Elucidate the influence of RFRP-3 on follicular development and steroid hormone synthe-sis in mice.Forty 6-week-old female KM mice were randomly allocated into four groups,namely the 0(control),100,500 and 2 000 ng/d treatment groups.Each group received daily intraperitoneal injections of RFRP-3 for a duration of 7 days.The ovarian coefficients were calculated and recorded by measuring the wet weight of the ovaries.The effects of RFRP-3 at different concentrations on follicular maturation in mice were investigated through HE staining.Western blot analysis was em-ployed to assess the expression of apoptosis-related proteins,steroidogenic enzymes,and oocyte se-cretion factors within the mouse ovaries.The concentrations of progesterone(P4)and estradiol(E2)in the serum of the mice were determined using radioimmunoassay techniques.The results re-vealed no significant differences in body weight between the experimental groups and the control.However,the ovarian coefficients in the 500 and 2 000 ng/d treatment groups were significantly re-duced when compared to the control(P＜0.01).A marked decrease in the numbers of secondary and antral follicles was observed in the 500 ng/d and 2 000 ng/d groups(P＜0.01),concomitant with a substantial increase in atretic follicles(P＜0.01).A significant decline in follicle count was also noted in these groups(P＜0.01).Compared to the control,the protein expression of Bax and Caspase3 was substantially upregulated in the 500 and 2 000 ng/d treatment groups(P＜0.05),while the expression of Bcl-2 protein was significantly downregulated(P＜0.01).Similarly,the protein expression of 3βHSD,StAR,CYP11a1,and CYP17a1 was significantly decreased at the 500 and 2 000 ng/d groups(P＜0.01).E2 concentrations were moderately reduced in the 500 ng/d group(P＜0.05)and significantly decreased in the 2000 ng/d group(P＜0.01),whereas P4 levels remained unchanged across all experimental groups.Notably,the expression of BMP15 and GDF9 at both the gene and protein levels was markedly attenuated in the 500 and 2 000 ng/d treatment groups compared to the control(P＜0.05).In conclusion,RFRP-3 appears to inhibit follicular de-velopment and steroid hormone secretion in mice by modulating the expression of apoptotic fac-tors,oocyte secretion factors,and steroidogenic enzymes within the ovaries.This study provides a theoretical foundation for understanding the regulatory mechanisms of RFRP-3 on mammalian follicular development and steroid hormone synthesis.

Objective To investigate the effec of oscillating field stimulation on the proliferation of endogenous neural stem cell(ENSC)and recovery of motor function after spinal cord injury(SCI)in rats.Methods The SCI model was established by using the improved Allen's strike method and was randomly divided into experimental group and control group,with 60 rats in each group.The experimental group received an oscillating field implant as the intervention,while the control group was equipped with the same device but without activation.After successful modeling,the spinal cord of rats was removed on the 3rd,7th and 14th day,respectively.The proliferation of ENSC was detected by neuroepithelial stem cell protein(Nestin)/5-bromodeoxyuridine(BrdU)immunofluorescence,and the expression of Wnt-3a protein in spinal cord was detected by immunohistochemistry.At the 4th,6th and 8th week,the spinal cord was stained with osmic acid to observe the formation of myelin sheath in rats,and Basso-Beattle-Bresnahan(BBB)functional score of each group of rats was performed before sampling.Results The immunofluorescence results showed that Nestin/BrdU double positive cells in the injured area could be seen in both groups on the 3rd day after SCI,reaching a peak on the 7th day,and gradually decreased on the 14th day,and still maintained at a high level.At the same time,number of Nestin/BrdU double positive cells in experimental group rats at each time point was more than that in control group.The immunohistochemical results showed that the expression level of Wnt-3a protein in experimental group rats at each time point was higher than that in control group(P＜0.05).At the 6th and 8th weeks,the number of myelin sheaths in experimental group rats was significantly higher than that in control group(P＜0.05).At the 4 th,6th and 8th weeks,the BBB scores of rats in experimental group were significantly higher than those in control group(P＜0.05).Conclusion The oscillating field stimulation can induce the proliferation of ENSC after SCI by activating Wnt/β-catenin signaling pathway,promote myelin regeneration,and improve motor function of rats.

Objective To investigate the effec of oscillating field stimulation on the proliferation of endogenous neural stem cell(ENSC)and recovery of motor function after spinal cord injury(SCI)in rats.Methods The SCI model was established by using the improved Allen's strike method and was randomly divided into experimental group and control group,with 60 rats in each group.The experimental group received an oscillating field implant as the intervention,while the control group was equipped with the same device but without activation.After successful modeling,the spinal cord of rats was removed on the 3rd,7th and 14th day,respectively.The proliferation of ENSC was detected by neuroepithelial stem cell protein(Nestin)/5-bromodeoxyuridine(BrdU)immunofluorescence,and the expression of Wnt-3a protein in spinal cord was detected by immunohistochemistry.At the 4th,6th and 8th week,the spinal cord was stained with osmic acid to observe the formation of myelin sheath in rats,and Basso-Beattle-Bresnahan(BBB)functional score of each group of rats was performed before sampling.Results The immunofluorescence results showed that Nestin/BrdU double positive cells in the injured area could be seen in both groups on the 3rd day after SCI,reaching a peak on the 7th day,and gradually decreased on the 14th day,and still maintained at a high level.At the same time,number of Nestin/BrdU double positive cells in experimental group rats at each time point was more than that in control group.The immunohistochemical results showed that the expression level of Wnt-3a protein in experimental group rats at each time point was higher than that in control group(P＜0.05).At the 6th and 8th weeks,the number of myelin sheaths in experimental group rats was significantly higher than that in control group(P＜0.05).At the 4 th,6th and 8th weeks,the BBB scores of rats in experimental group were significantly higher than those in control group(P＜0.05).Conclusion The oscillating field stimulation can induce the proliferation of ENSC after SCI by activating Wnt/β-catenin signaling pathway,promote myelin regeneration,and improve motor function of rats.

Elucidate the influence of RFRP-3 on follicular development and steroid hormone synthe-sis in mice.Forty 6-week-old female KM mice were randomly allocated into four groups,namely the 0(control),100,500 and 2 000 ng/d treatment groups.Each group received daily intraperitoneal injections of RFRP-3 for a duration of 7 days.The ovarian coefficients were calculated and recorded by measuring the wet weight of the ovaries.The effects of RFRP-3 at different concentrations on follicular maturation in mice were investigated through HE staining.Western blot analysis was em-ployed to assess the expression of apoptosis-related proteins,steroidogenic enzymes,and oocyte se-cretion factors within the mouse ovaries.The concentrations of progesterone(P4)and estradiol(E2)in the serum of the mice were determined using radioimmunoassay techniques.The results re-vealed no significant differences in body weight between the experimental groups and the control.However,the ovarian coefficients in the 500 and 2 000 ng/d treatment groups were significantly re-duced when compared to the control(P＜0.01).A marked decrease in the numbers of secondary and antral follicles was observed in the 500 ng/d and 2 000 ng/d groups(P＜0.01),concomitant with a substantial increase in atretic follicles(P＜0.01).A significant decline in follicle count was also noted in these groups(P＜0.01).Compared to the control,the protein expression of Bax and Caspase3 was substantially upregulated in the 500 and 2 000 ng/d treatment groups(P＜0.05),while the expression of Bcl-2 protein was significantly downregulated(P＜0.01).Similarly,the protein expression of 3βHSD,StAR,CYP11a1,and CYP17a1 was significantly decreased at the 500 and 2 000 ng/d groups(P＜0.01).E2 concentrations were moderately reduced in the 500 ng/d group(P＜0.05)and significantly decreased in the 2000 ng/d group(P＜0.01),whereas P4 levels remained unchanged across all experimental groups.Notably,the expression of BMP15 and GDF9 at both the gene and protein levels was markedly attenuated in the 500 and 2 000 ng/d treatment groups compared to the control(P＜0.05).In conclusion,RFRP-3 appears to inhibit follicular de-velopment and steroid hormone secretion in mice by modulating the expression of apoptotic fac-tors,oocyte secretion factors,and steroidogenic enzymes within the ovaries.This study provides a theoretical foundation for understanding the regulatory mechanisms of RFRP-3 on mammalian follicular development and steroid hormone synthesis.

Objective:To explore the osteogenic activity of in vitro cultured preosteoblasts treated by magnesium loaded hydrogel core-shell microspheres.Methods:A microfluidics chip device was designed and made to fabricate magnesium loaded hydrogel core-shell microspheres to precisely control the stable release of magnesium ions in vitro,the mechanism of controllable magnesium ion release in the promotion of osteogenic activity of MC3T3-E1 preosteoblasts was systematically investigated.Results:The magnesium loaded hydro-gel microspheres(PLGA/MgO-alginate microspheres)prepared by the microfluidics chip exhibited to be monodisperse with special core-shell structure and could control the stable release of magnesium ions at a concentration of 50 ppm in vitro.The in vitro study showed that it significantly enhanced the activity and proliferation ability of MC3T3-E1 preosteoblasts,promoted their osteogenic differ-entiation and mineralization ability,and increased ALP activity.Conclusion:Magnesium loaded hydrogel core-shell microspheres can promote the osteogenic activity of MC3T3-E1 osteoblasts in vitro.

Medication during pregnancy is widespread, but there are few reports on its fetal safety. Recent studies suggest that medication during pregnancy can affect fetal morphological and functional development through multiple pathways, multiple organs, and multiple targets. Its mechanisms involve direct ways such as oxidative stress, epigenetic modification, and metabolic activation, and it may also be indirectly caused by placental dysfunction. Further studies have found that medication during pregnancy may also indirectly lead to multi-organ developmental programming, functional homeostasis changes, and susceptibility to related diseases in offspring by inducing fetal intrauterine exposure to too high or too low levels of maternal-derived glucocorticoids. The organ developmental toxicity and programming alterations caused by medication during pregnancy may also have gender differences and multi-generational genetic effects mediated by abnormal epigenetic modification. Combined with the latest research results of our laboratory, this paper reviews the latest research progress on the developmental toxicity and functional programming alterations of multiple organs in offspring induced by medication during pregnancy, which can provide a theoretical and experimental basis for rational medication during pregnancy and effective prevention and treatment of drug-related multiple fetal-originated diseases.

Ginsenoside Rc,a dammarane-type tetracyclic triterpenoid saponin primarily derived from Panax ginseng,has garnered significant attention due to its diverse pharmacological properties.This review outlined the sources,putative biosynthetic pathways,extraction,and quantification techniques,as well as the pharmacokinetic properties of ginsenoside Rc.Furthermore,this study explored the pharmaco-logical effects of ginsenoside Rc against metabolic syndrome(MetS)across various phenotypes including obesity,diabetes,atherosclerosis,non-alcoholic fatty liver disease,and osteoarthritis.It also highlighted the impact of ginsenoside Rc on multiple associated signaling molecules.In conclusion,the anti-MetS effect of ginsenoside Rc is characterized by its influence on multiple organs,multiple targets,and multiple ways.Although clinical investigations regarding the effects of ginsenoside Rc on MetS are limited,its proven safety and tolerability suggest its potential as an effective treatment option.

Purpose: This study aimed to investigate the factors associated with chemoresistance to neoadjuvant chemotherapy (NACT) followed by radical hysterectomy (RH) and construct a nomogram to predict the chemoresistance in patients with locally advanced cervical squamous carcinoma (LACSC). Materials and Methods: This retrospective study included 516 patients with International Federation of Gynecology and Obstetrics (2003) stage IB2 and IIA2 cervical cancer treated with NACT and RH between 2007 and 2017. Clinicopathologic data were collected, and patients were assigned to training (n=381) and validation (n=135) sets. Univariate and multivariate analyses were performed to analyze factors associated with chemoresistance to NACT. A nomogram was built using the multivariate logistic regression analysis results. We evaluated the discriminative ability and accuracy of the model using a concordance index and a calibration curve. The predictive probability of chemoresistance to NACT was defined as > 34%. Results: Multivariate analysis confirmed menopausal status, clinical tumor diameter, serum squamous cell carcinoma antigen level, and parametrial invasion on magnetic resonance imaging before treatment as independent prognostic factors associated with chemoresistance to NACT. The concordance indices of the nomogram for training and validation sets were 0.861 (95% confidence interval [CI], 0.822 to 0.900) and 0.807 (95% CI, 0.807 to 0.888), respectively. Calibration plots revealed a good fit between the modelpredicted probabilities and actual probabilities (Hosmer-Lemeshow test, p=0.597). Furthermore, grouping based on the nomogram was associated with progression-free survival. Conclusion We developed a nomogram for predicting chemoresistance in LACSC patients treated with RH. This nomogram can help physicians make clinical decisions regarding primary management and postoperative follow-up of the patients.

Objective: To establish a standard operation procedure (SOP) for ribosome genotyping (ribotyping) on Clostridioides (C.) difficile, supplement and verify ribotyping typing library, so as to improve the comparability of data between different laboratories and to develop surveillance network of C. difficil in China. Methods: Molecular typing of 54 reference strains from the United States and Europe of C. difficile were performed by using the SOP referencing correspondence from abroad and from our laboratory with a BioNumerics 7.6 software to estimate the reference library of types of C. difficile. Identification of 374 clinical and animal isolates of C. difficile from 13 cities in China between 2010 and 2018, to supplement the library information. Kappa test was used to evaluate the consistency. Results: Results of capillary electrophoresis of reference strains appeared clear and stable, which guaranteed the clustering results being fast and accurate. Results from the supplementary typing showed that there were 84 types of isolates, of which 25 RT types were consistent with reference strains from abroad, while 58 RT types were different from referenced types. In the 40 referenced types, 15 RT types were not found in this study. In the consistency evaluation, the Kappa value was 0.891 and (P<0.01), showing the two Molecular typing as consistent and with close resemblance. Conclusions The result of capillary electrophoresis by applying SOP for ribotyping on C. difficile base on QIAxcel capillary electrophoresis system, appeared clear and stable. The standardized library seemed more easily used for comparability and data sharing between the laboratories.

Objective: To investigate the effect of cinobufacin combined with As₂O₃ on the angiogenesis of subcutaneous transplantation tumor in colorectal carcinoma BALB/C nude mice. Methods: The colorectal carcinoma transplantation tumor model of BALB/C nude mice was established and divided into 4 groups, 5 mice in each group. The growth state of nude mice was observed by injecting the corresponding reagents into the tumor in As₂O₃ group, cinobufacin group, cinobufacin combined As₂O₃ group and the blank control group (replaced by phosphate buffer), respectively. The nude mice were killed two weeks later, and the tumor tissues, liver and kidney tissues and orbital vein blood were taken. The tumor volume inhibition rate and mass inhibition rate of nude mice were calculated. The histomorphology of tumor, liver and kidney and blood routine were detected. The expressions of vascular endothelial growth factor (VEGF), epidermal growth factor receptor (EGFR), fibroblast growth factor-basic (b-FGF) and CD105 in transplanted tumor tissues were detected by using immunohistochemistry method, and the microvascular density (MVD) of transplanted tumor in nude mice was evaluated. Western blot method was used to detect the protein expression levels of VEGF, EGFR and b-FGF. Results: After 2 weeks of administration, the tumor volume and tumor mass in As₂O₃ group, cinobufacin group and cinobufacin combined As₂O₃ group were lower than those in the blank control group. The volume inhibition rate was 16%, 17%, 72%, and the mass inhibition rate was 31%, 33%, 78%, respectively, and the difference was statistically significant (all P < 0.01); there was no statistical difference between As₂O₃ group and cinobufacin group in the tumor volume and tumor mass (P > 0.05), and cinobufacin combined As₂O₃ group had the most obvious therapeutic effects (all P < 0.05). The immunohistochemistry method showed that the expressions of VEGF, EGFR, b-FGF and MVD were decreased in As₂O₃ group, cinobufacin group and cinobufacin combined As₂O₃ group compared with the blank control group, and there were statistical differences of the four groups (all P < 0.05). There was no statistical difference of the marker changes in As₂O₃ group and cinobufacin group (all P > 0.05), and cinobufacin combined As₂O₃ group had the most significant decrease in the marker changes, and there was a significant difference compared with the other groups (all P < 0.05). Western blot showed that compared with the blank control group, the other three groups could downregulate the protein expressions of VEGF, EGFR and b-FGF in the transplanted tumor of nude mice. And the decreasing expression of each protein in cinobufacin combined As₂O₃ group was the most significant. Pathomorphology examination showed that the histomorphology of liver and kidney of the four groups of nude mice was normal. Blood routine examination showed that compared with the blank control group, the white blood cell count of nude mice in other groups was decreased, and the difference was statistically significant (all P < 0.05). The white blood cell count of cinobufacin combined As₂O₃ group was decreased most significantly; there was no statistically significant difference in hemoglobin and platelet count among the four groups (all P > 0.05). Conclusions Cinobufacin and As₂O₃ show synergistic effects in the tumor angiogenesis and inhibition of transplantation tumor growth of colorectal cancer BALB/C nude mice. Moreover, cinobufacin and As₂O₃ have no obvious toxicity to the hepatic, kidney and hematopoietic tissues.