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.

ObjectiveTo investigate the effect of galvanic vestibular stimulation on stroke patients with lateropulsion. MethodsFrom February to October, 2023, 30 stroke patients with lateropulsion in the First Affiliated Hospital of Nanjing Medical University were divided into control group (n = 15) and experimental group (n = 15) randomly. The control group received routine rehabilitation and sham galvanic vestibular stimulation, and the experimental group received routine rehabilitation and true galvanic vestibular stimulation, for two weeks. They were assessed with Scale for Contraversive Pushing (SCP), subjective visual vertical (SVV), Line Cancellation Test (LCT), Star Cancellation Test (SCT), Berg Balance Scale (BBS), Holden Functional Ambulation Category (FAC) and Barthel Index (BI) before and after treatment. ResultsAll the indexes improved in both groups after intervention (|t| > 2.461, Z > 3.000, P < 0.05), except the SVV orientation, SVV certainty and SCT in the control group; while the SCP, SVV certainty, LCT and FAC were better in the experimental group than in the control group (|t| > 2.189, Z = -2.862, P < 0.05), and the differences before and after intervention were better in the experimental group than in the control group (|t| > 2.382, P < 0.05), except LCT. SCP was correlated with SVV orientation, SVV certainty, SCT, BBS, BI and FAC (|r| > 0.381, P < 0.05). ConclusionGalvanic vestibular stimulation can improve the lateropulsion, balance, walking function and activities of daily living in stroke patients, which may be related to improvement for spatial cognitive function, especially vertical perception.

Objective:To explore effects of transcutaneous auricular vagus nerve stimulation(taVNS)with task-oriented training(TOT)on upper extremity function and brain plasticity in post stroke hemiplegia patients. Method:Forty patients with stroke hemiplegia were randomly assigned to the experiment group(n=20)and the control group(n=20).The experiment group received taVNS combined with TOT,while the control group received only TOT.At baseline and at the end of treatment,the patients were assessed with motor evoked po-tentials(MEP),the Fugl-Meyer assessment upper extremity(FMA-UE),the Hongkong version of functional test for the hemiplegic upper extremity(FTHUE),the action research arm test(ARAT),the modified Barthel index(MBI),and the fatigue severity scale(FSS).The side effects were monitored during the treatment. Result:There were no significant differences in FMA-UE,FTHUE,ARAT,MBI and FSS between the two groups at baseline(P＞0.05).After treatment,all the functional outcomes except FSS in both two groups in-creased significantly compared with baseline(P＜0.05),and the FMA-UE、FTHUE、ARAT、MBI(occupational abili-ty)scores in the experiment group were significantly higher than those in the control group(P＜0.05).There were no significant differences in the latency,amplitude and induced rate of motor evoked potential(MEP)at baseline between the two groups(P＞0.05).Compare with the control group after treatment,the latency of con-tralesional MEP in the experiment group was significantly shorter than that in the control group(P＜0.05),and the induced rates of ipsilesional MEP were significantly higher(P＜0.05),but there were no significant differenc-es in contralesional amplitudes between the two groups.There was a significant correlation between the chang-es of contralesional MEP latency and the changes of FMA-UE(r=-0.705),FTHUE(r=-0.458),MBI in occu-pation(r=-0.654)in the experiment group(P＜0.05). Conclusion:taVNS combined with TOT can improve upper extremity motor function in patients with stroke hemiplegia by modulating plasticity of motor cortex.

Objective:To explore the clinical application value of the cognitive assessment scale for stroke patients(CASP)in post-stroke cognitive impairment(PSCI)based on the translation and reliability and validity testing of the original CASP in order to improve the cognitive assessment system for stroke patients.Method:The original version of CASP was translated into Chinese,and a total of 82 stroke patients were as-sessed using the Mini-Mental State Examination(MMSE),the Montreal Cognitive Assessment(MoCA)and the Chinese version of CASP(C-CASP).Reliability analysis was conducted using Cronbach's a coefficient and inter-group correlation coefficient(ICC).Structure validity and criterion validity were analyzed to assess the validi-ty.The stroke patients were stratified according to cognitive function,lesion location and injury nature using independent sample t-tests.Result:The reliability analysis showed the Cronbach's a coefficient was 0.792,and the inter-rater reliability ICC was 0.999 and 0.998,while the ICC was 0.985 and 0.982 for retest reliability.The structure validity anal-ysis extracted one factor with a cumulative contribution rate of 52.89％.The correlation coefficients of C-CASP with MMSE,MoCA and ADL were 0.867,0.848 and 0.597.The area under the curve(AUC)for C-CASP was 0.938,with an optimal cutoff score of 32,sensitivity of 94.5％,and specificity of 77.8％.The total scores and item scores of C-CASP in the patients with cognitive impairment were significantly lower than those in the patients with non-cognitive impairment(P＜0.05).The item scores of the"bisection of a horizontal line"in C-CASP were significantly lower in the patients with right hemisphere lesions compared to the patients with left hemisphere lesions(P＜0.05).Additionally,the patients with ischemic stroke scored significantly lower than the patients with hemorrhagic stroke on the items of"naming"and"comprehension"in C-CASP(P＜0.05).Conclusions:C-CASP is a reliable and valid screening tool for assessing cognitive function in stroke patients.C-CASP has a broader cognitive threshold,which is more suitable than MMSE for the assessment of cogni-tive function in stroke patients.

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:To explore the clinical application value of the cognitive assessment scale for stroke patients(CASP)in post-stroke cognitive impairment(PSCI)based on the translation and reliability and validity testing of the original CASP in order to improve the cognitive assessment system for stroke patients.Method:The original version of CASP was translated into Chinese,and a total of 82 stroke patients were as-sessed using the Mini-Mental State Examination(MMSE),the Montreal Cognitive Assessment(MoCA)and the Chinese version of CASP(C-CASP).Reliability analysis was conducted using Cronbach's a coefficient and inter-group correlation coefficient(ICC).Structure validity and criterion validity were analyzed to assess the validi-ty.The stroke patients were stratified according to cognitive function,lesion location and injury nature using independent sample t-tests.Result:The reliability analysis showed the Cronbach's a coefficient was 0.792,and the inter-rater reliability ICC was 0.999 and 0.998,while the ICC was 0.985 and 0.982 for retest reliability.The structure validity anal-ysis extracted one factor with a cumulative contribution rate of 52.89％.The correlation coefficients of C-CASP with MMSE,MoCA and ADL were 0.867,0.848 and 0.597.The area under the curve(AUC)for C-CASP was 0.938,with an optimal cutoff score of 32,sensitivity of 94.5％,and specificity of 77.8％.The total scores and item scores of C-CASP in the patients with cognitive impairment were significantly lower than those in the patients with non-cognitive impairment(P＜0.05).The item scores of the"bisection of a horizontal line"in C-CASP were significantly lower in the patients with right hemisphere lesions compared to the patients with left hemisphere lesions(P＜0.05).Additionally,the patients with ischemic stroke scored significantly lower than the patients with hemorrhagic stroke on the items of"naming"and"comprehension"in C-CASP(P＜0.05).Conclusions:C-CASP is a reliable and valid screening tool for assessing cognitive function in stroke patients.C-CASP has a broader cognitive threshold,which is more suitable than MMSE for the assessment of cogni-tive function in stroke patients.

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.

Objective:To evaluate the effectiveness and safety of phacoemulsification cataract extraction combined with intraocular lens implantation (PEI) plus goniosynechilysis (GSL) and goniotomy (GT) for advanced primary angle-closure glaucoma (PACG).Methods:An observational case series study was performed.Fifty eyes of 50 patients with advanced PACG were enrolled in Zhongshan Ophthalmic Center from August 2020 to June 2021.All the patients received PEI+ GSL+ GT and were followed up for over 6 months, with a mean follow-up of 7.5 (6, 10) months.Intraocular pressure (IOP) was measured with a Goldmann applanation tonometer.Best corrected visual acuity (BCVA) was examined with an ETDRS chart and converted to logarithm of the minimum angle of resolution (LogMAR) units for analysis.Types and number of anti-glaucoma medications applied before and after surgery, and the surgical complications were collected.Success rate of surgery was calculated.Complete surgical success was defined as an IOP of 5-18 mmHg (1 mmHg=0.133 kPa) with a reduction of 20% from baseline without anti-glaucoma medication, no vision-threatening complications, no loss of light perception, and no reoperation.Qualified success was defined as an IOP of 5-18 mmHg with a reduction of 20% from baseline with or without anti-glaucoma medication, no vision-threatening complications, no loss of light perception, and no reoperation.This study adhered to the Declaration of Helsinki.This research protocol was approved by an Ethics Committee of Zhongshan Ophthalmic Center (No.2021KYPJ177). Written informed consent was obtained from each subject before entering the cohort.Results:The mean preoperative IOP was (28.81±7.81)mmHg, and the IOP at the end of follow-up was (13.41±4.10)mmHg, showing a statistically significant decrease ( t=12.260, P<0.001). The postoperative IOP was decreased by 13.80 (9.10, 19.40)mmHg, with a percentage decrease of 51.1% (38.6%, 67.1%). The mean preoperative and postoperative BCVA was (0.92±0.11) LogMAR and (0.88±0.10) LogMAR, respectively, and no significant difference was found ( t=-0.560, P=0.580). The number of anti-glaucoma medications was reduced from 2 (1, 3) before operation to 0 (0, 0) after operation.The complete success rate of surgery was 80% (40/50), and the qualified success rate was 94% (47/50). Surgical complications mainly included hyphema in 7 eyes, IOP spike in 7 eyes, and corneal edema in 3 eyes.No vision-threatening complication occurred. Conclusions:PEI+ GSL+ GT is preliminarily effective and safe for advanced PACG by reducing IOP and application of anti-glaucoma medications with few complications.