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ObjectiveTo compare and evaluate the improvement degree of spermatogenic dysfunction mice at different recovery periods after cyclophosphamide modeling. MethodsForty-eight male ICR mice aged 4-5 weeks with the body weight of approximately 18-20 g were randomly divided into three control groups and three model groups, with 8 mice in each group. Each mouse of three model groups was intraperitoneally injected with 60 mg/kg cyclophosphamide continuously from the 1st to 7th day of the experiment, while each mouse of three control groups was intraperitoneally injected with the corresponding volume of normal saline. Then these mice were continued to be fed for another 7, 14 and 21 days after cyclophosphamide injection, respectively. A corresponding control group was set for each model group. The mice in each group were sacrificed after blood collection through orbital veins at corresponding time points. Testis, epididymis and seminal vesicle were taken and weighed, and their reproductive organ indexes were calculated. Histopathological changes of testis and epididymis were compared after HE staining.Sperm quality analysis was used to determine sperm-related indexes. Serum reproductive hormone content, testicular oxidative stress level and testicular signature enzyme activity were detected by ELISA and related kits.Results Compared with the control group, on the 7th, 14th and 21st day after cyclophosphamide treatment, the testicular index of mice in the model group decreased significantly (P<0.01). The epididymis index decreased significantly on the 7th and 14th day, and the seminal vesicle index decreased obviously on the 7th and 21st day (P<0.05). And the histopathological damage of testis and epididymis of the model group gradually alleviated over time. On the 7th and 14th day after cyclophosphamide treatment, the sperm count of the model group declined remarkably (P<0.01), the serum testosterone (T) level reduced (P<0.05), the malonaldehyde (MDA) content of testis increased significantly (P<0.01), the content of reduced glutathione (GSH) and superoxide dismutase (SOD) decreased obviously (P<0.05),the lactic dehydrogenase (LDH) activity of testis reduced obviously (P<0.05), the gamma-glutamyl transpeptidase (γ-GT) activity increased significantly (P<0.05), the latter two of which are important testicular signature enzymes. Therein on the 7th day after cyclophosphamide treatment, the sperm motility decreased significantly (P<0.001), the rate of sperm malformation increased obviously (P<0.05), the serum levels of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) increased notably (P<0.01). Nevertheless on the 21st day after cyclophosphamide treatment, the sperm-related indexes, the content of serum reproductive hormone, the level of testicular oxidative stress and the activity of testicular signature enzyme did not change significantly (P＞0.05). ConclusionThe reproductive toxicity in mice was more apparent on the 7th day after intraperitoneal injection with 60 mg/kg cyclophosphamide for seven days, at which time the more desirable spermatogenic dysfunction model of mice could be established. However, with the prolongation of the recovery period, the indexes of spermatogenic dysfunction in mice gradually recovered and approached the normal level on the 21st day after cyclophosphamide treatment.
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【Objective】 To investigate the effects of hsa_circ_0045943 targeting miR-106a on the biological characteristics of gastric cancer cells and its mechanism. 【Methods】 Human gastric cancer cells MKN-45， AGS and gastric mucosal epithelial cells GES-1 were cultured； circ_0045943 was detected by real-time polymerase chain reaction. The overexpression and silencing of circ_0045943 adenovirus vectors OE-circRNA and sh-circRNA together with their negative controls OE-NC and sh-NC were constructed and transfected； CCK-8 method was used to detect the proliferation activity of AGS cells after overexpression and silencing of circ_0045943； TUNEL method was used to detect the cell apoptosis； transwell assay was used to detect the cell migration and invasion； and would healing assay was used to detect the cell migration. Starbase database screened the binding site of miR-106a and circ_0045943. Real-time PCR was used to detect the expression of miR-106a， and the expression of circ_0045943 and the changes of miR-106a after the treatment of OE-circRNA and sh-circRNA. 【Results】 Real-time PCR showed that the expression of circ_0045943 decreased in gastric cancer cells MKN-45 and AGS compared to GES-1 （Pboth<0.001）. CCK-8 showed that the absorbance value of AGS cells in OE-circRNA group was lower than that in sh-circRNA group （P24 h<0.01， P48 h<0.001， and P72 h<0.001）. TUNEL showed the number of apoptotic AGS cells increased after overexpression of circ_0045943， but decreased after silencing of circ_0045943. Transwell assay showed that the migration and invasion of AGS cells were lower in OE-circRNA group than in sh-circRNA group （Pboth<0.001）. The wound healing assay showed that the migration rate of AGS cells in OE-circRNA group was the lowest， but was high in sh-circRNA group （P<0.001）. Starbase retrieved that circ_0045943 and miR-106a had complementary binding sequences. Real-time PCR showed that miR-106a was highly expressed in gastric cancer cells （P<0.001）， and the expressions of circ_0045943 and miR-106a significantly differed after treatment with OE-circRNA and sh-circRNA （Pboth<0.001）. With the increase or decrease of circ_0045943， the expression of miR-106a changed in the opposite direction. 【Conclusion】 Circ_0045943 has low expression in gastric cancer， and promoting or inhibiting circ_0045943 expression may regulate the proliferation， apoptosis， migration and invasion of gastric cancer cells by targeting miR-106a.
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Just as "the market failures" in business or "the government failures" in government department, the non - profit organizations failures is displaying on kinds of forms. The reason that the non - profit organizations failed includ: seeking profit, the serious insufficiency of the organization funds, the morals out of control, the unscientific achievements evaluation system. The countermeasure settle of non - profit organizations failures include: to establish the information publishing system and the corresponding punishing system, perfecting the organization management system, forming the multiplicated financeing mechanism leading by government, perfecting the moral restraint mechanism which the autonomy and the heteronomy unifies.