Objective:To investigate the clinical and pathological significance of the DICER1 mutation in follicular thyroid carcinoma (FTC).Methods:Sixty-eight cases of primary FTC resected between 2009 and 2023 were retrieved from The Affiliated Hospital of Qingdao University, Qingdao, China. Sanger sequencing was performed to identify DICER1 and TERT promoter mutations in all cases. Cases with DICER1 or TERT promoter mutations were subject to additional examination of potential mutations in KRAS, HRAS, and NRAS. The clinical and pathological features of DICER1-mutant FTCs were then analyzed. The relationship between DICER1 mutations and TERT-promoter/RAS mutations was also assessed.Results:DICER1 mutations were detected in 16 of the 68 FTC cases (23.5%), with 11 near E1813 at exon 25, 6 near D1709 at exon 24, and 1 in the splice region of exon 25. Two cases harbored two (distinct) mutations. All patients with DICER1-mutant FTC were female. Compared with patients with DICER1-wild-type FTC, those with DICER1-mutant were much younger, and had a higher proportion of minimally invasive subtype. Nine FTCs with DICER1 mutations were subject to further sequencing on adjacent non-cancerous tissues or lymph node tissues, but no mutations were detected. TERT-promoter or RAS hotspot mutations were not identified in any of the DICER1-mutant cases. However, TERT-promoter mutation was found in 6 DICER1-wild-type cases (8.8%, 6/68), with 3 cases also having RAS hotspot mutations and exhibiting highly aggressive biological behaviors.Conclusion:DICER1 mutations may occur in FTCs and appear mutually exclusive with RAS and TERT-promoter mutations, warranting further study as RAS-like mutations.

Background and aims:Environmental pollutants,particularly organochlorine insecticides like endosulfan(ENDO),are increasingly linked to liver toxicity and related diseases.Despite its widespread historical use,the mechanisms underlying ENDO-induced liver damage remain poorly understood.This study aims to elucidate the cellular and molecular mechanisms of ENDO-induced hepatotoxicity.Methods:C57BL/6 mice were exposed to ENDO for two weeks.Single-cell RNA sequencing(scRNA-seq)was subsequently performed on mouse livers to explore ENDO-induced hepatotoxicity at the single-cell level.Differentially expressed genes(DEGs)across cell types and treatments were identified and then subjected to pathway enrichment to uncover key biological processes affected by ENDO.Transcription factor(TF)regulatory network,pseudotime trajectory,and cellular communication analysis were used to explore the molecular and cellular changes after ENDO exposure.Results:ENDO not only caused direct hepatocyte injury but also activated hepatic stellate cells and lymphocytes,triggering inflammatory responses with upregulation of multiple key chemokines and cytotoxic genes.Additionally,ENDO exposure led to the recruitment and activation of myeloid cells,contributing to the inflammatory milieu.An increase in intercellular communication and changes to the hepatic microenvironment,especially the interaction between activated hepatic stellate cells and CD8+T cells were observed,further implicating these processes in ENDO-induced liver damage.Conclusions:This study provides new insights into the cellular and molecular mechanisms underlying liver injury induced by organochlorine insecticides like ENDO.Key genes and pathways involved in ENDO-associated liver toxicity have been identified at a single-cell resolution.These findings suggest that altered cellular communications and inflammatory responses may play pivotal roles in the pathogenesis of ENDO-induced liver injury.

Objective:To investigate the clinical and pathological significance of the DICER1 mutation in follicular thyroid carcinoma (FTC).Methods:Sixty-eight cases of primary FTC resected between 2009 and 2023 were retrieved from The Affiliated Hospital of Qingdao University, Qingdao, China. Sanger sequencing was performed to identify DICER1 and TERT promoter mutations in all cases. Cases with DICER1 or TERT promoter mutations were subject to additional examination of potential mutations in KRAS, HRAS, and NRAS. The clinical and pathological features of DICER1-mutant FTCs were then analyzed. The relationship between DICER1 mutations and TERT-promoter/RAS mutations was also assessed.Results:DICER1 mutations were detected in 16 of the 68 FTC cases (23.5%), with 11 near E1813 at exon 25, 6 near D1709 at exon 24, and 1 in the splice region of exon 25. Two cases harbored two (distinct) mutations. All patients with DICER1-mutant FTC were female. Compared with patients with DICER1-wild-type FTC, those with DICER1-mutant were much younger, and had a higher proportion of minimally invasive subtype. Nine FTCs with DICER1 mutations were subject to further sequencing on adjacent non-cancerous tissues or lymph node tissues, but no mutations were detected. TERT-promoter or RAS hotspot mutations were not identified in any of the DICER1-mutant cases. However, TERT-promoter mutation was found in 6 DICER1-wild-type cases (8.8%, 6/68), with 3 cases also having RAS hotspot mutations and exhibiting highly aggressive biological behaviors.Conclusion:DICER1 mutations may occur in FTCs and appear mutually exclusive with RAS and TERT-promoter mutations, warranting further study as RAS-like mutations.

Objective:To evaluate the efficacy and safety of anaprazole (40 mg and 60 mg) and compared with rabeprazole (20 mg) in the treatment of reflux esophagitis (RE).Methods:This multicenter, randomized, double-blinded, positive drug parallel controlled study was led by the First Affiliated Hospital of Naval Medical University (Shanghai Changhai Hospital) and a total of 24 clinical trial institutions nationwide including the First Affiliated Hospital of Nanchang University, Yichun People′s Hospital, Meihekou Central Hospital, the First Affiliated Hospital of Gannan Medical University, and Jinhua Central Hospital, participated in this research. A total of 156 patients with RE (Los Angeles grade A to D) were enrolled and randomly divided into 3 groups, anaprazole 40 mg group, anaprazole 60 mg group and rabeprazole 20 mg group, using a random number table in a ratio of 1∶1∶1. Patients in the above 3 groups were treated with the appropriate trial medication once per day for 4 or 8 weeks. The endoscopic healing rates were evaluated by Blinded Independent Central Review (BICR) and investigators. In addition, the improvement in the severity of individual symptoms (daytime reflux, daytime heartburn, nighttime reflux, nighttime heartburn) and medication safety were also evaluated. The endoscopic healing rates and 95% confidence intervals (95% CI) at week-8 and -4 were calculated by groups, as well as the difference in the healing rates and their 95% CI among groups. The chi-square test was used for statistical analysis. Results:A total of 153 subjects were included in the full analysis set (FAS), 144 in the per-protocol analysis set (PPS) and 151 in the safety set (SS). In the FAS, after 8 weeks of treatment, the endoscopic healing rates of anaprazole 40 mg group, anaprazde 60 mg group and raberazole 20 mg group blindly assessed by BICR were 86.0% (43/50), 86.5% (45/52) and 86.3% (44/51), respectively, and the 95% CI were 76.4% to 95.6%, 77.3% to 95.8% and 76.8% to 95.7%, respectively.The endoscopic healing rates of anaprazole 40 mg group, anaprazde 60 mg group and raberazole 20 mg group blindly evaluated by investigators were 88.0% (44/50), 90.4% (47/52) and 86.3% (44/51), respectively, and the 95% CI were 79.0% to 97.0%, 82.4% to 98.4% and 76.8% to 95.7%, respectively. The endoscopic healing rates were similar among groups. In the FAS, the differences in healing rates(95% CI) assessed by BICR and investigators between anaprazole 40 mg, anaprazole 60 mg and rabeprazole 20 mg group were -0.3%(-13.7% to 13.2%), 0.6%(-12.3% to 13.6%), respectively and 1.7%(-11.3% to 14.8%), 3.9%(-8.5% to 16.3%), respectively. The results of the PPS were consistent with those of the FAS. After 8 weeks of treatment, the severity scores of individual symptoms (daytime reflux, daytime heartburn, nighttime reflux, nighttime heartburn) decreased in all groups. The differences between post-treatment and baseline in anaprazole 40 mg group, anaprazole 60 mg group and rabeprazole 20 mg group were -1.54±1.00, -1.91±1.00, -1.51±0.76, -1.45±0.71; -1.30±0.94, -1.59±0.96, -1.33±0.65, -1.42±0.60; and -1.74±0.85, -1.76±0.93, -1.45±0.66, -1.66±0.79, respectively. The incidence of treatment emergent adverse event of anaprazole 40 mg group, anaprazole 60 mg group and rabeprazole 20 mg group were 57.1% (28/49), 48.1% (25/52) and 60.0% (30/50), respectively, and the incidence of treatment related adverse event were 18.4% (9/24), 25.0% (13/52) and 24.0% (12/50), respectively. There were no statistically significant differences in the incidence of treatment emergent adverse event and treatment related adverse event among 3 groups (all P>0.05). Conclusion:The efficacy and safety of anaprazole 40, 60 mg/d, and rabeprazole 20 mg/d in the treatment of RE are comparable.

Objective:To evaluate the efficacy and safety of anaprazole (40 mg and 60 mg) and compared with rabeprazole (20 mg) in the treatment of reflux esophagitis (RE).Methods:This multicenter, randomized, double-blinded, positive drug parallel controlled study was led by the First Affiliated Hospital of Naval Medical University (Shanghai Changhai Hospital) and a total of 24 clinical trial institutions nationwide including the First Affiliated Hospital of Nanchang University, Yichun People′s Hospital, Meihekou Central Hospital, the First Affiliated Hospital of Gannan Medical University, and Jinhua Central Hospital, participated in this research. A total of 156 patients with RE (Los Angeles grade A to D) were enrolled and randomly divided into 3 groups, anaprazole 40 mg group, anaprazole 60 mg group and rabeprazole 20 mg group, using a random number table in a ratio of 1∶1∶1. Patients in the above 3 groups were treated with the appropriate trial medication once per day for 4 or 8 weeks. The endoscopic healing rates were evaluated by Blinded Independent Central Review (BICR) and investigators. In addition, the improvement in the severity of individual symptoms (daytime reflux, daytime heartburn, nighttime reflux, nighttime heartburn) and medication safety were also evaluated. The endoscopic healing rates and 95% confidence intervals (95% CI) at week-8 and -4 were calculated by groups, as well as the difference in the healing rates and their 95% CI among groups. The chi-square test was used for statistical analysis. Results:A total of 153 subjects were included in the full analysis set (FAS), 144 in the per-protocol analysis set (PPS) and 151 in the safety set (SS). In the FAS, after 8 weeks of treatment, the endoscopic healing rates of anaprazole 40 mg group, anaprazde 60 mg group and raberazole 20 mg group blindly assessed by BICR were 86.0% (43/50), 86.5% (45/52) and 86.3% (44/51), respectively, and the 95% CI were 76.4% to 95.6%, 77.3% to 95.8% and 76.8% to 95.7%, respectively.The endoscopic healing rates of anaprazole 40 mg group, anaprazde 60 mg group and raberazole 20 mg group blindly evaluated by investigators were 88.0% (44/50), 90.4% (47/52) and 86.3% (44/51), respectively, and the 95% CI were 79.0% to 97.0%, 82.4% to 98.4% and 76.8% to 95.7%, respectively. The endoscopic healing rates were similar among groups. In the FAS, the differences in healing rates(95% CI) assessed by BICR and investigators between anaprazole 40 mg, anaprazole 60 mg and rabeprazole 20 mg group were -0.3%(-13.7% to 13.2%), 0.6%(-12.3% to 13.6%), respectively and 1.7%(-11.3% to 14.8%), 3.9%(-8.5% to 16.3%), respectively. The results of the PPS were consistent with those of the FAS. After 8 weeks of treatment, the severity scores of individual symptoms (daytime reflux, daytime heartburn, nighttime reflux, nighttime heartburn) decreased in all groups. The differences between post-treatment and baseline in anaprazole 40 mg group, anaprazole 60 mg group and rabeprazole 20 mg group were -1.54±1.00, -1.91±1.00, -1.51±0.76, -1.45±0.71; -1.30±0.94, -1.59±0.96, -1.33±0.65, -1.42±0.60; and -1.74±0.85, -1.76±0.93, -1.45±0.66, -1.66±0.79, respectively. The incidence of treatment emergent adverse event of anaprazole 40 mg group, anaprazole 60 mg group and rabeprazole 20 mg group were 57.1% (28/49), 48.1% (25/52) and 60.0% (30/50), respectively, and the incidence of treatment related adverse event were 18.4% (9/24), 25.0% (13/52) and 24.0% (12/50), respectively. There were no statistically significant differences in the incidence of treatment emergent adverse event and treatment related adverse event among 3 groups (all P>0.05). Conclusion:The efficacy and safety of anaprazole 40, 60 mg/d, and rabeprazole 20 mg/d in the treatment of RE are comparable.

Mitochondria as a signaling platform play crucial roles in deciding cell fate. Many classic anticancer agents are known to trigger cell death through induction of mitochondrial damage. Mitophagy, one selective autophagy, is the key mitochondrial quality control that effectively removes damaged mitochondria. However, the precise roles of mitophagy in tumorigenesis and anticancer agent treatment remain largely unclear. Here, we examined the functional implication of mitophagy in the anticancer properties of magnolol, a natural product isolated from herbal

Objective To evaluate the influence of fluoride on mitochondrial membrane potential of neuroblastoma SH-SY5Y cells,and on the expression levels of mitochondrial proteins mitofusion 1 (Mfn1) and fission 1 (Fis1).Methods A stable and feasible culture method of SH-SY5Y cells in vitro was established with different concentration of sodium fluoride [0.0 (control),0.4,2.0 and 4.0 mmol/L],and various periods exposure of 6,12,24,48 h;the mitochondrial membrane potential of SH-SY5Y cells was detected by mitochondrial membrane potential assay kit (JC-1);and the expression levels of Mfn1 and Fis1 proteins were detected by Western blotting.Results Compared with the control group (1.63 ± 0.18,1.13 ± 0.15,1.30 ± 0.02) for various periods exposure (6,12,48 h),the red/green fluorescence ratios of the mitochondrial membrane potential of SH-SY5Y cells exposed to 2.0 and 4.0 mmol/L of sodium fluoride were decreased significantly (1.01 ± 0.10,0.80 ± 0.04;0.75 ± 0.13,0.62 ± 0.10;0.82 ± 0.01,0.56 ± 0.04,P ＜ 0.05);compared with the control group (0.93 ± 0.03,1.05 ± 0.07,1.17 ± 0.04) for various periods exposure,the expression levels of mitochondrial Mfn1 protein were decreased significantly in 0.4,2.0,4.0 mmol/L sodium fluoride groups (6,12,48 h:0.75 ± 0.02,0.65 ± 0.05,0.57 ± 0.06;0.83 ± 0.06,0.79 ± 0.06,0.69 ±0.06;0.98 ± 0.05,0.73 ± 0.07,0.62 ± 0.09,P ＜ 0.05).Compared with the control group (0.90 ± 0.05) for exposure time 12 h,the expression levels of Fis1 protein were increased significantly in 2.0,4.0 mmol/L sodium fluoride groups (1.14 ± 0.06,1.23 ± 0.06,P ＜ 0.05).Conclusions The mitochondrial membrane potential and the expression levels of mitofusion 1 and fission 1 of SH-SY5Y cells treated with fluoride are abnormal,which might be associated with the theory of nerve cell damage from high oxidative stress.

Objective:To establish an HPLC method for the determination of ginsenoside Rg1 ,ginsenoside Re and ginsenoside Rb1 in Xueluotong capsules. Methods:A column of Waters Symmetry C18 ( 250 mm × 4. 6 mm,5 μm) at the temperature of 35 ℃ was used to separate the target components, the mobile phase consisted of acetonitrile-water with gradient elution, the flow rate was 1. 0 ml ·min-1 , the detection wavelength was 203 nm and the injection volume was 10 μl. Results:The linear range of ginsenoside Rg1 was 0. 055-2. 732 μg(r=0. 9998), and the average recovery was 107. 23% with RSD of 1. 17%(n=6). The linear range of ginsenoside Re was 0. 341-8. 542 μg(r=0. 9999), and the average recovery was 101. 63% with RSD of 3. 52%(n=6). The linear range of gin-senoside Rb1 was 0. 717-14. 336 μg(r=0. 9997), and the average recovery was 100. 63% with the RSD of 3. 79%(n=6). Conclu-sion:The method is simple, accurate and reliable, which can be used for the quality control of Xueluotong capsules.

Background and purpose:Ethanol has been reported to stimulate progression of breast cancer, yet the underlying mechanism is not fully understood. This study aimed to investigate effects of ethyl alcohol (EtOH) on the calcium-activated neutral protease (CANP)-cyclin E/focal adhesion kinase (FAK) signaling and cell migration in breast cancer cells, as well as the role of epidermal growth factor receptor (EGFR) in the EtOH-stimulated effects, in order to assess the signaling mechanism(s) underlying how EtOH enhances cancer progression.Methods:Human breast cancer cell line MCF-7 was employed as a model system, with MCF-10A mammary epithelial cells as control. In vitro wound healing assay was carried out to evaluate EtOH-induced cell migration. The effects of EtOH or epidermal growth factor on the proteolysis of cyclin E/FAK were detected by Western blot. EGFR inhibitor (EGFR-I) and a speciifc inhibitor for CANP, Calpeptin, were applied to pretreat cultured cells to explore their inlfuences on the cell migration and cyclin E/FAK proteolysis triggered by EtOH.Results:Treatment of model cells with EtOH (0.3%) stimulated significant proteolysis of cyclin E/FAK in a dose-/time-dependent manner and increased migration (+47.30%,P<0.05) in MCF-7 breast cancer cells, but had no signiifcant effect on migration in MCF-10A cells. Pretreatment with Calpeptin (10 μmol/L) signiifcantly reduced EtOH (0.3%)- or EGFR (10 ng/mL)-induced cyclin E/FAK truncation. EGFR-I (3 μmol/L) pro-foundly reduced EtOH-indcued CANP dependent proteolysis of CANP1 and cyclin E/FAK as well as cell migration (-53.00%,P<0.01).Conclusion:EtOH signiifcantly stimulates activation of CANP via EGFR pathway, resulting in proteolysis of cyclin E/FAK and migration in MCF-7 breast cancer cells, suggesting EGFR-CANP signaling to be a potential target for suppression of metastasis in breast cancer.

Objective To observe the effects of fluorosis on the levels of endogenous cystathionine beta-synthase (CBS) and hydrogen sulfide (H2S) in rats.Methods According to body weight,forty-eight Sprague-Dawley rats (body weight 105-180 g) were divided into three groups by a random number table(16 rats in each group,half male).Fluorine contents of the feed in control group,low-fluoride group and high-fluoride group were 9.80,15.40 and 23.80 mg/kg.After 6 months of fluorine exposure,the fluorine contents of urine and bone were determined by the method of fluorine ion-selective electrode ; H2S levels in serum and brain and the activity of CBS in brain were detected by methylene blue; and protein expression of CBS was detected by Western blotting.Results Compared with control group,dental fluorosis was found in rats of low-fluoride and high-fluoride groups.The differences of fluorine contents of urine and femur were statistically significant between groups(F =65.16,67.93,all P ＜ 0.05).The urinary and femoral fluorine in low-fluoride groups [(5.25 ± 0.45)mg/L,(1 196.54 ± 72.78)mg/kg] and high-fluoride groups[(13.17 ± 0.98)mg/L,(2 656.61 ± 170.12)mg/kg] were higher than those of control groups [(3.64 ± 0.20)mg/L,(870.71 ± 71.51)mg/kg,all P ＜ 0.05],and the increases were in a dose-dependent fashion(all P ＜ 0.01).The differences of H2S contents in serum and brain were statistically significant(F =4.83,1 456.13,all P ＜ 0.05).The H2S content in serum was higher in high-fluoride group [(17.64 ± 2.38) μ mol/L] than that of the control group [(10.29 ± 0.74) μ mol/L,P ＜ 0.01].The H2S contents in brain were higher in the low-fluoride [(364.74 ± 2.06)μmol/L] and high-fluoride groups [(513.43 ± 4.18) μmol/L] than those of the control group[(314.94 ± 0.72)μmol/L,all P ＜ 0.01],and the increase was in a dose-dependent fashion (P ＜ 0.01).The difference of CBS activity was statistically significant between groups (F =760.63,P ＜ 0.01).The CBS activities were lower in low-fluoride [(438.90 ± 2.83) mmol· kg-1· min-1] and high-fluoride groups [(529.83 ± 2.37)mmol· kg-1· min-1] than those of the control group [(596.33 ± 2.75) mmol · kg-1· min-1,all P ＜ 0.01],whereas the protein expression of CBS in brain in high-fluoride group (1.49 ± 0.08) was higher than that of the control group (1.19 ± 0.06,P ＜ 0.05).Conclusion Chronic fluorosis can affect the levels of endogenous CBS and H2,S,and the increases are in a dose-dependent fashion in addition to CBS activity.