Objective:To evaluate the role of necroptosis in paclitaxel-induced cognitive dysfunction in mice.Methods:Thirty SPF healthy male C57BL/6N mice, aged 6-8 weeks, weighing 20-25 g, were divided into 3 groups ( n=10 each) using a random number table method: vehicle control group (Veh group), paclitaxel group (PTX group), and paclitaxel+ a specific inhibitor of necroptosis Necrostatin-1 group (P+ N group). In PTX group and P+ N group, paclitaxel 10 mg/kg (diluted to 5 mg/ml in anhydrous ethanol and castor oil [1∶1], and further diluted to 1 mg/ml in 0.9% normal saline before use) was intraperitoneally injected daily for 7 consecutive days to induce cognitive dysfunction. P+ N group received an intraperitoneal injection of Necrostatin-1 6.5 mg/kg (diluted to 10 mg/ml in dimethyl sulfoxide, and further diluted to 1 mg/ml in 0.9% normal saline before use) at 2 h before paclitaxel administration every other day, 4 times in total. Veh group received the equal volume of solvent at the matched time points as previously described in P+ N group. After establishment of the model, spontaneous locomotor activity was assessed using the open field test, followed by the novel object recognition test and the Morris water maze to evaluate the cognitive function. The animals were sacrificed after the end of the Morris water maze test, and the hippocampal tissues were collected for determination of the expression of necroptosis-related proteins receptor-interacting serine/threonine-protein kinase 1 (RIPK1), RIPK3, mixed lineage kinase domain-like protein (MLKL), and phospho-MLKL (p-MLKL) (by Western blot analysis) and contents of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) (double immunofluorescence staining) and for observation of the localization of programmed necrosis cells (using enzyme-linked immunosorbent assay). Results:There were no significant differences in the total distance traveled and mean movement speed in the open field test or swimming speed in the Morris water maze test among the three groups ( P>0.05). Compared to Veh group, the time spent in the central zone in the open field and time spent in the original platform quadrant were significantly shortened, the discrimination index was decreased, the escape latency was prolonged, the number of crossing the original platform was reduced, the expression of RIPK1, RIPK3, MLKL and p-MLKL was up-regulated, the contents of TNF-α and IL-1β were increased, and the number of RIPK1-positive neurons was increased in PTX group ( P<0.05). Compared to PTX group, the time spent in the central zone in the open field test and time spent in the original platform quadrant were significantly prolonged, the discrimination index was increased, the escape latency was shortened, the number of crossing the original platform was increased, the expression of RIPK1, RIPK3, MLKL and p-MLKL was down-regulated, the contents of TNF-α and IL-1β were decreased, and the number of RIPK1-positive neurons was decreased in P+ N group ( P<0.05). Conclusions:Necroptosis in hippocampal neurons can lead to neuroinflammation, thus contributeing to paclitaxel-induced cognitive dysfunction in mice.

Objective To explore the experiences of children with Crohn's disease and their parents regarding the exclusion diet,and to provide a basis for formulating personalized dietary guidance programs.Methods A total of 12 children with Crohn's disease and their parents,hospitalized in the Department of Gastroenterology at a tertiary children's hospital in Guangzhou from June to December 2023,were selected as research subjects using objective sampling.Semi-structured interviews were conducted,and the data were analyzed and refined using Colaizzi's seven-step analysis method.Results Totally 3 themes and 14 sub-themes were extracted.①Lack of cognition and trust in Crohn's disease exclusion diet(unfamiliarity with the contents of the diet,misunderstanding of the diet's preparation,inadequate response to daily exclusion diet practices,parents' distrust in the exclusion diet).②The practical challenges of the Crohn's disease exclusion diet(the challenge of personal dietary preferences,the challenge of family meal preparation,the challenge of school feeding,food intolerance,feelings of monotony and weariness following the exclusion diet).③Innovations in practicing the Crohn's disease exclusion diet(managing taste fatigue,managing visual fatigue,innovative cooking methods,prioritizing exclusive enteral nutrition followed by the exclusion diet,overcoming the desire for universal food).Conclusion Children with Crohn's disease and their parents exhibit insufficient cognition and trust in the exclusion diet and face various challenges in practice.Clinical medical staff should adopt personalized coping strategies tailored to the specific circumstances of each child.

Objective:To investigate the clinicohistologic and molecular pathological characteristics of invasive stratified mucin-producing carcinoma(ISMC).Methods:The clinicopathological data,immunohistochemistry,alcian blue/periodic acid-Schiff(AB/PAS)staining,molecular detection and PD-L1 expressions of 11 cases of ISMC and 4 cases of stratified mucin-producing intraepithelial lesion(SMILE)in the pathological database of Anqing Medical Center of Anhui Medical University/Anqing Municipal Hospital and the first Affiliated hospital of Wannan Medical School/Yijishan Hostital between January of 2018 and March of 2024 were retrospectively analyzed.Results:ISMC patients often presented with irregular vaginal bleeding.Morphologically,the cells containing mucus in the cytoplasm were arranged in stratified layers,surrounded by a palisade,and the tumor cells might be signet ring-shaped or have clear cytoplasm.ISMC might occur in both pure and mixed forms.ISMC had highly invasive biological characteristics.CK7,p16,p40 and(or)p63 were expressed focally or positively in the form of palisades around the cancer nests.AB-PAS staining was positive.HPV test results:HPV16/18 was positive in ISMC(1/4);HPV16/18 was positive before surgery(4/6);no HPV was detected in SMILE;all ISMC cases expressed PD-L1.Eight ISMC patients were successfully followed up for 4-39 months(average 20.50 months),and four SMILE patients for 1-25 months(average 8.25 months).All followed-up patients survived;however,one ISMC patient developed multi-organ metastases post-surgery.Conclusion:ISMC has unique morphological characteristics and immunophenotypes,indicated by high invasiveness and poor prognosis.All ISMC are PD-L1 positive,suggesting that these patients may benefit from PD-L1 immunotherapy.

Objective:To explore the effectiveness, safety, and cost among urinary follicle-stimulating hormone (u-FSH), recombinant FSH (r-FSH)α, and r-FSHβ in the early follicular phase prolonged protocol for patients under 35 years old with normal ovarian function.Methods:It was a retrospective cohort study. Patients under 35 years old with normal ovarian function who underwent early follicular phase prolonged protocol for ovulation stimulation and using in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) for fertilization in Reproductive Health Hospital of the Third Affiliated Hospital of Zhengzhou University from January 2018 to December 2023 were recruited, including the fresh and frozen-thawed embryo transfer (FET) cycles. Patients were divided into u-FSH group, r-FSHα group, and r-FSHβ group. A total of 1 048 ovarian stimulation cycles were included, with 150 cycles, 490 cycles and 408 cycles in the three groups respectively. A total of 710 FET cycles with fresh cycle cancellation were included, with 95 cycles, 320 cycles and 295 cycles in the three groups respectively. The baseline data, pregnancy outcomes, safety, and cost were compared among the three groups. The main observation indicators were cumulative pregnancy rate and cumulative live birth rate (CLBR). A binary logistic regression model was used to control confounding factors, and to analyze the relationship between three ovulation inducing medicine and CLBR. Results:The difference in the number of oocytes retrieved among the u-FSH group, r-FSHα group, and r-FSHβ group was statistically significant [13.0 (10.0, 16.0), 14.0 (11.0, 18.0), 15.0 (11.0, 19.0), respectively, P=0.012], and the difference in the number of 2PN embryos was statistically significant [9.0 (6.0, 12.0), 10.0 (7.0, 13.0), 10.0 (7.0, 13.0), respectively, P=0.046]. There were no statistically significant differences in the number of available embryos, available embryo rate, the number of high-quality embryos, high-quality embryo rate, available blastocyst formation rate, fresh cycle clinical pregnancy rate, live birth rate in fresh cycle, cumulative pregnancy rate of frozen embryos with fresh cycle cancellation, CLBR of frozen embryos with fresh cycle cancellation, cumulative clinical pregnancy rate, CLBR, moderate to severe ovarian hyperstimulation syndrome incidence, ectopic pregnancy rate, multiple pregnancy rate and neonatal malformation rate among the three groups (all P>0.05). In terms of economy, the u-FSH group had the lowest total gonadotropin cost for each patient, while the r-FSHα group had the highest. The differences among the three groups were statistically significant [u-FSH group 4 429.08 (3 198.78, 5 044.23) yuan, r-FSHα group 6 023.72 (5 433.75, 7 529.65) yuan, r-FSHβ group 5 480.00 (4 550.90, 6 437.86) yuan, P<0.001]. Binary logistic regression analysis was conducted, using u-FSH as a control. The CLBR of the r-FSHα group and r-FSHβ group showed no statistically significant difference compared with the u-FSH group (a OR=0.95, 95% CI: 0.57-1.58, P=0.838; a OR=0.89, 95% CI: 0.54-1.48, P=0.654). Conclusion:For patients under 35 years old with normal ovarian function undergoing long protocol ovarian stimulation, the effectiveness and safety of the three ovarian-stimulating medicine are similar, but u-FSH has economic advantages.

Objective:To investigate the effect of atosiban on hemodynamic parameters of uterine arteries and clinical effect evaluation in patients with previous implantation failure undergoing frozen-thawed embryo transfer.Methods:A retrospective cohort study was conducted to analyze 298 cycles of FET in the Department of Reproductive Medicine of the Second Affiliated Hospital of Zhengzhou University from January 2021 to June 2023. Patients were categorized into atosiban group ( n=149) and control group ( n=149) according to whether administered atosiban or not. The related indicators and clinical outcomes were compared between the two groups. Hemodynamic parameters of the uterine arteries, including bilateral uterine artery peak systolic velocity/diastolic velocity (S/D), pulsatility index (PI), resistance index (RI), and serum levels of prostaglandin F2α (PGF2α) and oxytocin were compared before and after atosiban treatment. Univariate and multivariate logistic regression analysis were applied to assess the effect of atosiban on pregnancy outcomes. The effect of atosiban on live birth rate was analyzed by age stratification. Results:The implantation rate [51.92% (135/260)], the clinical pregnancy rate [67.11% (100/149)] and the live birth rate [59.06% (88/149)] in atosiban group were significantly higher than those in control group [41.13% (102/248), P=0.015; 51.01% (76/149), P=0.005; 40.27% (60/149), P=0.001]; and the early miscarriage rate [9.00% (9/100)] was lower than that of control group [19.74% (15/76), P=0.040]. Multivariate logistic regression analysis showed that atosiban was an independent influencing factor of live birth rate ( OR=2.236, 95% CI: 1.371-3.646, P=0.001). The post-treatment right uterine artery blood flow S/D [4.61 (4.00, 5.36)], PI [1.81 (1.58, 2.05)], RI [0.79 (0.75, 0.82)], and left uterine artery blood flow S/D [4.62 (3.83, 5.61)], PI (1.84±0.38), RI [0.79 (0.74, 0.82)] were all lower than those before treatment [right S/D 4.93 (4.06, 6.04), P<0.001; PI 1.93 (1.60, 2.17), P=0.001; RI 0.80 (0.76, 0.83), P<0.001; left S/D 5.05 (4.20, 6.32), P<0.001; PI 1.95±0.43, P<0.001; RI 0.81 (0.76, 0.84), P<0.001]. Besides, the levels of PGF2α [97.01 (85.15, 109.93) ng/L] and oxytocin [41.18 (37.16, 46.78) ng/L] after treatment in atosiban group were significantly lower than those before treatment [119.71 (108.85, 129.99) ng/L, P<0.001; 51.87 (46.44, 55.54) ng/L, P<0.001). Moreover, the endometrial peristalsis waves in atosiban group were significantly less after treatment [1.00 (0.00, 2.00) times/min] than before treatment [2.00 (1.00, 3.00) times/min], and the difference was statistically significant ( P<0.001). Conclusion:Atosiban can improve uterine artery blood flow and reduce endometrial peristalsis waves in women with previous implantation failure, which increases endometrial blood perfusion. Additionally, it can also reduce the levels of PGF2α and oxytocin, and optimize the pregnancy outcome of the frozen-thawed embryo transfer.

Objective:To investigate the therapeutic effects and underlying mechanisms of intrauterine perfusion of umbilical cord blood mononuclear cells (UCB-MNCs) on thin endometrium.Methods:SPF-grade Kunming mice aged 6-8 weeks were selected. A mouse model of thin endometrium was established by infusing 95% ethanol into the uterine cavity for a duration of 5 min. Using a completely randomized grouping method, 40 female mice with regular estrous cycles were randomly divided into four groups: untreated group (no intervention, n=10), sham-operated group (operation without modeling, n=10), experimental group (intrauterine infusion of UCB-MNCs during estrus after one estrous cycle post-modeling, n=10) and negative control group (intrauterine infusion of saline during estrus after one estrous cycle post-modeling, n=10). Following the administration of UCB-MNCs or physiological saline, all groups' uterine tissues were collected two estrous cycles later during their respective estrus phases. Hematoxylin-eosin staining was used to assess endometrial morphology, measure thickness, and count glands. Western blotting and reverse transcription real-time quantitative polymerase chain reaction were utilized to measure the relative protein and mRNA expression levels of leukemia inhibitory factor (LIF), vascular endothelial growth factor (VEGF), integrin (ITG) α V, ITG β 3 and proliferating cell nuclear antigen (PCNA) in the endometrium across different groups for intergroup comparisons. Results:The endometrial thickness and the number of glands in the untreated group [(507.32±85.66) μm, 18.67±6.66] showed no statistically significant differences compared with those in the sham-operated group [(502.78±73.26) μm, 19.33±7.73, all P>0.05]. The experimental group showed significantly increased endometrial thickness [(347.71±82.24) μm vs. (118.85±29.19) μm, P<0.001] and gland number (15.00±2.65 vs. 2.00±2.00, P=0.030) compared with the negative control group. There was no statistically significant difference in the relative protein and mRNA expression levels of LIF, VEGF, ITG α v, ITG β 3, and PCNA in the endometrium between the untreated group and the sham-operated group (all P>0.05). The relative protein and mRNA expression levels of endometrial LIF, VEGF, ITG α V, ITG β 3 and PCNA of the experimental group were all significantly higher than those in the negative control group (all P<0.05). Conclusion:Intrauterine perfusion with UCB-MNCs may promote endometrial regeneration and repair, as well as improve endometrial receptivity, through the upregulation of the expression levels of PCNA, LIF, VEGF, and ITG α V, ITGβ 3.

Objective:To evaluate the role of necroptosis in paclitaxel-induced cognitive dysfunction in mice.Methods:Thirty SPF healthy male C57BL/6N mice, aged 6-8 weeks, weighing 20-25 g, were divided into 3 groups ( n=10 each) using a random number table method: vehicle control group (Veh group), paclitaxel group (PTX group), and paclitaxel+ a specific inhibitor of necroptosis Necrostatin-1 group (P+ N group). In PTX group and P+ N group, paclitaxel 10 mg/kg (diluted to 5 mg/ml in anhydrous ethanol and castor oil [1∶1], and further diluted to 1 mg/ml in 0.9% normal saline before use) was intraperitoneally injected daily for 7 consecutive days to induce cognitive dysfunction. P+ N group received an intraperitoneal injection of Necrostatin-1 6.5 mg/kg (diluted to 10 mg/ml in dimethyl sulfoxide, and further diluted to 1 mg/ml in 0.9% normal saline before use) at 2 h before paclitaxel administration every other day, 4 times in total. Veh group received the equal volume of solvent at the matched time points as previously described in P+ N group. After establishment of the model, spontaneous locomotor activity was assessed using the open field test, followed by the novel object recognition test and the Morris water maze to evaluate the cognitive function. The animals were sacrificed after the end of the Morris water maze test, and the hippocampal tissues were collected for determination of the expression of necroptosis-related proteins receptor-interacting serine/threonine-protein kinase 1 (RIPK1), RIPK3, mixed lineage kinase domain-like protein (MLKL), and phospho-MLKL (p-MLKL) (by Western blot analysis) and contents of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) (double immunofluorescence staining) and for observation of the localization of programmed necrosis cells (using enzyme-linked immunosorbent assay). Results:There were no significant differences in the total distance traveled and mean movement speed in the open field test or swimming speed in the Morris water maze test among the three groups ( P>0.05). Compared to Veh group, the time spent in the central zone in the open field and time spent in the original platform quadrant were significantly shortened, the discrimination index was decreased, the escape latency was prolonged, the number of crossing the original platform was reduced, the expression of RIPK1, RIPK3, MLKL and p-MLKL was up-regulated, the contents of TNF-α and IL-1β were increased, and the number of RIPK1-positive neurons was increased in PTX group ( P<0.05). Compared to PTX group, the time spent in the central zone in the open field test and time spent in the original platform quadrant were significantly prolonged, the discrimination index was increased, the escape latency was shortened, the number of crossing the original platform was increased, the expression of RIPK1, RIPK3, MLKL and p-MLKL was down-regulated, the contents of TNF-α and IL-1β were decreased, and the number of RIPK1-positive neurons was decreased in P+ N group ( P<0.05). Conclusions:Necroptosis in hippocampal neurons can lead to neuroinflammation, thus contributeing to paclitaxel-induced cognitive dysfunction in mice.

Objective To explore the experiences of children with Crohn's disease and their parents regarding the exclusion diet,and to provide a basis for formulating personalized dietary guidance programs.Methods A total of 12 children with Crohn's disease and their parents,hospitalized in the Department of Gastroenterology at a tertiary children's hospital in Guangzhou from June to December 2023,were selected as research subjects using objective sampling.Semi-structured interviews were conducted,and the data were analyzed and refined using Colaizzi's seven-step analysis method.Results Totally 3 themes and 14 sub-themes were extracted.①Lack of cognition and trust in Crohn's disease exclusion diet(unfamiliarity with the contents of the diet,misunderstanding of the diet's preparation,inadequate response to daily exclusion diet practices,parents' distrust in the exclusion diet).②The practical challenges of the Crohn's disease exclusion diet(the challenge of personal dietary preferences,the challenge of family meal preparation,the challenge of school feeding,food intolerance,feelings of monotony and weariness following the exclusion diet).③Innovations in practicing the Crohn's disease exclusion diet(managing taste fatigue,managing visual fatigue,innovative cooking methods,prioritizing exclusive enteral nutrition followed by the exclusion diet,overcoming the desire for universal food).Conclusion Children with Crohn's disease and their parents exhibit insufficient cognition and trust in the exclusion diet and face various challenges in practice.Clinical medical staff should adopt personalized coping strategies tailored to the specific circumstances of each child.

Objective To observe the clinical efficacy of belimumab combined with glucocorticoids in the treatment of children with systemic lupus erythematosus(SLE).Methods A total of 64 children with SLE were randomly divided into the observation group and the control group,with 32 cases in each group.The control group was treated with oral prednisone tablets combined with hydroxychloroquine sulfate tablets,while the observation group was treated with belimumab on the basis of treatment in the control group.After 6 months of treatment,the clinical efficacy,systemic lupus erythematosus disease activity index(SLEDAI)scores and laboratory indicators[complement C3,complement C4,immunoglobulin IgG(IgG),B lymphocytes,white blood cell count(WBC),erythrocyte sedimentation rate(ESR),serum C-reactive protein(CRP),24-hour urine protein quantification(24 h UP),serum creatinine(SCr)and blood urea nitrogen(BUN)]were compared between the two groups.Results After 6 months of treatment,the total effective rate was higher in the observation group than that of the control group(93.8%vs.75.0%,P＜0.05).Compared with before treatment,both groups showed lower SLEDAI scores,IgG,B lymphocytes,ESR,CRP,SCr,24 h UP and BUN after treatment,while C3,C4 levels and WBC were higher.Moreover,the improvement of all indicators was better in the observation group than that in the control group(P＜0.05).Conclusion Belimumab combined with glucocorticoids can improve the clinical efficacy,control disease activity and enhance immune function in children with SLE.

Objective:To explore the effectiveness, safety, and cost among urinary follicle-stimulating hormone (u-FSH), recombinant FSH (r-FSH)α, and r-FSHβ in the early follicular phase prolonged protocol for patients under 35 years old with normal ovarian function.Methods:It was a retrospective cohort study. Patients under 35 years old with normal ovarian function who underwent early follicular phase prolonged protocol for ovulation stimulation and using in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) for fertilization in Reproductive Health Hospital of the Third Affiliated Hospital of Zhengzhou University from January 2018 to December 2023 were recruited, including the fresh and frozen-thawed embryo transfer (FET) cycles. Patients were divided into u-FSH group, r-FSHα group, and r-FSHβ group. A total of 1 048 ovarian stimulation cycles were included, with 150 cycles, 490 cycles and 408 cycles in the three groups respectively. A total of 710 FET cycles with fresh cycle cancellation were included, with 95 cycles, 320 cycles and 295 cycles in the three groups respectively. The baseline data, pregnancy outcomes, safety, and cost were compared among the three groups. The main observation indicators were cumulative pregnancy rate and cumulative live birth rate (CLBR). A binary logistic regression model was used to control confounding factors, and to analyze the relationship between three ovulation inducing medicine and CLBR. Results:The difference in the number of oocytes retrieved among the u-FSH group, r-FSHα group, and r-FSHβ group was statistically significant [13.0 (10.0, 16.0), 14.0 (11.0, 18.0), 15.0 (11.0, 19.0), respectively, P=0.012], and the difference in the number of 2PN embryos was statistically significant [9.0 (6.0, 12.0), 10.0 (7.0, 13.0), 10.0 (7.0, 13.0), respectively, P=0.046]. There were no statistically significant differences in the number of available embryos, available embryo rate, the number of high-quality embryos, high-quality embryo rate, available blastocyst formation rate, fresh cycle clinical pregnancy rate, live birth rate in fresh cycle, cumulative pregnancy rate of frozen embryos with fresh cycle cancellation, CLBR of frozen embryos with fresh cycle cancellation, cumulative clinical pregnancy rate, CLBR, moderate to severe ovarian hyperstimulation syndrome incidence, ectopic pregnancy rate, multiple pregnancy rate and neonatal malformation rate among the three groups (all P>0.05). In terms of economy, the u-FSH group had the lowest total gonadotropin cost for each patient, while the r-FSHα group had the highest. The differences among the three groups were statistically significant [u-FSH group 4 429.08 (3 198.78, 5 044.23) yuan, r-FSHα group 6 023.72 (5 433.75, 7 529.65) yuan, r-FSHβ group 5 480.00 (4 550.90, 6 437.86) yuan, P<0.001]. Binary logistic regression analysis was conducted, using u-FSH as a control. The CLBR of the r-FSHα group and r-FSHβ group showed no statistically significant difference compared with the u-FSH group (a OR=0.95, 95% CI: 0.57-1.58, P=0.838; a OR=0.89, 95% CI: 0.54-1.48, P=0.654). Conclusion:For patients under 35 years old with normal ovarian function undergoing long protocol ovarian stimulation, the effectiveness and safety of the three ovarian-stimulating medicine are similar, but u-FSH has economic advantages.