Objective To explore the application value of anhydrous ethanol as an alternative to methanol in the preparation of chromosomal specimens from peripheral blood lymphocytes, and to establish a set of quantitative analytical methods for objectively evaluating the effectiveness of specimen preparation. Methods Residual blood samples from routine laboratory slide preparation were used for lymphocyte culture. The standard slide preparation method was employed. The fixative in the control group was methanol and glacial acetic acid (3∶1). Four experimental groups were set up based on the ratio of anhydrous ethanol to glacial acetic acid in the fixative (volume ratios of 3∶1, 5∶1, 7∶1, and 9∶1 for experimental groups 1, 2, 3, and 4, respectively). A chromosomal analysis was conducted using an automated chromosome scanning/image analysis system to evaluate the morphology and dispersion of metaphase chromosomes in both control and experimental groups. Comparisons were made between the control and experimental groups regarding the dic + r aberration rate, ace aberration rate, chromosomal aberration rate, chromosome dispersion index, chromosome overlapping ratio, and dispersion index/overlapping ratio. Results Microscopic evaluation revealed that the preparation quality of experimental groups 1 and 2 was comparable to the control group. No statistically significant differences were observed in dic + r aberration rate between each of the experimental groups and the control (P > 0.05). All experimental groups except group 4 showed no significant differences in ace aberration rate and chromosome aberration rate compared with the control group (P > 0.05). Experimental groups 1 and 2 showed no significant differences in chromosome dispersion index, overlapping ratio, and dispersion index/overlapping ratio compared with the control group (P > 0.05). Conclusion A mixture of anhydrous ethanol and glacial acetic acid at a 5∶1 ratio is recommended for use as a fixative in the preparation of chromosomal specimens from peripheral blood lymphocytes. A quantitative index system for assessing the quality of chromosomal specimens was established, enabling objective evaluation of slide preparation effectiveness.

Objective To investigate the relationship between serum trimethylamine oxide(TMAO),endo-thelial cell specific molecule 1(Endocan)and cardiac function and pregnancy outcome in patients with hyper-tensive disorders of pregnancy(HDP).Methods A total of 182 patients with HDP admitted to Handan Ma-ternal and Child Health Hospital from January 2021 to June 2023(HDP group)and 98 healthy pregnant women admitted to this hospital during the same period(control group)were selected as research subjects.Serum TMAO,Endocan and left ventricular cardiac function indexes[left ventricular ejection fraction(LVEF),left ventricular end-diastolic volume(LVEDV)and left ventricular end-systolic volume(LVESV)]were compared between the two groups.According to pregnancy outcome,HDP patients were divided into poor outcome group(78 cases)and good outcome group(104 cases).Spearman correlation analysis was used to analyze the correlation between serum TMAO and Endocan and cardiac function indexes in HDP patients,and multi-factor Logistic regression was used to analyze the influencing factors of adverse pregnancy outcomes in HDP patients.The predictive value of serum TMAO and Endocan for adverse pregnancy outcomes in HDP patients was analyzed by receiver operating characteristic(ROC)curve.Results Compared with control group,serum TMAO,Endocan,LVEDV and LVESV were increased in HDP group,and LVEF was decreased(P＜0.05).Serum TMAO and Endocan in HDP patients were negatively correlated with LVEF(P＜0.05),and positively correlated with LVEDV and LVESV(P＜0.05).The incidence of adverse pregnancy outcomes in 182 HDP patients was 42.86％(78/182).Preeclampsia(PE),severe preeclampsia(SPE),24 h urine protein increase,LVEDV increase,LVESV increase,TMAO increase,Endocan increase were independent risk factors for adverse pregnancy outcomes in HDP patients,and LVEF increase was protective factor(P＜0.05).The area under the curve of serum TMAO combined with Endocan in predicting adverse pregnancy outcomes in HDP patients was 0.880,which was greater than 0.793 and 0.788 predicted by serum TMAO and Endocan a-lone.Conclusion The increase of serum TMAO and Endocan levels in HDP patients are relate to the decrease of cardiac function and adverse pregnancy outcomes,and the combined detection of the two has high predictive value for adverse pregnancy outcomes in HDP patients.

Objective To explore the relationship between serum pregnancy-specific protein 1(PSG1),stress-induced protein 2(Sestrin 2),growth arrest-specific protein 6(Gas6)and uterine artery blood flow pa-rameters and fetal growth restriction(FGR)in patients with gestational hypertension(GH).Methods A to-tal of 485 GH patients admitted to Handan Maternal and Child Health Hospital from January 2020 to October 2023 were selected as the research objects and divided into the occurrence group(81 cases)and the non-occur-rence group according to whether FGR occurred.The correlations between serum PSG1,Sestrin 2,Gas6 and uterine artery blood flow parameters[pulse index(PI),resistance index(RI),ratio of peak systolic velocity to end diastolic velocity(S/D)]was analysed,as well as the related influencing factors of FGR in GH patients.In addition,a Nomogram model based on the influencing factors was constructed to analyze the predictive value.Results The serum PSG1 level in occurrence group was significantly lower than that in non-occurrence group,and the serum Sestrin 2,Gas6 levels and PI,RI,S/D values were significantly higher than those in non-occurrence group(P＜0.05).Pearson correlation results showed that serum PSG1 was negatively correlated with the uterine artery blood flow parameters PI,RI,and S/D,and the levels of serum Sestrin 2 and Gas6 were positively correlated with the uterine artery blood flow parameters PI,RI,and S/D(P＜0.05).Gestational di-abetes mellitus,umbilical cord abnormalities,high Sestrin 2,high Gas6,high PI,high RI,and high S/D were independent risk factors for the occurrence of FGR in GH patients(P＜0.05),and increased PSG1 level was protective factor for the occurrence of FGR in GH patients(P＜0.05).Receiver operating characteristic(ROC)curve analysis showed that the area under the curve(AUC)of the Nomogram prediction model con-structed based on the influencing factors for predicting the occurrence of FGR in GH patients was 0.982,and the sensitivity and specificity were 0.943 and 0.938,respectively.The internal verification of the Bootstrap method shows that the Bias-corrected prediction curve basically coincides with the Ideal line,and the consis-tency index(C-index)was 0.964,indicating that the model was relatively stable.The decision curve shows that the threshold probability of this model was 0.01-1.00 and the net return rate was above 0.Conclusion Ser-um PSG1,Sestrin 2 and Gas6 in GH patients are closely related to uterine artery blood flow parameters and FGR,and the three are the influencing factors for the occurrence of FGR in GH patients.The constructed No-mogram model has a good predictive efficacy for FGR.

Since the birth of the first "test-tube baby" in Chinese mainland in 1988, assisted reproductive technology (ART) in China has matured significantly. The number of ART cycles has surpassed one million, and the number of assisted reproductive institutions and practitioners has attained a significant scale, contributing to the establishment of a fertility-friendly society. However, due to the complexity of the ART process, the diversity of personnel backgrounds, and the profound impact of any error that may occur, there is an urgent need to establish an efficient and effective safety management model for error prevention. This paper aims to outline the key processes and steps involved in the implementation of ART, explore control measures for these critical processes, and delve into error prevention strategies for gamete and embryo laboratories through the creation and utilization of a "gamete safety checklist".

Objective:To investigate the role of serotonergic (5-HT) neurons activation in the dorsal raphe nucleus (DRN) in heat-induced seizures and its influence on breathing patterns in Scn1a + /- mice. Methods:24-day-old male Scn1a + /- mice were used for experiments, and sudden unexpected death in epilepsy(SUDEP) model was established through heat induction for 5 consecutive days. (1) Fluoxetine intervention experiment: 20 male mice were randomly divided into model group( n=10) and fluoxetine group ( n=10) according the weight-matched method. The fluoxetine group received intraperitoneal injection of fluoxetine (10 mg/kg) 45 min before heat induction each day for 5 consecutive days, while the model group received equal volume of 0.9% NaCl solution. (2) Chemogenetic activation experiment: 20 male mice were randomly divided into vector control group( n=10) and chemogenetic activation group ( n=10) according the weight-matched method. Empty vector or rAAV-TPH2-hM3d(Gq)-EGFP-WPREs was stereotaxically injected into DRN 14 d prior to seizure induction, and deschloroclozapine (5 mg/kg) was intraperitoneally injected 30 min before heat induction. Seizure characteristics and survival were assessed through video monitoring, and respiratory parameters were monitored. Immunofluorescence was used to detect colocalization of tryptophan hydroxylase 2(TPH2) and c-Fos in DRN. Statistical analysis was performed using SPSS 24.0 and GraphPad Prism 9.0. The independent sample t-test or Mann-Whitney test was used for inter-group comparison. Results:(1) In the fluoxetine intervention experiment: the survival rates between the model group and fluoxetine group showed no statistically significant difference ( χ2=2.23, P>0.05). As for the frequency of grade Ⅳ seizures, the model group (1.50(1.25, 2.35)min) demonstrated higher frequency than the fluoxetine group (0.43(0.20, 0.67)min) ( t=-3.40, P<0.05).With respect to respiratory parameters, the model group demonstrated shorter expiratory time ((0.10±0.02) s) and inspiratory time ((0.15±0.02) s) compared to the fluoxetine group ((0.16±0.05) s, (0.19±0.04) s) ( t=-3.47, -3.73, both P<0.01). The respiratory rate in the model group ((269.96±44.84) times/min) was significantly higher compared to the fluoxetine group ((195.04±52.37) times/min) ( t=3.44, P<0.01). The tidal volume in the model group ((0.10±0.02) mL) was significantly lower than the fluoxetine group ((0.13±0.04) mL) ( t=-2.19, P<0.05).The number of TPH2+ /c-Fos+ co-expressing cells in the model group (11.00±4.00) was lower than that in the fluoxetine group (33.00±8.39)( t=-4.16, P<0.05). (2) In the chemogenetic activation experiment: compared to the vehicle group, the chemogenetic activation group demonstrated significantly enhanced survival rates ( χ2=5.83, P<0.05). As for the frequency of grade Ⅴ seizures, the vehicle group (2.11(1.62, 3.44) times/min) showed higher frequency compared to the chemogenetic activation group (0.81(0.00, 1.62) times/min) ( t=18.00, P<0.05). In terms of respiratory parameters, the vehicle group showed shorter expiratory time ((0.10±0.01) s) and inspiratory time ((0.14±0.01) s) compared to the chemogenetic activation group ((0.12±0.01) s, (0.15±0.01) s) ( t=-2.78, -2.50, both P<0.05). The respiratory rate in the vehicle group ((208.37±9.73) times/min) was significantly higher than the chemogenetic activation group ((191.85±8.83) times/min) ( t=3.98, P<0.01). The tidal volume in the vehicle group ((0.09±0.01) mL) was significantly lower than the chemogenetic activation group ((0.12±0.02) mL) ( t=-4.77, P<0.001).The number of TPH2+ /c-Fos+ co-expressing cells in the vehicle group (9.00±3.46) was lower than that in the chemogenetic activation group (43.00±11.02)( t=-5.20, P<0.01). Conclusion:Specific activation of serotonergic neurons in the DRN can ameliorate heat-induced epileptic symptoms, improve respiratory function, and prolong survival time in Scn1a + /- mice.

Objective:To evaluate the efficacy and safety of baricitinib combined with ruxolitinib cream in the treatment of progressive nonsegmental vitiligo.Methods:Clinical data were retrospectively collected from patients with progressive nonsegmental vitiligo in Boao Super Hospital. All the patients were treated with oral baricitinib daily (2 mg/day for patients weighing ≤ 50 kg; 4 mg/day for those > 50 kg) in combination with topical application of ruxolitinib cream twice daily for 24 consecutive weeks. Disease severity was assessed using the facial vitiligo area scoring index (F-VASI) and total body VASI (T-VASI) at baseline, week 12, and week 24. Adverse reactions were monitored throughout the treatment course.Results:Six patients with progressive nonsegmental vitiligo were collected, including 3 males and 3 females, aged 26 - 42 years, with the disease duration ranging from 0.5 to 25 years. At week 12, 3 patients achieved a 50% ~ < 75% improvement in facial vitiligo lesions (F-VASI 50), 1 patient achieved F-VASI 75 (75% ~ < 90% improvement), and 1 patient achieved T-VASI 50; at week 24, 4 patients achieved F-VASI 50, 1 patient achieved F-VASI 75, 1 patient achieved F-VASI 90 (≥ 90% improvement), and 3 patients achieved T-VASI 50. During the treatment, upper respiratory infection occurred in 1 patient, acne in 1 patient, pruritus in 2 patients, elevation of total cholesterol levels in 2 patients, and increase of high-density lipoprotein levels in 2 patients. No severe adverse events were observed during the treatment.Conclusion:The combination therapy with baricitinib and ruxolitinib cream may have potential efficacy and safety in the treatment of progressive nonsegmental vitiligo.

OBJECTIVE: To explore the effect of light music therapy on delirium in intensive care unit (ICU) patients undergoing invasive mechanical ventilation, and provide evidence-based support for clinical prevention of delirium. METHODS: A prospective randomized controlled trial was conducted. 140 patients with invasive mechanical ventilation admitted to the department of respiratory and critical care medicine of First Affiliated Hospital of Guangxi Medical University from January 2024 to January 2025 were enrolled. The patients were divided into intervention group and control group using a random number table method. The control group received routine treatment and nursing care, while the intervention group received light music therapy three times a day for 30 minutes each time for 7 consecutive days. The confusion assessment method-ICU (CAM-ICU) was used to evaluate delirium, and the incidence of delirium within 7 days was statistically analyzed. Richmond agitation-sedation score (RASS), critical care pain observation tool (CPOT) score, mechanical ventilation duration, the length of ICU stay, and ICU stay expenses were record. RESULTS: 129 cases were ultimately included, including 64 cases in the control group and 65 cases in the intervention group. There was no statistically significant difference in baseline data between the two groups, indicating comparability. The incidence of delirium in the intervention group was significantly lower than that in the control group (27.7% vs. 51.6%, χ ² = 7.687, P = 0.006). There was no significantly difference in RASS score between the two groups before enrollment (P = 0.840). After intervention, the RASS score in the intervention group significantly decreased, from 2.00 points on the 1st day of enrollment to 0.00 points on the 7th day, while the control group only decreased from 2.00 points to 1.50 points. The decreasing trend of the intervention group was more pronounced, especially on the 3rd day (P = 0.047) and the 7th day (P =0.005), with significant differences between the groups. The time effect (F = 18.929, P < 0.001), group effect (F = 6.655, P = 0.011), and time group interaction effect (F = 7.372, P < 0.001) of the two groups of RASS score were significant, suggesting that light music therapy has better timeliness and sustainability in improving patients' sedation status. There was no significantly difference in CPOT score between the two groups before enrollment (P = 0.902). After intervention, the CPOT score in the intervention group rapidly decreased from 3.00 points before enrollment to 1.00 points on the 1st day, and continued until the 7th day, while the control group showed a slower decrease from 2.50 points to 2.00 points and only dropped to 1.00 points on the 7th day. There were significant differences on 1st day and 3rd day between two groups (both P < 0.05). The time effect (F = 28.125, P < 0.001), group effect (F = 11.580, P = 0.001), and time group interaction effect (F = 4.048, P = 0.020) of the two groups of CPOT score were significant, indicating that light music therapy has better pain control, but the interaction effect is low, indicating that the impact of the intervention on the CPOT score was mainly concentrated in the early stage (1-3 days), and the long-term effect may be influenced by other factors. Compared with the control group, the intervention group showed a significant reduction in mechanical ventilation time (days: 10.57±2.94 vs. 11.95±3.74, P = 0.021) and the length of ICU stay (days: 14.91±4.37 vs. 17.53±4.83, P = 0.002). The ICU hospitalization expenses of the intervention group was slightly lower than that of the control group [ten thousand yuan: 22.431 (12.473, 28.489) vs. 29.362 (11.996, 41.389)], but the difference was not statistically significant (P = 0.086). CONCLUSIONS Light music therapy can effectively reduce the incidence of delirium in patients undergoing invasive mechanical ventilation, improve consciousness and pain perception, shorten mechanical ventilation time and hospital stay, and has significant clinical promotion value high-quality studies.
Humans ; Delirium/prevention & control* ; Intensive Care Units ; Respiration, Artificial ; Music Therapy ; Prospective Studies ; Male ; Female ; Middle Aged ; Critical Care ; Aged

Objective:To explore the repair mechanism of bone marrow stromal cells(BMSCs)transfected with miR-21 in rat sciatic nerve injuries.Methods:In vitro,rat BMSCs were transfected with miR-21 agonist(agomir-21)or blocker(antagomir-21)and their respective empty vectors,respectively,and divided into Control group,agomir-21 group,agomir-NC group,antagomir-21 group and antagomir-NC group.A 12 mm rat sciatic nerve injury model contai-ning decellularized nerve area was prepared,and the cells in each group were labeled with Hoechst33342 for postopera-tive implantation of decellularized nerve area.At 14 d after operation,the recovery of neuromotor function was evaluated by footprint test and the wet weight ratio of gastrocnemius.Immunofluorescence staining was applied to detect the expression of neurofilament protein 200(NF200),myelin basic protein(MBP),S100 calcium binding protein β(S100β)and glial fibrillary acidic protein(GFAP)within the regenerating nerve.Results:Compared with Control group,the sciatic nerve function index(SFI)and the wet weight ratio of gastrocnemius were significantly increased in agomir-21 group(P<0.05);and the density of regenerated nerve fibers and the expression of myelin marker proteins were increased,and the differentiation rate of BMSCs into Schwann cell-like cells(SCLCs)was elevated significantly(P<0.05).Conclusion:miR-21 significantly promotes peripheral nerve regeneration by enhancing the differentiation of BMSCs into SCLCs in vivo.

Since the birth of the first "test-tube baby" in Chinese mainland in 1988, assisted reproductive technology (ART) in China has matured significantly. The number of ART cycles has surpassed one million, and the number of assisted reproductive institutions and practitioners has attained a significant scale, contributing to the establishment of a fertility-friendly society. However, due to the complexity of the ART process, the diversity of personnel backgrounds, and the profound impact of any error that may occur, there is an urgent need to establish an efficient and effective safety management model for error prevention. This paper aims to outline the key processes and steps involved in the implementation of ART, explore control measures for these critical processes, and delve into error prevention strategies for gamete and embryo laboratories through the creation and utilization of a "gamete safety checklist".

Objective:To investigate the role of serotonergic (5-HT) neurons activation in the dorsal raphe nucleus (DRN) in heat-induced seizures and its influence on breathing patterns in Scn1a + /- mice. Methods:24-day-old male Scn1a + /- mice were used for experiments, and sudden unexpected death in epilepsy(SUDEP) model was established through heat induction for 5 consecutive days. (1) Fluoxetine intervention experiment: 20 male mice were randomly divided into model group( n=10) and fluoxetine group ( n=10) according the weight-matched method. The fluoxetine group received intraperitoneal injection of fluoxetine (10 mg/kg) 45 min before heat induction each day for 5 consecutive days, while the model group received equal volume of 0.9% NaCl solution. (2) Chemogenetic activation experiment: 20 male mice were randomly divided into vector control group( n=10) and chemogenetic activation group ( n=10) according the weight-matched method. Empty vector or rAAV-TPH2-hM3d(Gq)-EGFP-WPREs was stereotaxically injected into DRN 14 d prior to seizure induction, and deschloroclozapine (5 mg/kg) was intraperitoneally injected 30 min before heat induction. Seizure characteristics and survival were assessed through video monitoring, and respiratory parameters were monitored. Immunofluorescence was used to detect colocalization of tryptophan hydroxylase 2(TPH2) and c-Fos in DRN. Statistical analysis was performed using SPSS 24.0 and GraphPad Prism 9.0. The independent sample t-test or Mann-Whitney test was used for inter-group comparison. Results:(1) In the fluoxetine intervention experiment: the survival rates between the model group and fluoxetine group showed no statistically significant difference ( χ2=2.23, P>0.05). As for the frequency of grade Ⅳ seizures, the model group (1.50(1.25, 2.35)min) demonstrated higher frequency than the fluoxetine group (0.43(0.20, 0.67)min) ( t=-3.40, P<0.05).With respect to respiratory parameters, the model group demonstrated shorter expiratory time ((0.10±0.02) s) and inspiratory time ((0.15±0.02) s) compared to the fluoxetine group ((0.16±0.05) s, (0.19±0.04) s) ( t=-3.47, -3.73, both P<0.01). The respiratory rate in the model group ((269.96±44.84) times/min) was significantly higher compared to the fluoxetine group ((195.04±52.37) times/min) ( t=3.44, P<0.01). The tidal volume in the model group ((0.10±0.02) mL) was significantly lower than the fluoxetine group ((0.13±0.04) mL) ( t=-2.19, P<0.05).The number of TPH2+ /c-Fos+ co-expressing cells in the model group (11.00±4.00) was lower than that in the fluoxetine group (33.00±8.39)( t=-4.16, P<0.05). (2) In the chemogenetic activation experiment: compared to the vehicle group, the chemogenetic activation group demonstrated significantly enhanced survival rates ( χ2=5.83, P<0.05). As for the frequency of grade Ⅴ seizures, the vehicle group (2.11(1.62, 3.44) times/min) showed higher frequency compared to the chemogenetic activation group (0.81(0.00, 1.62) times/min) ( t=18.00, P<0.05). In terms of respiratory parameters, the vehicle group showed shorter expiratory time ((0.10±0.01) s) and inspiratory time ((0.14±0.01) s) compared to the chemogenetic activation group ((0.12±0.01) s, (0.15±0.01) s) ( t=-2.78, -2.50, both P<0.05). The respiratory rate in the vehicle group ((208.37±9.73) times/min) was significantly higher than the chemogenetic activation group ((191.85±8.83) times/min) ( t=3.98, P<0.01). The tidal volume in the vehicle group ((0.09±0.01) mL) was significantly lower than the chemogenetic activation group ((0.12±0.02) mL) ( t=-4.77, P<0.001).The number of TPH2+ /c-Fos+ co-expressing cells in the vehicle group (9.00±3.46) was lower than that in the chemogenetic activation group (43.00±11.02)( t=-5.20, P<0.01). Conclusion:Specific activation of serotonergic neurons in the DRN can ameliorate heat-induced epileptic symptoms, improve respiratory function, and prolong survival time in Scn1a + /- mice.