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 therapeutic potential of octaarginine (R8)-modified essential oil from Fructus Alpiniae zerumbet (EOFAZ) lipid microspheres (EOFAZ@^R8LM) for cardiovascular therapy. METHODS: EOFAZ@^R8LM was developed by leveraging the volatilization of EOFAZ and integrating it with the oil phase of LM, followed by surface modification with cell-penetrating peptide R8 to target the site of vascular endothelial injury. The therapeutic effects of this formulation in alleviating lipopolysaccharide-induced vascular endothelial inflammation were evaluated by assessing mitochondrial membrane potential (MMP), intracellular reactive oxygen species (ROS) levels, as well as inflammatory factors interleukin-6 (IL-6) and interleukin-1β (IL-1β) levels. RESULTS: EOFAZ@^R8LM effectively delivered EOFAZ to the site of injury and specifically targeted the mitochondria in vascular endothelial cells, thereby ameliorating mitochondrial dysfunction through regulation of MMP and reduction of intracellular ROS levels. Moreover, it attenuated the expression levels of IL-6 and IL-1β, exerting protective effects on the vascular endothelium. CONCLUSION Our findings highlight the significant therapeutic potential of EOFAZ@^R8LM in cardiovascular therapy, providing valuable insights for developing novel dosage forms utilizing EOFAZ for effective treatment against cardiovascular diseases.

Cholesterol (CH) plays a crucial role in enhancing the membrane stability of drug delivery systems (DDS). However, its association with conditions such as hyperlipidemia often leads to criticism, overshadowing its influence on the biological effects of formulations. In this study, we reevaluated the delivery effect of CH using widely applied lipid microspheres (LM) as a model DDS. We conducted comprehensive investigations into the impact of CH on the distribution, cell uptake, and protein corona (PC) of LM at sites of cardiovascular inflammatory injury. The results demonstrated that moderate CH promoted the accumulation of LM at inflamed cardiac and vascular sites without exacerbating damage while partially mitigating pathological damage. Then, the slow cellular uptake rate observed for CH@LM contributed to a prolonged duration of drug efficacy. Network pharmacology and molecular docking analyses revealed that CH depended on LM and exerted its biological effects by modulating peroxisome proliferator-activated receptor gamma (PPAR-γ) expression in vascular endothelial cells and estrogen receptor alpha (ERα) protein levels in myocardial cells, thereby enhancing LM uptake at cardiovascular inflammation sites. Proteomics analysis unveiled a serum adsorption pattern for CH@LM under inflammatory conditions showing significant adsorption with CH metabolism-related apolipoprotein family members such as apolipoprotein A-V (Apoa5); this may be a major contributing factor to their prolonged circulation in vivo and explains why CH enhances the distribution of LM at cardiovascular inflammatory injury sites. It should be noted that changes in cell types and physiological environments can also influence the biological behavior of formulations. The findings enhance the conceptualization of CH and LM delivery, providing novel strategies for investigating prescription factors' bioactivity.

Objective To observe the value of multi-slice spiral CT(MSCT)post-processing technologies for diagnosing otosclerosis.Methods Clinical data and original axial plain MSCT of 47 patients with otosclerosis(92 ears)and 65 patients with non-otosclerosis hearing impairment(79 ears)were retrospectively enrolled.MSCT post-processing images,including multi-planar reformation(MPR)of stapes and cochleas and curved planar reformation(CPR)of ossicular chains were obtained.The diagnostic value of original MSCT images alone and raw data of MSCT combing with post-processing images for diagnosing otosclerosis were compared.Results Otosclerosis was correctly diagnosed in 66 ears according to original MSCT images alone,but in 89 ears combined with MSCT post-processing images.The sensitivity of original MSCT images alone and combined with MSCT post-processing images was 71.74％and 96.74％,respectively,and the diagnostic accuracy was 81.29％and 96.49％,respectively,those of the latter were both higher than of the former(both P＜0.05),which had specificities being not significantly different(92.41％vs.96.20％,P＞0.05).Conclusion Combining with post-processing technologies could increase the sensitivity and accuracy of MSCT for diagnosing otosclerosis.

Objective:To evaluate the clinical efficacy of Bushen Huoxue Shujin Decoction combined with thunder fire moxibustion in the treatment of lumbar intervertebral disc herniation with kidney deficiency and blood stasis syndrome.Methods:Randomized controlled trial. A total of 200 lumbar intervertebral disc herniation patients from Dongzhimen Hospital of Beijing University of Chinese Medicine from January to December 2021 were selected as observation objects and the computer random method was used to divide 66 patients into combination group, 67 in the control group 1, and 67 in the control group 2. The Control group 1 was given conventional western medicine and thunder fire moxibustion, the control group 2 was given conventional western medicine and Bushen Huoxue Shujin Decoction, and the combined group was given conventional western medicine and Bushen Huoxue Shujin Decoction and thunder fire moxibustion. All the groups were treated with 14 days as a course of treatment, a total of 3 courses. TCM syndrome scores were performed before and after treatment, and lumbar joint activity was measured by using a muscle state testing analyzer; the levels of NF-κB, prostaglandin E2 (PGE2) and hypoxia-inducible factor-2α (HIF-2α) were determined by ELISA; adverse reactions during treatment were recorded and clinical efficacy was evaluated.Results:The total effective rate of the combination group was 92.42%(61/66), the control group 1 was 71.64% (48/67), and the control group 2 was 74.63% (50/67), with a statistically significant difference among the three groups ( χ2=10.28, P=0.006). After treatment, the scores of waist and leg pain, lumbar stiffness, lower limb numbness, and tongue dullness in the combination group were significantly lower than those in the control group 1 and control group 2 ( F values were 15.25, 12.21, 11.77, 14.49, respectively, P<0.01); the range of motion of lumbar joint forward flexion, backward extension, lateral flexion, and lateral rotation in the combination group were significantly greater than those in the control group 1 and control group 2 ( F values were 19.66, 29.50, 33.33, and 24.54, respectively, P<0.01); the levels of serum NF-κB [(41.29±5.91)ng/L vs. (49.97±5.98)ng/L, (50.92±6.02)ng/L, F=47.00], PGE2 [(67.09±8.08)ng/L vs. (80.22±9.92)ng/L, (78.17±9.09)ng/L, F=40.27], HIF-2α[(16.95±3.46) ng/L vs. (20.83±3.98)ng/L, (19.67±3.89)ng/L, F=18.38] in combination group were significantly lower than those in the control group 1 and control group 2 ( P<0.01). During the treatment period, the incidence of adverse reactions in the combination group was 15.15% (10/66), control group 1 was 8.96% (6/67), and control group 2 was 10.45% (7/67), there was no statistically significant difference between the three groups ( χ2=1.36, P=0.506). Conclusion:The combination of Bushen Huoxue Shujin Decoction and thunder fire moxibustion can improve the TCM syndrome and range of motion of patients with kidney deficiency and blood stasis syndrome of lumbar intervertebral disc herniation, inhibit the expression of inflammatory factors, improve clinical efficacy, and have good safety.

OBJECTIVE：To optimize the p reparation technology of citronellol submicroemulsion. METHODS ：The content of citronellol in Citronellol submicroemulsion was determined by HPLC. Citronellol submicroemulsion by high-speed shearing dispersion-high pressure homogenization method ，with centrifugation stability constant （ke） and particle size were used as evaluation indexes. Its formulation and preparation technology were optimized and validated. Drug-loading amount and encapsulation rate of the preparation were detected. RESULTS ：The linear range of citronellol were 4-64 μg/mL（R 2＝0.999 9）. RSDs of precision ，stability（24 h）and reproducibility tests were all lower than 3％. The recoveries were 97.64％-101.97％（RSD＝ 2.28％，n＝3），97.71％-99.50％（RSD＝1.29％，n＝3），96.87％-101.48％（RSD＝2.86％，n＝3）. The optimal formulation included that total weight of soybean oil and medium chain triglycerides （1 ∶ 1，g/g）was 3.75 g，1.2％ soybean phospholipid was 0.6 g， cholesterol was 0.06 g，citronellol was 1.25 g，0.6 ％ sodium oleate was 0.3 g，15-hydroxystearic acid polyethylene glycol ester was 0.75 g，poloxamer 188 was 0.75 g，water added to 50 mL. After prepared by optimal technology at 4 ℃ which contained shearing speed of 13 000 r/min，lasting for 5 min， primary emulsion was adjusted to pH 7 with dilute hydro- chloric acid ，and homogenized with 600 Bar high pressure for 1434412440@qq.com 5 min. The parameters of Citronellol submicroemulsion accor- ding to optimal formulation and technology contained mean particle size of （91.05±0.26）nm，PDI of （0.20±0.01）， Zeta-potential of （－30.86±0.39）mV，average content of 649511230@qq.com citronellol（100.21±0.01）％，the drug-loading amount was （2.481 7 ± 0.000 7） mg/mL，the encapsulation rate was （99.27 ± 0.03）％ . CONCLUSIONS ：The optimal formulation and technology is stable and feasible.

Objective:To investigate the application of magnetic resonance imaging (MRI) in clinical staging and classification of adult Japanese encephalitis.Methods:The clinical data and craniocerebral MRI findings of 35 adult patients with Japanese encephalitis admitted in General Hospital of Ningxia Medical University from August to September 2018 were analyzed retrospectively. The MRI imaging characteristics were compared among patients with different stages and types, the apparent diffusion coefficient (ADC value) of thalamic lesions in patients of different stages was analyzed. SPSS 23.0 software was used to analyze the data.Results:Six moderate cases all had lesions involving the thalamus, and the number of intracranial lesions was <2. In 11 severe cases, 8 had lesions involving thalamus and 5 had lesions involving hippocampus; the number of intracranial lesions was <2 in 3 cases, the number of intracranial lesions was 2-4 in 3 cases, and the number of intracranial lesions was >4 in 5 cases. In 18 cases critical cases, the lesions involved thalamic in 14 cases, hippocampus in 14 cases, cerebral cortex in 14 cases, cerebral feet in 9 cases, basal ganglia area in 6 cases, and brain stem in 2 cases, respectively; 2 cases had the intracranial lesions <2, 6 cases had intracranial lesions 2-4, 10 cases had intracranial lesions >4. In 11 preliminary stage patients, 9 cases had DWI high signal and 2 had FLAIR slightly high signal; in 19 extreme stage patients, 16 cases had DWI high signal, 11 cases had FLAIR slightly high signal, 3 cases had T1WI high signal and 6 cases had T2WI high signal. In 5 recovery stage patients, 1 case had DWI slightly high signal, 5 cases had FLAIR high signal, and 1 case had T2WI high signal. The ADC values of thalamic lesions in recovery and extreme patients were higher than those in the preliminary stage ( q=3.931 and 4.012, P<0.05). The ADC value of thalamic lesions in the recovery period was higher than that in the extreme period ( q=3.372, P<0.05). Conclusions:The number of lesions and the range of involvement are associated with disease severity in adult Japanese encephalitis. The DWI sequence is easy to detect at early stage, and the FLAIR sequence shows a long time span of lesions; and the DWI and FLAIR sequences are of great significance for the early clinical staging and classification of adult encephalitis patients. At the same time, the ADC value shows a trend of disease progresses, which can be used as a supplement for the clinical staging in adult encephalitis patients.

Objective To explore the diagnostic value of the i-Scan for detection of polypoid lesions in right hemicolon during colonoscopy. Methods A total of 200 patients who underwent colonoscopy in Beijing Shijitan Hospital from January 2015 to December 2015 were enrolled. After completion of the first colonoscopy in right hemicolon, a second withdrawal was performed, using white light mode ( white light group, n=93) and i-Scan mode ( i-scan group, n=96) to detect polypoid lesions in the proximal colon. The detection rates of polyp and adenoma were compared between the two groups. Results During the twice withdrawal, compared with white light group, more polyps and adenomas were detected in i-Scan group (1. 469 VS 1. 011, P=0. 028; 0. 979 VS 0. 624,P=0. 039). The proportion of patients with more polyps and adenomas in the i-Scan group was significantly higher than that in the white light group [ 37. 5％( 36/96) VS 22. 6％ ( 21/93) , P=0. 025;24. 0％ ( 23/96) VS 11. 8％ ( 11/93) ,P=0. 030] . i-Scan mode detected more small polyps with diameter<5 mm [ 84. 0％ ( 42/50 ) VS 58. 3％ ( 14/24 ) , P=0. 016 ] . However, there were no differences between the two groups in the size, location, and morphology of the detected adenomas ( all P>0. 05) . The polyp detection rates of the i-Scan group and white light group were 61. 5％ (59/96) and 48. 4％ (45/93), respectively (P=0. 071), and the adenoma detection rates were 47. 9％ (46/96) and 35. 5％ (33/93), respectively (P=0. 083). Conclusion I-Scan mode can increase the detection rate of polyps and adenomas in right hemicolon, and improve detection of polypoid lesions and bsmall polyps in patients with multiple polyps and adenomas.

OBJECTIVE:To prepare aspirin phospholipid complex (ASP-PC) and conduct the characterization. METHODS:Using the combination rate of ASP and PC as index,single factor test was used to screen the preparation method of ASP-PC,PC type,solvent type,reaction time,reaction temperature,solvent volume and drug-lipid ratio. The verification test was conducted. UV spectrophotometry,Thermogravimetric analysis,X-ray diffraction and Fourier transform infrared spectroscopy were used for the characterization of ASP-PC. RESULTS:Magnetic stirring-condensing reflux method was adopted,drug-soybean phospholipids ratio was 1:3 (mol/mol),solvent was tetrahydrofuran,reacting for 3 h under 58 ℃. The average combination rate of prepared ASP-PC was 83.52％(RSD=1.16％,n=3). Compared with ASP,physical mixture of ASP and PC,UV spectrum showed that ASP-PC had no new absorption peak. Thermogravimetric analysis,X-ray diffraction and Fourier transform infrared spectroscopy showed the ASP and PC in ASP-PC were interacted;and ASP-PC changed little in quality within 0-300 ℃. CONCLUSIONS:ASP-PC can be successfully prepared,in which,ASP and PC were combined successfully;while there are still trace amounts of ASP in the form of crystals.

Objective To investigate the value of NICE classification on real-time diagnosis for neoplastic and non-neoplastic colorectal polyps. Methods The histological type of 107 polypoid lesions from 50 patients detected by colonoscopy was predicted by 4 endoscopic physicians using NICE classification, including 1 chief physician, 2 associate chief physician, and 1 attending physician. Taking pathological result as gold standard, the real-time diagnostic accuracy of NICE classification for neoplastic colorectal polyps was evaluated, and the differences of diagnostic accuracy between the 4 endoscopic physicians were compared. Results Among the 107 polypoid lesions, 49 were neoplastic polyps and 58 were non-neoplastic polyps. The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of NICE classification on determination of the histological type of polyps were 81. 6% (40/49), 70. 0% (40/58), 74. 8% ( 80/107) , 70. 0%( 40/58) , and 81. 6%( 40/49) , respectively. There was no statistical difference on the prediction ability between the 4 physicians using NICE classification. Conclusion NICE classification is a feasible method for real-time prediction of histology type of colorectal polyps, and it is undemanding to the qualifications of physicians.