Anti-proliferative and apoptotic effects of hull-less pumpkin extract on human papillary thyroid carcinoma cell line / 대한해부학회지

Papillary thyroid carcinoma (PTC) is one of the most common cancers of the endocrine system. Previous studies have shown that the extract of hull-less pumpkin seed (HLPS) has a significant anti-cancer effect. The aim of this study was to evaluate the effect of this plant extract on the proliferation of PTC cells. In this study, an extract of this plant was prepared by soxhlet extraction method and analyzed by Gas Chromatography-Mass Spectrometry. The cytotoxicity of PTX and plant extract was investigated using the methylthiazol tetrazolium (MTT) method. For careful investigation of morphological alteration, we used hematoxylin and eosin and Giemsa stinging. Based on MTT assay test, the IC 50 value of paclitaxel (PTX) was significantly less than the hydro-alcoholic extract of HLPS at all of the incubation time. Our results of histological staining showed that HLPS and PTX induced significant morphological alteration in the PTC cultured cell that consistent with cell death. Comparing the groups treated by PTX or HLPS with control group showed significant differences. It seems that HLPS extract has an apparent effect on treatment of PTC, at least in laboratory condition, albeit for realistic decision about the effect of HLPS on PTC, more molecular investigations are necessary.

Anti-proliferative and apoptotic effects of hull-less pumpkin extract on human papillary thyroid carcinoma cell line / 대한해부학회지

Papillary thyroid carcinoma (PTC) is one of the most common cancers of the endocrine system. Previous studies have shown that the extract of hull-less pumpkin seed (HLPS) has a significant anti-cancer effect. The aim of this study was to evaluate the effect of this plant extract on the proliferation of PTC cells. In this study, an extract of this plant was prepared by soxhlet extraction method and analyzed by Gas Chromatography-Mass Spectrometry. The cytotoxicity of PTX and plant extract was investigated using the methylthiazol tetrazolium (MTT) method. For careful investigation of morphological alteration, we used hematoxylin and eosin and Giemsa stinging. Based on MTT assay test, the IC 50 value of paclitaxel (PTX) was significantly less than the hydro-alcoholic extract of HLPS at all of the incubation time. Our results of histological staining showed that HLPS and PTX induced significant morphological alteration in the PTC cultured cell that consistent with cell death. Comparing the groups treated by PTX or HLPS with control group showed significant differences. It seems that HLPS extract has an apparent effect on treatment of PTC, at least in laboratory condition, albeit for realistic decision about the effect of HLPS on PTC, more molecular investigations are necessary.

From dentistry students' points of view: do anatomy classes we took actually boost our learning during clinic? / 대한해부학회지

The aim of medical education is to teach the essence of practical skills alongside with the theoretical knowledge. Teaching anatomy, as the center of medical education, should be leading to use this knowledge as a skill during clinical period. According to the rising numbers of dentistry faculties' experiences, inappropriate education results in misguidance during clinic. Thus, this study was conducted to find about the pre-clinical and clinical dentistry students' points of view on the helpfulness of anatomy classes in achieving clinical goals. Present descriptive cross-sectional study evaluated Guilan University of Medical Sciences' pre-clinical and clinical dentistry students' opinions on the effectiveness of anatomy classes during their clinical period in 2017. The sampling method used here was census via questionnaire and scoring was according to Likert scaling system. Analyses showed that anatomy of the nervous system was the most assistive course, which helped dentistry students during their clinical period (P<0.001). The least scored course was visceral organs and that means they did not use most of their learnings from classes with this topic (P<0.001). They also stated that other important factors such as using cadavers and moulages in practical sessions, teaching clinical skills theoretically before practical sessions and performing group activities are crucial for them to recall important details of the relevant courses during clinical period. Results of this study suggests that alongside with the various topics of anatomy courses, other factors like professors' characteristics and their teaching methods are also of important factors helping the dentistry students throughout clinic.

Relation between apel gene -141t>g polymorphism and idiopathic male infertility

Background: Infertility is defined as a lack of conception in a coupIe having unprotectes intercourse, for one year. Despite enormous progresses in the understanding of human reproductive physiology, the underlying causes of male infertility remains undefined in about 50.0% of cases, which are referred to as idiopathic infertility

Human apurinic/apyrimidinic endonuclease [ApEl] is a multifunctional protein that has an important role in the base excision repair [BER] pathway. ApEl -141 T>G polymorphism is located in the promoter region

The aim of this study was to investigate the relation between ApEl -141T>G polymorphism and idiopathic male infertility

Methods: hi this case-control study, Samples were collected from 90 patients diagnosed with idiopathic male infertility and also from 60 control subjects. Collected samples were genotyped by allele-specific PCR [AS-PCR]

Results: There was significantly association between -141TX/] polymorphism and idiopathic male infertility [p=0.022]

Conclusion: The polymorphism 141T>G can be associated with male infertility. However, further studies are needed to confirm the results

Oxytocin can decrease germ cells dapsgtotic index in testis under acute ischemia reperfaasion in a rat model

Background: Ischemia reperhs on [IR] is the main pathology of torsion of testis I and it is a common urologic emergency. There is some evidence that shows oxytocin I [OT] plays role in ischemia reperhsion. Objective: To evaluate this hypothesis that OT can decrease germ cell apoptotic index in testis under acute ischemia reperfusion in arat model. Materials and Methods: 20 adult rats were randomly divided into four groups: I Control, IR, OT and IF.+ OT [OTA]. Testicular ischemia was achieved by 720" torsion of the left testis for 2 hr. Then, torsion was removed and reperfusion was performed. Immediately after induction of reperfusion 0.03 pgkg OT were administered intraperitoneally to the IR+ OT. Three hours after surgery left testis I was removed and evaluations were made by Johnson's score, ELISA, immunohistochemistry and histomorphometry for study of maturity of spermatogenesis, endocrine profiles, apoptosis and quantitative studies, respectively. Results: The results showed in addition tissue edema and congestion, a significant reduced in Johnson's score were detected in IR group in comparison with controls [p=0.01], and apoptotic index increased significantly @=0.001]. Administration of OT in OT+IR group, increased Johnson's score but it was not statistically significant. Germinal epithelium thickness was increased significantly [p=0.03], although apoptotic index decreased significantly in comparison with the IR group [p=0.04]. However there was not significant difference in serum levels of I testosterone, FSH and LH innone of groups [p=0.07].Conclusion: These results suggested that OT can decrease apoptotic index and: improves complication of a cute ischemic reperftlsion in testis in a rat model

Relationship between seminal melatonin with sperm parameters, DNA Fragmentation and nuclear maturity in Intra-Cytoplasmic sperm injection candidates

Objective: Melatonin, the chief secretory product of the pineal gland, regulates dynamic physiological adaptations that occur in seasonally breeding mammals as a response to changes in daylight hours. Because of the presence of melatonin in semen and the membrane melatonin receptor in spermatozoa, the impact of melatonin on the regulation of male infertility is still questionable. The aim of this study was to determine the effects of endogenous melatonin on human semen parameters [sperm concentration, motility and normal morphology], DNA fragmentation [DF] and nuclear maturity

Materials and Methods: In this clinical prospective study, semen samples from 75 infertile men were routinely analyzed and assessed for melatonin and total antioxidant capacity [TAC] levels using the enzyme-linked immunosorbent assay [ELISA] and colorimetric assay kits, respectively. DF was examined by the sperm chromatin dispersion [SCD] test. Acidic aniline blue staining was used to detect chromatin defects in the sperm nuclei

Results: There was no significant correlation between seminal plasma melatonin and TAC with sperm parameters and nuclear maturity. However, we observed a positive significant correlation between DF and melatonin level [r=0.273, P<0.05]

Conclusion: Melatonin in seminal plasma is positively correlated with damaged sperm DNA of infertile patients. The mechanism of this phenomenon needs further study

Vitrification and subsequent in vitro maturation of mouse preantral follicles in presence of growth factors

Cryopreservation of ovarian tissue or follicles has been proposed as an alternative method for fertility preservation. Although successful vitrification of follicles has been reported in several mammalian species, the survival rate is generally low. The aim of this study was to investigate the effects of fibroblast growth factor [FGF] and epidermal growth factor [EGF] on in vitro preantral follicle development after vitrification. In this experimental study, preantral follicles with diameter of 150-180 microm were mechanically isolated from ovaries of 18-21 days old NMRI mice. Follicles were vitrified and warmed, then cultured in alpha-minimal essential medium [alpha-MEM] without growth factor supplementation as control group [group I], while supplemented with 20 ng/ml FGF [group II], 20 ng/ml EGF [group III], and 20 ng/ml FGF +20 ng/ml EGF [group IV]. After 12 days, human chorionic gonadotrophin [hCG]/EGF was added to culture medium, and after 18-20 hours, the presence of cumulus oocyte complexes [COCs] and oocyte maturation were assessed. The chi-square [chi2] test was used to analyze survival and ovulation rates of the follicles. Our results showed that the rate of metaphase II [MII] oocytes in FGF group increased in comparison with control and other treatment groups [p<0.027], but there was no difference between control with EGF and EGF+FGF groups in oocyte maturation rate [p>0.05]. There was a significant decrease in survival rate of follicles in EGF+FGE group in comparison with other groups [p<0.008]. After in vitro ovulation induction, the follicles in EGF group showed a higher ovulation rate [p<0.008] than those cultured in other groups. FGF has beneficial effect on oocyte maturation, and EGF increases COCs number in vitro. Combination of EGF and FGE decreases the number of survived follicles

impact of different time intervals between hCG priming and oocyte retrieval on ART outcomes

Abnormal oocyte morphology has been associated with the hormonal environment to which the gametes are exposed. In this study, we evaluated the oocytes morphology, fertilization rate, embryos quality, and implantation rate resulted of retrieved oocytes in different times after human chorionic gonadotrophin [HCG] administration. A total of 985 metaphase II oocytes were retrieved 35, 36, 37 and 38 h after the injection of HCG as groups 1, 2, 3, and 4 respectively. Oocyte morphology was divided into [I] normal morphology, [II] extracytoplasmic abnormalities, [III] cytoplasmic abnormalities and [IV] intracytoplasmic vacuoles and in each group, oocytes were evaluated according to this classification. Extracytoplasmic abnormalities were encountered in 17.76% and 31.1% of these oocytes [groups 3 and 4 respectively, p=0.007] in comparison with 12.23% group 2. Cytoplasmic abnormalities in group 4 were higher than other groups. 23.88% [p=0.039] and 43.25% [p=0.089] of resulted 2PN [two pronucleus] from groups 3 and 4 showed grade Z3 respectively in comparison to group 2 [16.44%]. Normal and various categories of abnormal oocytes did not differ regarding fertilization and cleavage rates [p=0.061]. However, group 4 showed significant difference in the rate of embryos fragmentation [grade III and IV embryo] in comparison with group 2 [40.96% vs. 24.93%, p=0.078]. The pregnancy rate was higher in G2 and G3 groups [28.5 and 24.13% respectively]. Oocyte retrieval time following HCG priming affected on oocyte morphology, 2PN pattern and embryos qualities subsequently. Both good quality embryo formation and pregnancy outcomes were noticeably higher when oocytes were retrieved 36 h after HCG priming in ART program

Evaluation of serial thawing-refreezing on human spermatozoa resistance using cryovials and straws

We designed this study to detect the cryoinjury rate on human sperm after serial freezing and thawing, taking into consideration the effects of using cryovials and straws. In this experimental study, semen specimens obtained from 15 subjects were divided into normozoospermic and oligozoospermic groups. Each of the normozoospermic and oligozoo spermic semen specimens were additionally divided into two groups: i. washed and ii. unwashed. Specimens were repeatedly freeze-thawed by using cryovials and straws with the fast liquid nitrogen vapor method, until no motile sperm remained. Sperm motility, recovery, and morphology rate were then determined after thawing, and compared between the groups while taking into consideration the effects of using cryovials and straws. Motile spermatozoa were observed in all normozoospermic samples up to thaw 6 with both cryovials and straws while in oligozoospermic specimens up to thaw 4 [straw] and thaw 3 [cryovial] in the freeze-thawing cycle. Normozoospermic sample analysis showed no significant difference in morphology rate. There was a significant increase in motility and recovery percentages for washed samples, which was observed with straws in compared to the unwashed groups. Oligozoospermic sample analysis indicated a significant increase in motility, recovery [p<0.01], and morphology [p<0.001] rates in washed specimens compared to unwashed specimens using straws. The importance of washing sperm was obvious for oligozoospermic specimens. Normozoospermic sperm resisted freezing longer than oligozoospermic sperm. Use of straws and cryovials made significant differences in motility, recovery, and morphology of sperm in each thaw. This difference was slightly higher for oligozoospermic specimens. Results indicated that the percentage of motility was higher for washed normozoospermic specimens in each thaw when straws were used, whereas the percentage of motility, recovery, and morphology were promoted after frozen oligozoospermic specimens were washed using straws.

effect of melatonin on the developmental potential and implantation rate of mouse embryos

Melatonin is a scavenger agent that has been used to promote in vitro embryo development. This study was designed to show the effects of melatonin on the quality and quantity rate of preimplantation mouse embryo development and pregnancy. In this experimental study, super ovulated, mated mice were killed by cervical dislocation to collect two-cell zygotes from the oviduct of pregnant 1 day NMRI mice. Zygotes were cultured to the hatching blastocyst stage and the numbers of embryos at different stages were recorded under an inverted microscope. The cleavage rates of two-cell zygotes were assayed until the blastocyst and hatching blastocyst stage in drops of T6 medium that contained either melatonin [1, 10, and 100×10[6], 10 and 100×10[9] M] or no melatonin. The cell numbers of blastocysts were determined by differential staining, implantation outcomes were studied, and development and pregnancy rate were compared by the Chi-square [development] and Fisher's exact [pregnancy rate] tests. The addition of 10 and 100 nM melatonin to the embryo culture media promoted the development of the two-cell stage embryos to blastocyst and hatching blastocysts [p<0.01] and caused a significant increase in total cell number [TCN], trophoectoderm [TE], and inner cell mass [ICM] of the blastocysts [p<0.01]. A difference was observed in the percentage of transferred embryos that were successfully implanted between the control and treatment groups [p<0.05]. The data indicate that 10 and 100 nM of melatonin positively impact mouse embryo cleavage rates, blastocyst TCN, and their implantation. Therefore, melatonin at low concentrations promotes an embryonic culture system in mice

effect of retinoic acid on in vitro maturation and fertilization rate of mouse germinal vesicle stage oocytes

Retinoids are recognized as important regulators of cell differention and tissue function, Previous studies, performed both in vivo and in vitro, indicate that retin-oids influence several reproductive events. In this study, we investigated the effect of all-trans retinoic acid [t-RA] on maturation and fertilization rate of immature oocytes [germinal vesicle]. Germinal vesicle [GV] oocytes were recovered from 4-6 week old female mice 48 hours after injection of 10 IU pregnant mare serum gonadotropin [PMSG]. Collected oocytes were divided into seven groups: control, sham and five experimental groups. t-RA at concentrations of 1, 2, 4, 6, 8 jjM were added to oocyte maturation medium in the experimental groups. The maturation rate was recorded after 24 hours of culture in a humidified atmosphere of 5% CO[2] at 37°C. Fertilization and developmental rates of matured oocytes were recorded after in vitro fertilization [IVF] and 24 hour culture. The rate of oocytes that developed to the metaphase II stage of maturation significantly increased with 2 and 4 microM t-RA compared to the control and sham groups [p<0.05]. In addition, the number of fertilized oocytes was significantly higher in 4 microM retinoic acid compared to the control [p<0.05], but the difference between the number of fertilized oocytes which developed to the 2-cell stage was not significant between the two groups. The results show that t-RA enhanced mouse oocyte maturation in vitro and improved fertilization and development rates in a dose dependent manner

Morphological identification of cell death in dorsal root ganglion neurons following peripheral nerve injury and repair in adult rat

Axotomy causes sensory neuronal loss. Reconnection of proximal and distal nerve ends by surgical repair improves neuronal survival. It is important to know the morphology of primary sensory neurons after the surgical repair of their peripheral processes. Animals [male Wistar rats] were exposed to models of sciatic nerve transection, direct epineurial suture repair of sciatic nerve, autograft repair of sciatic nerve, and sham operated. After 1 and 12 weeks of the surgery, the number of L5 dorsal root ganglion [DRG] and ultrastructure of L4-L5 DRG neurons was evaluated by fluorescence and electron microscopy, respectively. Nerve transection caused sensory neuronal loss and direct epineurial suture but no autograft repair method decreased it. Evaluation of morphology of the neurons showed classic features of apoptosis as well as destructive changes of cytoplasmic organelles such as mitochondria, rough endoplasmic reticulum and Golgi apparatus in primary sensory neurons. These nuclear and cytoplasmic changes in primary sensory neurons were observed after the surgical nerve repair too. The present study implies that the following peripheral nerve transection apoptosis as well as cytoplasmic cell death contributes to neuronal cell death and reconnection of proximal and distal nerve ends dose not prevent these processes

Morphometeric study of primary sensory neurons following surgical repair of sciatic nerve

Peripheral nerve transection causes neuonal cell death. Effects of surgical nerve repair on survival of primary sensory neurons was to be studied. This is a morphometeric study of dorsal root ganglion neurons of adult rats by Hoechst staining conducted at Neuroscience laboratory and Cellular and Molecular Center, Medical faculty, Iran University of Medical Sciences. Models of direct epineurial suture and using of nerve guidance channel of polyvinilydin flourid [PVDF] were used and neuronal number and volume of dorsal root ganglion of L5 were evaluated by morphometeric method. Direct epineurial suture technique decreased sensory neuronal loss and both direct epineurial suture and using of nerve guidance channel decreased reduction of the ganglion volume. Present study showed distal segment of peripheral nerve is essential for neuronal survival, but surgical nerve repair could not completely prevent from neuronal cell death

Immcinohistochemical study of spinal motor neurons following sciatic nerve repair in fldult rat

Epineural suture and autologous graft are two routine techniques in peripheral nerve surgery. However, their efficiency can be highly limited depending on the type of lesion and the gap between two nerve stumps and because of deficient proper nerve donors. So much interest has been focused on the development of alternative instruments for bridging the nerve gaps. In the present study, we have used charged polyvinelidene fluoride [PVDF] tube filled with nerve growth factor [NGF] and collagen gel as a substitute for nerve autograft and compared the results with other current surgical techniques. We studied the changes of spinal motoneurons to evaluate the effect of repairing techniques. In this study, 30 male Wistar rats weighing 200-250 g were divided randomly in five groups: axotomy, epineural suture, autograft, nerve guidance channel, and sham operation. In all experimental groups, the left sciatic nerve was transected at mid-thigh level. The nerve was not repaired in axotomy group. In epineural suture group, it was sutured end-to-end. In autograft group, a 10 mm piece of nerve was rotated 180° and sutured again in the nerve gap. Finally, in nerve guidance channel group, a piece of PVDF tube containing NGF7s [100 ng/ml] and collagen gel [1.28 mg/ml] was replaced in the gap. After one week, one month, and two months, L4-6 segments of spinal cord were removed and 5 urn paraffin sections were prepared for bax immunohistochemical study. In all groups contralateral spinal cord was used as the control. The proportion of Bax-positive apoptotic motoneurons was studied in all groups to evaluate the efficiency of different repairing techniques. Mean percentage of Bax-positive neurons to the total number of motor neurons in left side was analyzed. One way ANOVA showed significant difference after two months. LSD post hoc test showed that mean percentage of Bax-positive neurons in axotomized group was significantly higher compared to other surgical groups [p<0.05]. The number of apoptotic neurons after one week, one month and two months in each type of surgical approach showed no significant difference between one week and one month and between one month and two months. Comparison of motoneuron population in left side [experimental] with right side [control] showed no significant differences after one week, but significant differences were seen [p<0.01] after one month and two months. In sham group, no Bax-positive neuron was found after one week, one month, and two months. A PVDF tube filled with NGF and collagen gel can be used as a proper substitute for autografts and protect motoneurons following peripheral nerve injury