Objective To develop and evaluate a nucleic acid amplification test for spectinomycin-resistant Neisseria gonorrhoeae(N.gonorrhoeae).Methods N.gonorrhoeae-specific primers NG1/NG2 and primers specific to the N.gonorrhoeae rpsE gene mutation(80_82 delTTA)were designed.Genomic nucleic acids of spectinomycin-sensitive and resistant N.gonorrhoeae,Escherichia coli,Pseudomonas aeruginosa,and Salmonella typhi were used as templates to be amplified by PCR and quantitative real-time PCR(qPCR).The sensitivity and specificity of the method were evaluated accordingly.Results The NG1/NG2 primers could effectively amplify specific fragments of N.gonorrhoeae,yielding negative results for the nucleic acid amplification test of the other types of bacteria tested.E64/E175R and E-87/E95R could effectively differentiate the wild type and mutant(80_82 delTTA)rpsE genes.In PCR reactions,the minimum limits of NG1/NG2,E64/E175R,and E87/E95R for the target genes were 414.8 copies,414.8 copies,and 4.1 copies/μL,respectively,while those for qPCR reactions were 41.5,41.5,and 4.1×10-2 copies/μL,respectively.Conclusion A nucleic acid amplification test for spectinomycin-resistant N.gonorrhoeae with high specificity and sensitivity was successfully established in this study,which is expected to provide support for the rapid diagnosis of N.gonorrhoeae infection and treatment decision-making in clinical settings.

Objective To investigate the role of a novel rpsE gene mutation in mediating high-level spectinomycin resistance in Neisseria gonorrhoeae and to evaluate its effect on the biological fitness of the bacteria.Methods Spectinomycin-containing medium was used to screen for Neisseria gonorrhoeae strains with spontaneous mutations that conferred spectinomycin resistance.Minimum inhibitory concentrations(MIC)were determined,and the rpsE gene was sequenced.Changes in the growth rates of spectinomycin-resistant strain were assessed using the drop plate method and growth curves.Additionally,in vitro competition experiments were conducted with spectinomycin at different concentrations to assess changes in the biological fitness of the spectinomycin-resistant strain.Results A Neisseria gonorrhoeae strain with high-level spectinomycin resistance mediated by a novel rpsE gene mutation(88_90delGTT)was successfully identified and designated NG-SPTR.Compare with the wild-type strain,the NG-SPTR exhibited reduced growth rate(optical density[OD]comparison,P<0.05).In addition,in vitro competition experiments showed a competitive index(CI)<1 in gonococcal base liquid(GCBL)without or with low-concentration spectinomycin(≤16 μg/mL).In the GCBL with 32 μg/mL spectinomycin,the CI value gradually increased from<1 before 18 h to>1 after 18 h.The mutant strain showed CI>1 in GCBL with spectinomycin concentrations≥64 μg/mL.Conclusion The rpsE gene mutation(88_90delGTT)mediates high-level spectinomycin resistance in Neisseria gonorrhoeae,and imposes a fitness cost on the bacteria.The biological fitness of the mutant strain is influenced by the concentration of spectinomycin.

Chronic obstructive pulmonary disease(COPD)is a polygenic disease influenced by the interaction of genetic and environmental factors,characterized by persistent airflow limitation and chronic respiratory symptoms.Traditional gene sequencing methods have revealed the genetic basis of COPD but are limited by the average data of cell populations,making it difficult to fully display the complexity of the disease.Single-cell RNA sequencing technology provides a new perspective,allowing researchers to analyze gene expression at the individual cell level,revealing cellular heterogeneity,which is of great significance for understanding the pathological process of COPD,discovering biomarkers,and personalized medicine.This article reviews the principles of single-cell sequencing technology and its application in COPD research.

AIM: To investigate the differences in varying stages of non-proliferative diabetic retinopathy(NPDR)using optical coherence tomography angiography(OCTA).METHODS: Cross-sectional study. A total of 77 cases(77 eyes)of diabetic patients were included, and they were divided into non-diabetic retinopathy(NDR; 23 eyes)and non-proliferative diabetic retinopathy(NPDR; 54 eyes)groups, further subdivided into mild NPDR(20 eyes), moderate NPDR(20 eyes), and severe NPDR(14 eyes). Foveal avascular zone(FAZ)area, superficial and deep capillary plexus densities(SSP and DSP), and visual acuity(LogMAR)were compared between NDR and NPDR groups. Furthermore, the visual acuity, FAZ area and levels of SSP and DSP were compared in different degrees of NPDR. Correlation analysis were conducted to elucidate relationships between FAZ area, visual acuity, SSP, DSP, and severity of the disease.RESULTS: Compared with the NDR group, the visual acuity(LogMAR)and macular FAZ area increased, while SSP and DSP were decreased in the NPDR group(P<0.05); there were significant differences in visual acuity, FAZ area and SSP and DSP levels in different degrees of NPDR(P<0.05). Visual acuity(LogMAR)and FAZ area displayed a positive correlation with the severity of disease, while SSP and DSP showed a negative correlation.CONCLUSION: With the progression of NPDR, the visual acuity(LogMAR)and FAZ area increased, and the SSP and DSP decreased.

Objective To establish and verify nomogram prediction model with portal vein thrombosis in patients with cirrhosis,so as to evaluate its clinical application value.Methods The clinical data of 464 patients with cirrhosis with or without PVT admitted by our hospital from January 2015 to October 2022 were collected,univariate and multivariate Logistic regression analysis were used to screen the independent risk factors for PVT in patients with liver cirrhosis,and a nomogram prediction model was established,and the nomogram pre-diction model was verified for discrimination,calibration and clinical validity.Results Univariate and multivariate Logistic regression a-nalysis showed that the history of splenectomy,portal vein diameter,D-dimer and C-reactive protein-to-albumin ratio(CAR)were independent risk factors for cirrhosis with portal vein thrombosis(P＜0.05),the above independent risk factors were incorporated and a nomogram prediction model was successfully established.The nomogram was verified in both the modeling and validation populations:the area under the receiver operating characteristic(ROC)curve were 0.846(95％CI:0.803-0.888)and 0.815(95％CI:0.743-0.887),respectively,and the prediction model had good discrimination.The P-values in Hosmer-Lemeshow test were 0.783 and 0.453,respectively,and the model fitted well.The calibration chart showed that the nomogram model showed good calibration for the pre-diction probability of liver cirrhosis with PVT.Decision curve analysis(DCA)showed that the nomogram had good application value.Conclusion By establishing and validating a simple and practical nomogram prediction model for the risk of PVT in patients with cirrho-sis,it can help clinicians identify and intervene high-risk patients early.

Objective To establish and evaluate a microbial sensitivity test method for Neisseria gonorrhoeae based on resazurin coloration.Methods Based on the broth microdilution method,resazurin was added as a live bacteria indicator.WHO G,a WHO gonococcal reference strain,was used to optimize the incubation time for resazurin-stained bacteria and the color change was visually observed to obtain the results.Agar dilution method(the gold standard)and resazurin-based microdilution assay were used to determine the minimum inhibitory concentration(MIC)of azithromycin,ceftriaxone,and spectinomycin for 3 reference strains and 32 isolates of Neisseria gonorrhoeae.The results were analyzed based on essential agreement(EA),which reflected the consistency of the MIC values,category agreement(CA),which reflected the consistency in the determination of drug resistance,intermediary,and sensitivity,very major error(VME),which reflected false sensitivity,and major error(ME),which reflected pseudo drug resistance,to evaluate the accuracy of resazurin-based microdilution assay as a microbial sensitivity test of of Neisseria gonorrhoeae.CA and EA rates≥90%and VME and ME rates≤3%were found to be the acceptable performance rates.Results The results obtained 6 hours after resazurin was added were consistent with those of the agar dilution method and the resazurin-based microdilution assay was established accordingly based on this parameter.The EA of resazurin-based microdilution assay for measuring the MIC results of azithromycin,ceftriaxone,and spectinomycin was 97.1%,91.5%,and 94.3%,respectively,and the CA was 88.6%,94.3%,and 94.3%,respectively.The VME was 0%for all three antibiotics,while the ME was 11.4%,5.7%,and 5.7%,respectively.Conclusion The resazurin-based microdilution assay established in this study showed good agreement with agar dilution method for measuring the MIC of antibiotics against Neisseria gonorrhoeae.Moreover,the sensitivity results of this method were highly reliable and could be easily obtained through naked eye observation.Nonetheless,the results of drug resistance should be treated with caution and the optimization of parameters should be continued.

Objective: The aim of this study was to investigate the effect of changes in nasal tip protrusion on the sense of upper lip protrusion in different populations and to provide a reference for the improvement of soft tissue beauty. Methods : Informed consent and portrait authorization were obtained from the model. A female model whose face met the criteria was selected, and a 3D model of her was obtained using a 3D stereo camera. Based on the original model, ZBRUSH2019 software was used to simulate changes in nose tip protrusion. Then, 9 segments of facial dynamic rotation videos were produced, and an electronic questionnaire was created through a questionnaire website to evaluate the effect of different nose tip protrusions on perceived upper lip protrusion. Randomly selected orthodontic patients, orthodontics professionals and general adults completed the electronic questionnaire. In the questionnaire, a Visual Analog Scale was used to evaluate the perceived degree of protrusion of the upper lip. The higher the score, the more prominent the upper lip of the model in the image. The questionnaire data were statistically analyzed using a generalized linear mixed model. Results : As the nasal tip position became progressively more retracted, the subjects' upper lip protrusion scores increased. Among male subjects, the results showed that the general population thought that the upper lip protrusion was higher than the patients did when the nasal tip protrusion was +6 mm and +4.5 mm (P = 0.023, P = 0.047). When the nasal tip protrusion was +6 mm, the scores of the general population were higher than those of the orthodontics professionals (P = 0.023). However, when the nasal tip variable was -6 mm, their score was lower than that of the patients (P = 0.003), and there was no significant difference in other retest distances between groups (P>0.05). Conclusion When the protrusion of the nasal tip decreased, the three groups experienced a visual illusion of upper lip protrusion. When the nasal tip protrusion is too large, the general population perceived the visual illusion of the upper lip protrusion as being more obvious.

Recent insights collectively suggest the important roles of lysyl oxidase (LysOX) in the pathological processes of several acute and chronic neurological diseases, but the molecular regulatory mechanisms remain elusive. Herein, we explore the regulatory role of LysOX in the seizure-induced ferroptotic cell death of neurons. Mechanistically, LysOX promotes ferroptosis-associated lipid peroxidation in neurons via activating extracellular regulated protein kinase (ERK)-dependent 5-lipoxygenase (Alox5) signaling. In addition, overexpression of LysOX via adeno-associated viral vector (AAV)-based gene transfer enhances ferroptosis sensitivity and aggravates seizure-induced hippocampal damage. Our studies show that pharmacological inhibition of LysOX with β-aminopropionitrile (BAPN) significantly blocks seizure-induced ferroptosis and thereby alleviates neuronal damage, while the BAPN-associated cardiotoxicity and neurotoxicity could further be reduced through encapsulation with bioresponsive amorphous calcium carbonate-based nanocarriers. These findings unveil a previously unrecognized LysOX-ERK-Alox5 pathway for ferroptosis regulation during seizure-induced neuronal damage. Suppressing this pathway may yield therapeutic implications for restoring seizure-induced neuronal injury.

Hybrid lipid‒nanoparticle complexes have shown attractive characteristics as drug carriers due to their integrated advantages from liposomes and nanoparticles. Here we developed a kind of lipid-small molecule hybrid nanoparticles (LPHNPs) for imaging and treatment in an orthotopic glioma model. LPHNPs were prepared by engineering the co-assembly of lipids and an amphiphilic pheophorbide a‒quinolinium conjugate (PQC), a mitochondria-targeting small molecule. Compared with the pure nanofiber self-assembled by PQC, LPHNPs not only preserve the comparable antiproliferative potency, but also possess a spherical nanostructure that allows the PQC molecules to be administrated through intravenous injection. Also, this co-assembly remarkably improved the drug-loading capacity and formulation stability against the physical encapsulation using conventional liposomes. By integrating the advantages from liposome and PQC molecule, LPHNPs have minimal system toxicity, enhanced potency of photodynamic therapy (PDT) and visualization capacities of drug biodistribution and tumor imaging. The hybrid nanoparticle demonstrates excellent curative effects to significantly prolong the survival of mice with the orthotopic glioma. The unique co-assembly of lipid and small molecule provides new potential for constructing new liposome-derived nanoformulations and improving cancer treatment.

Objective:To explore the effect of health education of integrated medical and nursing in the follow-up management of atrial fibrillation patients with radiofrequency ablation.Methods:From September 2019 to August 2020, convenience sampling was used to select 112 atrial fibrillation patients with radiofrequency ablation in the Affiliated Brain Hospital of Nanjing Medical University as the research object. According to the random number table method, the patients were divided into the experimental group and the control group, each with 56 cases. The control group conducted health education conducted by specialist nurses for follow-up management, and the experimental group carried out the health education of integrated medical and nursing on the basis of the control group for follow-up management. The awareness rate of health-related knowledge, health-related behaviors, quality of life and the incidence of complications were compared between the two groups of patients.Results:After 6 months of follow-up, the awareness rate of health-related knowledge, the total score of health-related behaviors, and the total score of Atrial Fibrillation Effect on Quality-of-Life (AFEQT) of the experimental group were higher than those of the control group, and the differences were statistically significant ( P<0.05) . After 6 months of follow-up, the incidence of complications in the experimental group was lower than that in the control group, and the difference was statistically significant ( P<0.05) . Conclusions:The health education of integrated medical and nursing helps to improve the health-related knowledge of atrial fibrillation patients with radiofrequency ablation, promotes the development of health-related behaviors, and has positive application value for preventing postoperative complications and improving the quality of life of patients.