The KCNQ potassium channels play a crucial role in modulating neural excitability, and their dysfunction is closely associated with epileptic disorders. While variants in KCNQ2 have been extensively studied, KCNQ3-related disorders have rarely been reported. With advances in next-generation sequencing technologies, an increasing number of cases of KCNQ3-related disorders have been identified. However, the correlation between genotype and phenotype remains poorly understood. In this study, we established a variant library consisting of 24 missense mutations in KCNQ3 and introduced these mutations into three different template types: KCNQ3, KCNQ3-A315T (Q3*), and KCNQ3-KCNQ2 tandem (Q3-Q2). We then analyzed the effects of these mutations on the KCNQ3 channel function using patch-clamp recording. The most informative parameter across all three backgrounds was the current density of the mutant channels. The current density patterns in the Q3* and Q3-Q2 backgrounds were similar, with most mutations resulting in an almost complete loss of function (LOF), they were concentrated in the pore-forming domain of KCNQ3. In contrast, mutations in the voltage-sensing domain or C-terminus did not show significant differences from the wild-type channel. Interestingly, these LOF mutations were typically associated with self-limited familial neonatal epilepsy, while neurodevelopmental disorders (NDD) were more closely associated with mutations that did not significantly differ from the wild-type. V_1/2, another important parameter of the electrophysiological properties, could not be accurately determined in the majority of KCNQ3 mutations due to its nearly complete LOF in the Q3* and Q3-Q2 backgrounds. Intriguingly, the V_1/2 of functional mutations were primarily leftward shifted, indicating a gain-of-function (GOF) effect, which was typically associated with NDD. In addition to previously reported mutations, we identified G553R as a novel GOF mutation. In the co-transfection background, parameters such as V_1/2 could be determined, but the dysfunctional effects of these mutations were mitigated by the co-expression of wild-type KCNQ3 and KCNQ2 subunits, resulting in no significant differences between most mutations and the wild-type channel. Furthermore, we applied KCNQ modulators to reverse the electrophysiological abnormalities caused by KCNQ3 variants. The LOF mutations were reversed by the application of Pynegabine (HN37), a KCNQ opener, while the GOF mutation responded well to Amitriptyline (AMI), a KCNQ inhibitor. These findings provide essential insights into the pathogenic mechanisms underlying KCNQ3-related disorders and may inform clinical decision-making.
KCNQ3 Potassium Channel/genetics* ; Humans ; Mutation, Missense/genetics* ; KCNQ2 Potassium Channel/genetics* ; Patch-Clamp Techniques ; HEK293 Cells ; Animals ; Phenylenediamines/pharmacology* ; Carbamates

Objective To explore the mediating effect of coping styles on the early postoperative cognitive appraisal of health and stigma in patients with radical prostatectomy,so as to provide a reference for an early intervention in postoperative stigma.Methods Convenience sampling was used to select 223 patients who received radical prostatectomy at 5 Tier-ⅢA hospitals in Guangzhou from November 2022 to December 2023.Data were collected using the demographic data questionnaire,social impact scale,brief health cognition appraisal scale and medical coping style scale.Pearson correlation analysis was performed to examine the correlations between the stigma,health cognition appraisal and coping styles.Structural equation modelling was employed to analyse the mediating effect of coping styles on the health cognition appraisals and stigma.Results A total of 210 patients completed the study.The score of stigma for the patients after the radical prostatectomy was(49.46±13.56).Stigma exhibited positive correlations with negative cognitive appraisals(threat and harm)and negative coping styles(avoidance and submission)(both P<0.01).There were negative correlations between stigma and challenging cognitive appraisal and confrontation coping style(both P<0.01).Submission coping style played a mediating role between the cognitive appraisal of health(harm,threat and challenge)and stigma(β=0.072,0.075,-0.121,all P<0.01),accounting for 14.06％,23.15％and 100.00％,respectively,of the total effect.Conclusion Submission coping style is the mediator between the early postoperative cognitive appraisals and stigma in patients with radical prostatectomy.Negative cognitive appraisal positively predicts stigma through the submission coping style,while challenging cognitive appraisal negatively predicts stigma through the submission coping style.Caregivers should assess the early coping styles of patients after surgery,guide patients to correctly cope with the disease,and help them to avoid submissive responses to the postoperative stress hence to reduce the sense of stigma.

Objective To explore the mediating effect of coping styles on the early postoperative cognitive appraisal of health and stigma in patients with radical prostatectomy,so as to provide a reference for an early intervention in postoperative stigma.Methods Convenience sampling was used to select 223 patients who received radical prostatectomy at 5 Tier-ⅢA hospitals in Guangzhou from November 2022 to December 2023.Data were collected using the demographic data questionnaire,social impact scale,brief health cognition appraisal scale and medical coping style scale.Pearson correlation analysis was performed to examine the correlations between the stigma,health cognition appraisal and coping styles.Structural equation modelling was employed to analyse the mediating effect of coping styles on the health cognition appraisals and stigma.Results A total of 210 patients completed the study.The score of stigma for the patients after the radical prostatectomy was(49.46±13.56).Stigma exhibited positive correlations with negative cognitive appraisals(threat and harm)and negative coping styles(avoidance and submission)(both P<0.01).There were negative correlations between stigma and challenging cognitive appraisal and confrontation coping style(both P<0.01).Submission coping style played a mediating role between the cognitive appraisal of health(harm,threat and challenge)and stigma(β=0.072,0.075,-0.121,all P<0.01),accounting for 14.06％,23.15％and 100.00％,respectively,of the total effect.Conclusion Submission coping style is the mediator between the early postoperative cognitive appraisals and stigma in patients with radical prostatectomy.Negative cognitive appraisal positively predicts stigma through the submission coping style,while challenging cognitive appraisal negatively predicts stigma through the submission coping style.Caregivers should assess the early coping styles of patients after surgery,guide patients to correctly cope with the disease,and help them to avoid submissive responses to the postoperative stress hence to reduce the sense of stigma.

Objective:To explore the correlation between the early neurological deterioration and the poor short-term outcome after intravenous thrombolysis with recombinant tissue Plasmin activator (rt PA) in patients with acute ischemic stroke.Methods:A retrospective analysis of 268 patients with acute ischemic stroke who received recombinant tissue Plasmin activator(rt-PA) intravenous thrombolytic therapy in the Department of Neurology of Civil Aviation General Hospital from January 2019 to October 2022 was performed. The modified Rankin scale(mRS) was used to evaluate short-term outcome at 14 days after thrombolysis, with a score of 0 to 2 defined as good outcome and a score of 3 to 5 or death defined as poor outcome. Clinical, laboratory and imaging data of the two groups were collected. Univariate analysis was used to compare the baseline data of the two groups, and multivariate Logistic regression analysis was used to determine the independent risk factors for poor short-term outcomes.Results:Univariate analysis showed that age, baseline NIHSS score, fasting blood sugar, glycated hemoglobin, the proportion of coronary heart disease, proportion of atrial fibrillation, proportion of paraventricular high signal (3 score), proportion of hemorrhage transformation, proportion of END, proportion of double circulation infarction, proportion of new cortical infarction, proportion of new internal capsule infarction, proportion of large atherosclerosis of TOAST classification, and proportion of cardiogenic embolism of TOAST classification in the good outcome group were higher than those in the poor outcome group(68.00 (59.00, 80.00) years old vs 64.00 (54.50, 72.00) years old, 9 (6, 13) points vs 5 (3, 7) points, 6.85 (5.25, 9.20) mmol/L vs 5.64 (4.90, 6.75) mmol/L, 6.40 (5.70, 7.75)% vs 6.00 (5.60, 6.80)%, 31.3% (31/99) vs 17.7% (30/169), 28.3% (28/99) vs 9.5% (16/169), 26.3% (26/99) vs 13.0% (22/169), 19.2% (19/99) vs 5.9%(10/169), 53.5% (53/99) vs 21.9% (37/169), 10.1% (10/99) vs 2.4% (4/169), 44.4% (44/99) vs 22.5% (38/169), 41.4% (41/99) vs 27.8% (47/169), 33.3% (33/99) vs 17.8% (30/169), 25.3% (25/99) vs 6.5% (11/169)), the proportion of small artery occlusion and others of TOAST classification ,platelets were lower than the poor outcome group (29.3%(29/99) vs 53.8%(91/169),12.1%(12/99) vs 21.8%(37/169),199(163,240)×10 9/L vs 221(179,259)×10 9/L). The differences were statistically significant(the statistical values were U=2.80, U=7.94, U=3.24, U=2.29, χ 2=6.53, χ 2=16.11, χ 2=7.45, χ 2=11.40, χ 2=28.02, χ 2=7.54, χ 2=14.18, χ 2=5.24, χ 2=34.44, and U=2.16, respectively; P values were 0.005, <0.001, 0.001, 0.022, 0.011, <0.001, 0.006, 0.001, <0.001, 0.006, <0.001, 0.002, <0.001, and 0.031, respectively). Multivariate Logistic analysis showed that only baseline NIHSS score ( OR=1.212, 95% CI :1.123-1.309, P<0.001) and END ( OR=3.397, 95% CI:1.763-6.546, P<0.001) were independently related to the poor short-term outcome after intravenous thrombolysis in acute ischemic stroke patients. Conclusions:END and High baseline NIHSS score were independent risk factors for poor short-term outcome after intravenous thrombolysis in acute ischemic stroke patients.

Objective To investigate the pathogenic mutations of BRCA1 and BRCA2 in patients with early-onset breast cancer(≤35 years) and explore the relationships between BRCA1/2 mutations and clinical features.Methods Seventy-four patients with early-onset breast cancer were enrolled,who were treated in Hospital 307 between September 2014 and June 2016.High-throughput sequencing was used to test the 49 exon sequences and adjacent sequences of BRCA1 and BRCA2.χ2 test was used to analyze the distribution of BRCA1/2 pathogenic mutations in each group that was set up according to clinical features.Results Fifteen mutations(20.27%) were identified,including 5(6.76%) in BRCA1 and 10(13.51%) in BRCA2.Eleven new pathogenic mutations were discovered,and BRCA1:c.5470_5477delTGCCCAAT was found in one patient.The frequency of BRCA1/2 mutations in the group with a family history of breast cancer or ovarian cancer was higher than in the group without a family history (40.91% vs 11.54%) (χ2=6.534,P=0.011).Conclusion BRCA1/2 pathogenic mutation is significant for early-onset breast cancer,especially for those with a family history of breast or ovarian cancer.The new mutations may be specific to Chinese people.BRCA1:c.5470_5477delTGCCCAAT may be the ancestor mutation among the Chinese.

Objective To investigate the distribution and resistant pattern of Enterobacter cloacae to antimicrobial agents for reasonable use of antibiotics .Methods E .cloacae strains were isolated from patients from January 2009 to December 2013 .The strains were identified by VITEK‐2 Compact System of BioMerieux and tested for antimicrobial susceptibility by Kirby‐Bauer disk diffusion method .The results were analyzed .Results A total of 397 nonduplicate E .cloacae strains were isolated during 5 years ,accounting for 5 % of the total gram‐negative isolates . The strains were mainly isolated from sputum (48 .9% ) , followed by secretions (30 .5% )and urine (7 .3% ) .The percentage of E .cloacae strains resistant to all the antibiotics tested was on decline except carbapenems ,cefoxitin and amoxicillin‐clavulanic acid .Carbapenems were relatively more active against E .cloacae strains .The E .cloacae strains showed higher resistance rate toβ‐lactams .Conclusions It is necessary to strengthen the monitoring of clinical isolates to guide rational use of antimicrobial agents .

OBJECTIVE:To study the effects of combined use of cefoperazone/sulbactam(CFS),meropenem(MRP)with le-vofloxacin (LVX) on mutant prevention concentration for Acinetobacter baumannii so as to provide a experimental foundation for preventing bacterial drug resistance. METHODS:In the test,there were groups of single use of CFS,MRP and LVX,and groups of combined use of CFS+LVX and MRP+LVX. Double dilution agar method was used to respectively detect minimal inhibitory con-centration (MIC) for standard Acinetobacter baumannii strains ATCC 19606 and 36 clinically isolated strains in all groups,and then the index of fraction inhibitory concentration(FIC)was calculated. Agar dilution method on plates was employed to detect mu-tant prevention concentration (MPC) for Acinetobacter baumannii in all groups,and then corresponding selection index (SI) was calculated. RESULTS:After combined use of CFS or MRP with LVX,synergistic effect accounted for 55%(20/36)and 50%(18/36). For standard strain ATCC 19606,after combined use of CFS+LVX and MRP+LVX,MPC reduced from 32.0 and 4.0 mg/L to 1.8 and 0.8 mg/L;For 36 clinically isolated strains,after combined use of CFS+LVX and MRP+LVX,MPC reduced from 8.0-32.0 and 1.0-2.0 mg/L to 0.25-1.0 and 0.25-0.5 mg/L. SI were all obviously reduced after combination. CONCLUSIONS:The combina-tion of CFS or MRP with LVX can reduce MPC for Acinetobacter baumannii more significantly than the single use,and narrow mu-tant selection window.

Aim To study the change of mutant pre- vention concentration (MPC) in carbapenem-resistant Acinetobacter baumannii (CRAB) treated with moxi- floxacin ( MFX ) and/ or cefoperazone/ sulbactam (CFS) in vitro, and provide a theoretical support for preventing the bacterial resistance. Methods To cal- culate the fractional inhibitory concentration (FIC) in- dex, the minimum inhibitory concentration (MIC) of 20 clinical isolates of CRAB treated with MFX and/ or CFS was determined by checkerboard microdilution as- say. In addition, to calculate the selection index (SI), the MPC of 20 clinical isolates of CRAB treated with MFX and/ or CFS was determined by agar plate di- lution assay. Results Our study showed that there was synergistic/ addictive action, rather than antago- nism action against clinical isolates of CRAB when treated with MFX + CFS. The SI of the 20 isolates trea- ted with MFX or CFS alone was 4 ~128 and 8 ~64 re- spectively, but reduced to 1 ~8 and 4 ~16 when trea- ted with MFX + CFS, which decreased by 2 ~16 and 2 ~4 times respectively compared with the single treat- ment. Conclusion These results suggest that the combination treatment of MFX + CFS against clinical isolates of CRAB might lower the MPC of the isolates treated with MFX/ CFS alone, narrow the mutant selec- tion window, and prevent the generation of drug - re- sistant mutants.

Objective To evaluate the ability of Vitek 2 Compact YST identification cards and pyrosequencing analysis for identifying the clinical isolates of yeast-like fungi.Methods Vitek 2 Compact YST identification cams and ITS1 region pyrosequencing analysis were used to identify the clinical isolates of yeast-like fungi at Jinan Military General Hospital in 2011.The strains which could not be identified to species by pyrosequencing analysis were identified again by the method of ITS1 region Sanger sequencing.The strains with inconsistent identified results by YST and pyrosequencing were identified again using API 20C.Results A total of 282 srtains were isolated from various clinical specimens for culturing in 2011.In addition to the three which could only be identified to similar species,other strains could be clearly identified to species by the method of ITS1 reverse pyrosequencing.Three strains which failed identified to species by pyrosequencing couldn't be distinguished fully by the method of Sanger sequencing.There were 7 strains unidentified,14 strains low discrimination(％ id ＜ 80％),and 6 strains misidentification using YST cams.The identification coincidence rate was 90.4％.The identification coincidence rates were various in different species of yeast-like fungi using YST cards.Conclusions The vast majority of clinical isolates of yeast-like fungi could be identified by ITS1 reverse pyrosequencing analysis.Vitek 2 Compact YST caRD basically meets the needs.But attention should be paid to the standard operating and regular internal quality control.For some rare species it is also needed to combine the source of specimen,colonies and characteristics to determine the identification results for the rare stains.

Objective To explore the distribution characteristics of intra/extracranial large-artery disturbance in patients with ischemic stroke (IS)/transient ischemic attack (TIA).Methods The medical history information of 169 IS/TIA patients was recorded.The results of laboratory examination,electrocardiogram,echocardiography,abdominal ultrasound and chest radiography were analyzed.At the same time,the intra/extra-cranial large arteries were examined to analyze the degree,distribution and pattern of their disturbance.Results Among them,there were intra/extra-cranial large-arterial stenosis (n =112,66.3％) and small-arterial disturbance (n =57,33.7％).Multiple vascular disturbance pattern was more common in patients with intra/extra-cranial large-arterial stenosis (n =71,63.4％).The incidence of intracranial large-arterial stenosis was significantly higher than that of extracranial large-arterial stenosis (53.8％ vs.35.5％,P ＜ 0.05).And middle cerebral artery was the most commonly affected artery,36 of those were culprit vessels accounting for 73.5％ (36/49).Conclusions The IS/TIA patients have a high incidence of intra/extra-cranial large-arterial stenosis.Multiple vascular disturbance is a more common pattern.The incidence of intracranial large-arterial stenosis is greater than that of extracranial and middle cerebral artery is the most common affected artery.