OBJECTIVE: To prepare decellularized nerve grafts from alpha-1, 3-galactosyltransferase (GGTA1) gene-edited pigs and explore their biocompatibility for xenotransplantation. METHODS: The sciatic nerves from wild-type pigs and GGTA1 gene-edited pigs were obtained and underwent decellularization. The alpha-galactosidase (α-gal) content in the sciatic nerves of GGTA1 gene-edited pigs was detected by using IB4 fluorescence staining and ELISA method to verify the knockout status of the GGTA1 gene, and using human sciatic nerve as a control. HE staining and scanning electron microscopy observation were used to observe the structure of the nerve samples. Immunofluorescence staining and DNA content determination were used to evaluate the degree of decellularization of the nerve samples. Fourteen nude mice were taken, and subcutaneous capsules were prepared on both sides of the spine. Decellularized nerve samples of wild-type pigs ( n=7) and GGTA1 gene-edited pigs ( n=7) were randomly implanted in the subcutaneous capsules. Blood was drawn at 1, 3, 5, and 7 days after implantation to detect neutrophil counting. RESULTS: IB4 fluorescence staining and ELISA detection showed that GGTA1 gene was successfully knocked out in the nerves of GGTA1 gene-edited pigs. HE staining showed that the structure of the decellularized nerve from GGTA1 gene-edited pigs was well preserved; the nerve basement membrane tube structure was visible under scanning electron microscopy; no cell nuclei was observed, and the extracellular matrix components was retained in the nerve grafts by immunofluorescence staining; and the DNA content was significantly reduced when compared with the normal nerves ( P<0.05). In vivo experiments showed that the number of neutrophils in the two groups were similar at 1, 3, and 7 days after implantation, with no significant difference ( P>0.05); only at 5 days, the number of neutrophils was significantly lower in the GGTA1 gene-edited pigs than in the wild-type pigs ( P<0.05). CONCLUSION The decellularized nerve grafts from GGTA1 gene-edited pigs have well-preserved nerve structure, complete decellularization, retain the natural nerve basement membrane tube structure and components, and low immune response after xenotransplantation through in vitro experiments.
Animals ; Transplantation, Heterologous ; Galactosyltransferases/genetics* ; Sciatic Nerve/immunology* ; Swine ; Tissue Engineering/methods* ; Humans ; Graft Rejection/prevention & control* ; Gene Editing ; Mice ; Mice, Nude ; Heterografts/immunology* ; Animals, Genetically Modified ; Tissue Scaffolds ; Decellularized Extracellular Matrix

Objective:To compare the outcomes of multiple scales, primarily the speech, spatial, and other qualities of hearing scale for parents（SSQ-P）, in children with ipsilateral vs. Contralateral cochleareimplantat ion（CRI）. Methods: A total of 69 children who received cochlear implantation surgery from April 1999 to June 2024 were included. Patients were divided into two groups based on whether the implantation was on the same side. General information such as gender, age, age at initial implantation and reimplantation was collected. The primary caregivers of the children were followed up by telephone using the categories of auditory performance（CAP）, speech intelligibility rating（SIR）, and SSQ-P questionnaires. Statistical methods including stepwise regression, linear regression, and permutation tests were employed to investigate if there were any statistically significant differences in the scores of CAP, SIR, SSQ-P total, SSQ-P speech perception, SSQ-P spatial hearing, and SSQ-P auditory quality dimensions between the ipsilateral and contralateral reimplantation groups. Results:Of the 69 children included, 62 were in the ipsilateral reimplantation group with a mean age of 11.1 years, and 7 were in the contralateral reimplantation group with a mean age of 11.7 years. Statistical analysis showed that patients in the contralateral reimplantation group had significantly lower SSQ-P total scores （P<0.05） and spatial hearing dimension scores （P<0.05） than those in the ipsilateral reimplantation group after controlling for the corresponding confounders. Conclusion:The effect of ipsilateral reimplantation of cochlear implants is superior to that of contralateral reimplantation in terms of overall auditory function and spatial hearing in daily life for children, but the mechanisms require further investigation.
Humans ; Cochlear Implantation ; Child ; Parents ; Speech Perception ; Male ; Cochlear Implants ; Female ; Hearing ; Surveys and Questionnaires ; Speech ; Child, Preschool

Objective To investigate the effects of early pulmonary rehabilitation training on lung function recovery,complication rates,and quality of life in patients who undergo thoracoscopic surgery.Methods A ran-domized controlled trial was conducted involving 132 patients who underwent thoracoscopic surgery between June 2020 and June 2023.Participants were randomly allocated to either an early pulmonary rehabilitation program(n=72)or a conventional rehabilitation program(n=60).The early intervention group commenced a staged rehabilitation protocol—including respiratory training and physical exercise—within 24 to 72 hours following surgery,whereas the conventional group received standard postoperative nursing care only.Outcomes measured included postoperative pulmonary function(FEV1,FVC),incidence of postoperative complications,length of hospi-tal stay,and quality of life scores(maximum score:40).These outcomes were compared between the two groups.Results On postoperative days 3 and 7,the early group showed significantly higher FEV1 values compared to the conventional group[(1.6±0.17)L vs.(1.3±0.21)L;(1.9±0.20)L vs.(1.6±0.11)L,respectively],as well as improved FVC measurements[(2.5±0.20)L vs.(2.1±0.14)L;(2.9±0.25)L vs.(2.3±0.23)L(all P＜0.05)].The early intervention group also demonstrated a significantly lower overall complication rate(14.3％vs.33.3％,x2=6.79,P=0.009),including reduced incidences of atelectasis(6.9％vs.20.0％,P=0.031)and pulmonary infection(4.2％vs.13.3％,P=0.044).Additionally,patients in the early group had a shorter average hospital stay[(8.4±1.2)days vs.(10.9±2.3)days,P=0.018]and reported higher quality of life scores[(35.6±3.7)vs.(30.8±4.5),P=0.032].No significant difference was observed between the two groups in terms of the overall incidence of adverse events(23.6％vs.31.7％,x2=1.07,P=0.301).Conclusions Early pulmonary rehabilitation significantly facilitates the recovery of lung function,reduces the incidence of postoperative complica-tions,enhances quality of life,and demonstrates a favorable safety profile among patients undergoing thoracoscopic surgery.Therefore,this intervention should be more broadly implemented in clinical practice to optimize postop-erative recovery outcomes.

Objective To explore the diagnostic value of artificial intelligence(AI)in retinal vein occlusion(RVO).Methods The fundus images from 9,000 patients who visited our ophthalmology department and underwent non-mydriat-ic fundus photography from July 2016 to June 2022 were selected.Among these images,700 fundus images of patients with RVO were screened.These fundus images were processed using AI in collaboration with School of Electrical Engineering and Automation of Henan Polytechnic University.Besides,the state-of-the-art two-stage algorithms(the Faster-RCNN model),advanced one-stage detection algorithms(the YOLOV4 and YOLOV5 models),and a modified YOLOV5 model spe-cifically designed for RVO were employed in this study to process the collected RVO data.In addition,a comparative analy-sis of the results was conducted.Moreover,the sensitivity and specificity of the YOLOV5 model for the diagnosis of RVO were evaluated based on the receiver operating characteristic(ROC)curve.Results The Faster-RCNN model demon-strated the highest detection accuracy;however,its parameter scale was significantly larger than that of other models,and its frame rate was only 8 frames per second,which did not meet real-time detection requirements.The detection accuracy of the modified YOLOV5 model was only 3％lower than that of the Faster-RCNN model,but it had a significantly smaller parameter scale compared with the Faster-RCNN model,achieving a frame rate of 22 frames per second higher than the Faster-RCNN model.The area under the ROC curve(AUC)plotted based on the Faster-RCNN,YOLOV4,YOLOV5,and modified YOLOV5 models was 0.946(95％CI:0.933-0.959),0.794(95％CI:0.772-0.816),0.864(95％CI:0.845-0.884),and 0.930(95％CI:0.915-0.944),respectively.The sensitivity and specificity of the modified YOLOV5 model for the diagnosis of RVO were 87.0％and 98.9％,respectively.Conclusion The modified YOLOV5 model,with its high detection accuracy,smaller parameter scale,and high sensitivity and specificity for the diagnosis of RVO,can be consid-ered a new AI-assisted diagnostic method for RVO.

Objective To investigate the effects of early pulmonary rehabilitation training on lung function recovery,complication rates,and quality of life in patients who undergo thoracoscopic surgery.Methods A ran-domized controlled trial was conducted involving 132 patients who underwent thoracoscopic surgery between June 2020 and June 2023.Participants were randomly allocated to either an early pulmonary rehabilitation program(n=72)or a conventional rehabilitation program(n=60).The early intervention group commenced a staged rehabilitation protocol—including respiratory training and physical exercise—within 24 to 72 hours following surgery,whereas the conventional group received standard postoperative nursing care only.Outcomes measured included postoperative pulmonary function(FEV1,FVC),incidence of postoperative complications,length of hospi-tal stay,and quality of life scores(maximum score:40).These outcomes were compared between the two groups.Results On postoperative days 3 and 7,the early group showed significantly higher FEV1 values compared to the conventional group[(1.6±0.17)L vs.(1.3±0.21)L;(1.9±0.20)L vs.(1.6±0.11)L,respectively],as well as improved FVC measurements[(2.5±0.20)L vs.(2.1±0.14)L;(2.9±0.25)L vs.(2.3±0.23)L(all P＜0.05)].The early intervention group also demonstrated a significantly lower overall complication rate(14.3％vs.33.3％,x2=6.79,P=0.009),including reduced incidences of atelectasis(6.9％vs.20.0％,P=0.031)and pulmonary infection(4.2％vs.13.3％,P=0.044).Additionally,patients in the early group had a shorter average hospital stay[(8.4±1.2)days vs.(10.9±2.3)days,P=0.018]and reported higher quality of life scores[(35.6±3.7)vs.(30.8±4.5),P=0.032].No significant difference was observed between the two groups in terms of the overall incidence of adverse events(23.6％vs.31.7％,x2=1.07,P=0.301).Conclusions Early pulmonary rehabilitation significantly facilitates the recovery of lung function,reduces the incidence of postoperative complica-tions,enhances quality of life,and demonstrates a favorable safety profile among patients undergoing thoracoscopic surgery.Therefore,this intervention should be more broadly implemented in clinical practice to optimize postop-erative recovery outcomes.

Objective To explore the diagnostic value of artificial intelligence(AI)in retinal vein occlusion(RVO).Methods The fundus images from 9,000 patients who visited our ophthalmology department and underwent non-mydriat-ic fundus photography from July 2016 to June 2022 were selected.Among these images,700 fundus images of patients with RVO were screened.These fundus images were processed using AI in collaboration with School of Electrical Engineering and Automation of Henan Polytechnic University.Besides,the state-of-the-art two-stage algorithms(the Faster-RCNN model),advanced one-stage detection algorithms(the YOLOV4 and YOLOV5 models),and a modified YOLOV5 model spe-cifically designed for RVO were employed in this study to process the collected RVO data.In addition,a comparative analy-sis of the results was conducted.Moreover,the sensitivity and specificity of the YOLOV5 model for the diagnosis of RVO were evaluated based on the receiver operating characteristic(ROC)curve.Results The Faster-RCNN model demon-strated the highest detection accuracy;however,its parameter scale was significantly larger than that of other models,and its frame rate was only 8 frames per second,which did not meet real-time detection requirements.The detection accuracy of the modified YOLOV5 model was only 3％lower than that of the Faster-RCNN model,but it had a significantly smaller parameter scale compared with the Faster-RCNN model,achieving a frame rate of 22 frames per second higher than the Faster-RCNN model.The area under the ROC curve(AUC)plotted based on the Faster-RCNN,YOLOV4,YOLOV5,and modified YOLOV5 models was 0.946(95％CI:0.933-0.959),0.794(95％CI:0.772-0.816),0.864(95％CI:0.845-0.884),and 0.930(95％CI:0.915-0.944),respectively.The sensitivity and specificity of the modified YOLOV5 model for the diagnosis of RVO were 87.0％and 98.9％,respectively.Conclusion The modified YOLOV5 model,with its high detection accuracy,smaller parameter scale,and high sensitivity and specificity for the diagnosis of RVO,can be consid-ered a new AI-assisted diagnostic method for RVO.

Objective To investigate the value of monocyte to lymphocyte ratio(MLR)and neutrophil percentage to albumin ratio(NPAR)in predicting diabetic macular edema(DME).Methods One hundred and one diabetic retinopathy patients admitted to the Third Affiliated Hospital of Xinxiang Medical University from January 2018 to February 2023 were selected as the research subjects,and they were divided into DME group(n=56)and non-DME group(n=45)based on fun-dus examination results.The general data such as gender,age,course of diabetes and laboratory indicators were collected by consulting medical records.Fasting elbow venous blood was collected early in the morning of the next day after the diagnosis of DME in both groups,the monocytes(MONO)count,lymphocyte(LYM)count,white blood cell(WBC)count,percentage of neutrophils(NEUT),plasma albumin(ALB),glycosylated haemoglobin(HbA1c)were measured by full automatic blood routine analyzer,and MLR,NPAR were calculated.General information and laboratory indexes of patients in the two groups were compared,and risk factors for DME were analyzed by multivariate logistic regression,and receiver operator characteristic(ROC)curve was applied to evaluate the predictive value of MLR and NPAR for DME.Results The course of diabetes,MONO count,NEUT,MLR,NPAR,WBC count,and HbA1c level of patients between the DME group were significantly higher than those in the non-DME group(P＜0.05);there was no statistically significant difference in gender,age,LYM count,and ALB level of patients between the two groups(P＞0.05).Multivariate logistic regression analysis showed that increased levels of WBC,MLR,and NPAR were independent risk factors for the occurrence of DME(P＜0.05).The ROC curve showed that the best cut-off value of MLR was 0.192,and the area under the curve(AUC)for the prediction of DME was 0.729(95％confidence interval:0.631-0.826),with a sensitivity of 58.9％and a specificity of 82.2％;while the best cut-off value of NPAR was 1.404,and the AUC for predicting DME occurrence was 0.884(95％confidence interval:0.820-0.949),with a sensitivity of 75.0％and a specificity of 91.1％;the AUC of MLR and NPAP for predicting the occurrence of DME was 0.906(95％confidence interval:0.851-0.906),with a sensitivity of 69.6％and a specificity of 93.3％.With MLR＞0.192 as positive and NPAR＞1.404 as positive,the parallel test of MLR and NPAR predicted the occurrence of DME with a sensitivity of 87.5％,a specificity of 71.1％,and an accuracy of 80.2％;while the tandem test of MLR and NPAR predicted the occurrence of DME with a sensitivity of 46.4％,a specificity of 97.8％,and an accuracy of 69.3％.Conclusion Increased levels of MLR and NPAR are independent risk factors for the occurrence of DME and have certain predictive value for DME.The predictive value of combined MLR and NPAR test for DME is higher than that of separate test,and parallel experiment is more helpful for the early prediction of DME.

Objective To investigate the expression and significance of Src homology 2 domain-containing protein tyrosine phosphatase 2(SHP2)and phosphorylated SHP2(P-SHP2)in oxygen-induced retinopathy(OIR).Methods Twenty clean-grade C57BL/6J(B6)7-day-old neonatal mice were randomly divided into normal control group and OIR group,with 10 mice in each group.The mice in the normal control group and the nursing mother mice were fed together in a normal oxygen and room temperature environment.Mice in the OIR group were placed in an oxygen chamber with a temperature of 22-25 ℃,humidity of(60±10)％and oxygen volume fraction stable at(75±5)％for five days together with the nursing mother mice,and then transferred to a normal oxygen environment.At 12,14,17 days of age,three mice were selected from the normal con-trol group and the OIR group,respectively;and the expressions of SHP2 and P-SHP2 proteins in the retinal tissues of mice in the two groups were detected by Western blot.Two 17-day-old mice were randomly selected from the normal control group and the OIR group,respectively;and the retinal histopathology of mice was observed by hematoxylin-eosin staining.Results In the normal control group,the cell structure in all layers of the retinal tissue of 17-day-old mice was orderly,the inner limiting mem-brane and endothelial nucleus were intact,and the shape was normal.In the OIR group,the intercellular structure in all layers of the retinal tissue of 17-day-old mice was disordered,and the vascular endothelial nucleus was seen to break through the reti-nal inner limiting membrane.There was no statistically significant difference in the relative expression of SHP2 protein in the retinal tissues of mice at different ages in the normal control group(F=2.052,P＞0.05).The relative expression level of P-SHP2 protein in the retinal tissues of mice at 14,17 days of age in the normal control group was significantly lower than that at 12 days of age(P＜0.05),and the relative expression level of P-SHP2 protein in the retinal tissues of mice at 17 days of age was significantly lower than that at 14 days of age(P＜0.05).The relative expression level of SHP2 protein in the retinal tissues of mice at 14 days of age in the OIR group was significantly higher than that at 12,17 days of age(P＜0.05).There was no statistically significant difference in the relative expression level of SHP2 protein in the retinal tissues between 17-day-old mice and 12-day-old mice in the OIR group(P＞0.05).The relative expression level of P-SHP2 protein in the retinal tissues of mice at 14 days of age in the OIR group was significantly higher than that at 12,17 days of age(P＜0.05).There was no statistically significant difference in the relative expression level of P-SHP2 protein in the retinal tissues between 17-day-old mice and 12-day-old mice in the OIR group(P＞0.05).The relative expression level of SHP2 protein in the retinal tissues of mice at 12,17 days of age in the OIR group was significantly lower than that in the normal control group(P＜0.05).There was no statistically significant difference in the relative expression level of SHP2 protein in the retinal tissues of mice at 14 days of age between the two groups(P＞0.05).The relative expression level of P-SHP2 protein in the retinal tissues of 12-day-old mice in the OIR group was significantly lower than that in the normal control group(P＜0.05),while the relative expression level of P-SHP2 protein in the retinal tissues of 17-day-old mice was significantly higher than that in the normal control group(P＜0.05).There was no statistically significant difference in the relative expression level of P-SHP2 protein in the retinal tissues of mice between the two groups at 14 days of age(P＞0.05).Conclusion SHP2 and P-SHP2 are expressed with tem-poral fluctuations in OIR.Hypoxia may promote conformational changes of SHP2 in OIR mice,and play an active role in phos-phorylation;and participate in and promote the occurrence and development of OIR.

AIM: To compare the effect of small-incision lenticule extraction(SMILE)with different corneal cap thicknesses on postoperative astigmatism and short-term visual quality of patients with myopic astigmatism.METHODS: A total of 54 patients(108 eyes)with myopic astigmatism who underwent SMILE from June 2020 to June 2022 in our hospital were selected for the prospective controlled study, and patients were randomly assigned into two groups, with 27 cases(54 eyes)each. The corneal cap thickness design was 110 μm for the group A and 120 μm for the group B, while other operation parameters were consistent. Additionally, the uncorrected visual acuity(UCVA), spherical equivalent(SE), stiffness parameter A1(SP-A1), visual quality and vector parameters at baseline, 1 d,1 wk and 1 mo after surgery were compared between two groups.RESULTS: There was a statistically significant difference in UCVA, SE, and SP-A1 between the two groups at various time points before and after surgery(all P<0.05), and UCVA in the group A was better than that in the group B at 1 d after surgery(P<0.05). There was no statistically significant difference in the results of astigmatism vector analysis between the two groups of patients(both P>0.05). The objective scattering index(OSI)of the group A was lower than that of the group B, while Strehl ratio(SR)of the group A was higher than that of the group B at 1 d after surgery(both P<0.05). There was no significant difference in modulation transfer function cutoff frequency(MTF cut off), contrast vision, visual symptoms and overall satisfaction, postoperative complications between the two groups(all P>0.05).CONCLUSION: SMILE procedures with both 110 μm and 120 μm corneal cap thicknesses are safe and effective in correcting myopic astigmatism without affecting postoperative SE, astigmatism, SP-A1 or contrast visual acuity. Whereas 110 μm corneal cap thickness results in faster early postoperative visual recovery and better early visual quality than 120 μm.

Objective To construct myeloid-specific Spi1 gene knockout mice and analyze their genotypes,so as to provide animal model basis for the study of pathological mechanism of diseases and drug targets.Methods Ac-cording to the principle of CRISPR/Cas9 technology and Cre/LoxP system,sgRNA and Donor vectors were de-signed and constructed.The transcript of Exon 2(Exon 2)was used as the knockout region,and Loxp elements were placed on both sides of Exon 2.Cas9 protein,sgRNA and Donor vector were mixed and microinjected into the fertilized eggs of C57BL/6J mice,the fertilized eggs were transplanted into the uterus of C57BL/6J pregnant female mice,and F0 generation was obtained after 19～20 days.Positive F0 mice were mated with C57BL/6J mice to ob-tain stable F1 Spi1flox/+mice.Spi1flox/+mice of F1 generation were selfed to obtain Spi1flox/flox mice.Spi1flox/flox mated with Lyz2-Cre+mice to obtain Spi1flox/+/Lyz2-Cre+mice,and then mated with Spi1flox/flox,the Spi1flox/flox/Lyz2-Cre+mice were myeloid-specific Spi1 gene knockout(KO)mice.Spi1flox/flox/Lyz2-cre-mice were used as wild-type(WT)mice.DNA of WT and KO mice was extracted,and the genotypes were identified by agarose gel electro-phoresis after PCR amplification.Western blot was used to detect the expression of spleen focus forming virus provi-ral integration oncogene,Spi-1/purine rich box-1(PU.1)in immune cells of WT and KO mice.Results The results of PCR identification showed that the genotype of mice with only 220 bp amplified by flox primer was Spi1flox/flox homozygote,and the genotype of mice with 700 bp amplified by Lyz2-Cre primer was Lyz2-Cre+.Western blot showed that compared with WT group,the protein PU.1 was not expressed in bone marrow-derived macropha-ges(BMDMs)and peritoneal macrophages(PM)in KO group(P<0.01).There was no significant difference of statistics in the expression level of PU.1 in T cells between KO mice and WT mice.The results of PCR and West-ern blot showed that myeloid-specific Spi1 KO mice were successfully constructed.Conclusion The myeloid-spe-cific Spi1 gene KO mice are successfully constructed and identified,which provides animal model basis for further revealing the potential mechanism of PU.1 inimmune regulation.