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 investigate the epidemic characteristics and trends of major natural focus diseases and vector-borne infectious diseases in Yantai City, and provide a basis for formulating reasonable prevention and control strategies.Methods:The descriptive analysis was conducted on the epidemic data of natural focal diseases and vector-borne infectious diseases in Yantai City collected from the Infectious Disease Surveillance and Reporting Management System, a subsystem of the China Disease Control and Prevention Information System, covering the period from January 1, 2019, to December 31, 2023.Results:From 2019 to 2023, a total of 2 746 cases of 12 kinds of natural focus diseases, vector-borne infectious diseases and zoonosis were reported in Yantai City. The top four diseases reported in incidence rate were severe fever with thrombocytopenia syndrome (SFTS, 3.13/100 000), brucellosis (2.57/100 000), hemorrhagic fever with renal syndrome (HFRS, 1.09/100 000), and scrub typhus (0.73/100 000). There were statistically significant differences in the reported incidence rate of the four major natural focus diseases and vector-borne infectious diseases between different years (χ 2 = 92.55, 34.88, 132.33, 39.95, P < 0.001). The four major natural focal diseases and vector-borne infectious diseases had reported cases throughout the year. SFTS cases peaked in June, accounting for 28.80% (320/1 111). Brucellosis cases were predominantly reported from March to August, accounting for 65.54% (599/914). HFRS cases peaked in November, accounting for 38.66% (150/388), while scrub typhus cases peaked in October, accounting for 69.88% (181/259). The top three districts/cities with the highest incidence of the four major natural focus diseases and vector-borne infectious diseases were Laizhou City (585 cases), Zhaoyuan City (377 cases), and Haiyang City (358 cases). The incidence rates of SFTS in women was higher than that in men (χ 2 = 13.92, P < 0.001), and the incidences rates of brucellosis and HFRS in men were higher than those in women (χ 2 = 123.09, 73.32, P < 0.001). The cases of the four diseases were mainly farmers, accounting for 76.06% (845/1 111), 78.12% (714/914), 81.44% (316/388), and 85.71% (222/259), respectively. Conclusions:From 2019 to 2023, the reported incidence rate of the four major natural focus diseases and vector-borne infectious diseases in Yantai City presents an overall upward trend, with obvious seasonal, regional and demographic characteristics. Targeted prevention and control measures should be developed for high-risk areas and populations based on the characteristics of different diseases.

Objective:To evaluate the value of D-dimer(D-D) levels in the diagnosis of thromboembolism in children with severe Mycoplasma pneumoniae pneumonia (SMPP).Methods:A case control study was conducted on 51 SMPP patients admitted to Qingdao Women and Children′s Hospital Affiliated to Qingdao University and Jining First People′s Hospital from January 1, 2021 to August 1, 2024.They were divided into a thrombus group (19 cases) and a non-thrombus group (32 cases) according to whether they had thromboembolism.The characteristics of the cases were analyzed.Logistic regression was used to analyze the relationship between increased D-D levels and the risk of thrombosis.The value of D-D levels in predicting thromboembolism in SMPP patients was evaluated by the receiver operating characteristic curve.Results:Of the 19 patients with thrombosis, 11 had with pulmonary thrombosis, 6 had cerebrovascular thrombosis, 3 had heart thrombosis, 1 had spleen artery thrombosis, and 3 had multiple site thrombosis.Compared with the non-thrombus group, the thrombus group had higher D-D levels[(18.5±4.9) mg/L vs.(3.1±0.8) mg/L, t=3.118, P=0.006], higher C-reactive protein levels[25.5(15.5, 92.7) mg/L vs.7.9(3.9, 21.4) mg/L, Z=3.292, P=0.001], higher lactate dehydrogenase levels[484(351, 743) U/L vs.347(285, 396) U/L, Z=2.770, P=0.006] and a higher proportion of pleural effusion[63.1%(12/19) vs.25.0%(8/32), χ2=7.282, P=0.009].Increased D-D levels were an independent risk factor for thromboembolism in SMPP patients ( P=0.005, OR=1.254, 95% CI: 1.069-1.472).When the D-D level was used for predicting thromboembolism in SMPP patients, its cut-off value was 4.46 mg/L, its Youden index was 0.707, its area under the curve was 0.893(95% CI: 0.807-0.979), its sensitivity was 89.5%, its specificity was 81.2%, and its negative predictive value was 92.9%. Conclusions:D-D levels have high value in predicting thromboembolism in SMPP patients, and it can help timely identify patients at high risk.

Objective:To investigate the epidemic characteristics and trends of major natural focus diseases and vector-borne infectious diseases in Yantai City, and provide a basis for formulating reasonable prevention and control strategies.Methods:The descriptive analysis was conducted on the epidemic data of natural focal diseases and vector-borne infectious diseases in Yantai City collected from the Infectious Disease Surveillance and Reporting Management System, a subsystem of the China Disease Control and Prevention Information System, covering the period from January 1, 2019, to December 31, 2023.Results:From 2019 to 2023, a total of 2 746 cases of 12 kinds of natural focus diseases, vector-borne infectious diseases and zoonosis were reported in Yantai City. The top four diseases reported in incidence rate were severe fever with thrombocytopenia syndrome (SFTS, 3.13/100 000), brucellosis (2.57/100 000), hemorrhagic fever with renal syndrome (HFRS, 1.09/100 000), and scrub typhus (0.73/100 000). There were statistically significant differences in the reported incidence rate of the four major natural focus diseases and vector-borne infectious diseases between different years (χ 2 = 92.55, 34.88, 132.33, 39.95, P < 0.001). The four major natural focal diseases and vector-borne infectious diseases had reported cases throughout the year. SFTS cases peaked in June, accounting for 28.80% (320/1 111). Brucellosis cases were predominantly reported from March to August, accounting for 65.54% (599/914). HFRS cases peaked in November, accounting for 38.66% (150/388), while scrub typhus cases peaked in October, accounting for 69.88% (181/259). The top three districts/cities with the highest incidence of the four major natural focus diseases and vector-borne infectious diseases were Laizhou City (585 cases), Zhaoyuan City (377 cases), and Haiyang City (358 cases). The incidence rates of SFTS in women was higher than that in men (χ 2 = 13.92, P < 0.001), and the incidences rates of brucellosis and HFRS in men were higher than those in women (χ 2 = 123.09, 73.32, P < 0.001). The cases of the four diseases were mainly farmers, accounting for 76.06% (845/1 111), 78.12% (714/914), 81.44% (316/388), and 85.71% (222/259), respectively. Conclusions:From 2019 to 2023, the reported incidence rate of the four major natural focus diseases and vector-borne infectious diseases in Yantai City presents an overall upward trend, with obvious seasonal, regional and demographic characteristics. Targeted prevention and control measures should be developed for high-risk areas and populations based on the characteristics of different diseases.

Objective:To evaluate the value of D-dimer(D-D) levels in the diagnosis of thromboembolism in children with severe Mycoplasma pneumoniae pneumonia (SMPP).Methods:A case control study was conducted on 51 SMPP patients admitted to Qingdao Women and Children′s Hospital Affiliated to Qingdao University and Jining First People′s Hospital from January 1, 2021 to August 1, 2024.They were divided into a thrombus group (19 cases) and a non-thrombus group (32 cases) according to whether they had thromboembolism.The characteristics of the cases were analyzed.Logistic regression was used to analyze the relationship between increased D-D levels and the risk of thrombosis.The value of D-D levels in predicting thromboembolism in SMPP patients was evaluated by the receiver operating characteristic curve.Results:Of the 19 patients with thrombosis, 11 had with pulmonary thrombosis, 6 had cerebrovascular thrombosis, 3 had heart thrombosis, 1 had spleen artery thrombosis, and 3 had multiple site thrombosis.Compared with the non-thrombus group, the thrombus group had higher D-D levels[(18.5±4.9) mg/L vs.(3.1±0.8) mg/L, t=3.118, P=0.006], higher C-reactive protein levels[25.5(15.5, 92.7) mg/L vs.7.9(3.9, 21.4) mg/L, Z=3.292, P=0.001], higher lactate dehydrogenase levels[484(351, 743) U/L vs.347(285, 396) U/L, Z=2.770, P=0.006] and a higher proportion of pleural effusion[63.1%(12/19) vs.25.0%(8/32), χ2=7.282, P=0.009].Increased D-D levels were an independent risk factor for thromboembolism in SMPP patients ( P=0.005, OR=1.254, 95% CI: 1.069-1.472).When the D-D level was used for predicting thromboembolism in SMPP patients, its cut-off value was 4.46 mg/L, its Youden index was 0.707, its area under the curve was 0.893(95% CI: 0.807-0.979), its sensitivity was 89.5%, its specificity was 81.2%, and its negative predictive value was 92.9%. Conclusions:D-D levels have high value in predicting thromboembolism in SMPP patients, and it can help timely identify patients at high risk.

Objective:To study the epidemic characteristics of scrub typhus in Yantai City from 2006 to 2022, and provide a scientific basis for prevention and control of scrub typhus in Yantai City.Methods:In the subsystem "Infectious Disease Monitoring System" of the "China Disease Prevention and Control Information System", data on the outbreak of scrub typhus in Yantai City from January 1, 2006 to December 31, 2022 were collected and processed using software such as SPSS 23.0 for descriptive statistics.Results:A total of 599 scrub typhus cases were reported in Yantai City from 2006 to 2022, with an average annual reported incidence of 0.51/100 000. The reported incidence showed an upward trend year by year from 2006 to 2014 (χ 2trend = 144.72, P < 0.001), and reached a peak in 2014 (1.04/100 000). There was another small peak in 2019 (0.91/100 000). The incidence of scrub typhus showed a clear seasonal unimodal distribution, with October being the peak, accounting for 72.62% (435/599). In addition to Changdao County, other 12 cities (districts) reported scrub typhus cases. The top 3 cities with average annual reported incidence were Zhaoyuan City (1.58/100 000), Laizhou City (1.47/100 000) and Laiyang City (0.75/100 000). In all reported cases, the male to female ratio was 1.00∶1.07 (289∶310). The cases were mainly concentrated in the age range of 50 to 79, accounting for 74.46% (446/599). There was a statistically significant difference in the number of patients in different age groups in different years ( H = 38.71, P = 0.001). Farmers accounted for 88.65% (531/599) of the affected population. Conclusions:The incidence of scrub typhus in Yantai City shows an increasing trend and is widely distributed. It is recommended to strengthen the diagnosis, monitoring and infection management of scrub typhus.

ObjectiveTo research the mechanism underlying the effect of raw and processed Aurantii Fructus Immaturus switched to Zhishi Shaoyaosan （ZSS） on constipation-predominant irritable bowel syndrome （C-IBS） rats via the brain-gut-microbiota axis. MethodEighty rats were randomly divided into the blank， model， positive drug （pinaverium bromide， 15.625 mg·kg^-1）， raw ZSS， stir-fried ZSS， bran-fried ZSS， charcoal-fried ZSS and finished ZSS groups （3.75 g·kg^-1）， with 10 rats in each group. Except for the blank group， which received intragastric administration of 0.9% sodium chloride solution at room temperature， all other groups were administered the ice solution at 0 to 4 ℃ （2 mL·d^-1， for a total of 14 d） to establish the C-IBS rat model. The fecal water content and the propulsion rate of small intestine were detected after 14 d of continuous drug administration. The levels of 5-hydroxytryptamine （5-HT）， vasoactive intestinal peptide （VIP）， neuro-peptide Y （NPY）， calcitonin gene-related peptide （CGRP）， substance P （SP）， diamine oxidase （DAO） and D-lactic acid （D-LA） were detected by enzyme linked immunosorbent assay （ELISA）. Hematoxylin-eosin （HE） staining was used to observe the changes in colonic pathological injury in each group. The expression levels of cyclic adenosine monophosphate （cAMP）， protein kinase A （PKA） and aquaporin-3 （AQP3） mRNA in colon tissues were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and the protein expressions of VIP and AQP3 in colon tissues were detected by Western blot. The content of short chain fatty acids （SCFAs） was determined by gas chromatography-mass spectrometry. ResultCompared with the blank group， the fecal water content and intestinal propulsion rate of rat in the model group were significantly decreased （P<0.01）， and the levels of 5-HT， VIP， CGRP and SP in serum were significantly increased. Simultaneously， the NPY levels significantly decreased （P<0.01）， the levels of DAO and D-LA in plasma were significantly increased （P<0.01）， and the mucosal epithelium of colon tissue was slightly damaged， with reduced goblet cells and significantly reduced luminal granules. The mRNA expression levels of AQP3， cAMP and PKA and the protein expression levels of AQP3 and VIP in colon tissue were significantly decreased （P<0.05， P<0.01）. The total amount of SCFAs in feces showed an obvious decreasing trend， with the contents of acetic acid， isobutyric acid， isovaleric acid， valeric acid and caproic acid decreased significantly， while the contents of propionic acid and butyric acid increased significantly （P<0.05， P<0.01）. Compared with the model group， the treatment groups increased the intestinal propulsion rate， improved the intestinal mucosal barrier function， and adjusted the level of serum brain-gut peptide in C-IBS rats （P<0.05， P<0.01）. The expression levels of AQP3， cAMP， PKA mRNA and VIP， AQP3 protein in colon tissue of rats in all treatment groups were increased. All the treatment groups had a significant downregulation of the content of SCFAs except for isobutyric acid in rat feces， and the effect of ZSS prepared by the bran-fried Aurantii Fructus Immaturus was superior than that of other ZSS. ConclusionThe raw and processed Aurantii Fructus Immaturus switched to ZSS could influence the brain-gut-microbiota axis to treat C-IBS rats and it is more reasonable to use bran-fried Aurantii Fructus Immaturus in ZSS.

Background Patients with schizophrenia of Yangming Fushi syndrome experience more severe symptoms,and a substantial proportion of patients derive inadequate benefit from antipsychotics and suffer from serious adverse effects,yet few studies have been conducted on the treatment of schizophrenia of Yangming Fushi syndrome with Zengye Chengqi decoction.Objective To explore the efficacy of Zengye Chengqi decoction combined with olanzapine in the treatment of schizophrenia of Yangming Fushi syndrome,in order to provide references for the treatment of schizophrenia with the combination of traditional Chinese and western medicine.Methods A total of 60 patients attending the Second Affiliated Hospital of Xinxiang Medical College from January 2022 to August 2023 and fulfilling the International Classification of Diseases(ICD-10)diagnostic criteria for schizophrenia were enrolled,and assigned into study group(n=30)and control group(n=30)using random number table methods.All patients were treated with olanzapine,and study group was given Zengye Chengqi decoction on this basis.Treatment for both groups lasted for 4 weeks.All participants were assessed using Positive and Negative Syndrome Scale(PANSS),Montreal Cognitive Assessment(MoCA)and Event-Related Potential P300 at baseline and end of treatment.The occurrence of adverse reactions was recorded at the end of treatment.Results Study group reported a higher treatment effective rate compared with control group(χ2=9.320,P=0.002).After treatment,study group detected a significant reduction in PANSS subscales and total scores(F=10.287,8.258,8.844,20.079,P<0.01),and a notable increase in scores of delayed recall and orientation domains from MoCA(F=4.463,22.255,P<0.05 or 0.01)when compared with control group,with statistical difference.For the P3 component of event-related potential,study group produced significantly larger amplitudes than control group(F=4.247,P<0.05).The incidence rate of abnormal liver function,constipation and increased body mass index(BMI)in study group was lower than those in control group(χ2=4.320,4.463,7.200,P<0.05 or 0.01).Conclusion Zengye Chengqi decoction combined with olanzapine are found to be effective in improving the psychotic symptoms and cognitive function and alleviating the adverse reactions of patients with schizophrenia of Yangming Fushi syndrome.

Objective:To study the clinical manifestations and genetic characteristics of neonatal-onset primary mitochondrial disease (PMD) caused by nuclear gene mutations.Methods:From May 2020 to March 2022, the clinical data, genetic results and follow-up information of neonates with PMD admitted to the Department of Neonatology of our two hospitals were retrospectively analyzed.Results:A total of 4 patients were enrolled, all with hyperlactatemia and metabolic acidosis. In case 1, the fetal cranial MRI showed agenesis of corpus callosum. In case 2, echocardiography after birth indicated hypertrophic cardiomyopathy. Whole exome sequencing found the following mutations: EARS2 nuclear gene c.1294C>T and c.971G>T variants, COA6 nuclear gene c.411_412insAAAG variant, ACAD9 nuclear gene c.1278+1G>A and c.895A>T variants, FOXRED1 nuclear gene c.1054C>T and c.3dup variants. Mitochondrial second-generation sequencing and multiplex ligation-dependent probe amplification showed no abnormalities. Cases 1 and 3 died during the neonatal period. Case 2 died at 2-year-and-2-month of age. Case 4 was followed up to 1 year of age with developmental delay.Conclusions:The main phenotypes of neonatal-onset PMD caused by nuclear gene mutations are hyperlactatemia, refractory metabolic acidosis and cardiomyopathy, which have a poor prognosis. Proactive genetic tests are helpful for early diagnosis.

Objective To investigate the serum levels of miR-665 and miR-144 in the elderly pa-tients with chronic heart failure(CHF)and their relationship with cardiac function.Methods A total of 120 elderly CHF patients who were diagnosed and treated in our hospital from March 2021 to March 2023 were collected and then divided into NYHA grade Ⅱ(n=39),Ⅲ(n=51)and Ⅳ(n=30)subgroups according to the results of NYHA classification.Another 120 elderly healthy volunteers during the same period were recruited as the control group.Clinical data and cardiac function indicators were collected,and the expression levels of miR-665 and miR-144 in se-rum were detected.Pearson correlation analysis was applied to analyze the relationship of miR-665 and miR-144 levels with cardiac function indicators.Results The CHF group had significantly de-creased LVEF,increased left ventricular end-diastolic diameter(LVEDD)and left ventricular end-systolic diameter(LVESD),and elevated serum levels of miR-665 and miR-144 than the control group(P＜0.01).Sequentially reduced LVEF and raised LVEDD and LVESD values and serum miR-665 and miR-144 levels were observed in the patients with NYHA grades Ⅱ,Ⅲ,and Ⅳ in turn(P＜0.01).Pearson correlation analysis showed that there was a positive correlation of the serum level of miR-665 with that of miR-144 in CHF patients(r=0.693,P=0.000),of the miR-665 and miR-144 levels with LVEDD(r=0.485,r=0.507,P＜0.01)and LVESD(r=0.539,r=0.494,P＜0.01),and a negative correlation of the serum levels with LVEF(r=-0.577,r=-0.591,P＜0.01).Conclusion The serum levels of miR-665 and miR-144 are elevated in elderly CHF patients,and are closely associated with their cardiac function.