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 mechanism of proliferation of gastric cancer cells induced byHelicobacter pylori(Hp) from the perspective of the mitochondrial unfolded protein response(UPRMT). Methods : C57BL6/J mice, human gastric cancer cells AGS and SGC-7901 cells were infected with Hp, mitochondrial proteins were extracted, and the expressions of UPRMT, including activation of transcription factor 5(ATF5), heat shock protein 60(mtHSP60), heat shock protein 70(mtHSP70) and mitochondrial protease(ClpP), were detected by Western blot. Immunohistochemistry was used to locate the expression sites of 4 proteins in mouse gastric tissue. AGS and SGC-7901 cells were infected with Hp, and cell proliferation was detected by RTCA and CCK-8. Finally, the expression of four indexes in gastric cancer tissues was analyzed by TCGA samples in the UALCAN database. Results : Hp infection could significantly promote the expression of ATF5, mtHSP60, mtHSP70 and ClpP in the mitochondria of gastric epithelial cells of C57BL6/J mice, AGS and SGC-7901 cells, and promote the proliferation of gastric cancer cells. When compared to normal gastric tissues in TCGA samples, ATF5, mtHSP60, mtHSP70, and ClpP were highly expressed in gastric cancer tissues, with statistically significant difference(allP<0.01). Conclusion Hp infection can induce UPRMTin gastric epithelial cells and gastric cancer cells, which in turn promotes cell proliferation.

Objective To investigate the therapeutic effect of traditional Chinese medicine (TCM) on portal vein thrombosis (PVT) in patients with liver cirrhosis and its medication characteristics. Methods A retrospective analysis was performed for 89 patients with liver cirrhosis and PVT who were hospitalized and treated in Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine, and according to whether TCM treatment was applied in combination, they were divided into TCM group with 59 patients and control group with 30 patients. Related data were collected for the two groups, including demographic data, laboratory examination, radiological examination, gastroscopy, history of surgery, portal hypertension-related complications, medication, and follow-up data. The independent samples t -test was used for comparison of normally distributed continuous data between two groups, and the Mann-Whitney U rank sum test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test or the Fisher's exact test was used for comparison of categorical data between two groups. An ordinal polytomous Logistic regression analysis was used for multivariate analysis. TCM Inheritance Computing Platform (V3.0) was used to perform a drug effect cluster analysis of TCM prescriptions. Results The multivariate logistic regression analysis showed that esophageal and gastric varices (odds ratio [ OR ]=3.144, 95% confidence interval [ CI ]: 1.221-8.094), PVT involving the portal vein (PV) and the superior mesenteric vein (SMV) ( OR =51.667, 95% CI : 3.536-754.859), PVT involving PV+spleen vein (SV)+SMV ( OR =13.271, 95% CI : 2.290-76.928), cavernous transformation of the portal vein ( OR =11.896, 95% CI : 1.172-120.696), and TCM intervention ( OR =0.348, 95% CI : 0.129-0.938) were influencing factors for the outcome of PVT in liver cirrhosis. Follow-up results showed that compared with the control group, the TCM group had a significantly lower progression rate (16.95% vs 56.67%, P < 0.001) and a significantly lower incidence rate of variceal rupture and bleeding (8.47% vs 33.33%, P < 0.001). Effective TCM drugs with a relatively high frequency of use included deficiency-tonifying drugs (359 times, 34.6%), blood-activating and stasis-resolving drugs (202 times, 19.5%), and diuresis-inducing and dampness-draining drugs (180 times, 17.3%); the TCM drugs with a relatively high frequency of use included Astragalus membranaceus (57 times, 8.7%), Angelica sinensis (50 times, 7.6%), and leech (48 times, 7.3%); TCM drug combinations with a relatively high frequency of use included Astragalus membranaceus+Angelica sinensis, Astragalus membranaceus+leech, Angelica sinensis+leech, and Astragalus membranaceus+Angelica sinensis+leech. Conclusion Qi-tonifying, blood-activating, and stasis-breaking drugs, such as Astragalus membranaceus, Angelica sinensis, and leech, can promote the stabilization or recanalization of PVT in liver cirrhosis and reduce the incidence rate of bleeding events due to portal hypertension.

Objective:To use metagenomic sequencing to compare the differences in intestinal microbiota species and metabolic pathways in patients with hepatitis B cirrhosis with or without ascites and further explore the correlation between the differential microbiota and clinical indicators and metabolic pathways.Methods:20 hepatitis B cirrhosis cases [10 without ascites (HBLC-WOA), 10 with ascites (HBLC-WA), and 5 healthy controls (HC)] were selected from the previously studied 16S rRNA samples. Metagenome sequencing was performed on the intestinal microbiota samples. The Kruskal-Wallis rank sum test and Spearman test were used to identify and analyse differential intestinal microbiota populations, metabolic pathways, and their correlations.Results:(1) The overall structure of the intestinal microbiota differed significantly among the three groups ( R = 0.19, P = 0.018). The HC group had the largest abundance of Firmicutes and the lowest abundance of Proteobacteria at the genus level. Firmicutes abundance was significantly decreased ( Pfdr < 0.01), while Proteobacteria abundance was significantly increased ( Pfdr < 0.01) in patients with cirrhosis accompanied by ascites; (2) LEfSe analysis revealed that 29 intestinal microbiota (18 in the HBLC-WA group and 11 in the HBLC-WOA group) played a significant role in the disease group. The unclassified Enterobacteriaceae and Klebsiella species in the HBLC-WA group and Enterobacteriaceae in the HBLC-WOA group were positively correlated with the Child-Turcotte-Pugh (CTP) score, prothrombin time, and international normalized ratio score and negatively correlated with albumin and hemoglobin levels ( P < 0.05). Escherichia and Shigella in the HBLC-WA group were positively correlated with CTP scores ( P < 0.05); (3) The correlation analysis results between the KEGG pathway and 29 specific intestinal microbiota revealed that Enterobacteriaceae and arachidonic acid, α-linolenic acid, glycerolipid metabolism, and fatty acid degradation were positively correlated in the lipid metabolism pathway, while most Enterobacteriaceae were positively correlated with branched-chain amino acid degradation and negatively correlated with aromatic amino acid biosynthesis in the amino acid metabolic pathway. Conclusion:A significant increment of Enterobacteriaceae in the intestines of HBLC-WA patients influenced hepatic reserve function and was associated with amino acid and lipid metabolic pathways. Therefore, attention should be paid to controlling the intestinal microbiota to prevent complications and improve the prognosis in patients with hepatitis B cirrhosis, especially in those with ascites.

We investigate the different expression of toxin gene mazF3,6,9 and antitoxin gene mazE3,6,9 in the drug-resistance Mycobacterium tuberculosis,we used quantitative real-time polymerase chin reaction method to detect the expression level of toxin gene mazF3,6,9 and antitoxin gene mazE3,6,9 in M.tuberculosis (20 mono-resistance strains,20 multidrug resistance strains and standard strain H37Rv).The differences of gene expression levels between groups were analyzed by oneway ANOVA.Contrasting with control group,toxin genes mazF6,9 were up-regulated expression levels both in mono-resistance (11.1519±22.31721;8.4306±17.97897) and multidrug resistance (4.6016±1.29018;6.9627±6.92948),had statistical significance (P＜0.01),mazF3 expression levels had statistical significance neither in mono-resistance nor in multidrug resistance (P＞0.05);antitoxin genes mazE3 was in down-expression level,and had statistical significance both in mono-resistance (0.3606±0.12527) and multidrug resistance (0.2016±0.16542) (P＜0.01),mazE6 had no statistical significance (P＞0.05)either in mono-resistance or multi drug resistance,mazE9 only in multidrug resistance(0.3989±0.37679) was in downexpression level,and has statistical significance (P＜0.001).The toxin gene mazF6,9 and antitoxin gene mazE3,9 may participate in the drug-resistance formation of M.tuberculosis.

To observe the effects of vitamin E (Vit E) on mercuric chloride (HgCl2)-induced renal interstitial fibrosis (RIF) in rats and discuss its antioxidative mechanism.

Objective: To investigate the characteristics of traditional Chinese medicine (TCM) syndromes and their elements in patients with posthepatitic cirrhosis by analyzing the relationships between signs and symptoms and biochemical parameters. Methods: A total of 440 patients with hepatitis B cirrhosis treated in Shanghai Public Health Center and Shuguang Hospital, Longhua Hospital and Central Hospital of Putuo District Affiliated to Shanghai University of Traditional Chinese Medicine during January 2002 to January 2006 were enrolled in this study. Signs and symptoms and biochemical information of patients were collected by using a self-designed questionnaire regarding the four examinations of TCM. Signs and symptoms were firstly analyzed to find the frequency of occurrence. Then, the patients were divided into two groups according to non-existent or existent sign and symptom and the correlations between the signs and symptoms which occurred most frequently and their biochemical parameters were analyzed. Results: Sixteen symptoms which occurred most frequently were fatigue, colored urine, liver palms, opaque complexion, string-like pulse, weakness at waist and knees, dry month and bitter taste in the mouth, profuse dreaminess and poor sleepiness, heaviness of limbs, abdominal distention, yellow eyes, fine pulse, impetuosity and susceptibility to rage, splenomegaly, poor appetite, and distension and fullness in the chest and hypochondrium. A previous study on syndrome differentiation of 900 patients with posthepatitic cirrhosis showed 4 syndrome patterns: internal accumulation of dampness-heat, liver-kidney yin deficiency, internal accumulation of blood stasis-heat, and liver depression and spleen deficiency. Further analysis showed that internal accumulation of dampness-heat syndrome was characterized by obvious hepatic inflammation, poor synthesis function and more asctites. Liver-kidney yin deficiency syndrome was characterized by low-level hepatic inflammation, poor synthesis function and more ascites. Internal accumulation of blood stasis-heat syndrome was characterized by low-level hepatic inflammation, poorer synthesis function, ascites and splenomegalia, and liver depression and spleen deficiency syndrome was characterized by slight hepatic inflammation, synthesis function injury, decreased internal portal vein diameter and less ascites. Conclusion: Different syndrome patterns have different pathological features, showing the complexity and polymorphism of syndrome construction.

OBJECTIVE: To investigate the effects of Fuzheng Huayu Recipe on renal interstitial fibrosis in rats induced by the toxic substances. METHODS: Forty Wistar rats were divided into 3 groups, including the normal group, model control and Fuzheng Huayu Recipe treated group. The renal fibrotic model was induced with intraperitoneal injection of dimethylnitrosamine (DMN) for 4 weeks and oral administration of food containing heavy metals such as Hg, etc for 8 weeks. From the 5th week, the rats were orally administered Fuzheng Huayu Recipe for 4 weeks. The renal tissue was stained with HE and periodic acid-silver metheramine, respectively. The serum creatinine and urea nitrogen were assayed with the kits, and the renal hydroxyproline (Hyp) contents in the homogenate of kidney tissue were measured with HCl hydrolysis. RESULTS: In the model group, the renal tubule was swollen, and the tubular epithelial cells were degenerated, necrosed and detached, with the infiltration of inflammatory cells; the tubular interstitium was expanded, characterized with the deposition of large collagens; the tubular basement membrane became thicker and wrinkled; and the renal Hyp content and serum creatinine and urea nitrogen levels increased remarkably as compared to the normal group. After the treatment with Fuzheng Huayu Recipe, the renal tubular inflammation and interstitial collagen deposition were alleviated, and the serum creatinine and urea nitrogen levels decreased significantly as compared to the model group. CONCLUSION: Administration of DMN and heavy metals can induce the renal interstitial fibrosis and impair the renal function. Fuzheng Huayu Recipe can improve the impaired renal functions and reverse the renal interstitial fibrosis.