Background:To compare the efficacy and safety of monthly administrations of gonadotropin releasing hormone (GnRH) agonists LY01005 and Zoladex ? in Chinese patients with premenopausal breast cancer. Methods:From October 2020 to November 2021, 188 premenopausal breast cancer patients were enrolled in 34 hospitals and randomized 1:1 to receive either LY01005 or Zoladex ? every 28 days for a total of three injections. All patients concomitantly received oral tamoxifen (TAM). The primary efficacy endpoint was cumulative probability of maintaining menopausal level [oestradiol (E2) ≤30 pg/ml] from day 29 to day 85. The second efficacy endpoint included changes in E2, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) compared with the baseline. Pharmacokinetics (PK), pharmacodynamics (PD), and safety were analyzed. The study also evaluated the pharmacokinetic and pharmacodynamic characteristics of LY01005. Results:A total of 188 patients were randomised and 187 patients received either LY01005 or Zoladex ?. Cumulative probabilities of maintaining menopausal level (E2≤30 pg/ml) from day 29 to day 85 were 93.1% for LY01005 and 86.3% for Zoladex ?. The between-group difference was 6.8% (95% CI: -2.3%, 15.9%) and primary efficacy in the LY01005 group was not inferior to that in the Zoladex ? group. Changes in E2, LH, and FSH levels compared with the baseline were equivalent between the two groups (E2: 89.34% to 90.23% vs. 82.11% to 85.02%; LH: 88.89% to 95.52% vs. 89.70% to 97.02%; FSH: 75.36% to 80.85% vs.73.07% to 80.24%, respectively). After three consecutive doses of LY01005, the LH and FSH levels of the subjects showed a transient increase after the first dose, reached a peak on the second day and then started to decrease. The LH and FSH reached a lower level and remained at or below that level until the 85th day. Both treatments were well-tolerated. Conclusion:LY01005 is as effective as Zoladex ? in suppressing E2 to menopausal levels in Chinese patients with premenopausal breast cancer, with a similar safety profile.

Objective To investigate the effect of Biejiajian Pill(BJJP)on malignant biological behavior of hepatocellular carcinoma Hep3B cells and its regulatory mechanism.Methods A total of 72 healthy SD rats were randomly divided into blank control(BC)group,low(0.55 g/kg),medium(1.10 g/kg)and high(2.20 g/kg)BJJP experimental group,and drug-containing serum was prepared.Hep3B cells were divided into BC group,normal rat serum treatment(NC)group,low dose BJJP(LBJJP)group,medium dose BJJP(MBJJP)group and high dose BJJP(HBJJP)group,empty plasmid(pcDNA3.1)group,and CKLF-like MARVEL transmembrane domain containing 6(CMTM6)overexpression(pcDNA3.1-CMTM6)group,and the NC+pcDNA3.1 group,MBJJP+pcDNA3.1 group,NC+pcDNA3.1-CMTM6 group and MBJJP+pcDNA3.1-CMTM6 group.The proliferation of hepatoma Hep3B cells was detected by CCK-8.The migration and invasion of hepatoma Hep3B cells were detected by Transwell assay.The expression levels of proliferation-related proteins proliferating cell nuclear antigen(PCNA),epithelial-mesenchymal transition(EMT)related proteins(E-cadherin,N-cadherin and Vimentin),and CMTM6 proteins in hepatoma Hep3B cells were detected by Western blotting experiments.Results Compared with those in BC group,there were no significant differences in the proliferation,migration and invasion abilities of Hep3B cells,or the expression levels of PCNA,EMT related proteins(E-cadherin,N-cadherin and Vimentin)and CMTM6 protein in NC group(P＞0.05).Compared with NC group,LBJJP,MBJJP and HBJJP drug-containing serum inhibited the proliferation,migration and invasion of Hep3B cells,downregulated the protein expression of PCNA;MBJJP and HBJJP upregulated the protein expression of E-cadherin.The protein expressions of N-cadherin,Vimentin and CMTM6 were downregulated,with significant differences(P＜0.05).Compared with pcDNA3.1 group,the protein expression of CMTM6,cell proliferation,migration,invasion,PCNA protein expression,N-cadherin protein expression,and Vimentin protein expression in Hep3B cells in pcDNA3.1-CMTM6 group were significantly upregulated,while the protein expression of E-cadherin was significantly downregulated(P＜0.05).Compared with NC+pcDNA3.1 group,the proliferation,migration and invasion abilities of Hep3B cells in MBJJP+pcDNA3.1 group were decreased,the expression levels of PCNA,Vimentin and N-cadherin protein were decreased,while the expression level of E-cadherin protein was increased.Compared with NC+pcDNA3.1-CMTM6 group,MBJJP+pcDNA3.1-CMTM6 group had the same results in the proliferation,migration and invasion abilities of Hep3B cells and the protein expression levels of PCNA,Vimentin,N-cadherin and E-cadherin.The differences were statistically significant(all P＜0.05).Conclusion BJJP may inhibit the proliferation,migration,invasion and EMT of hepatoma Hep3B cells by regulating the expression of CMTM6.

Objective:To evaluate the impact of oral melatonin on clinical pregnancy outcomes in elderly infer-tile patients with diminished ovarian reserve(DOR)after in vitro fertilization and embryo transfer(IVF-ET).Methods:Sixty-two elderly infertile patients with DOR who underwent IVF at the Reproductive Medicine Center of Qinghai Province People's Hospital from January 1,2022 to June 30,2023 were selected.They were divided into intervention group(24 cases)and control group(38 cases)according to whether they received oral melatonin pretreatment before ovulation induction.Follicular fluid without diluent or blood contamination was collected from the first large follicle on the day of oocyte retrieval,and melatonin levels and 8-hydroxy-2'-deoxyglycoside(8-OHdG)levels in the follicular gluid were measured.Ovulation promotion indicators and pregnancy outcomes were compared and analyzed between the two groups.Results:The concentration of melatonin in follicular fluid in the intervention group was significantly higher than that in the control group(31.75 ng/L vs.10.40 ng/L,P＜0.001),the concentration of 8-OHdG was lower than that of the control group(133.61 ng/L vs.145.30 ng/L,P=0.004);the implantation rate in the intervention group was significantly higher than that in the control group(56.10％vs.26.87％,P=0.002),and the biochemical pregnancy rate in the intervention group was significantly higher than that in the control group(12.50％vs.0,P=0.050).There were no significant differences in normal fertilization rate,high-quality embryo rate,clinical pregnancy rate and cumulative live birth rate between the two groups(P＞0.05).Conclusions:Oral melatonin can increase its level in follicular fluid,which is beneficial in improving endom-etrial receptivity,and has a certain positive effect on improving IVF-ET outcomes.

Abstract Hyperuricemia (HUA) is a metabolic disorder syndrome caused by purine metabolism dysregulation, and its prevalence increases year by year. The development and progression of HUA are accompanied by significant alterations in the composition of intestinal microbiota, making probiotics a potential and safe method to reduce serum uric acid. Probiotics ameliorate HUA through three pathways: competing with intestinal epithelial cells for purine absorption to decrease uric acid synthesis, inhibiting xanthine oxidase activity through modulation of inflammatory cytokines to reduce the conversion of purine to uric acid, as well as restoring and maintaining an orderly state of the gut microbiota to facilitate normal uric acid excretion. This article reviews the role of probiotics in ameliorating HUA, so as to provide the reference for the application of probiotics in the prevention and intervention of HUA.

Objective: To evaluate the effect of bioactive peptides combined with probiotics on serum uric acid (SUA) in patients with hyperuricemia (HUA), so as to provide the evidence for prevention and treatment of HUA. Methods: The patients with HUA aged 18 to 65 years were selected and randomly divided into an intervention group and a control group. The patients in the intervention group received bioactive peptides combined with probiotics for 28 days at a dose of 3 g/d, while the patients in the control group received an equal dose of placebos. Demographic information, body mass index (BMI), blood pressure and blood lipid were collected through questionnaire surveys, physical examination and laboratory tests. SUA levels were detected before and after 14 days and 28 days of interventions. The differences of SUA levels between the two groups were compared using generalized estimation equation. Results: Totally 108 patients with HUA were recruited, including 54 patients in the intervention group and 53 patients in the control group (1 dropout). Before interventions, there were no statistically significant differences in gender, age, course of HUA, exercise duration, frequency of alcohol consumption, frequency of meat broth consumption, BMI, prevalence of hypertension and prevalence of dyslipidemia between the two groups (all P>0.05). After 14 days of interventions, the SUA levels of the patients in the intervention group decreased by 3.00 μmol/L, while those in the control group increased by 7.00 μmol/L. After 28 days of interventions, the SUA levels of the patients in the intervention group and the control group decreased by 26.00 μmol/L and 16.00 μmol/L, respectively. However, there was no statistically significant interaction between the intervention time and group (both P>0.05). Subgroup analysis showed that after 28 days of interventions, the decrease in SUA levels in the patients aged 55 years and older and without hypertension in the intervention group was greater than those in the control group (both P<0.05). Conclusions Bioactive peptides combined with probiotics showed no significant difference in reducing SUA levels in patients with HUA compared to the control group. The effect was more significant for patients aged 55 years and older and without hypertension.

Objective: To investigate the association between plant-based diet and different types of obesity, so as to provide references for obesity prevention. Methods: Residents aged 35-75 years from 33 counties (cities, districts) in Zhejiang Province were selected as study subjects using a multistage stratified random sampling method between April and December 2024. Demographic information and living behaviors were collected using questionnaire surveys. Height, weight and waist circumference were measured, and body mass index (BMI) was calculated. BMI ≥28.0 kg/m2 was defined as obesity, waist circumference ≥90 cm in males or ≥85 cm in females was defined as central obesity, and individual with obesity who also had central obesity was defined as having compound obesity. Food intake over a 3-day period was collected using the consecutive 3-day 24-hour dietary recall method. The plant diet index (PDI), healthful plant diet index (HPDI), and unhealthful plant diet index (UPDI) were calculated, and categorized into quintiles (Q1-Q5) based on their distribution. Association between the PDI, PDI, UPDI and different types of obesity were analyzed using multivariable logistic regression models. Results: A total of 4 882 individuals were surveyed, including 2 233 males (45.74%) and 2 649 females (54.26%). The average age was (55.42±12.14) years. There were 537 individuals of obesity, 1 718 individuals of central obesity, and 500 individuals of compound obesity, with detection rates of 11.00%, 35.19%, and 10.24%, respectively. Multivariable logistic regression analysis showed that, after adjusting for demographic information and living behaviors, compared with Q1 group, HPDI Q5 group showed a 29.6% lower risk of obesity (OR=0.704, 95%CI: 0.525-0.943) and a 32.1% lower risk of compound obesity (OR=0.679, 95%CI: 0.502-0.918). Conversely, the UPDI Q5 group exhibited a 39.5% higher risk of obesity (OR=1.395, 95%CI: 1.032-1.886) and a 39.8% higher risk of compound obesity (OR=1.398, 95%CI: 1.025-1.907). No statistically significant association was found between PDI and obesity, central obesity, and compound obesity (all P>0.05). As HPDI increased, the risks of obesity and compound obesity showed decreasing trends; as UPDI increased, the risks of obesity and compound obesity showed increasing trends (all Ptrend<0.05). Conclusion A healthful plant-based diet is associated with reduced risks of obesity and compound obesity, while an unhealthful plant-based diet is associated with increased risks of obesity and compound obesity.

Objective To investigate the effect of Biejiajian Pill(BJJP)on malignant biological behavior of hepatocellular carcinoma Hep3B cells and its regulatory mechanism.Methods A total of 72 healthy SD rats were randomly divided into blank control(BC)group,low(0.55 g/kg),medium(1.10 g/kg)and high(2.20 g/kg)BJJP experimental group,and drug-containing serum was prepared.Hep3B cells were divided into BC group,normal rat serum treatment(NC)group,low dose BJJP(LBJJP)group,medium dose BJJP(MBJJP)group and high dose BJJP(HBJJP)group,empty plasmid(pcDNA3.1)group,and CKLF-like MARVEL transmembrane domain containing 6(CMTM6)overexpression(pcDNA3.1-CMTM6)group,and the NC+pcDNA3.1 group,MBJJP+pcDNA3.1 group,NC+pcDNA3.1-CMTM6 group and MBJJP+pcDNA3.1-CMTM6 group.The proliferation of hepatoma Hep3B cells was detected by CCK-8.The migration and invasion of hepatoma Hep3B cells were detected by Transwell assay.The expression levels of proliferation-related proteins proliferating cell nuclear antigen(PCNA),epithelial-mesenchymal transition(EMT)related proteins(E-cadherin,N-cadherin and Vimentin),and CMTM6 proteins in hepatoma Hep3B cells were detected by Western blotting experiments.Results Compared with those in BC group,there were no significant differences in the proliferation,migration and invasion abilities of Hep3B cells,or the expression levels of PCNA,EMT related proteins(E-cadherin,N-cadherin and Vimentin)and CMTM6 protein in NC group(P＞0.05).Compared with NC group,LBJJP,MBJJP and HBJJP drug-containing serum inhibited the proliferation,migration and invasion of Hep3B cells,downregulated the protein expression of PCNA;MBJJP and HBJJP upregulated the protein expression of E-cadherin.The protein expressions of N-cadherin,Vimentin and CMTM6 were downregulated,with significant differences(P＜0.05).Compared with pcDNA3.1 group,the protein expression of CMTM6,cell proliferation,migration,invasion,PCNA protein expression,N-cadherin protein expression,and Vimentin protein expression in Hep3B cells in pcDNA3.1-CMTM6 group were significantly upregulated,while the protein expression of E-cadherin was significantly downregulated(P＜0.05).Compared with NC+pcDNA3.1 group,the proliferation,migration and invasion abilities of Hep3B cells in MBJJP+pcDNA3.1 group were decreased,the expression levels of PCNA,Vimentin and N-cadherin protein were decreased,while the expression level of E-cadherin protein was increased.Compared with NC+pcDNA3.1-CMTM6 group,MBJJP+pcDNA3.1-CMTM6 group had the same results in the proliferation,migration and invasion abilities of Hep3B cells and the protein expression levels of PCNA,Vimentin,N-cadherin and E-cadherin.The differences were statistically significant(all P＜0.05).Conclusion BJJP may inhibit the proliferation,migration,invasion and EMT of hepatoma Hep3B cells by regulating the expression of CMTM6.

AIM:This study aims to investigate the mechanisms through which Astragaloside IV(AS)pre-vents and treats osteoporosis by regulating intestinal flora.METHODS:Thirty 3-month-old female Sprague-Dawley(SD)rats were selected for the study.Ten rats were randomly assigned to a sham group,while the remaining twenty underwent bilateral ovariectomy(OVX)to simulate osteoporosis.Following the modeling,the twenty OVX rats were randomly divid-ed into two groups:the OVX group and the AS treatment group,which received continuous gavage for 12 weeks.Bone mineral density(BMD)of the femur and lumbar vertebrae was measured using dual-energy X-ray absorptiometry(DXA).Hematoxylin-eosin(HE)staining was employed to assess the microstructure of the femur and colonic mucosa,while immu-nohistochemistry was used to measure the expression of collagen type Ⅰ alpha 1 chain(COL1A1)protein in the femur.Ad-ditionally,RT-qPCR was utilized to analyze the mRNA expression of bone formation-related indicators,including alkaline phosphatase(ALP),COL1A1,and Runt-related transcription factor 2(RUNX2).Fresh fecal samples were collected from the rats for 16S rDNA sequencing to detect changes in intestinal microbiota composition.RESULTS:Compared to the sham group,OVX rats exhibited a significant increase in body weight and a marked decrease in femur and lumbar ver-tebrae bone density.HE staining revealed trabecular bone fractures with a disrupted reticular structure in the OVX group,along with the presence of numerous cavities and fat vacuoles in the bone marrow.The colonic mucosa showed signs of vil-lous shedding and mild crypt atrophy.Immunohistochemistry results demonstrated a substantial reduction in brown-yellow granules and COL1A1 expression in the OVX group.Conversely,in the AS group,there was a reduction in body weight and a significant increase in bone density of the femur and lumbar vertebrae.The trabecular architecture appeared more or-ganized,with less severe fractures compared to the OVX group.In the AS group,the number of cavities and fat vacuoles in the bone marrow was also reduced,and the colonic mucosa exhibited improved villous structure and less crypt atrophy.Immunohistochemical analysis indicated that AS treatment significantly enhanced COL1A1 expression.Furthermore,after AS intervention,the mRNA expression levels of ALP,COL1A1,and RUNX2 were notably increased.16S rDNA sequenc-ing revealed a significant increase in the abundance of Firmicutes,Proteobacteria,f_Pseudonocardiaceae,f_Marinifilace-ae,f_Oscillospiraceae,f_Ruminococcaceae,and f_Peptostreptococcaceae,while p_Euryarchaeota,Bacteroidetes,and f_Muribaculaceae showed significant reductions.Overall,OVX led to increased diversity in the species distribution of in-testinal microbiota,whereas AS treatment helped recalibrate the aforementioned phyla(families)and reduce diversity.CONCLUSION:Astragaloside IV can increase bone density in OVX rats,improve bone microstructure,promote bone formation,and prevent colonic mucosal damage by regulating the relative abundance of intestinal flora.

Objective:To evaluate the impact of oral melatonin on clinical pregnancy outcomes in elderly infer-tile patients with diminished ovarian reserve(DOR)after in vitro fertilization and embryo transfer(IVF-ET).Methods:Sixty-two elderly infertile patients with DOR who underwent IVF at the Reproductive Medicine Center of Qinghai Province People's Hospital from January 1,2022 to June 30,2023 were selected.They were divided into intervention group(24 cases)and control group(38 cases)according to whether they received oral melatonin pretreatment before ovulation induction.Follicular fluid without diluent or blood contamination was collected from the first large follicle on the day of oocyte retrieval,and melatonin levels and 8-hydroxy-2'-deoxyglycoside(8-OHdG)levels in the follicular gluid were measured.Ovulation promotion indicators and pregnancy outcomes were compared and analyzed between the two groups.Results:The concentration of melatonin in follicular fluid in the intervention group was significantly higher than that in the control group(31.75 ng/L vs.10.40 ng/L,P＜0.001),the concentration of 8-OHdG was lower than that of the control group(133.61 ng/L vs.145.30 ng/L,P=0.004);the implantation rate in the intervention group was significantly higher than that in the control group(56.10％vs.26.87％,P=0.002),and the biochemical pregnancy rate in the intervention group was significantly higher than that in the control group(12.50％vs.0,P=0.050).There were no significant differences in normal fertilization rate,high-quality embryo rate,clinical pregnancy rate and cumulative live birth rate between the two groups(P＞0.05).Conclusions:Oral melatonin can increase its level in follicular fluid,which is beneficial in improving endom-etrial receptivity,and has a certain positive effect on improving IVF-ET outcomes.

AIM:This study aims to investigate the mechanisms through which Astragaloside IV(AS)pre-vents and treats osteoporosis by regulating intestinal flora.METHODS:Thirty 3-month-old female Sprague-Dawley(SD)rats were selected for the study.Ten rats were randomly assigned to a sham group,while the remaining twenty underwent bilateral ovariectomy(OVX)to simulate osteoporosis.Following the modeling,the twenty OVX rats were randomly divid-ed into two groups:the OVX group and the AS treatment group,which received continuous gavage for 12 weeks.Bone mineral density(BMD)of the femur and lumbar vertebrae was measured using dual-energy X-ray absorptiometry(DXA).Hematoxylin-eosin(HE)staining was employed to assess the microstructure of the femur and colonic mucosa,while immu-nohistochemistry was used to measure the expression of collagen type Ⅰ alpha 1 chain(COL1A1)protein in the femur.Ad-ditionally,RT-qPCR was utilized to analyze the mRNA expression of bone formation-related indicators,including alkaline phosphatase(ALP),COL1A1,and Runt-related transcription factor 2(RUNX2).Fresh fecal samples were collected from the rats for 16S rDNA sequencing to detect changes in intestinal microbiota composition.RESULTS:Compared to the sham group,OVX rats exhibited a significant increase in body weight and a marked decrease in femur and lumbar ver-tebrae bone density.HE staining revealed trabecular bone fractures with a disrupted reticular structure in the OVX group,along with the presence of numerous cavities and fat vacuoles in the bone marrow.The colonic mucosa showed signs of vil-lous shedding and mild crypt atrophy.Immunohistochemistry results demonstrated a substantial reduction in brown-yellow granules and COL1A1 expression in the OVX group.Conversely,in the AS group,there was a reduction in body weight and a significant increase in bone density of the femur and lumbar vertebrae.The trabecular architecture appeared more or-ganized,with less severe fractures compared to the OVX group.In the AS group,the number of cavities and fat vacuoles in the bone marrow was also reduced,and the colonic mucosa exhibited improved villous structure and less crypt atrophy.Immunohistochemical analysis indicated that AS treatment significantly enhanced COL1A1 expression.Furthermore,after AS intervention,the mRNA expression levels of ALP,COL1A1,and RUNX2 were notably increased.16S rDNA sequenc-ing revealed a significant increase in the abundance of Firmicutes,Proteobacteria,f_Pseudonocardiaceae,f_Marinifilace-ae,f_Oscillospiraceae,f_Ruminococcaceae,and f_Peptostreptococcaceae,while p_Euryarchaeota,Bacteroidetes,and f_Muribaculaceae showed significant reductions.Overall,OVX led to increased diversity in the species distribution of in-testinal microbiota,whereas AS treatment helped recalibrate the aforementioned phyla(families)and reduce diversity.CONCLUSION:Astragaloside IV can increase bone density in OVX rats,improve bone microstructure,promote bone formation,and prevent colonic mucosal damage by regulating the relative abundance of intestinal flora.