AIM:This study aims to investigate the impact of TWIK-1 channels on abnormal pacemaker activi-ties induced by hypokalemia and to elucidate the underlying mechanisms.METHODS:The gene sequences encoding hu-man TWIK-1,specific TWIK-1 shRNA and TWIK-1-T118i mutant were synthesized and subsequently subcloned into lenti-viral vectors.To knock down the TWIK-1 gene in human induced pluripotent stem cell-derived cardiomyocytes(hiPSC-CMs),the cells were transduced with lentivirus carrying the specific TWIK-1 shRNA sequences.For the overexpression of TWIK-1 or the TWIK-1-T118i mutant in HL-1 mouse cardiomyocytes,the cells received lentiviral transduction containing the respective gene sequences.Patch-clamp techniques were employed to assess the effects of 1 mmol/L extracellular K+on the membrane potentials and whole-cell currents of the cardiomyocytes.RESULTS:Under conditions of 1 mmol/L extra-cellular K+,depolarization of membrane potentials was observed in the hiPSC-CMs and the HL-1 mouse cardiomyocytes ex-pressing human TWIK-1 channel,leading to the induction of abnormal pacemaker activities.This phenomenon could be reversibly abolished by the removal of extracellular Na+or inhibited through TWIK-1 knockdown.In contrast,the mem-brane potentials of HL-1 mouse cardiomyocytes expressing human TWIK-1-T118i mutant hyperpolarized,with no occur-rence of abnormal pacemaker activities.The hiPSC-CMs exhibiting abnormal pacemaker activities at 1 mmol/L extracellu-lar K+demonstrated TWIK-1-like Na+leak currents,which were blocked by quinine,a non-selective blocker of TWIK-1.CONCLUSION:The TWIK-1 channels play a critical role in the development of hypokalemia-induced abnormal pace-maker activities in human cardiomyocytes by facilitating Na+leak currents.

[Purpose]To analyze the trends of cancer incidence and mortality from 1991 to 2020 in Jiashan County of Zhejiang Province,and to predict trends from 2021 to 2035.[Methods]Cancer cases and deaths from 1991 to 2020 in Jiashan County were collected.The crude inci-dence rate and mortality rate,age-standardized rate(ASR)were calculated.The annual percentage change(APC)and average annual percentage change(AAPC)were calculated by Joinpoint model to analyze the trend of cancer incidence and mortality.The age-period-cohort model was used to predict the trends of cancer incidence and mortality from 2021 to 2035 and the estimated annual percentage change(EAPC)was calculated.[Results]The age-standardized incidence rates by Chinese standard population(ASIRC)in Jiashan were 159.06/105,187.25/105,and 254.37/105 during 1991-2000,2001-2010,and 2011-2020,respectively.The trends in young and middle-aged women and people above 65 years old were more predominant.Lung,stomach,colorectal,liver and female breast cancers were major incident cancers for the period from 1991 to 2020.The cancer incidence showed an increasing trend from 1991 to 2020(AAPC=2.17％,P＜0.05)and is expect-ed to continue to increase over 2021 to 2035(EAPC=3.03％,P＜0.05).The age-standardized mor-tality rates by Chinese standard population(ASMRC)in Jiashan were 119.74/105,117.79/105,and 100.11/105 during 1991-2000,2001-2010,and 2011-2020,respectively.Lung,liver,stomach,colorectal,esophageal,and pancreatic cancers were leading causes of cancer deaths from 1991 to 2020.The cancer mortality showed a decreasing trend from 1991 to 2020(AAPC=-1.00％,P＜0.05)and is expected to continue to decline over 2021 to 2035(EAPC=-1.67％,P＜0.05).[Conclusion]The mortality rate from cancers has been gradually declined in Jiashan County,while the incidence rate has increased significantly and will continue to increase in the next 15 years.High incidence of lung cancer,liver cancer,gastric cancer and colorectal cancer makes it crucial to pay close attention to key groups and strengthen the control of risk factors of high inci-dence cancer.

AIM:This study aims to investigate the impact of TWIK-1 channels on abnormal pacemaker activi-ties induced by hypokalemia and to elucidate the underlying mechanisms.METHODS:The gene sequences encoding hu-man TWIK-1,specific TWIK-1 shRNA and TWIK-1-T118i mutant were synthesized and subsequently subcloned into lenti-viral vectors.To knock down the TWIK-1 gene in human induced pluripotent stem cell-derived cardiomyocytes(hiPSC-CMs),the cells were transduced with lentivirus carrying the specific TWIK-1 shRNA sequences.For the overexpression of TWIK-1 or the TWIK-1-T118i mutant in HL-1 mouse cardiomyocytes,the cells received lentiviral transduction containing the respective gene sequences.Patch-clamp techniques were employed to assess the effects of 1 mmol/L extracellular K+on the membrane potentials and whole-cell currents of the cardiomyocytes.RESULTS:Under conditions of 1 mmol/L extra-cellular K+,depolarization of membrane potentials was observed in the hiPSC-CMs and the HL-1 mouse cardiomyocytes ex-pressing human TWIK-1 channel,leading to the induction of abnormal pacemaker activities.This phenomenon could be reversibly abolished by the removal of extracellular Na+or inhibited through TWIK-1 knockdown.In contrast,the mem-brane potentials of HL-1 mouse cardiomyocytes expressing human TWIK-1-T118i mutant hyperpolarized,with no occur-rence of abnormal pacemaker activities.The hiPSC-CMs exhibiting abnormal pacemaker activities at 1 mmol/L extracellu-lar K+demonstrated TWIK-1-like Na+leak currents,which were blocked by quinine,a non-selective blocker of TWIK-1.CONCLUSION:The TWIK-1 channels play a critical role in the development of hypokalemia-induced abnormal pace-maker activities in human cardiomyocytes by facilitating Na+leak currents.

[Purpose]To analyze the trends of cancer incidence and mortality from 1991 to 2020 in Jiashan County of Zhejiang Province,and to predict trends from 2021 to 2035.[Methods]Cancer cases and deaths from 1991 to 2020 in Jiashan County were collected.The crude inci-dence rate and mortality rate,age-standardized rate(ASR)were calculated.The annual percentage change(APC)and average annual percentage change(AAPC)were calculated by Joinpoint model to analyze the trend of cancer incidence and mortality.The age-period-cohort model was used to predict the trends of cancer incidence and mortality from 2021 to 2035 and the estimated annual percentage change(EAPC)was calculated.[Results]The age-standardized incidence rates by Chinese standard population(ASIRC)in Jiashan were 159.06/105,187.25/105,and 254.37/105 during 1991-2000,2001-2010,and 2011-2020,respectively.The trends in young and middle-aged women and people above 65 years old were more predominant.Lung,stomach,colorectal,liver and female breast cancers were major incident cancers for the period from 1991 to 2020.The cancer incidence showed an increasing trend from 1991 to 2020(AAPC=2.17％,P＜0.05)and is expect-ed to continue to increase over 2021 to 2035(EAPC=3.03％,P＜0.05).The age-standardized mor-tality rates by Chinese standard population(ASMRC)in Jiashan were 119.74/105,117.79/105,and 100.11/105 during 1991-2000,2001-2010,and 2011-2020,respectively.Lung,liver,stomach,colorectal,esophageal,and pancreatic cancers were leading causes of cancer deaths from 1991 to 2020.The cancer mortality showed a decreasing trend from 1991 to 2020(AAPC=-1.00％,P＜0.05)and is expected to continue to decline over 2021 to 2035(EAPC=-1.67％,P＜0.05).[Conclusion]The mortality rate from cancers has been gradually declined in Jiashan County,while the incidence rate has increased significantly and will continue to increase in the next 15 years.High incidence of lung cancer,liver cancer,gastric cancer and colorectal cancer makes it crucial to pay close attention to key groups and strengthen the control of risk factors of high inci-dence cancer.

Objective To evaluate the relationship between perioperative myocardial injury(PMI)and serum N/OFQ levels in elderly patients with coronary heart disease.Methods Totally 120 elderly patients who underwent hip fracture surgery under general anesthesia from January 2022 to May 2023 were included,including 60 patients with coronary heart disease(CHD group)and 60 patients without coronary heart disease(control group).The venous blood of patients was collected 10 minutes before anesthesia induction(T0),12 hours after surgery(T1)and 24 hours after surgery(T2)to detect the content of N/OFQ and high-sensitivity myocardial troponin I(hs-cTnI)in serum.Record perioperative adverse cardiovascular events(PACE)and the use of vasoactive drugs during surgery.Results Compared with the control group,the N/OFQ and hs cTnI levels at T0 and T1 in the CHD group were significantly increased(P<0.05).There was a positive correlation between N/OFQ and hs-cTnI levels at T1 and T2 in CHD and control group(P<0.05).The use of PACE and intraoperative vasoactive drugs in the CHD group was higher than that in the control group(P<0.05).Conclusion There is a correlation between the increased N/OFQ content and PMI in elderly patients with coronary heart disease after surgery,which may become an early predictive indicator of PMI.

Objective By population survey,to explore the epidemiological characteristics of gastric precancerous lesions in Lishui District of Nanjing and provide objective basis for the prevention and treatment of early gastric cancer.Methods From July 2021 to December 2022,21 977 patients who received endoscopy and/or 13C-UBT in Lishui District People's Hospital and 6 medical community units in Nanjing City were retrospectively analyzed for demography characteristics,detection rate of gastric precancerous lesions,and H.Pylori infection rate.Results(1)590 cases of gastric precancerous lesions were detected(detection rate 2.68%);(2)The total detection rate of precancerous lesions and three pathological types in males were all higher than those in females(all P<0.001);(3)The minimum age for the total detection rate of precancerous lesions in males and the mini-mum age for each pathological type were lower than in females(P<0.001,0.009,0.005,0.002);(4)The popu-lation total H.pylori infection rate was 23.10%,the H.pylori infection rate in patients with precancerous lesions was higher than that in non-precancerous lesions(P<0.001),both H.pylori infection rate of male and female in precancerous lesions were all higher than those of non-precancerous lesions of the same sex(all P<0.001),in addition,the H.pylori infection rate of male whether in precancerous or non-precancerous lesions was higher than that of female(all P<0.001);(5)The precancerous lesions detection rate in male,female,and the overall age range of 20～29 to 70～79 years is positively correlated with age growth(P<0.001),and rapidly decreases after the age of 79,the of H.pylori infection rate was also positively correlated with age growth(P<0.001),and the trend of age change(P<0.001)was parallel to the precancerous lesions detection rate.Conclusions The detec-tion rate of gastric precancerous lesions in this region is above the average level in China;the total H.pylori infec-tion rate is at a relatively low level in China;the H.pylori infection rate is parallel to the age trend of the detection rate of gastric precancerous lesions,and increases with age.

Objective:To explore the effect of clinical conventional fractionated dose radiation on the expression levels of immunogenic cell death (ICD) related proteins in patients with nasopharyngeal carcinoma (NPC).Methods:A total of 38 newly-treated NPC patients admitted to the Affiliated Cancer Hospital of Guizhou Medical University from November 2020 to December 2021 were enrolled, all of whom received induction chemotherapy and concurrent chemoradiotherapy, and another 20 healthy volunteers were selected as controls for a prospective study. The contents of ICD related proteins, namely calreticulin (CRT), high mobility group box 1 protein (HMGB-1) and heat shock protein 70 (HSP70) and the proportion of dendritic cell (DC) in the peripheral blood of patients were detected before treatment, after induction chemotherapy and after concurrent chemoradiotherapy, respectively. The correlation between the above indicators, general clinical data and short-term efficacy was analyzed by statistical methods such as t-test and analysis of variance (ANOVA). Results:The levels of HSP70 and HMGB-1 in peripheral blood of NPC patients before treatment were higher than those of healthy controls (both P<0.05). After concurrent chemoradiotherapy, the content of CRT was significantly higher than that before treatment ( P<0.05), whereas the difference before and after induction chemotherapy and the difference before and after concurrent chemoradiotherapy were not significantly correlated with the short-term efficacy of NPC patients. HSP70 level was significantly decreased after concurrent chemoradiotherapy ( P<0.001). There were no significant differences in the content of HMGB-1 after induction chemotherapy and concurrent chemoradiotherapy (both P>0.05). Conclusion:NPC patients receiving TPF regimen (docetaxel+cisplatin+fluorouracil) for induction chemotherapy and sequential cisplatin concurrent chemotherapy may induce ICD in NPC cells, and CRT has potential value in reflecting the clinical efficacy of NPC.

BACKGROUND: The transforming growth factor β1 (TGF-β1)-induced epithelial-mesenchymal transition (EMT) has been proven associated with the pathogenesis of asthmatic airway remodeling, in which the Wnt/β-catenin pathway plays an important role, notably with regard to TGF-β1. Recent studies have shown that 1α, 25-dihydroxyvitamin D3(1α, 25(OH)2D3) inhibits TGF-β1-induced EMT, although the underlying mechanism have not yet been fully elucidated. METHODS: Alveolar epithelial cells were exposed to 1α, 25(OH)2D3, ICG-001, or a combination of both, followed by stimulation with TGF-β1. The protein expression of E-cadherin, α-smooth muscle actin, fibronectin, and β-catenin was analyzed by western blotting and immunofluorescence analysis. The mRNA transcript of Snail was analyzed using RT-qPCR, and matrix metalloproteinase 9 (MMP-9) activity was analyzed by gelatin zymogram. The activity of the Wnt/β-catenin signaling pathway was analyzed using the Top/Fop flash reporters. RESULTS: Both 1α, 25(OH)2D3 and ICG-001 blocked TGF-β1-induced EMT in alveolar epithelial cells. In addition, the Top/Fop Flash reporters showed that 1α, 25(OH)2D3 suppressed the activity of the Wnt/β-catenin pathway and reduced the expression of target genes, including MMP-9 and Snail, in synergy with ICG-001. CONCLUSION 1α, 25(OH)2D3 synergizes with ICG-001 and inhibits TGF-β1-induced EMT in alveolar epithelial cells by negatively regulating the Wnt/β-catenin signaling pathway.

Objective:To explore the relationship between rs3758391 polymorphism of SIRT1 gene and depressive symptoms, and to further understand the role of SIRT1 gene in major depressive disorder. Methods:A total of 323 patients with major depressive disorder were retrospectively collected from the Jinhua Second Hospital, Wenzhou Kangning Hospital and Shanghai Mental Health Center, Shanghai Jiao Tong University School of Medicine. A total of 347 healthy subjects were also recruited. Depressive symptoms were evaluated by using the Hamilton Depression Scale (HAMD), and rs3758391 polymorphism was genotyped by using the TaqMan SNP genotyping Assay. The effect of rs3758391 polymorphism on the expression of SIRT1 mRNA in brain was analyzed by BRAINEAS database, and the difference of depressive symptom severity among three genotypes at rs3758391 polymorphism was compared by multivariate analysis of variance. Results:The frequencies of C and T alleles of rs3758391 polymorphism in SIRT1 gene were 18.7% and 81.3% in the case group, and 14.3% and 85.7% in the control group, respectively. The allelic distribution frequencies between the two groups were significantly different (χ2=4.86, P=0.03). There were significant differences in mood, cognitive impairment and HAMD scores among patients with different genotypes of rs3758391 polymorphism (P<0.05). The results of eQTL analysis showed that rs3758391 polymorphism was significantly correlated with the expression of SIRT1 gene in occipital cortex (OCTX) (P=0.003). Conclusion:rs3758391 polymorphism of SIRT1 gene may be a risk factor for major depressive disorder in Chinese Han population, and is associated with the severity of depressive symptoms, especially with emotional symptoms and cognitive impairment.

Objective To explore the effect of rat nerve growth factor combined with early rehabilitation intervention in the treatment of hearing injury caused by cytomegalovirus (CMV) infection.Methods 106 cases of hearing impairment caused by cytomegalovirus infection diagnosed in our hospital from January 2012 to February 2018 were randomly divided into observation group and control group,53 cases in each group.The control group were treated only with ganciclovir and comprehensive rehabilitation therapy,while the treatment group was treated with nerve growth factor on the basis of the control group.Before and after treatment,the children in both groups underwent multi-frequency steady-state evoked potential test,cytomegalovirus antibody DNA fluorescence quantification,and their mothers underwent cytomegalovirus antibody test.The total effective rate of the two groups and the effective rate of the observation group with different degrees of hearing impairment were compared,and the therapeutic effect of the observation group with congenital and acquired CMV infection was compared.Results The total effective rate was 80.7％ in the observation group,which was higher than 65.9％ in the control group (P ＜ 0.05).The effective rate of mild to moderate hearing loss in observation group was higher than that in severe hearing impairment (P ＜0.05).The efficacy of the rat nerve growth factor in the treatment of hearing loss after CMV infection was higher than that of hearing loss caused by congenital cytomegalovirus infection (P ＜ 0.05).Conclusions Rat nerve growth factor combined with early comprehensive rehabilitation intervention is an effective method to treat acoustic nerve injury caused by CMV infection.The effect of rat nerve growth factor on mild and moderate hearing impairment was better than that of severe hearing impairment.The therapeutic effect of acquired CMV infection on hearing damage was better than that of by congenital infection.