Objective:To investigate the hepatoprotective effect of N-acetylcysteine(NAC)on rats with liver injury induced by cisplatin and its effect on intestinal flora and the expression of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and nuclear factor-kappa B(NF-κB).Methods:Male Sprague-Dawley rats were randomly divided into control group(CG),cisplatin group(CP),and NAC group.The rats in the NAC group were given NAC 15 mg/kg by gavage for 8 consecutive days.At half an hour after intragastric administration on the fifth day,all rats except those in the NC group were given intraperitoneal injection of 8 mg/kg cisplatin to induce acute liver injury.An automatic biochemical analyzer was used to measure the content of aspartate aminotransferase(AST),alanine aminotransferase(ALT),alkaline phosphatase(ALP),and total bilirubin(TBIL);liver index was calculated for the rats;Western blot was used to measure the relative expression levels of NF-κB,IL-6,and TNF-α in liver tissue;the 16S rDNA technique was used to measure and analyze the amplification information of the V3-V4 regions of each sample.Results:Compared with the NC group,the CP group had significant increases in the content of AST,ALT,ALP,and TBIL,while NAC reversed the abnormal liver function caused by cisplatin.Compared with the NC group,the CP group had a sig-nificant increase in liver index(P=0.000),while the NAC group had a significant reduction in liver index compared with the CP group(P=0.007).Compared with the NC group,the CP group had signifi-cant increases in the expression levels of IL-6,TNF-α,and NF-κB,while the NAC group showed reductions in the expression of these genes,with significant differences in the expression of IL-6 and TNF-α(P=0.006 and 0.000).Compared with the NC group,the CP group had a significant increase in the α-diversity index of intesti-nal flora,while compared with the CP group,the NAC group tended to have a reduction in the α-diversity index of intestinal flora.Com-pared with the CP group at the phylum level,the NAC group had an increase in the abundance of Actinobacteria and a reduction in the abundance of Firmicutes.Compared with the CP group at the genus level,the NAC group had a reduction in the abundance of Rumino-coccaceae and increases in the abundance of Bifidobacterium and Allobaculum.Conclusion:NAC can alleviate acute liver injury caused by cisplatin,possibly by downregulating the expression of IL-6,TNF-α,and NF-κB and regulating the abundance and diver-sity of intestinal flora.

Objective:To compare the efficacy of Ilizarov ring external fixation and unilateral rail external fixation in the treatment of infected bone defects following surgery for tibial fractures.Methods:A retrospective cohort study was conducted to analyze the clinical data of 50 patients with infected bone defects after surgery for tibial fractures, who were admitted to the 940th Hospital of the Joint Logistics Support Force of the PLA from August 2019 to November 2021, including 37 males and 13 females, aged 19-59 years [(42.2±8.8)years]. After debridement and osteotomy, 28 patients were treated with Ilizarov ring external fixation (Ilizarov group) and 22 with unilateral rail external fixation (unilateral fixation group). All the patients in the two groups had previously undergone internal fixation with plates or Kirschner wires for tibial fracture before bone transport. Bone transport started at one week for three stages after successful infection control and osteotomy and was conducted. The following parameters were compared between the two groups: frame-wearing time and healing index after bone transport, self-rating anxiety scale (SAS) grade at 6 months after bone transport, Paley score and Association for the Study and Application of the Method of Ilizarov (ASAMI) score at the last follow-up, Hospital for Special Surgery (HSS) knee score and Baird-Jackson ankle score on admission, after external fixator removal and at the last follow-up, and incidence of postoperative complications.Results:All the patients were followed up for 28-36 months [(32.5±1.6)months]. There were no significant differences in frame-wearing time or healing index between the two groups after bone transport ( P>0.05). At 6 months after bone transport, the SAS grade in the unilateral fixation group (13 patients with mild anxiety, 8 with moderate anxiety, and 1 with severe anxiety) was better than that in the Ilizarov group (6 patients with mild anxiety, 19 with moderate anxiety, 3 with severe anxiety) ( P<0.01). No significant differences were found in the Paley score or ASAMI score between the two groups at the last follow-up ( P>0.05). There were no significant differences in HSS knee score or Baird-Jackson ankle score between the two groups on admission, after external fixator removal or at the last follow-up ( P>0.05). No significant differences were observed in the incidence of pin tract infection, poor healing, infection in the bone elongation area, or re-fracture between the two groups ( P>0.05). The incidence of postoperative axial deviation was 0 in the Ilizarov group, lower than 18% in the unilateral fixation group (4/22) ( P<0.05). Conclusion:Although Ilizarov ring external fixation and unilateral rail external fixation demonstrate comparable efficacy in the treatment of infected bone defects after surgery for tibial fractures, the former provides superior mechanical stability and postoperative axial deviation correction, while the latter offers advantages in reducing psychological burden and enhancing treatment tolerance.

Objective To test the hypothesis that backward crawling and forward crawling share similar inter-joint coordination patterns,thus providing potential evidence for the application of backward crawling in rehabilitation training.Methods The acceleration signals in the X,Y,and Z directions for 9 joints(including bilateral wrists,elbows,shoulders,knees,and hips)in 9 volunteers during forward and backward crawling were collected using a custom signal acquisition system,and the pressure signals were also recorded when the palms contacted the ground.The collected acceleration signals were preprocessed,segmented into cycles,and vectorized.Based on the pressure signals,a single crawling cycle was divided into support phase and swing phase.In addition,principal component analysis was applied to extract inter-joint coordination in limbs at various scales(sagittal,coronal,and transverse planes).Pearson correlation coefficients of inter-joint coordination patterns were compared between forward and backward crawling in support period,swing period,and full cycle.Results The correlation coefficients for coordination patterns in the full cycle at the transverse plane scale were 0.813 5(PC1)and 0.837 5(PC2),and the correlation coefficient of the support period PC2 was 0.901 8.At the sagittal plane scale,the correlation coefficient of the support period PC1 was 0.948 5.Conclusion The study provides preliminary evidence that limb motion coordination patterns during backward crawling are similar to those observed during forward crawling.Future research will further explore the effects of backward crawling on functional rehabilitation in individuals with motor impairments.

Objective To investigate the clinical and genetic characteristics of infantile early-onset Moyamoya disease associated with ring finger protein 213(RNF213)gene variants.Methods This study included retrospective,consecutive case series of infantile early-onset Moyamoya disease admitted to the Department of Pediatric Neurology,Wuhan Children's Hospital,Tongji Medical College,Huazhong University of Science and Technology,between June 2020 and June 2024.General and clinical data were collected,including sex,age,chief complaint,admission time,history of present illness,personal history,family history,neurological physical examination,auxiliary investigations,diagnosis/treatment course,and follow-up status.Genetic testing results of the patients and their parents were analyzed.Mutation sites were screened and annotated through bioinformatics analysis,and suspicious variants were validated by Sanger sequencing.The molecular evolutionary genetics analysis software MEGA(v11.0)was used for amino acid sequence alignment and conservation analysis of identified missense mutations.Based on the protein structure databases,the three-dimensional structure of the missense proteins was obtained through cryo-electron microscopy.The identified missense mutations were mapped onto this structure,and PyMOL(v2.0.6)was used for protein structure visualization and hydrogen bond analysis at variant sites.Results(1)Three infants with early-onset Moyamoya disease were enrolled(1 male,2 females).Initial symptoms included epileptic seizures,transient ischemic attacks,or acute cerebral infarction.Genetic testing revealed RNF213 missense mutations in all three patients:patient 1 carried p.Arg4810Lys and p.Thr1727Met(both maternally inherited);patient 2 carried p.Arg4810Lys(paternally inherited);patient 3 carried a de novo p.Lys4136Gln mutation(both parents were wild-type).All patients underwent indirect cerebral revascularization.Postoperatively,epileptic seizures and cerebral ischemic events gradually resolved,the modified Rankin scale scores at the last follow-up were 2,1,and 2,respectively.(2)The amino acid residues threonine 1727(THR1727),lysine 4136(LYS4136),and arginine 4810(ARG4810)in the RNF213 protein were absolutely conserved across all three patients.Compared to wild-type ARG4810,the p.Arg4810Lys mutation occurred within an α-helix and reduced internal hydrogen bond interactions by one.The p.Thr1727Met mutation reduced hydrogen bonding by one compared to wild-type THR1727.The p.Lys4136Gln mutation reduced hydrogen bonding by one relative to wild-type LYS4136 and altered residues connecting two α-helices.Conclusions Infantile early-onset Moyamoya disease has insidious onset,severe progression,and diverse clinical manifestations,making it prone to misdiagnosis.RNF213 gene p.Arg4810Lys,p.Thr1727Met and p.Lys4136Gln mutations contribute to the pathogenesis of Moyamoya disease by affecting molecular mechanisms underlying its protein function.Early genetic testing for RNF213 variants aids clinical diagnosis.Indirect cerebral revascularization may benefit the affected infants.These findings require further validation through large-scale,prospective,multicenter studies.

Objective To explore the resistance mechanism of tigecycline insensitivity of multidrug-resistant Acine-tobacter baumannii,and provide reference for clinical rational antimicrobial use as well as prevention and control of healthcare-associated infection.Methods 22 strains of tigecycline insensitive multidrug-resistant Acinetobacter bau-mannii(TIS-MDR-AB)and 22 strains of tigecycline sensitive multidrug-resistant Acinetobacter baumannii(TS-MDR-AB)isolated clinically from the First Affiliated Hospital of Guangzhou Medical University from April 2022 to May 2023 were collected.Efflux pump phenotype inhibition assay was performed using efflux pump inhibitor car-bonyl cyanide m-chlorophenylhydrazone(CCCP).The main efflux pump genes(adeB,adeG,adeJ),as well as tigecycline-resistant gene tet(X),were screened by polymerase chain reaction(PCR)technique,and their mRNA expression levels were detected by real-time fluorescence quantitative PCR.Mutations in the efflux pump regulatory gene adeRS were analyzed by Sanger sequencing analysis.Results The detection rates of efflux pump genes adeB,adeG and adeJ were all above 95％in two MDR-AB groups,and tet(X)gene was not detected.Efflux pump inhibi-tor assay showed that the minimum inhibitory concentration(MIC)of TIS-MDR-AB strains decreased after adding CCCP,3 strains showed positive efflux pump phenotype.The mRNA expression level of MDR-AB adeB in the TIS-MDR-AB group was higher than that in the TS-MDR-AB group(P＜0.01),while the expression of adeG and adeJ genes was no statistically different.Multiple mutations were found in the adeR and adeS genes,the adeS of 2 strains was inserted ISAba1,and 3 strains were inserted ISAba13.Conclusion The overexpression of adeABC in the efflux pump system may play an important role in the mechanism of reduced sensitivity of MDR-AB to tigecycline,and its overexpression may be related to the insertion sequence or mutation in the efflux pump regulatory gene adeRS.

Objective To test the hypothesis that backward crawling and forward crawling share similar inter-joint coordination patterns,thus providing potential evidence for the application of backward crawling in rehabilitation training.Methods The acceleration signals in the X,Y,and Z directions for 9 joints(including bilateral wrists,elbows,shoulders,knees,and hips)in 9 volunteers during forward and backward crawling were collected using a custom signal acquisition system,and the pressure signals were also recorded when the palms contacted the ground.The collected acceleration signals were preprocessed,segmented into cycles,and vectorized.Based on the pressure signals,a single crawling cycle was divided into support phase and swing phase.In addition,principal component analysis was applied to extract inter-joint coordination in limbs at various scales(sagittal,coronal,and transverse planes).Pearson correlation coefficients of inter-joint coordination patterns were compared between forward and backward crawling in support period,swing period,and full cycle.Results The correlation coefficients for coordination patterns in the full cycle at the transverse plane scale were 0.813 5(PC1)and 0.837 5(PC2),and the correlation coefficient of the support period PC2 was 0.901 8.At the sagittal plane scale,the correlation coefficient of the support period PC1 was 0.948 5.Conclusion The study provides preliminary evidence that limb motion coordination patterns during backward crawling are similar to those observed during forward crawling.Future research will further explore the effects of backward crawling on functional rehabilitation in individuals with motor impairments.

Objective To investigate the clinical and genetic characteristics of infantile early-onset Moyamoya disease associated with ring finger protein 213(RNF213)gene variants.Methods This study included retrospective,consecutive case series of infantile early-onset Moyamoya disease admitted to the Department of Pediatric Neurology,Wuhan Children's Hospital,Tongji Medical College,Huazhong University of Science and Technology,between June 2020 and June 2024.General and clinical data were collected,including sex,age,chief complaint,admission time,history of present illness,personal history,family history,neurological physical examination,auxiliary investigations,diagnosis/treatment course,and follow-up status.Genetic testing results of the patients and their parents were analyzed.Mutation sites were screened and annotated through bioinformatics analysis,and suspicious variants were validated by Sanger sequencing.The molecular evolutionary genetics analysis software MEGA(v11.0)was used for amino acid sequence alignment and conservation analysis of identified missense mutations.Based on the protein structure databases,the three-dimensional structure of the missense proteins was obtained through cryo-electron microscopy.The identified missense mutations were mapped onto this structure,and PyMOL(v2.0.6)was used for protein structure visualization and hydrogen bond analysis at variant sites.Results(1)Three infants with early-onset Moyamoya disease were enrolled(1 male,2 females).Initial symptoms included epileptic seizures,transient ischemic attacks,or acute cerebral infarction.Genetic testing revealed RNF213 missense mutations in all three patients:patient 1 carried p.Arg4810Lys and p.Thr1727Met(both maternally inherited);patient 2 carried p.Arg4810Lys(paternally inherited);patient 3 carried a de novo p.Lys4136Gln mutation(both parents were wild-type).All patients underwent indirect cerebral revascularization.Postoperatively,epileptic seizures and cerebral ischemic events gradually resolved,the modified Rankin scale scores at the last follow-up were 2,1,and 2,respectively.(2)The amino acid residues threonine 1727(THR1727),lysine 4136(LYS4136),and arginine 4810(ARG4810)in the RNF213 protein were absolutely conserved across all three patients.Compared to wild-type ARG4810,the p.Arg4810Lys mutation occurred within an α-helix and reduced internal hydrogen bond interactions by one.The p.Thr1727Met mutation reduced hydrogen bonding by one compared to wild-type THR1727.The p.Lys4136Gln mutation reduced hydrogen bonding by one relative to wild-type LYS4136 and altered residues connecting two α-helices.Conclusions Infantile early-onset Moyamoya disease has insidious onset,severe progression,and diverse clinical manifestations,making it prone to misdiagnosis.RNF213 gene p.Arg4810Lys,p.Thr1727Met and p.Lys4136Gln mutations contribute to the pathogenesis of Moyamoya disease by affecting molecular mechanisms underlying its protein function.Early genetic testing for RNF213 variants aids clinical diagnosis.Indirect cerebral revascularization may benefit the affected infants.These findings require further validation through large-scale,prospective,multicenter studies.

Objective:To compare the efficacy of Ilizarov ring external fixation and unilateral rail external fixation in the treatment of infected bone defects following surgery for tibial fractures.Methods:A retrospective cohort study was conducted to analyze the clinical data of 50 patients with infected bone defects after surgery for tibial fractures, who were admitted to the 940th Hospital of the Joint Logistics Support Force of the PLA from August 2019 to November 2021, including 37 males and 13 females, aged 19-59 years [(42.2±8.8)years]. After debridement and osteotomy, 28 patients were treated with Ilizarov ring external fixation (Ilizarov group) and 22 with unilateral rail external fixation (unilateral fixation group). All the patients in the two groups had previously undergone internal fixation with plates or Kirschner wires for tibial fracture before bone transport. Bone transport started at one week for three stages after successful infection control and osteotomy and was conducted. The following parameters were compared between the two groups: frame-wearing time and healing index after bone transport, self-rating anxiety scale (SAS) grade at 6 months after bone transport, Paley score and Association for the Study and Application of the Method of Ilizarov (ASAMI) score at the last follow-up, Hospital for Special Surgery (HSS) knee score and Baird-Jackson ankle score on admission, after external fixator removal and at the last follow-up, and incidence of postoperative complications.Results:All the patients were followed up for 28-36 months [(32.5±1.6)months]. There were no significant differences in frame-wearing time or healing index between the two groups after bone transport ( P>0.05). At 6 months after bone transport, the SAS grade in the unilateral fixation group (13 patients with mild anxiety, 8 with moderate anxiety, and 1 with severe anxiety) was better than that in the Ilizarov group (6 patients with mild anxiety, 19 with moderate anxiety, 3 with severe anxiety) ( P<0.01). No significant differences were found in the Paley score or ASAMI score between the two groups at the last follow-up ( P>0.05). There were no significant differences in HSS knee score or Baird-Jackson ankle score between the two groups on admission, after external fixator removal or at the last follow-up ( P>0.05). No significant differences were observed in the incidence of pin tract infection, poor healing, infection in the bone elongation area, or re-fracture between the two groups ( P>0.05). The incidence of postoperative axial deviation was 0 in the Ilizarov group, lower than 18% in the unilateral fixation group (4/22) ( P<0.05). Conclusion:Although Ilizarov ring external fixation and unilateral rail external fixation demonstrate comparable efficacy in the treatment of infected bone defects after surgery for tibial fractures, the former provides superior mechanical stability and postoperative axial deviation correction, while the latter offers advantages in reducing psychological burden and enhancing treatment tolerance.

Objective To explore the resistance mechanism of tigecycline insensitivity of multidrug-resistant Acine-tobacter baumannii,and provide reference for clinical rational antimicrobial use as well as prevention and control of healthcare-associated infection.Methods 22 strains of tigecycline insensitive multidrug-resistant Acinetobacter bau-mannii(TIS-MDR-AB)and 22 strains of tigecycline sensitive multidrug-resistant Acinetobacter baumannii(TS-MDR-AB)isolated clinically from the First Affiliated Hospital of Guangzhou Medical University from April 2022 to May 2023 were collected.Efflux pump phenotype inhibition assay was performed using efflux pump inhibitor car-bonyl cyanide m-chlorophenylhydrazone(CCCP).The main efflux pump genes(adeB,adeG,adeJ),as well as tigecycline-resistant gene tet(X),were screened by polymerase chain reaction(PCR)technique,and their mRNA expression levels were detected by real-time fluorescence quantitative PCR.Mutations in the efflux pump regulatory gene adeRS were analyzed by Sanger sequencing analysis.Results The detection rates of efflux pump genes adeB,adeG and adeJ were all above 95％in two MDR-AB groups,and tet(X)gene was not detected.Efflux pump inhibi-tor assay showed that the minimum inhibitory concentration(MIC)of TIS-MDR-AB strains decreased after adding CCCP,3 strains showed positive efflux pump phenotype.The mRNA expression level of MDR-AB adeB in the TIS-MDR-AB group was higher than that in the TS-MDR-AB group(P＜0.01),while the expression of adeG and adeJ genes was no statistically different.Multiple mutations were found in the adeR and adeS genes,the adeS of 2 strains was inserted ISAba1,and 3 strains were inserted ISAba13.Conclusion The overexpression of adeABC in the efflux pump system may play an important role in the mechanism of reduced sensitivity of MDR-AB to tigecycline,and its overexpression may be related to the insertion sequence or mutation in the efflux pump regulatory gene adeRS.

Objective To investigate the influencing mechanism of micro ribonucleic acid(miR)-375 targeting phos-phatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)pathway on high glucose-induced proliferation and angiogenesis in human retinal endothelial cells(hRECs).Methods The hRECs were cultured in vitro,and transfection and dual lucifer-ase assay were performed on them.These hRECs were divided into the control group,high glucose group,high glucose+miR-375 group,and high glucose+miR-375+LM22B-10 group.The Cell Counting Kit-8 was used to detect the cell prolifera-tion ability,the angiogenesis assay was used to detect the vascular formation ability,real-time quantitative PCR was used to detect the miR-375 and PI3K mRNA expressions in hRECs,and Western blot was used to detect the PI3K and p-AKT/AKT protein expressions in hRECs.Results At 48 h and 72 h after the cultivation,compared with the control group,the pro-liferation viability,PI3K and p-AKT/AKT protein expressions,vascular formation ability,and PI3K mRNA expression in hRECs significantly increased,and the miR-375 expression in hRECs significantly decreased in the high glucose group,high glucose+miR-375 group and high glucose+miR-375+LM22B-10 group(all P＜0.05).Compared with the high glucose group,the proliferation viability,PI3K mRNA and protein expressions,p-AKT/AKT protein expression and vascular forma-tion ability in hRECs were significantly reduced,and miR-375 expression significantly increased in the high glucose+miR-375 group(all P＜0.05).Compared with the high glucose+miR-375 group,the proliferation viability,vascular formation a-bility and p-AKT/AKT protein expression in hRECs significantly increased in the high glucose+miR-375+LM22B-10 group(all P＜0.05);there was no significant difference in the miR-375 and PI3K mRNA(all P＞0.05).After transfected with miR-375 mimic and wt-PI3K-pGL4,the relative luciferase activity in hRECs significantly decreased compared with transfec-tion with miR-375 NC and mut-PI3K-pGL4(all P＜0.05).Conclusion The targeted inhibition of the PI3K/AKT pathway by miR-375 can suppress the high glucose-induced proliferation and angiogenesis of hRECs,alleviating DR.