Objective To identify potential biomarkers for proliferative diabetic retinopathy(PDR)using proteomics and transcriptomics data.Methods In this study,the proteomics dataset(PXD046630)and two transcriptomics datasets(GSE60436 and GSE102485)were derived from the aqueous humor samples and fibrovascular membranes of PDR patients,respectively.Differentially expressed genes(DEGs)were identified via R software,specifically the limma and edgeR pack-ages.The shared DEGs between PXD046630 and GSE60436 were analyzed via protein-protein interaction(PPI),Gene On-tology(GO)enrichment,and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses.The key DEGs were validated in GSE102485 via receiver operating characteristic(ROC)curve analysis.A quantitative polymerase chain reaction(qPCR)assay was used to confirm the mRNA of these candidate biomarkers in human retinal microvascular endothelial cells(HRMECs)cultured in high glucose and low oxygen conditions.Results A total of 59 shared DEGs and 26 hub genes were identified from the PXD046630 and GSE60436 datasets.KEGG analysis revealed that six pathways,inclu-ding extracellular matrix-receptor interaction,proteoglycans in cancer,and complement and coagulation cascades,were enriched in 12 key DEGs.Fibronectin 1(FN1),tissue inhibitor of metalloproteinase 3(TIMP3),complement factor H(CFH),decorin(DCN),and lipoprotein receptor-related protein-2(LRP2)were identified as potential biomarkers on the basis of their AUC values being greater than 0.900(CI≥95％).The mRNA expression levels of FN1,CFH,and LRP2 were significantly increased in HRMECs cultured in high glucose and low oxygen conditions.Conclusion FN1,CFH,and LRP2 are potential biomarkers for PDR,and further studies are needed to explore their roles and therapeutic potential in PDR.

Objective:To anatomically and phenotypically characterize the insular cortex(IC)-nucleus tractus soli-tarii(NTS)neural pathway.Methods:Adult male Sprague-Dawley rats were divided into three experimental cohorts for neural circuit tracing.Anterograde labeling was achieved by injecting anterograde self-complementary adeno-associated viruses(scAAVs)into the IC.Retrograde tracing involved NTS injections of either retrograde scAAVs or FluoroGold(FG),combined with immunofluorescence histochemical staining to identify IC-originating projection neurons.For postsynaptic neurochemical phenotype characterization,IC was injected with AAV2/1-CaMKII-Cre,while a mixture of AAV2/9-Syn-DIO-mCherry and AAV2/9-VGAT1-EGFP was injected into the NTS.The rats were allowed to survive for one week following scAAVs or FG injection or four weeks after recombinase-dependent systems injection.Then the rats were sacrificed,and serial brain sections were prepared for immunofluorescence histochemical staining(brain section containing FG)and subsequent fluorescence/confocal microscopic analysis.Results:(1)Anterograde viral tracing re-vealed dense axonal terminals from the IC projecting to the medial subnucleus of the NTS,while retrograde tracing re-vealed that IC neurons projecting to the NTS were predominantly localized within the dysgranular layer;(2)IC-NTS projection neurons were exclusive glutamatergic(100%,n=3);(3)NTS neurons receiving IC inputs were mainly lo-calized in the medial subnucleus,and were predominantly GABAergic(79.8±3.2%,n=3).Conclusion:The pres-ent results indicate that a descending pathway from excitatory neurons of the IC terminates onto inhibitory neurons of the NTS,which might represent a potential neuromodulatory target for visceral pain disorders.

Objective To identify potential biomarkers for proliferative diabetic retinopathy(PDR)using proteomics and transcriptomics data.Methods In this study,the proteomics dataset(PXD046630)and two transcriptomics datasets(GSE60436 and GSE102485)were derived from the aqueous humor samples and fibrovascular membranes of PDR patients,respectively.Differentially expressed genes(DEGs)were identified via R software,specifically the limma and edgeR pack-ages.The shared DEGs between PXD046630 and GSE60436 were analyzed via protein-protein interaction(PPI),Gene On-tology(GO)enrichment,and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses.The key DEGs were validated in GSE102485 via receiver operating characteristic(ROC)curve analysis.A quantitative polymerase chain reaction(qPCR)assay was used to confirm the mRNA of these candidate biomarkers in human retinal microvascular endothelial cells(HRMECs)cultured in high glucose and low oxygen conditions.Results A total of 59 shared DEGs and 26 hub genes were identified from the PXD046630 and GSE60436 datasets.KEGG analysis revealed that six pathways,inclu-ding extracellular matrix-receptor interaction,proteoglycans in cancer,and complement and coagulation cascades,were enriched in 12 key DEGs.Fibronectin 1(FN1),tissue inhibitor of metalloproteinase 3(TIMP3),complement factor H(CFH),decorin(DCN),and lipoprotein receptor-related protein-2(LRP2)were identified as potential biomarkers on the basis of their AUC values being greater than 0.900(CI≥95％).The mRNA expression levels of FN1,CFH,and LRP2 were significantly increased in HRMECs cultured in high glucose and low oxygen conditions.Conclusion FN1,CFH,and LRP2 are potential biomarkers for PDR,and further studies are needed to explore their roles and therapeutic potential in PDR.

Objective:To explore the value of magnetic resonance diffusion kurtosis imaging (DKI) in the assessment of the condition and prognosis of neonatal acute bilirubin encephalopathy (ABE).Methods:A retrospective selection was made of 196 neonates with acute hyperbilirubinemia who were hospitalized in the Second Affiliated Hospital of Shantou University Medical College from June 2021 to September 2023 as the research subjects. According to the presence or absence of brain injury, they were divided into the ABE group ( n=112) and the non-ABE group ( n=84). Based on the neonatal Bilirubine-induced Neurological Dysfunction (BIND) scoring system, children in the ABE group were divided into the mild group ( n=50, score 1-3 points), the moderate group ( n=33, score 4-6 points), and the severe group ( n=29, score 7-9 points). The clinical data and DKI parameters among each group were analyzed. Univariate and multivariate analyses were used to evaluate the influencing factors of prognosis in children with ABE. The receiver operating characteristic (ROC) curve was used to analyze the diagnostic value of DKI parameters for the prognosis of children with ABE. Results:The birth weight and gestational age in the ABE group were significantly lower than those in the non-ABE group, and the peak value of total bilirubin (TBIL) was significantly higher than that in the non-ABE group (all P<0.05). There was no statistically significant difference in fractional anisotropy (FA) values and mean diffusivity (MD) values among each group of children (all P>0.05). The mean kurtosis (MK) values, axial kurtosis (KA) values, and radial kurtosis (KR) values of children with ABE in the severe group were significantly higher than those in the other groups (all P<0.05). After Spearman correlation analysis, the FA value and MD value of children with ABE were not correlated with the severity of the disease (all P>0.05), while the MK value, KA value and KR value were positively correlated with the severity of the disease (all P<0.05). The patients were followed up for 12 months. Among them, 87 cases had a normal prognosis and 25 cases had a poor prognosis, including 2 cases of cerebral palsy, 5 cases of hearing loss, 4 cases of movement disorders, 12 cases of cerebral palsy combined with hearing loss, and 2 cases of movement disorders combined with hearing loss. The results of univariate analysis showed that there were statistically significant differences in birth weight, peak TBIL, BIND score, MK value, KA value, and KR value between the two groups of children with different prognoses (all P<0.05). The results of Cox multivariate regression analysis showed that birth weight, peak TBIL, BIND score, MK value, KA value, and KR value were independent influencing factors for poor prognosis in children with ABE (all P<0.05). The results of the ROC curve showed that the area under the curve and specificity of the MK value in predicting the poor prognosis of children with ABE were significantly higher than those of the KA and KR values (all P<0.05). Conclusions:The DKI parameters MK value, KA value, and KR value are sensitive indicators reflecting the severity of brain injury and predicting prognosis in children with ABE, among which the MK value has the highest predictive value.

Objective:To anatomically and phenotypically characterize the insular cortex(IC)-nucleus tractus soli-tarii(NTS)neural pathway.Methods:Adult male Sprague-Dawley rats were divided into three experimental cohorts for neural circuit tracing.Anterograde labeling was achieved by injecting anterograde self-complementary adeno-associated viruses(scAAVs)into the IC.Retrograde tracing involved NTS injections of either retrograde scAAVs or FluoroGold(FG),combined with immunofluorescence histochemical staining to identify IC-originating projection neurons.For postsynaptic neurochemical phenotype characterization,IC was injected with AAV2/1-CaMKII-Cre,while a mixture of AAV2/9-Syn-DIO-mCherry and AAV2/9-VGAT1-EGFP was injected into the NTS.The rats were allowed to survive for one week following scAAVs or FG injection or four weeks after recombinase-dependent systems injection.Then the rats were sacrificed,and serial brain sections were prepared for immunofluorescence histochemical staining(brain section containing FG)and subsequent fluorescence/confocal microscopic analysis.Results:(1)Anterograde viral tracing re-vealed dense axonal terminals from the IC projecting to the medial subnucleus of the NTS,while retrograde tracing re-vealed that IC neurons projecting to the NTS were predominantly localized within the dysgranular layer;(2)IC-NTS projection neurons were exclusive glutamatergic(100%,n=3);(3)NTS neurons receiving IC inputs were mainly lo-calized in the medial subnucleus,and were predominantly GABAergic(79.8±3.2%,n=3).Conclusion:The pres-ent results indicate that a descending pathway from excitatory neurons of the IC terminates onto inhibitory neurons of the NTS,which might represent a potential neuromodulatory target for visceral pain disorders.

Objective:To explore the value of magnetic resonance diffusion kurtosis imaging (DKI) in the assessment of the condition and prognosis of neonatal acute bilirubin encephalopathy (ABE).Methods:A retrospective selection was made of 196 neonates with acute hyperbilirubinemia who were hospitalized in the Second Affiliated Hospital of Shantou University Medical College from June 2021 to September 2023 as the research subjects. According to the presence or absence of brain injury, they were divided into the ABE group ( n=112) and the non-ABE group ( n=84). Based on the neonatal Bilirubine-induced Neurological Dysfunction (BIND) scoring system, children in the ABE group were divided into the mild group ( n=50, score 1-3 points), the moderate group ( n=33, score 4-6 points), and the severe group ( n=29, score 7-9 points). The clinical data and DKI parameters among each group were analyzed. Univariate and multivariate analyses were used to evaluate the influencing factors of prognosis in children with ABE. The receiver operating characteristic (ROC) curve was used to analyze the diagnostic value of DKI parameters for the prognosis of children with ABE. Results:The birth weight and gestational age in the ABE group were significantly lower than those in the non-ABE group, and the peak value of total bilirubin (TBIL) was significantly higher than that in the non-ABE group (all P<0.05). There was no statistically significant difference in fractional anisotropy (FA) values and mean diffusivity (MD) values among each group of children (all P>0.05). The mean kurtosis (MK) values, axial kurtosis (KA) values, and radial kurtosis (KR) values of children with ABE in the severe group were significantly higher than those in the other groups (all P<0.05). After Spearman correlation analysis, the FA value and MD value of children with ABE were not correlated with the severity of the disease (all P>0.05), while the MK value, KA value and KR value were positively correlated with the severity of the disease (all P<0.05). The patients were followed up for 12 months. Among them, 87 cases had a normal prognosis and 25 cases had a poor prognosis, including 2 cases of cerebral palsy, 5 cases of hearing loss, 4 cases of movement disorders, 12 cases of cerebral palsy combined with hearing loss, and 2 cases of movement disorders combined with hearing loss. The results of univariate analysis showed that there were statistically significant differences in birth weight, peak TBIL, BIND score, MK value, KA value, and KR value between the two groups of children with different prognoses (all P<0.05). The results of Cox multivariate regression analysis showed that birth weight, peak TBIL, BIND score, MK value, KA value, and KR value were independent influencing factors for poor prognosis in children with ABE (all P<0.05). The results of the ROC curve showed that the area under the curve and specificity of the MK value in predicting the poor prognosis of children with ABE were significantly higher than those of the KA and KR values (all P<0.05). Conclusions:The DKI parameters MK value, KA value, and KR value are sensitive indicators reflecting the severity of brain injury and predicting prognosis in children with ABE, among which the MK value has the highest predictive value.

Objective To analyze the epidemiological characteristics of mobile colistin resistance(mcr)genes in global Klebsiella pneumoniae(K.pneumoniae).Methods The genomes of K.pneumoniae were downloaded from the NCBI genome database by the As-pera software.After quality filtering using the CheckM v1.1.3 and Quest 5.0.2 software,the genomes were annotated using the Prokka v1.13.All the mcr gene sequences were downloaded from the NCBI website and a database was built using the makeblastdb command.Then,a self-made Perl script program was used to extract the nucleotide sequences of all genes from the annotation file as Query,and the local BLASTN analysis was performed to obtain mcr-positive strains.The gene sequence files and Profiles files of 7 housekeeping genes of K.pneumoniae were downloaded from the PubMLST website as the database,and the nucleotide sequences of the genes were extracted as Query using a self-made Perl script program.The local BLASTN analysis was implemented to determine the sequence type(ST)of each genome.Meta information of each strain,including isolation source,sample type,country,and date,were extracted in batch from the GenBank file of the downloaded K.pneumoniae genomes using a self-written Perl program to analyze the distribution characteristics of mcr-positive strains.The distribution differences of the mcr between different ST types were compared by the Chi-square test.Results Among the 11 429 global K.pneumoniae genomes included in this study,229 mcr genes were detected from 207 strains.Six variants of mcr were identified,mainly mcr-1(87/229,38.0％),mcr-8(59/229,25.8％),and mcr-9(59/229,25.8％).76 STs were identified from 207 strains,with ST15(21/207,10.1％),ST43(17/207,8.2％),ST11(16/207,7.7％),and ST 147(16/207,7.7％)being the predominant.Among 87 mcr-1 positive strains,31 STs were found,with ST43(17/87,19.5％)and ST15(10/87,11.5％)being the main ones.Among 59 mcr-8 positive strains,17 STs were identified,with ST43(17/59,28.8％)and ST11(9/59,15.3％)being the predominant.Among 59 mcr-9 positive strains,27 STs were detected,with ST147(11/59,18.6％)and ST274(11/59,18.6％)being the main ones.There were statistical differences in the variants of mcr genes carried by different ST types.The mcr-positive K.pneumoniae came from 28 countries across five continents worldwide,led by China(68/207,32.9％)and Thailand(45/207,21.7％),which were mainly from the human body(100/207,48.3％)and had a concentrated outbreak time from 2015 to 2018.Conclusion Among the global K.pneumoniae,the prevalence of mcr is mainly domina-ted by mcr-1,mcr-8,and mcr-9.The dominant clones of mcr-1 are mainly ST15 and ST43,while those of mcr-8 are mainly ST11 and ST43.The popularity of mcr-9 is mainly based on ST147 and ST274.Strengthening the monitoring of such bacteria may play an impor-tant role in preventing and controlling nosocomial infections.

Enzyme-driven micro/nanomotors consuming in situ chemical fuels have attracted lots of attention for biomedical applications. However, motor systems composed by organism-derived organics that maximize the therapeutic efficacy of enzymatic products remain challenging. Herein, swimming proteomotors based on biocompatible urease and human serum albumin are constructed for enhanced antitumor therapy via active motion and ammonia amplification. By decomposing urea into carbon dioxide and ammonia, the designed proteomotors are endowed with self-propulsive capability, which leads to improved internalization and enhanced penetration in vitro. As a glutamine synthetase inhibitor, the loaded l-methionine sulfoximine further prevents the conversion of toxic ammonia into non-toxic glutamine in both tumor and stromal cells, resulting in local ammonia amplification. After intravesical instillation, the proteomotors achieve longer bladder retention and thus significantly inhibit the growth of orthotopic bladder tumor in vivo without adverse effects. We envision that the as-developed swimming proteomotors with amplification of the product toxicity may be a potential platform for active cancer treatment.

Objective:To evaluate the clinical efficacy of microneedle radiofrequency in the treatment of moderate acne vulgaris.Methods:From August 2018 to August 2020, 393 patients (192 males and 201 females, aged 15-38 years) with moderate acne were enrolled in the Department of Dermatology, the First Affiliated Hospital of Shantou University Medical College and Shantou Chaonan Minsheng Hospital, including 201 patients in experimental group and 192 patients in control group. In the experimental group, microneedle radiofrequency therapy was used once every 2 weeks for 3 times in total. The control group adopted the fire needle, once every 2 weeks, a total of 3 times. The efficacy of both groups was evaluated at week 8.Results:A total of 378 patients were actually completed: 196 patients in the experimental group, and 182 patients in the control group. At the eighth week of follow-up, the total effective rate was 81.12% in the experimental group and 70.43% in the control group. The efficacy of the two groups was statistically different (χ 2=4.42, P<0.05). Conclusions:The efficacy of microneedle radiofrequency therapy in the treatment of moderate acne vulgaris is better than that of fire needle, with good tolerance, short recovery period, few adverse reactions and high compliance, which has clinical promotion value.

Excessive theta (θ) frequency oscillation and synchronization in the basal ganglia (BG) has been reported in elderly parkinsonian patients and animal models of levodopa (L-dopa)-induced dyskinesia (LID), particularly the θ oscillation recorded during periods when L-dopa is withdrawn (the off L-dopa state). To gain insight into processes underlying this activity, we explored the relationship between primary motor cortex (M1) oscillatory activity and BG output in LID. We recorded local field potentials in the substantia nigra pars reticulata (SNr) and M1 of awake, inattentive resting rats before and after L-dopa priming in Sham control, Parkinson disease model, and LID model groups. We found that chronic L-dopa increased θ synchronization and information flow between the SNr and M1 in off L-dopa state LID rats, with a SNr-to-M1 flow directionality. Compared with the on state, θ oscillational activity (θ synchronization and information flow) during the off state were more closely associated with abnormal involuntary movements. Our findings indicate that θ oscillation in M1 may be consequent to abnormal synchronous discharges in the BG and support the notion that M1 θ oscillation may participate in the induction of dyskinesia.