreens is recommended. The aim of this study was assessment of SPF accuracy in some Iranian sunscreens. Among healthy individuals with skin types I to III, 25 volunteers enrolled, but finally ten persons for two sunscreens "X" and "Y" completed the study. Candidates were tested during three days using phototherapy UV 800 K with UV B lamps, UV skin-tester and UV-meter [Waldman Company, Germany]. Minimal erythemal dose in unprotected areas of the skin [MEDup] was initially measured and then in protected areas [MEDp] was determined, according to the MEDup amount and national sunscreens' SPF label [SPF30]. Volunteer was exposed to his own MEDp and also 85% of it, after 24 hours, the two areas were examined for erythema. The final test result declared "correct", for the tested cream, if the accuracy of the SPF was confirmed in at least 90% of volunteers. The accuracy of test was confirmed according to SPF assessment standard protocol. All the candidates were male with mean age of 33.5 +/- 8.51 yrs. The MED mean was 73/09 +/- 15/6 mj/cm2. Final results declared "correct" for both products. In addition, the standard lotion test result was correct for all candidates. According to the results, we can assure people and physicians about reliability of labeled SPF, at least in some of the national products

Treatment of vitiligo with blister grafting technique

Surgical treatments of vitiligo are punch grafting, blister grafting, flip-top transplantation, split skin grafting, etc aiming at rebuilding of melanocytic population in those patients who do not respond to medical treatment The objective of this study was determination of efficacy of blister grafting technique in the treatment of vitiligo. This study was done on 10 patients with vitiligo of face and /or distal extremities who had received different medical treatments including PUVA and had not responded, and their diseases were stable. Blister in recipient site was created by cryotherapy and in donor site by using vacuum device. Then donor site blister was transferred to the recipient site and bath sites were covered by dressing. 10 patients [8 females and 2 males] with mean age of 31.2 +/- 11.4 years entered the study. After 1-6 weeks, first signs of repigmentation were observed and after 4 months complete repigmentation occured in 7 patients [70%] In two patients, a repigmentation of more than 50% was observed while in one patient no pigmentation was seen which was related to errors in surgical technique. Blister grafting surgery in limited patches of vitiligo which have not responded to medical treatments gives excellent results of prolonged repigmentation without any scar formation

Serum IgE level in chronic lupoid leishmaniasis

It appears that chronic lupoid leishmaniasis is the result of type 2 predominant T helper response to parasite and a defect in the down regulation of IL-4 production during infection. The objective of this study was to evaluate the underlying immune status in these patients and their predominant T helper activity we considered serum IgE as an indicator of TH2 activity and IL-4 production as it has been shown in atopic diathesis. In 34 cases of chronic lupoid leishmaniasis serum IgE level was measured and compared with 34 control cases of age and sex matched healthy individuals without atopic diathesis. P< 0.05 was considered statistically significant. There were 21 females and 13 males with a mean age of 14.35 +/- 8.3 years in the patients group. The mean age of the control group was 16.11 +/- 8.4 [P>0.05 and matched]. Nine patients had atopic diathesis. Mean serum IgE level in patients and in the control group was 102.6 +/- 22.4 i.u/ml and 135.6 +/- 24.9 i.u/ml, respectively [P>0.05 with no significant difference]. Mean serum IgE level in patients without atopic diathesis [25 cases] was 66.8 +/- 113 i.u/ml which was significantly lower than the control group [P<0.05]. In this study, serum IgE level in cases with chronic lupoid leishmaniasis was lower than the control group and it seems that in these patients, there is not an underlying Th2 over activity as it is seen in atopic diathesis

Serologic evaluation of HIV-1 and syphilis in patients with clinical anogenital wart

Genital infection with human papilloma virus [HPV] is the most common sexually transmitted disease in the United States. Patients having genital warts are at excessive risk of HIV and syphilis. Uninformed patients with high risk behaviors can help spread this condition in society. Therefore, CDC recommends HIV and syphilis screening for those with a sexual transmitted disease or a sexual risk factor. But this strategy is rarely considered in Iran. The aim of this study was to evaluate the cost-effectiveness of HIV and syphilis screening tests in patients with genital wart. In this case-control study, 100 patients with genital wart referring to Dermatology Department of Imam-Reza Hospital [August 2005 - August 2007] were included as the case group. This study was approved by the local ethics. The Control group was the same in number, age average, sex and marital status. All 200 patients studied considering HIV or syphilis infection. Finally, collected data were statistically analyzed by SPSS software, using chi[2] and student T test. Male to female ratio was 0.87. Majority of patients aged 30-39 years. Two patients were HIV positive and only one had Syphilis, without significant difference with the control group [Pv =0.29, Pv =0.50]. Screening is not cost effective in patients with asymptomatic anogenital warts. Since both HIV positive cases were intravenous drug abusers, this study emphasized on HIV and syphilis screening in patients with high risk behavior

[ assessment of the awareness of senior students about the effects of sunlight on the skin and necessity and methods of sun protection]

Nearly ninety percent [90%] of skin cancers occur in sun exposed areas. In this study the awareness of senior students of Mashhad University [Ferdowsi] about the effects of sunlight on the skin plus necessity and methods of sun protection is evaluated. In this descriptive study in 2004 at Mashhad University [Ferdowsi], 802 senior students of 9 majors enrolled. This study was approved by the local committee of Medical Ethics. The standard questionnaires including 24 questions distributed among students. The students divided into groups according to their score in the 24 awareness questions: weak [score

Acute hemorrhagic edema of infancy: a case report

Acute hemorrhagic edema of infancy [AHEI] is a rare skin disorder which was first described by Snow in 1913.Clinical manifestations are usually limited to cutaneous lesions and is most commonly observed in infants under the age of 1 year. Usually, there is a history of recent upper respiratory tract infection and antibiotic administration. Clinical presentation of this disease includes petechia and ecchymosis on the head, face and distal extremities. Herein, a typical case of AHEI in a 10-month-old infant is reported. Differential diagnoses of AHEI is discussed

Hereditary benign telangiectasia: Report of 6 cases in a family

Hereditary benign telangiectasia is an uncommon idiopathic skin disorder characterized by generalized telangiectases and angiomatous lesions of the skin. The diagnosis should be suspected in patients with such cutaneous lesions, positive family history, no associated bleeding problems, and no mucosal involvement. We present six cases of hereditary benign telangiectasia in three generations of one family