Indirect co-culture of testicular cells with bone marrow mesenchymal stem cells leads to male germ cell-specific gene expressions

Objective: Non-obstructive azoospermia is mostly irreversible. Efforts to cure this type of infertility have led to the application of stem cells in the reproduction field. In the present study, testicular cell-mediated differentiation of male germ-like cells from bone marrow-derived mesenchymal stem cells [BM-MSCs] in an in vitro indirect co-culture system is investigated

Materials and Methods: In this experimental study, mouse BM-MSCs were isolated and cultured up to passage three. Identification of the cells was evaluated using specific surface markers by flow-cytometry technique. Four experimental groups were investigated: control, treatment with retinoic acid [RA], indirect co-culture with testicular cells, and combination of RA and indirect co-culture with testicular cells. Finally, following differentiation, the quantitative expression of germ cell-specific markers including Dazl, Piwil2 and Stra8 were evaluated by real-time polymerase chain reaction [PCR]

Results: Molecular analysis revealed a significant increase in Dazl expression in the indirect co-culture with testicular cells group in comparison to the control group. Quantitative expression level of Piwil2 was not significantly changed in comparison to the control group. Stra8 expression was significantly higher in RA group in comparison to other groups

Conclusion: Indirect co-culture of BM-MSCs in the presence of testicular cells leads to expression of male germ cell-specific gene, Dazl, in the induced cells. Combination of co-culture with testicular cells and RA did not show any positive effect on the specific gene expressions

Di-[2-ethylhexyl] phthalate-induced hippocampus-derived neural stem cells proliferation

The brain and spinal cord have a limited capacity for self-repair under damaged conditions. One of the best options to overcome these limitations involves the use of phytochemicals as potential therapeutic agents. In this study, we have aimed to investigate the effects of di-[2-ethylhexyl] phthalate [DEHP] on hippocampus-derived neural stem cells [NSCs] proliferation to search phytochemical candidates for possible treatment of neurological diseases using endogenous capacity. In this experimental study, neonatal rat hippocampus-derived NSCs were cultured and treated with various concentrations of DEHP [0, 100, 200, 400 and 600 micro M] and Cirsium vulgare [C. vulgare] hydroethanolic extract [0, 200, 400, 600, 800 and 1000 micro g/ml] for 48 hours under in vitro conditions. Cell proliferation rates and quantitative Sox2 gene expression were evaluated using MTT assay and real-time reverse transcription polymerase chain reaction [RT-PCR]. We observed the highest average growth rate in the 400 micro M DEHP and 800 micro g/ml C. vulgare extract treated groups. Sox2 expression in the DEHP-treated NSCs significantly increased compared to the control group. Gas chromatography/mass spectrometry [GC/ MS] results demonstrated that the active ingredients that naturally occurred in the C. vulgare hydroethanolic extract were 2-ethyl-1-hexanamine, n-heptacosane, 1-cyclopentanecarboxylic acid, 1-heptadecanamine, 2,6-octadien-1-ol,2,6,10,14,18,22-tetracosahexaene, and DEHP. DEHP profoundly stimulated NSCs proliferation through Sox2 gene overexpression. These results provide and opportunity for further use of the C. vulgure phytochemicals for prevention and/or treatment of neurological diseases via phytochemical mediated-proliferation of endogenous adult NSCs

Relationship between meiotic spindles visualization and intracytoplasmic sperm injection outcomes in human oocytes / 대한해부학회지

In assisted reproductive techniques, the operator attempts to select morphologically best embryos to predict embryo viability. Development of polarized light microscope, which evaluates the oocytes' spindles according to birefringence of living cells, had been helpful in oocyte selection. The aim of this study is evaluating the relationship between meiotic spindles visualization and intracytoplasmic sperm injection (ICSI) outcomes in human oocytes. In this study, 264 oocytes from 24 patients with an average age of 30.5±7.5 years with infertility duration of 1 to 10 years were collected. The oocytes were randomly allocated to the control injection group (n=126) and the oocyte imaging group (spindle-aligned group) (n=138). In the spindle-aligned group, the meiotic spindle was identified by means of polarized light microscope to align the spindle at 6 or 12 o'clock. Then the spindle-aligned group was divided into three sub-groups based on spindle morphology: fine, average, and (poor). After ICSI, embryos were checked every 24 hours and scored; 72 hours later, high-grade embryos were transferred intravaginally to uterus. This study showed that the fertilization rate in the spindle-aligned group was higher than the control group (P<0.05). After cleavage, a positive correlation was observed between spindle morphology and embryo morphology. Among the sub-groups of spindle-aligned group, the embryos' morphology of the fine group was better than the other subgroups and embryos of the poor group had lower quality and more fragmentation. The results revealed that the selection of oocytes based on meiotic spindle imaging can significantly improve the rate of fertilization and embryo cleavage and certainly increase the rate of implantation.

Applied anatomy, today's requirement for clinical medicine courses / 대한해부학회지

Anatomy as an indispensable part of the medical curricula, offering impeccable knowledge, prepares the students to enter the practical atmosphere. The aim of this study was to evaluate the clinical application of anatomy courses of the medical students in Zanjan University of Medical Sciences. This cross-sectional study was conducted in 2015 with census sampling on all clinical students (trainees and interns). To collect feedback from students, the questionnaire designed by researchers was used. The Likert rating scale of very high, high, medium, low, and very low was considered and scores of 5 (very high) to 1 (very low) were applied. Data were analyzed by SPSS software. Among the courses of anatomy, trunk anatomy has the greatest impact on clinical courses of medical students (P<0.001). Subjects of muscular system, lymphatic system, vascular system, and nervous system were of significant clinical application during clinical periods; however, no significant clinical application observed for skeletal system (P<0.05). Teaching clinical tips by professors can help improve the performance of medical students in clinical education. In addition, using three-dimensional anatomical software is suggested as well.

Effect of supraphysiological dose of Nandrolone Decanoate on the testis and testosterone concentration in mature and immature male rats: a time course study

Background: Most studies on anabolic-androgenic steroids abuse have been done in adult rats, but few data are available to immature

Objective: This study was conducted to assay the effect of Nandrolone Decanoate [ND] on the testis and testosterone concentration in male immature rats compare with mature ones in short and long time

Materials and Methods: 40 mature rats were divided into 4 groups: group A [short term] and group B [long-term] received 10 mg/kg/day ND interaperitoneally for 35 and 70 days, respectively. Group C [control] without any treatment, and group D [vehicle] received dimethyl sulfoxide [DMSO] solution in two periods 35 and 70 days. 40 immature rats were divided into 4 groups same as mature ones. After surgery body weight, testis size, histomorphometry of testis, and serum testosterone level were evaluated

Results: Our results showed that ND decreased the number of Leydig cells in group B [39.9 +/- 919], group A [43.4 +/- 120], and long term [40.6 +/- 299] immature rats, which could result in a reduction of testosterone concentration significantly in all experimental groups except short term mature group. Number of sertoli cells, testis size, and diameter of seminiferous tubules decreased in the long-term immature group. Eventually, the number of sperm was decreased in mature and immature groups, but a severe depletion of sperm was occurred in both mature and immature in long time in comparison to the control group [p< 0.05]

Conclusion: This time course study showed that supraphysiological dose of ND may negatively affect the number of Leydig cells, sperm cell, and testosterone concentration of immature rats in the same matter of mature rats. However, the number of sertoli cell, testis size, and seminferous diameter were decreased only in the long immature rats

Expression of surface markers and myogenic potential of rat bone marrow- and adipose-derived stem cells: a comparative study / 대한해부학회지

In recent years, examination and comparison of the biological characteristics of bone marrow- and adipose-derived mesenchymal stem cells (MSCs) from various perspectives have come into the focus of stem cell research, as these cells should be well characterized in order to utilize them in future cellular therapies. Therefore, in the present study, surface protein markers and the skeletal myogenic differentiation potential of rat bone marrow- and adipose-derived MSCs were examined. The expression of CD44, CD45, CD73, and CD90 on bone marrow- and adipose-derived MSCs was characterized using flow cytometry. Subsequently, the stem cells were differentiated into myogenic lineages, and the expression of the skeletal myogenic markers MyoD1, Myog, and Myh2 was studied in cells using real time polymerase chain reaction and immunofluorescence. Our results reveal that the pattern of CD marker expression differs between these 2 types of MSCs to some extent, whereas no significant difference was observed with respect to their myogenic differentiation potential. Therefore, we concluded that despite the differences observed in the biological features of these 2 types of MSCs, their myogenic potential appears to be similar, and that adipose-derived stem cells may be useful in skeletal muscle tissue engineering, due to their easy isolation and capacity for rapid expansion in a short time span.

[Surveying ergonomic factors of backpack in Tehranian primary school children]

Children are in their developing years and their bones and physical structure has not yet fully developed to a point of resiliency that can withstand stress, pressure, and a modicum of physical abuse and about 90% of students are carrying backpacks. Unsuitable backpacks cause shoulder pain, back pain, low back pain and skeletal malformation. Carrying backpacks heavier than 10% of body weight is not recommended. Thus, the objective of this study was to survey adaption between student backpacks and their anthropometric dimension. In this descriptive- analytical study, 50 primary school children [10 females and 40 males] cooperated in anthropometric measurement and its comparison with their backpack dimension. 212 female school children were contributed in comparison between their backpack weight and 10% body weight. Body dimensions were measured with VICON Analyze motion system [460V, oxford, UK] and ergonomics factors for their backpacks were recorded by a checklist. Weight measured with a digital scale and BMI was calculated based on WHO software. Significant difference was shown between shoulder breath-backpack width up, waist breath - backpack width down, shoulder width-shoulder strap width and back height-backpack height. 8% backpack handles, 44%shoulder straps and 34% backpack behinds had good padding. Two percent of backpacks had waist strap and no backpacks had chest strap. 8% of shoulder straps were S shaped and 4% of backpacks were rolling. Just 28.3% backpack weights have been under 10% of body weight. Ergonomics knowledge regarding backpacks is very important for selecting and buying the right product. However, the backpack producers should also be' informed to produce ergonomics backpack. The reduction of extra items in children backpacks by parents is beneficial, but effective management programs by ministry of education is also needed for informing school administrative persons and school children about the health problem of carring un ergonomic backpacks