Objective:To investigate the expression of miR-383(Micro RNA-383)in osteosarcoma cells and to verify whether upregulation of miR-383 can enhance the therapeutic efficacy of bortezomib against osteosarcoma.Methods:Fluorescence in situ hybridization (FISH) was used to detect the expression of miR-383 in osteosarcoma and normal bone tissues. Real-time quantitative polymerase chain reaction (qRT-PCR) was employed to measure the expression of miR-383 in different osteosarcoma cell lines (SaoS-2, HOS, U-2OS, and MG63)and the osteoblast cell line hFOB 1.19.The proliferative capacity of osteosarcoma cells treated with 5 nmol/L and 10 nmol/L bortezomib was assessed using the cell counting kit-8 (CCK-8) with dimethyl sulfoxide (DMSO) as a control. The activity of caspase-3 was also measured. HOS and MG63 cells were treated with DMSO, bortezomib, miR-383 mimics, or negative controls, and the proliferative capacity and apoptosis levels were re-evaluated using CCK-8 and flow cytometry, respectively.Results:FISH results showed that the level of miR-383-5p in osteosarcoma tissues was significantly lower than that in normal bone tissues ( P<0.05). qRT-PCR results indicated that miR-383 levels in osteosarcoma cells (MG63, HOS, Saos-2, U-2OS) were lower than those in osteoblasts (hFOB1.19), with significant differences among different osteosarcoma cell lines(all P<0.05).The lowest levels of miR-383 were observed in HOS and MG63 cells. CCK-8 and caspase-3 activity assays revealed that among the cells treated with DMSO and two doses of bortezomib, HOS and MG63 cells had higher baseline proliferative capacity. Compared with DMSO-treated control cells, cells treated with 5 nmol/L and 10 nmol/L bortezomib exhibited inhibited proliferation (all P<0.05) and increased caspase-3 activity (all P<0.05). The effect of 10 nmol/L bortezomib was stronger than that of 5 nmol/L (all P<0.05). Compared with negative control-transfected cells, osteosarcoma cells (MG63 and HOS) with overexpressed miR-383 showed inhibited proliferation and increased apoptosis levels (all P<0.05). After bortezomib treatment, osteosarcoma cells (MG63 and HOS)with overexpressed miR-383 exhibited reduced proliferative capacity and enhanced apoptosis levels (all P<0.05). Conclusions:miR-383 exerts anticancer effects in osteosarcoma by inhibiting cell proliferation. Its overexpression significantly enhances the therapeutic efficacy of bortezomib, offering a new direction for the treatment strategies of osteosarcoma.

Objective:To investigate the expression of miR-383(Micro RNA-383)in osteosarcoma cells and to verify whether upregulation of miR-383 can enhance the therapeutic efficacy of bortezomib against osteosarcoma.Methods:Fluorescence in situ hybridization (FISH) was used to detect the expression of miR-383 in osteosarcoma and normal bone tissues. Real-time quantitative polymerase chain reaction (qRT-PCR) was employed to measure the expression of miR-383 in different osteosarcoma cell lines (SaoS-2, HOS, U-2OS, and MG63)and the osteoblast cell line hFOB 1.19.The proliferative capacity of osteosarcoma cells treated with 5 nmol/L and 10 nmol/L bortezomib was assessed using the cell counting kit-8 (CCK-8) with dimethyl sulfoxide (DMSO) as a control. The activity of caspase-3 was also measured. HOS and MG63 cells were treated with DMSO, bortezomib, miR-383 mimics, or negative controls, and the proliferative capacity and apoptosis levels were re-evaluated using CCK-8 and flow cytometry, respectively.Results:FISH results showed that the level of miR-383-5p in osteosarcoma tissues was significantly lower than that in normal bone tissues ( P<0.05). qRT-PCR results indicated that miR-383 levels in osteosarcoma cells (MG63, HOS, Saos-2, U-2OS) were lower than those in osteoblasts (hFOB1.19), with significant differences among different osteosarcoma cell lines(all P<0.05).The lowest levels of miR-383 were observed in HOS and MG63 cells. CCK-8 and caspase-3 activity assays revealed that among the cells treated with DMSO and two doses of bortezomib, HOS and MG63 cells had higher baseline proliferative capacity. Compared with DMSO-treated control cells, cells treated with 5 nmol/L and 10 nmol/L bortezomib exhibited inhibited proliferation (all P<0.05) and increased caspase-3 activity (all P<0.05). The effect of 10 nmol/L bortezomib was stronger than that of 5 nmol/L (all P<0.05). Compared with negative control-transfected cells, osteosarcoma cells (MG63 and HOS) with overexpressed miR-383 showed inhibited proliferation and increased apoptosis levels (all P<0.05). After bortezomib treatment, osteosarcoma cells (MG63 and HOS)with overexpressed miR-383 exhibited reduced proliferative capacity and enhanced apoptosis levels (all P<0.05). Conclusions:miR-383 exerts anticancer effects in osteosarcoma by inhibiting cell proliferation. Its overexpression significantly enhances the therapeutic efficacy of bortezomib, offering a new direction for the treatment strategies of osteosarcoma.

Objective:To explore the changes in the functional composition of genital tract microbiota in patients with recurrent implantation failure (RIF) and its relationship with cytokines related to immune regulation.Methods:This was a case-control study. The study included patients from the Reproductive Medicine Center of Peking University Third Hospital, enrolled from August 2020 to January 2021. The enrolled patients were divided into control group ( n=20) and the RIF group ( n=15). The study examined the expression of flora and cytokines in vaginal and uterine lavage fluid by 16S rRNA sequencing and MSD electrochemiluminescence. Spearman analysis was used to observe the correlation between flora and differently expressional cytokines. Results:Compared with control group, there were statistically significant differences in the Shannon index of vaginal microbiota and uterine lavage fluid microbiota in the RIF group ( P=0.032, P=0.015). There was a statistically significant difference in β diversity of the bacterial community in uterine lavage fluid ( P=0.020). The expression level of monocyte chemotactic protein-4 (MCP-4) in the RIF group [(3.39±1.23) ng/L] was significantly higher than that in control group [(1.07±0.31) ng/L, P=0.044], while the expression level of interleukin (IL)-17A [(1.12±0.29) ng/L] was significantly lower than that in control group [(2.70±0.52) ng/L, P=0.040]. The expression levels of interferon (IFN)-γ [(4.70±2.10) ng/L], IL-10 [(0.20±0.10) ng/L], tumor necrosis factor-α [(0.84±0.13) ng/L], and IL-5 [(0.35±0.05) ng/L] in the RIF group in the uterine lavage were significantly higher than those in control group [(0.97±0.30) ng/L, P=0.049; (0.05±0.01) ng/L, P=0.009; (0.56±0.07) ng/L, P=0.045; (0.23±0.03) ng/L, P=0.029]. Correlation analysis revealed significant correlations between vaginal flora and MCP-4 ( r=-0.42, P=0.025), and in the microbial community of uterine cavity lavage fluid, there was a significant positive correlation between the Muribaculaceae genus and IL-5 ( r=0.51, P=0.017). Conversely, the Vulcaniibacterium genus, Cupriavidus genus, and Anoxybacillus genus exhibited a significantly negative correlation with IL-5 ( r=-0.55, P=0.010; r=-0.62, P=0.003; r=-0.45, P=0.041). Additionally, the genera Anoxybacillus, Geobacillus, Thermus, and Meiothermus showed a significantly negative correlation with IFN-γ ( r=-0.43, P=0.015; r=-0.38, P=0.035; r=-0.39, P=0.029; r=-0.38, P=0.035). Conclusion:The reproductive tract microbiota and corresponding cytokines in RIF patients are disrupted, and there is a correlation between the microbiota and cytokines.

Objective:To explore the changes in the functional composition of genital tract microbiota in patients with recurrent implantation failure (RIF) and its relationship with cytokines related to immune regulation.Methods:This was a case-control study. The study included patients from the Reproductive Medicine Center of Peking University Third Hospital, enrolled from August 2020 to January 2021. The enrolled patients were divided into control group ( n=20) and the RIF group ( n=15). The study examined the expression of flora and cytokines in vaginal and uterine lavage fluid by 16S rRNA sequencing and MSD electrochemiluminescence. Spearman analysis was used to observe the correlation between flora and differently expressional cytokines. Results:Compared with control group, there were statistically significant differences in the Shannon index of vaginal microbiota and uterine lavage fluid microbiota in the RIF group ( P=0.032, P=0.015). There was a statistically significant difference in β diversity of the bacterial community in uterine lavage fluid ( P=0.020). The expression level of monocyte chemotactic protein-4 (MCP-4) in the RIF group [(3.39±1.23) ng/L] was significantly higher than that in control group [(1.07±0.31) ng/L, P=0.044], while the expression level of interleukin (IL)-17A [(1.12±0.29) ng/L] was significantly lower than that in control group [(2.70±0.52) ng/L, P=0.040]. The expression levels of interferon (IFN)-γ [(4.70±2.10) ng/L], IL-10 [(0.20±0.10) ng/L], tumor necrosis factor-α [(0.84±0.13) ng/L], and IL-5 [(0.35±0.05) ng/L] in the RIF group in the uterine lavage were significantly higher than those in control group [(0.97±0.30) ng/L, P=0.049; (0.05±0.01) ng/L, P=0.009; (0.56±0.07) ng/L, P=0.045; (0.23±0.03) ng/L, P=0.029]. Correlation analysis revealed significant correlations between vaginal flora and MCP-4 ( r=-0.42, P=0.025), and in the microbial community of uterine cavity lavage fluid, there was a significant positive correlation between the Muribaculaceae genus and IL-5 ( r=0.51, P=0.017). Conversely, the Vulcaniibacterium genus, Cupriavidus genus, and Anoxybacillus genus exhibited a significantly negative correlation with IL-5 ( r=-0.55, P=0.010; r=-0.62, P=0.003; r=-0.45, P=0.041). Additionally, the genera Anoxybacillus, Geobacillus, Thermus, and Meiothermus showed a significantly negative correlation with IFN-γ ( r=-0.43, P=0.015; r=-0.38, P=0.035; r=-0.39, P=0.029; r=-0.38, P=0.035). Conclusion:The reproductive tract microbiota and corresponding cytokines in RIF patients are disrupted, and there is a correlation between the microbiota and cytokines.

Objectives:To investigate whether adjuvant treatment with growth hormone (GH) can improve the pregnancy outcomes of patients with fresh embryo transfer after repeated implantation failures (RIF).Methods:This retrospective cohort study was performed in patients who underwent in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) fresh transfer cycle and were expected RIF from January 2016 to December 2020 in Reproductive Medicine Center, Department of Obstetrics and Gynecology, Peking University Third Hospital. Using SPSS26.0, a 1∶1 propensity score matching was conducted based on age, body mass index (BMI), infertility factors, and antral follicle count with a caliper value of 0.015. The data obtained were then divided into GH treatment group ( n=552) and non-GH treatment group ( n=552). The controlled ovarian stimulation outcomes, implantation rate, clinical pregnancy rate, and live birth rate were compared between the two groups. Results:The implantation rate, the clinical pregnancy rate and the live birth rate of RIF patients in GH treatment group and non-GH treatment group were not significantly different (all P>0.05). Furthermore, analysis of various groups based on ovulation induction protocols similarly indicated no statistically significant differences in live birth rate, implantation rate, and clinical pregnancy rate between the two groups (all P>0.05). Logistic regression also indicated that GH adjuvant therapy has no effect on the live birth rate in RIF patients ( OR=1.035, 95% CI: 0.720-1.487, P=0.854). Conclusion:Adjuvant GH treatment may not improve the live birth rate of fresh embryo transfer in RIF patients.

Objectives:To investigate whether adjuvant treatment with growth hormone (GH) can improve the pregnancy outcomes of patients with fresh embryo transfer after repeated implantation failures (RIF).Methods:This retrospective cohort study was performed in patients who underwent in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) fresh transfer cycle and were expected RIF from January 2016 to December 2020 in Reproductive Medicine Center, Department of Obstetrics and Gynecology, Peking University Third Hospital. Using SPSS26.0, a 1∶1 propensity score matching was conducted based on age, body mass index (BMI), infertility factors, and antral follicle count with a caliper value of 0.015. The data obtained were then divided into GH treatment group ( n=552) and non-GH treatment group ( n=552). The controlled ovarian stimulation outcomes, implantation rate, clinical pregnancy rate, and live birth rate were compared between the two groups. Results:The implantation rate, the clinical pregnancy rate and the live birth rate of RIF patients in GH treatment group and non-GH treatment group were not significantly different (all P>0.05). Furthermore, analysis of various groups based on ovulation induction protocols similarly indicated no statistically significant differences in live birth rate, implantation rate, and clinical pregnancy rate between the two groups (all P>0.05). Logistic regression also indicated that GH adjuvant therapy has no effect on the live birth rate in RIF patients ( OR=1.035, 95% CI: 0.720-1.487, P=0.854). Conclusion:Adjuvant GH treatment may not improve the live birth rate of fresh embryo transfer in RIF patients.

Female reproductive tract microbiota refers to the collection of microorganisms in female reproductive tract, its dynamics, function and interactions with host have been investigated in depth. In recent years, the advancement of next-generation sequencing and metagenomic sequencing and other technologies has transformed researches on reproductive tract microbial community, unveiling its possible impact on reproduction function. Furthermore, abnormal reproductive tract microbiota may predispose to infertility, miscarriage, premature rupture of membranes, premature delivery and other diseases. Here we review the current literature that focuses on female reproductive tract microbiota and highlight its possible role on female fertility.

Female reproductive tract microbiota refers to the collection of microorganisms in female reproductive tract, its dynamics, function and interactions with host have been investigated in depth. In recent years, the advancement of next-generation sequencing and metagenomic sequencing and other technologies has transformed researches on reproductive tract microbial community, unveiling its possible impact on reproduction function. Furthermore, abnormal reproductive tract microbiota may predispose to infertility, miscarriage, premature rupture of membranes, premature delivery and other diseases. Here we review the current literature that focuses on female reproductive tract microbiota and highlight its possible role on female fertility.

Achalasia of cardia caused by neuromuscular dysfunction at esophagus-stomach junction is a functional disease of esophageal dynamic dysfunction. It is characterized by absence of peristalsis of esophageal body and failure of the lower esophageal sphincter to relax. The methods of therapy include botulinum toxin injection, stent placement, laparoscopic Heller myotomy and balloon dilatation. However,these methods have some shortages,such as easy to recur, causing larger trauma,etc. . With the development of technology of endoscopy,a new method peroral endoscopic myotomy ( POEM)is widely used in clinical practice,and it can used in some special patients such asⅢ type achalasia,pediatric and elderly patients,as well as sigmoid-type achalasia. This article reviewed the current status and progress of POEM in treatment of achalasia of cardia.