Objective To investigate the role of miRNA-30a-3p on inhibiting the proliferation,invasion and metastasis of HCC cells by targeting Caspase 1 involved in pyroptosis.Methods The qRT-PCR and immunohistochemical staining were used to detect the expressions of miRNA-30a-3p and Caspase 1 in HCC cells.SMMC-7721 cells were transfected with miR-30a-3p agonists,inhibitors and Caspase 1-specific inhibitors.Western blot was obtained to detect the expression of EMT-related proteins (N-cadherin,vimentin,snail,MMP-2) and Caspase 1,IL-18 and IL-1β.Plate clone assay,CCK-8 kit and Transwell were carried out to detect the proliferation and immigration of HCC cells.Results Caspase 1 was highly expressed (t =17.54,P ＜ 0.05) in HCC tissues.Overexpression of miR-30a-3p inhibited HCC cells proliferation and metastasis,while miR-30a-3p inhibition increased the proliferation and metastasis of HCC cells.Overexpression of miR-30a-3p decreased the expression of Caspase 1 (t =12.73,P ＜ 0.05) and inhibited the induction of pyroptosis,inhibiting the expression of IL-18 (t =7.32,P ＜ 0.05) and IL-1 β (t =7.32,P ＜0.05).When miRNA-30a-3p was inhibited,the cell viability of HCC was increased (F1 =9.57,P ＜0.05).When miRNA-30a-3p and Caspase 1 were inhibited together,the cell viability of HCC decreased (F2 =10.66,P ＜ 0.05).Conclusion MiRNA-30a-3p regulate cell pyroptosis through Caspase 1 pathway,inhibiting the proliferation,invasion and metastasis of HCC cells.

Objective To explore the role of miR-137 in the proliferation and migration of hepatocellular carcinoma (HCC) cells by regulating Notch1 and mediating autophagy.Methods The human SMMC7721 hepatoma cell line was transfected with miR-137 mimics,miR-137 inhibitor and Notch1 interfering RNA (siRNA),and divided into normal control group (NC group),miR-137 mimics group,miR-137 inhibitor group,Notch1 siRNA group.The expression levels of miR-137 and Notch1 mRNA after the transfection were detected by RT-PCR in SMMC7721 cells.Transwell experiments were performed to analyze the effect of miR-137 and Notch1 on the migration and invasion of SMMC7721 cells.The expression levels of β-catenin and vimentin in SMMC7721 cells were detected by immunohistochemistry.The number of autophagosomes was detected by double labeled adenovirus.Western blot was utilized to detect the expression of Notch1,E-Cadherin,N-Cadherin,vimentin,P62,and LC3.Results The results of RT-PCR showed that the relative expression level of Notch1 in miR-137 inhibitor group (5.71 ± 0.45) was significantly higher than that in miR-137 mimics group (0.21 ± 0.06) with statistical significance (P ＜ 0.05).The Transwell experiments showed that there were fewer invasive metastatic hepatoma cells in miR-137 mimics group (66.00 ± 4.55) and Notch1 siRNA group (88.00 ± 6.78) than that in the miR-137 inhibitor group (515.00 ±35.12) (P ＜0.05).The expression levels of β-catenin in miR-137 mimics group and Notch1 siRNA group were significantly increased and the expression level of vimentin was decreased (P ＜ 0.05).The results of autophagy double labeled adenovirus test showed that the number of autophagosomes in miR-137 mimics group (5.50 ± 3.70) was significantly fewer than that in miR-137 inhibitor group (32.75 ± 4.11),and the difference was statistically significant (P ＜ 0.05).The expression levels of Notch1,N-cadherin,vimentin,and LC3 protein in miR-137 mimics group were much lower than that in miR-137 inhibitor group and NC group,and the expression levels of E-Cadherin and P62 protein were greatly increased.The expression level of Notch1,N-cadherin,and LC3 protein in Notch1 siRNA group were significantly lower than that in NC group,and the expression levels of E-cadherin and P62 protein were much higher than that in NC group.Conclusion MiR-137 can inhibit the proliferation,migration and invasion of HCC cells by inhibiting the expression of Notch1 and autophagy,which may become a new target for the treatment of HCC.

Objective To investigate the effect of Liensinine on lipopolysaccharide ( LPS)-induced acute lung injury (ALI) in mice. Methods BALB/c mice were randomly divided into six group: control group, LPS group, LPS+Liensinine (2 mg/kg, 4 mg/kg, 8 mg/kg) groups, and dexamethasone group. Acute lung injury in mice was induced by nasal instillation of LPS. After 12 h, the pathological changes of lung tissue were observed. The levels of TNF-α, IL-6 and IL-1β in bronchoalveolar lavage fluid (BALF) were detected by ELISA. The number of neutrophils in BALF was detected using Wright-Giemsa staining. Total protein content was detected by BCA protein quantification assay. The pulmonary capillary permeability was examined with Evans blue. The MPO activity, MDA content, SOD activity, and GSH content in lung homogenate supernatant were detected by spectrophotometry. The content of ROS in lung tissue was detected by flow cytometry. Results The LPS group showed inflammatory cell infiltration, thickening of bronchial alveolar wall and pulmonary congestion in the lung tissue, while Liensinine improved the lung injury. In the LPS group, the contents of TNF- α, IL-6 and IL-1β in BALF were significantly increased, the number of neutrophils and the content of total protein were significantly increased, pulmonary capillary permeability, MPO activity and MDA content were increased, SOD activity and GSH content were decreased, the content of ROS was increased; while the Liensinine group reduced the contents of TNF-α, IL-6, IL-1β in BALF, reduced the number of neutrophils and total protein content, decreased the pulmonary capillary permeability, attenuated MPO activity and MDA contents and increased SOD activity and GSH content, and reduced ROS content in the LPS-challenged lung tissue. Conclusions Liensinine protects mice from LPS-induced acute lung injury by its anti-inflammatory and anti-oxidative activities.

Objective:To explore the effect of miR-223-3p regulating FOXO3a-mediated autophagy in hepatic injury-reperfusion injury (LIRI).Methods:The model of hepatic ischemia-reperfusion injury (IR) was established in C57BL6 mice. According to different reperfusion timepoints, mice were randomly divided into 2 h, 6 h, 12 h and 24 h group. For sham group, there was no intraoperative clamping of hepatic pedicle. Murine hepatic AML12 cells were treated with miR-223-3p mimics, miR-223-3p inhibitor and FOXO3a interfering RNA. A hypoxic 1 h reoxygenation 6 h model was established. And miRNA-NC, miR-223-3p mimics, miR-223-3p inhibitor and siRNA-NC and FOXO3a siRNA groups were assigned. Hepatic injury and apoptosis were detected by hematoxylin eosin or TdT-mediated nick end labeling (HE/TUNEL) at different timepoints. The changes of proliferating cell nuclear antigen (PCNA) and Caspase-3 in hepatocytes were detected by immunohistochemistry. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were employed for detecting the expressions of miR-223-3p, FOXO3a, LC3, p62 and Caspase-3 in hepatocytes.Results:The results of HE/TUNEL indicated that reperfusion injury and apoptosis of hepatic tissue were most severe in 12 h group. In hepatic ischemia-reperfusion model, RT-PCR results showed that the expressions of miR-223-3p and FOXO3a were higher in IR group than those in sham group (1.00±0), the expression level of miR-223-3p mRNA peaked at 12 h (9.13±2.12) after reperfusion and FOXO3a was the highest at 6 h (5.23±0.90, P<0.05). Western blot showed that the expression of FOXO3a peaked at 6 h post-reperfusion and the expressions of LC3 and caspase-3 were the highest at 12 h. ( P<0.05). In the model of cell hypoxia and reoxygenation, RT-PCR indicated that the expression of FOXO3a mRNA decreased in miR-223-3p mimics group (0.45±0.21) as compared with miRNA-NC group (1.00±0). In contrast, miR-223-3p inhibitor group increased (2.73±0.53, P<0.05). Western blot indicated that FOXO3a protein expression was highest in miR-223-3p inhibitor and miR-223-3p mimics groups whereas LC3 and Caspase-3 were the highest in miR-223-3p mimics group ( P<0.05). The expression of FOXO3a was higher in siRNA-NC group than that in FOXO3a siRNA group while the expressions of Caspase-3 and LC3 were the higher in FOXO3a siRNA group. Conclusions:FOXO3a has protective effect on hepatic ischemia-reperfusion injury. It may be related to its inhibition of autophagy and apoptosis and miR-223-3p promotes injury through a down-regulation of FOXO3a-mediated autophagy. It suggests that miR-223-3p and FOXO3a are negatively correlated and may be potential gene therapeutic targets for hepatic injury.

Objective To investigate the effect of miRNA-30a-3p on the proliferation,invasion and metastasis of liver cancer cells by targeting Atg3-mediated autophagy pathway.Methods The immunohistochemical staining was used to detect content of miRNA-30a-3p and Atg3 and their correlation in human hepatocellular carcinoma.Liver cancer cells were cultured in vitro and hunger environment was used to induce autophagy.RFP-GFP-LC3 double-labeled adenovirus infected hepatoma cells were used to detect autophagosomes in hepatoma cells.The expressions of autophagy-related proteins (autophagocytosis associated protein (Atg3),polyubiquitin-binding protein p62,autophagy microtubule-associated protein light chain 3 (LC3)) and EMT-related proteins (N-cadherin,vimentin,snail,ZO-1) were detected by Western blot.Platelet cloning assay and transwell assay were carried out to detect the proliferation,invasion and metastasis of carcinoma cell.CCK-8 kit was used to detect hepatocarcinoma cells' viability.Results The expression of miRNA-30a-3p was down-regulated.The expression of Atg3,E-cadherin and N-cadherin in miRNA-30a-3p high-expressed hepatocellular carcinoma was lower than that in miRNA-30a-3p low-expressed hepatocellular carcinoma.Increasing the expression of miRNA-30a-3p in hepatocellular carcinoma cells can decrease the expression of Atg3 and LC3,increase the expression of p62 and inhibit the formation of autophagosomes;otherwise,Atg3 and LC3 were increased,p62 was decreased and the formation of autophagosomes was promoted.Inhibition of Atg3 expression could decrease the expression of EMT-related proteins.When miRNA-30a-3p was inhibited,the cell viability of HCC was increased at each time point (F1 =10.314,P ＜0.05).When miRNA-30a-3p and Atg3 were inhibitor together,the cell viability of HCC was decreased at each time point(F2 =6.599,P ＜ 0.05).Conclusion miRNA-30a-3p can inhibit Atg3-mediated autophagy pathway and reduce cell autophagy activity,thus inhibiting the proliferation,invasion and metastasis of hepatocarcinoma cells.

BackgroundThe schizophrenia is majorly treated with drug and through physical therapy. However， both treatments would lead to adverse reactions， which could affect therapy adherence and treatment efficacy. Previous studies have shown that aerobic exercise can help alleviate cognitive function in patients with Alzheimer's disease and depressive disorder. At present， little research has been done on such alleviation in schizophrenia patients. ObjectiveTo explore the effect of aerobic exercise on cognitive function in male patients with chronic schizophrenia， so as to provide references for relevant treatments. MethodsA total of 76 male patients with chronic schizophrenia hospitalized in the Fourth People's Hospital of Wuhu between December 2022 and April 2023 were selected as the study subjects and， in accordance with random number table， divided into study group （n=36） and control group （n=40）. Both groups received conventional drug treatment. On this basis， the study group received a 60-minute aerobic exercise 5 times a week for 8 weeks as intervention. Before and after intervention， assessment of cognitive function was performed by using MATRICS Consensus Cognitive Battery （MCCB） and Stroop Color Word Test （SCWT）. ResultAfter intervention， compared with the control group， the study group spent less time on finishing the Trail Making Test and scored higher in both the spatial span test and maze test （Z=-2.070， -2.306， -2.375， P<0.05）. Repeated measure ANOVA results showed that the time main effect of Hopkins Verbal Learning Test score was statistically significant in the two groups after intervention （F=39.067， P<0.05）. So was the interaction effect between the time and group of the Brief Visuospatial Memory Test and Verbal Fluency Test scores （F=10.092， 9.252， P<0.05）. After intervention， the scores of the Brief Visuospatial Memory Test and Verbal Fluency Test in the study group were higher than those in the control group （t=6.689， 4.249， P<0.05）. As for the study group itself， the scores were higher than those before intervention （t=23.746， 23.842， P<0.05）. After intervention， the numbers of correct reading in color test and word test in the study group were more than those in the control group （Z=-2.358， -2.771， P<0.05）. The interaction effect between the time and group of the reaction time in color test， word test and color word interference test were statistically significant in both groups （F=23.383， 19.888， 19.662， P<0.05）. After intervention， the reaction time in color test and color word interference test of the study group was shorter than those of the control group （t=4.895， 6.163， P<0.05）. As for the study group itself， the reaction time were shorter than before intervention （t=54.318， 42.425， 42.141， P<0.01）. ConclusionAerobic exercise may help alleviate the cognitive problems in male patients with chronic schizophrenia in terms of information processing speed， working memory， reasoning/problem solving ability， word learning and memorizing， visual learning and memorizing， and executive function. ［Funded by Wuhu Science and Technology Plan Project （number， 2021jc2-3）］

Objective:To investigate the intervention effect and its mechanism of apocynin, an inhibitor of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX) on silicosis induced by silica (SiO 2) in rats. Methods:In October 2021, 24 SPF SD male rats were divided into control group, silicosis model group and apocynin intervention group according to random number table method, with 8 rats in each group. SiO 2 was exposed by one-time intratracheal instillation. The rats in the apocynin intervention group were intraperitoneally injected with apocynin 50 mg/kg, 3 times a week, on the second day after treatment. The rats were sacrificed 28 days later, and lung coefficients were calculated after lung tissues were weighed. Hematoxylin-eosin staining and Masson staining were used to observe the lung histopathological changes in each group, respectively. The levels of NOX, reactive oxygen species (ROS), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) in lung tissue were detected. The expressions of interleukin-1 beta (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were determined by Enzyme-Linked Immunosorbent Assay (ELISA). The level of hydroxyproline (HYP) was detected by alkaline hydrolysate. The expressions of transforming growth factor beta 1 (TGF-β1), E-cadherin (E-cad) and α-smooth muscle actin (α-SMA) in lung tissue were detected by Western blotting. Results:Compared with the control group, the body weight of silicosis model group was decreased, the lung tissue showed obvious inflammatory infiltration and fibrosis, and the levels of lung coefficient, IL-1β, IL-6, TNF-α and TGF-β1 were significantly increased ( P<0.05). Compared with the silicosis model group, the lung tissue injury in the apocynin intervention group was significantly improved, the lung coefficient, NOX, ROS, MDA, IL-1β, IL-6, TNF-α and TGF-β1 levels were decreased, and the activity of GSH-Px was increased ( P<0.05). Compared with the silicosis model group, the expressions of HYP and α-SMA were decreased and the level of E-cad was increased in the apocynin intervention group ( P<0.05) . Conclusion:Apocynin may alleviate SiO 2-induced fibrosis in silicosis rats by reducing oxidative stress, the release of inflammatory factors and inhibiting the process of epithelial-mesenchymal transition.

Objective:To investigate the intervention effect and its mechanism of apocynin, an inhibitor of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX) on silicosis induced by silica (SiO 2) in rats. Methods:In October 2021, 24 SPF SD male rats were divided into control group, silicosis model group and apocynin intervention group according to random number table method, with 8 rats in each group. SiO 2 was exposed by one-time intratracheal instillation. The rats in the apocynin intervention group were intraperitoneally injected with apocynin 50 mg/kg, 3 times a week, on the second day after treatment. The rats were sacrificed 28 days later, and lung coefficients were calculated after lung tissues were weighed. Hematoxylin-eosin staining and Masson staining were used to observe the lung histopathological changes in each group, respectively. The levels of NOX, reactive oxygen species (ROS), glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) in lung tissue were detected. The expressions of interleukin-1 beta (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) were determined by Enzyme-Linked Immunosorbent Assay (ELISA). The level of hydroxyproline (HYP) was detected by alkaline hydrolysate. The expressions of transforming growth factor beta 1 (TGF-β1), E-cadherin (E-cad) and α-smooth muscle actin (α-SMA) in lung tissue were detected by Western blotting. Results:Compared with the control group, the body weight of silicosis model group was decreased, the lung tissue showed obvious inflammatory infiltration and fibrosis, and the levels of lung coefficient, IL-1β, IL-6, TNF-α and TGF-β1 were significantly increased ( P<0.05). Compared with the silicosis model group, the lung tissue injury in the apocynin intervention group was significantly improved, the lung coefficient, NOX, ROS, MDA, IL-1β, IL-6, TNF-α and TGF-β1 levels were decreased, and the activity of GSH-Px was increased ( P<0.05). Compared with the silicosis model group, the expressions of HYP and α-SMA were decreased and the level of E-cad was increased in the apocynin intervention group ( P<0.05) . Conclusion:Apocynin may alleviate SiO 2-induced fibrosis in silicosis rats by reducing oxidative stress, the release of inflammatory factors and inhibiting the process of epithelial-mesenchymal transition.

ObjectiveTo explore the possible mechanism of Tongfeng Decoction （痛风汤） for preventing and treating acute gouty arthritis. MethodsSixty Wistar male rats were divided into normal group， model group， colchicine group and low-， medium- and high-dose Tongfeng Decoction groups according to the random number table， 10 rats in each group. Tongfeng Decoction of 11.34， 22.68 and 45.36 g/kg were given by gavage to low-， medium- and high-dose Tongfeng Decoction groups， colchicine 3.15×10^-4 g/（kg·d） to the colchicine group， and normal saline 10 ml/（kg·d） to the normal group and model group respectively for 7 consecutive days. After 1 hour of gavage on day 5， rats in all groups except the normal group were modelled as acute gouty arthritis in the ankle joint of the right hind limb with the modified Coderre's method； rats in the normal group were injected with 0.2 ml of normal saline at the same location. The swelling degree of the ankle joint was measured before modelling and after 6 h， 12 h， 24 h and 48 h of modelling， respectively. HE staining was used to observe the histopathological and morphological changes in the synovial tissue of the ankle joint； immunoblotting and RT-qPCR were used to detect NOD-like receptor protein 3 （NLRP3）， apoptosis-associated speck-like protein （ASC）， cysteine aspartate protease 1 （Caspase-1）， and interleukin 1β （IL-1β） protein and mRNA expression levels in ankle synovial tissues， respectively； immunohistochemistry was used to detect the positive expression of Caspase-1 and IL-1β in ankle synovial tissues； ELISA was used to detect the tumour necrosis factor alpha （TNF-α）， IL-1β， and interleukin in serum； immunofluorescence staining was used to observe the formation of neutrophil extracellular traps （NETs） in the synovial tissues of the ankle joints. ResultsCompared with the normal group， rats in the model group showed elevated joint swelling at all time points， significantly increased inflammatory cell infiltration and the number of synovial cell layers in the synovial tissues of the ankle joints， elevated NLRP3， Caspase-1， ASC and IL-1β protein and mRNA expression， elevated positive expression of Caspase-1 and IL-1β， increased formation of NETs， and elevation of TNF-α， IL-6， and IL-1β in serum （P＜0.05）. Compared with the model group， there was an improvement in synovial cell proliferation and inflammatory cell infiltration of the ankle joint in the colchicine group， high- and medium-dose Tongfeng Decoction groups. In the colchicine group and the high-， medium- and low-dose Tongfeng Decoction groups， the degree of joint swelling， positive expression of Caspase-1， IL-1β and formation of NETs in the synovial tissue of the ankle joint， and TNF-α， IL-6 and IL-1β in serum reduced 24 h and 48 h after modelling； in colchicine group and high-dose Tongfeng Decoction group， NLRP3， ASC， Caspase-1， IL-1β protein and mRNA expression in the synovial tissues of ankle joints all reduced （P＜0.05）. The colchicine group and high-dose Tongfeng Decoction group were superior to the low-dose Tongfeng Decoction group in reducing ASC， Caspase-1， IL-1β protein and mRNA expression in the ankle synovial tissues， the positive expression of Caspase-1， as well as TNF-α， IL-6， and IL-1β level in serum （P＜0.05）. ConclusionTongfeng Decoction showed effectiveness for the prevention and treatment of acute gouty arthritis， and its mechanism may be related to the inhibition of the assembly and activation of NLRP3 inflammatory vesicles in ankle synovial tissues， the inhibition of the release of inflammatory factors， and the formation of NETs.

Objective:To determine the total and age-specific cut-off values of total prostate specific antigen (tPSA) and the ratio of free PSA divided total PSA (fPSA/tPSA) for screening prostate cancer in China.Methods:Based on the Chinese Colorectal, Breast, Lung, Liver, and Stomach cancer Screening Trial (C-BLAST) and the Tianjin Common Cancer Case Cohort (TJ4C), males who were not diagnosed with any cancers at baseline since 2017 and received both tPSA and fPSA testes were selected. Based on Cox regression, the overall and age-specific (＜60, 60-＜70, and ≥70 years) accuracy and optimal cut-off values of tPSA and fPSA/tPSA ratio for screening prostate cancer were evaluated with time-dependent receiver operating characteristic curve (tdROC) and area under curve (AUC). Bootstrap resampling was used to internally validate the stability of the optimal cut-off value, and the PLCO study was used to externally validate the accuracy under different cut-off values.Results:A total of 5 180 participants were included in the study, and after a median follow-up of 1.48 years, a total of 332 prostate cancer patients were included. In the total population, the tdAUC of tPSA and fPSA/tPSA screening for prostate cancer were 0.852 and 0.748, respectively, with the optimal cut-off values of 5.08 ng/ml and 0.173, respectively. After age stratification, the age specific cut-off values of tPSA in the <60, 60-<70, and ≥70 age groups were 3.13, 4.82, and 11.54 ng/ml, respectively, while the age-specific cut-off values of fPSA/tPSA were 0.153, 0.135, and 0.130, respectively. Under the age-specific cut-off values, the sensitivities of tPSA screening for prostate cancer in males <60, 60-70, and ≥70 years old were 92.3%, 82.0%, and 77.6%, respectively, while the specificities were 84.7%, 81.3%, and 75.4%, respectively. The age-specific sensitivities of fPSA/tPSA for screening prostate cancer were 74.4%, 53.3%, and 55.9%, respectively, while the specificities were 83.8%, 83.7%, and 83.7%, respectively. Both bootstrap's internal validation and PLCO external validation provided similar results. The combination of tPSA and fPSA/tPSA could further improve the accuracy of screening.Conclusion:To improve the screening effects, it is recommended that age-specific cut-off values of tPSA and fPSA/tPSA should be used to screen for prostate cancer in the general risk population.