ObjectiveCandidate genes related to the regulation of litter size traits in English Springer Spaniel breeding bitches are explored, and the genetic mechanism underlying fertility in this breed is investigated, in order to provide reference molecular markers for genomic selection of high fecundity. MethodsWhole genome resequencing was performed on English Springer Spaniel breeding bitches that had given birth to at least 3 litters, and the bitches were divided into a high-litter-size group and a low-litter-size group according to the average litter size. Selection signal analysis was used to obtain the intersection of fixation index (F_st) and nucleotide diversity (Pi) signals as highly selected regions, and candidate genes were screened based on gene annotation and functional enrichment analysis. ResultsThe average litter size in the high-litter-size group (7.41±1.27) was significantly higher than that in the low-litter-size group (3.82±1.20) (P＜0.05), and the total number of live offspring in the high-litter-size group (7.06±1.10) was extremely significantly higher than that in the low-litter-size group (3.67±1.11) (P＜0.01). A total of 3 155 706 SNPs were detected in the two groups, 63.09% of which were located in intergenic regions, 33.96% in intronic regions, and 0.38％, 0.57％, and 0.09％ in exonic regions, 3' untranslated regions (UTRs), and 5'UTRs, respectively. Among the SNPs in exonic regions, 5 256 were nonsynonymous variants, accounting for 43.55%. A total of 1 752 differential genes were identified after annotation screening. Gene ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG) enrichment analyses identified 13 candidate genes that may affect reproductive performance and litter size traits, including WDR35, SMAD7, RPGR, RERGL, PGRMC2, LOC482182, GIMD1, COX7B2, COX16, BMPR2, BMP6, BICD1, and SLC9C1. Their functions mainly involve reproductive hormone regulation, embryonic development, GTPase activation, and oocyte apoptosis. ConclusionEnglish Springer Spaniel breeding bitches have undergone significant artificial selection for litter size traits. These 13 candidate genes play key roles in oocyte maturation and regulation during early pregnancy, providing a new molecular basis for elucidating the genetic mechanism of canine reproductive traits.

OBJECTIVE To prepare vancomycin hydrochloride(VH)-loaded polydopamine(PDA)nanoparticles(VH@PDA nanoparticles),and study their antimicrobial effect in combination with photothermal therapy.METHODS Using PDA as the carrier,VH was loaded to prepare VH@PDA nanoparticles(PDA nanoparticles were prepared using the same method).The nanoparticles were characterized with laser particle size analyzer,transmission electron microscope,and UV visible absorption spectrometer;the drug loading capacity and encapsulation efficiency of the nanoparticles were determined.The near-infrared laser irradiation was adopted to determine their photothermal ability.Taking Staphylococcus aureus as the research object,the bactericidal properties of the nanoparticles in combination with photothermal therapy in vitro were clarified through plate colony coating method,live/dead staining analysis,crystal violet staining.Using L929 cells as the research object,the effects of VH@PDA nanoparticles at different mass concentrations(0,3.125,6.25,12.5,25,50 and 100 μg/mL)on cell viability were investigated to assess their biocompatibility.RESULTS VH@PDA nanoparticles were successfully loaded with VH,exhibiting uniform particle size(approximately 300 nm),distinct pore size,and a spherical structure.The drug loading capacity was 11.34%,the encapsulation efficiency was 32.00%,the photothermal conversion efficiency reached 23.55%,and they demonstrated stable photothermal performance.The antibacterial effect results of the combined photothermal therapy demonstrated that without near-infrared laser irradiation,the antibacterial effects of VH,PDA nanoparticles,and VH@PDA nanoparticles were not significant.However,when subjected to near-infrared laser irradiation,VH@PDA nanoparticles exhibited a pronounced antibacterial effect.The results of the cell experiments revealed that after treatment with VH@PDA nanoparticles at various mass concentrations,the cell viability rates remained above 80%.CONCLUSIONS VH@PDA nanoparticles are successfully prepared,which exhibit stable photothermal properties,significant antibacterial effects when combined with photothermal therapy,and good biocompatibility.

Objective This study aims to explore the impact of changes in psychological status on non-suicidal self-inju-ry(NSSI)among junior high school students,in order to provide a theoretical basis for promoting their mental health development and to formulate more effective prevention and intervention measures.Methods A longitudinal study design was employed.Baseline data were collected in 2021(T1)from 652 first-year junior high school students selected through stratified cluster ran-dom sampling from two middle schools in a city.In 2023(T2),585 valid follow-up cases were obtained(effective response rate:89.72％).Chi-square tests,binary logistic regression,and ROC curve analysis were used to examine the effects of dynamic changes in psychological stress,internet addiction,depression,anxiety,and insomnia on NSSI(significance level α=0.05).Results ① Univariate analysis showed that the detection rate of NSSI significantly decreased from 22.05％(129/585)at T1 to 8.55％(50/585)at T2(x2=41.164,P＜0.001),with an average annual decline of 6.75％.Gender differences:the detec-tion rate decreased from 14.68％(43/293)to 5.80％(17/293)in boys(x2=18.577,P＜0.001),and from 29.45％(86/292)to 11.30％(33/292)in girls(x2=5.658,P=0.017),with a narrowing gender gap(T1:14.77％→ T2:5.70％).The NSSI detection rates between boys and girls were statistically significant at both time points(x2=18.577,5.658).② Com-parison between the persistent group and the NSSI-remission group showed that both decreased insomnia severity(OR=3.525,95％CI:1.230-10.105)and increased insomnia severity(OR=5.431,95％CI:1.895-15.570)were associated with an in-creased risk of NSSI persistence or onset(P＜0.01).③ Predictive efficacy:When scores of GAD-7+PHQ-9+ISI all in-creased by≥2 points,AUC=0.709,with a sensitivity of 44.0％-67.7％for predicting NSSI.When PHQ-9+ISI scores both increased by≥2 points,AUC=0.705,with a sensitivity of 50.2％-67.8％.Conclusion This study reveals a high natural re-mission rate of NSSI(82.17％,106/129 in the remission group)during junior high school,but also identifies gender heteroge-neity and persistent risks.Accumulated psychological stress,aggravated internet addiction,and worsened emotional disorders(anxiety/depression)are factors associated with NSSI persistence or onset.Clinical interventions should focus on monitoring dy-namic psychological indicators and early threshold identification.Although these results provide valuable insights,future research is needed to further explore the interaction mechanisms among these factors and how to effectively translate these findings into practical prevention and intervention measures.

Kidney transplantation is widely recognized as the optimal treatment for children with end-stage renal disease(ESRD),offering significant improvements in growth,development,and long-term quality of life compared with prolonged dialysis.However,kidney transplantation in low-age(＜5 years old)and low-weight(＜15 kg)children presents significant clinical challenges due to their delicate vas-cular structures,limited surgical space,and complex perioperative management.This report presents two cases of kidney transplantation in low-age,low-weight children performed at Peking University People's Hospital.Case 1:a 2-year-3-month-old boy(8.8 kg),presenting a preoperative serum creatinine of 248μmol/L post-dialysis and the estimated glomerular filtration rates(eGFR)of 35.17 mL/(min·1.73 m2).Case 2:a 3-year-8-month-old girl(11.25 kg),presenting a preoperative creatinine of 281 μmol/L post-dialysis and the eGFR of 22.63 mL/(min·1.73 m2).Both recipients underwent transplantation via the extraperitoneal approach,with end-to-side anastomosis of the donor renal artery and vein to the recipient's common iliac artery and vein,respectively.The ureters were anastomosed to the bladder using the tunnel technique,and double-J stents were placed intraoperatively.The surgeries were uneventful,and both pa-tients exhibited rapid recovery of renal function.Postoperatively,serum creatinine levels decreased to 26μmol/L(Case 1)and 39 μmol/L(Case 2)by the third day,with the eGFR reaching 245.23 mL/(min·1.73 m2)and 164.12 mL/(min·1.73 m2),respectively.No complications,such as vascular thrombosis,ureteral stenosis,or abdominal compartment syndrome were observed during follow-up.A comprehensive literature review was conducted to contextualize these cases within global advancements in pediatric renal transplantation.Current evidence highlights the growing adoption of kidney transplantation for low-age,low-weight children,though debates persist regarding optimal surgical strategies(specifical-ly,the intraperitoneal versus extraperitoneal approaches).This case report underscores the feasibility of the extraperitoneal approach in overcoming anatomical limitations of low-weight pediatric recipients,with distinct advantages including reduced gastrointestinal complications and enhanced accessibility for post-operative ultrasound monitoring.Furthermore,mean arterial pressure(MAP)and central venous pressure(C VP)were systematically monitored intraoperatively to ensure optimal renal blood perfusion and graft viability.Our single-center experience provides valuable insights into surgical strategy selection and peri-operative management for this high-risk population.Nevertheless,larger multicenter studies are warranted to validate long-term outcomes and refine standardized protocols.

Kidney transplantation is widely recognized as the optimal treatment for children with end-stage renal disease(ESRD),offering significant improvements in growth,development,and long-term quality of life compared with prolonged dialysis.However,kidney transplantation in low-age(＜5 years old)and low-weight(＜15 kg)children presents significant clinical challenges due to their delicate vas-cular structures,limited surgical space,and complex perioperative management.This report presents two cases of kidney transplantation in low-age,low-weight children performed at Peking University People's Hospital.Case 1:a 2-year-3-month-old boy(8.8 kg),presenting a preoperative serum creatinine of 248μmol/L post-dialysis and the estimated glomerular filtration rates(eGFR)of 35.17 mL/(min·1.73 m2).Case 2:a 3-year-8-month-old girl(11.25 kg),presenting a preoperative creatinine of 281 μmol/L post-dialysis and the eGFR of 22.63 mL/(min·1.73 m2).Both recipients underwent transplantation via the extraperitoneal approach,with end-to-side anastomosis of the donor renal artery and vein to the recipient's common iliac artery and vein,respectively.The ureters were anastomosed to the bladder using the tunnel technique,and double-J stents were placed intraoperatively.The surgeries were uneventful,and both pa-tients exhibited rapid recovery of renal function.Postoperatively,serum creatinine levels decreased to 26μmol/L(Case 1)and 39 μmol/L(Case 2)by the third day,with the eGFR reaching 245.23 mL/(min·1.73 m2)and 164.12 mL/(min·1.73 m2),respectively.No complications,such as vascular thrombosis,ureteral stenosis,or abdominal compartment syndrome were observed during follow-up.A comprehensive literature review was conducted to contextualize these cases within global advancements in pediatric renal transplantation.Current evidence highlights the growing adoption of kidney transplantation for low-age,low-weight children,though debates persist regarding optimal surgical strategies(specifical-ly,the intraperitoneal versus extraperitoneal approaches).This case report underscores the feasibility of the extraperitoneal approach in overcoming anatomical limitations of low-weight pediatric recipients,with distinct advantages including reduced gastrointestinal complications and enhanced accessibility for post-operative ultrasound monitoring.Furthermore,mean arterial pressure(MAP)and central venous pressure(C VP)were systematically monitored intraoperatively to ensure optimal renal blood perfusion and graft viability.Our single-center experience provides valuable insights into surgical strategy selection and peri-operative management for this high-risk population.Nevertheless,larger multicenter studies are warranted to validate long-term outcomes and refine standardized protocols.

Objective:To observe the clinical efficacy of gentle moxibustion at different temperatures in treating people with diarrhea-predominant bowel syndrome(IBS-D)due to spleen deficiency.Methods:A total of 108 IBS-D patients were divided into two groups using the random number table method,with 54 participants in each group.Moxibustion group 1 received gentle moxibustion at(43±1)℃at bilateral Tianshu(ST25)and Zusanli(ST36),lasting 30 min each session;moxibustion group 2 received gentle moxibustion at(37±1)℃at the same points.Both groups received the intervention 3 times weekly for a total of 18 sessions.Abdominal pain intensity,stool form,pattern-based efficacy,quality of life,and mental health assessments were performed at weeks 0,3,6,and 8.Results:The total effective rate for abdominal pain intensity was 87.8%in moxibustion group 1 versus 51.1%in moxibustion group 2,and the difference was statistically significant(P<0.05).When the treatment finished,abdominal pain intensity,the Bristol score,IBS-symptom severity scale(IBS-SSS)score,self-rating anxiety scale(SAS)score,and self-rating depression scale(SDS)score dropped significantly in both groups(P<0.05),and the IBS-quality of life(IBS-QOL)score increased markedly(P<0.05).Between-group comparisons demonstrated that abdominal pain intensity,the Bristol general score,IBS-SSS score,traditional Chinese medicine(TCM)pattern score,and SDS score were significantly lower in moxibustion group 1 than in moxibustion group 2 at treatment week 6(P<0.05),and the IBS-QOL score was notably higher in moxibustion group 1(P<0.05).Conclusion:Whether at 43℃or 37℃,gentle moxibustion at Tianshu(ST25)and Zusanli(ST36)can improve abdominal pain,stool form,and quality of life,reduce disease severity,and mitigate TCM pattern in IBS-D patients;43℃gentle moxibustion performs better than 37℃gentle moxibustion in improving abdominal pain,stool form,disease severity,TCM pattern,quality of life,anxiety,and depression in IBS-D.

Objective:To construct a VP8-mRNA vaccine using human rotavirus spike protein VP8 domain as the immunogen and analyze its immunogenicity in mice.Methods:The VP8-mRNA sequence was designed, optimized, and synthesized. The VP8 gene of rotavirus G1P[8] type was used to construct the plasmid pUC57-VP8-Kan-SapⅠ, which was then sequenced. The plasmid confirmed by sequencing was subjected to large-scale amplification and extraction, followed by linearization, in vitro transcription, and capping. The purified capped products were encapsulated with lipid nanoparticles using a microfluidic control apparatus. The encapsulated VP8-mRNA vaccine was administered intramuscularly to mice at 10, 15, and 20 μg. Serum samples were collected for antibody detection by ELISA. Cellular immune responses were detected by flow cytometry and ELISPOT. Statistical analysis was performed using one-way or two-way analysis of variance and Tukey-Kramer test. Results:The encapsulated VP8-mRNA vaccine was rounded and spherical, with a particle size of about 100 nm, a polymer dispersion index of 0.088, and an encapsulation rate of 92.3%. Two doses of VP8-mRNA vaccine immunization could induce a good immune response in mice. The level of IgG antibody induced after immunization in the 15 μg group was comparable to that of the 20 μg group, and there was no statistical difference ( P>0.05), but the antibody levels in the two groups were significantly higher than that in the 10 μg group ( P<0.000 1). VP8-mRNA vaccine could induce neutralizing antibodies against rotavirus G1 and G9 types. The highest level of neutralizing antibodies against rotavirus type G1 was observed in the 15 μg group, which was significantly higher than that in the 10 μg group ( P<0.05). All immunization groups exhibited good neutralizing ability against rotavirus G9 type. The results of ELISPOT showed that lymphocytes from mice in each vaccine group were able to secrete IFN-γ when stimulated with VP8 peptide. Flow cytometry showed that the proportions of CD8 + T cell subsets in the vaccine groups were higher than that in the control group. Conclusion:The VP8-mRNA vaccine has good immunogenicity in mice and can induce good humoral and T-cell immune responses.

Objective This study aims to explore the impact of changes in psychological status on non-suicidal self-inju-ry(NSSI)among junior high school students,in order to provide a theoretical basis for promoting their mental health development and to formulate more effective prevention and intervention measures.Methods A longitudinal study design was employed.Baseline data were collected in 2021(T1)from 652 first-year junior high school students selected through stratified cluster ran-dom sampling from two middle schools in a city.In 2023(T2),585 valid follow-up cases were obtained(effective response rate:89.72％).Chi-square tests,binary logistic regression,and ROC curve analysis were used to examine the effects of dynamic changes in psychological stress,internet addiction,depression,anxiety,and insomnia on NSSI(significance level α=0.05).Results ① Univariate analysis showed that the detection rate of NSSI significantly decreased from 22.05％(129/585)at T1 to 8.55％(50/585)at T2(x2=41.164,P＜0.001),with an average annual decline of 6.75％.Gender differences:the detec-tion rate decreased from 14.68％(43/293)to 5.80％(17/293)in boys(x2=18.577,P＜0.001),and from 29.45％(86/292)to 11.30％(33/292)in girls(x2=5.658,P=0.017),with a narrowing gender gap(T1:14.77％→ T2:5.70％).The NSSI detection rates between boys and girls were statistically significant at both time points(x2=18.577,5.658).② Com-parison between the persistent group and the NSSI-remission group showed that both decreased insomnia severity(OR=3.525,95％CI:1.230-10.105)and increased insomnia severity(OR=5.431,95％CI:1.895-15.570)were associated with an in-creased risk of NSSI persistence or onset(P＜0.01).③ Predictive efficacy:When scores of GAD-7+PHQ-9+ISI all in-creased by≥2 points,AUC=0.709,with a sensitivity of 44.0％-67.7％for predicting NSSI.When PHQ-9+ISI scores both increased by≥2 points,AUC=0.705,with a sensitivity of 50.2％-67.8％.Conclusion This study reveals a high natural re-mission rate of NSSI(82.17％,106/129 in the remission group)during junior high school,but also identifies gender heteroge-neity and persistent risks.Accumulated psychological stress,aggravated internet addiction,and worsened emotional disorders(anxiety/depression)are factors associated with NSSI persistence or onset.Clinical interventions should focus on monitoring dy-namic psychological indicators and early threshold identification.Although these results provide valuable insights,future research is needed to further explore the interaction mechanisms among these factors and how to effectively translate these findings into practical prevention and intervention measures.

Objective:To observe the clinical efficacy of gentle moxibustion at different temperatures in treating people with diarrhea-predominant bowel syndrome(IBS-D)due to spleen deficiency.Methods:A total of 108 IBS-D patients were divided into two groups using the random number table method,with 54 participants in each group.Moxibustion group 1 received gentle moxibustion at(43±1)℃at bilateral Tianshu(ST25)and Zusanli(ST36),lasting 30 min each session;moxibustion group 2 received gentle moxibustion at(37±1)℃at the same points.Both groups received the intervention 3 times weekly for a total of 18 sessions.Abdominal pain intensity,stool form,pattern-based efficacy,quality of life,and mental health assessments were performed at weeks 0,3,6,and 8.Results:The total effective rate for abdominal pain intensity was 87.8%in moxibustion group 1 versus 51.1%in moxibustion group 2,and the difference was statistically significant(P<0.05).When the treatment finished,abdominal pain intensity,the Bristol score,IBS-symptom severity scale(IBS-SSS)score,self-rating anxiety scale(SAS)score,and self-rating depression scale(SDS)score dropped significantly in both groups(P<0.05),and the IBS-quality of life(IBS-QOL)score increased markedly(P<0.05).Between-group comparisons demonstrated that abdominal pain intensity,the Bristol general score,IBS-SSS score,traditional Chinese medicine(TCM)pattern score,and SDS score were significantly lower in moxibustion group 1 than in moxibustion group 2 at treatment week 6(P<0.05),and the IBS-QOL score was notably higher in moxibustion group 1(P<0.05).Conclusion:Whether at 43℃or 37℃,gentle moxibustion at Tianshu(ST25)and Zusanli(ST36)can improve abdominal pain,stool form,and quality of life,reduce disease severity,and mitigate TCM pattern in IBS-D patients;43℃gentle moxibustion performs better than 37℃gentle moxibustion in improving abdominal pain,stool form,disease severity,TCM pattern,quality of life,anxiety,and depression in IBS-D.

Objective:To construct a VP8-mRNA vaccine using human rotavirus spike protein VP8 domain as the immunogen and analyze its immunogenicity in mice.Methods:The VP8-mRNA sequence was designed, optimized, and synthesized. The VP8 gene of rotavirus G1P[8] type was used to construct the plasmid pUC57-VP8-Kan-SapⅠ, which was then sequenced. The plasmid confirmed by sequencing was subjected to large-scale amplification and extraction, followed by linearization, in vitro transcription, and capping. The purified capped products were encapsulated with lipid nanoparticles using a microfluidic control apparatus. The encapsulated VP8-mRNA vaccine was administered intramuscularly to mice at 10, 15, and 20 μg. Serum samples were collected for antibody detection by ELISA. Cellular immune responses were detected by flow cytometry and ELISPOT. Statistical analysis was performed using one-way or two-way analysis of variance and Tukey-Kramer test. Results:The encapsulated VP8-mRNA vaccine was rounded and spherical, with a particle size of about 100 nm, a polymer dispersion index of 0.088, and an encapsulation rate of 92.3%. Two doses of VP8-mRNA vaccine immunization could induce a good immune response in mice. The level of IgG antibody induced after immunization in the 15 μg group was comparable to that of the 20 μg group, and there was no statistical difference ( P>0.05), but the antibody levels in the two groups were significantly higher than that in the 10 μg group ( P<0.000 1). VP8-mRNA vaccine could induce neutralizing antibodies against rotavirus G1 and G9 types. The highest level of neutralizing antibodies against rotavirus type G1 was observed in the 15 μg group, which was significantly higher than that in the 10 μg group ( P<0.05). All immunization groups exhibited good neutralizing ability against rotavirus G9 type. The results of ELISPOT showed that lymphocytes from mice in each vaccine group were able to secrete IFN-γ when stimulated with VP8 peptide. Flow cytometry showed that the proportions of CD8 + T cell subsets in the vaccine groups were higher than that in the control group. Conclusion:The VP8-mRNA vaccine has good immunogenicity in mice and can induce good humoral and T-cell immune responses.