Objective To explore the mechanism of 3-methyladenine(3-MA)for alleviating early diabetic renal injury.Methods Mouse models of streptozotocin(STZ)-induced diabetes mellitus were randomized into model group and 3-MA treatment group for daily treatments with normal saline and 10 mg/kg 3-MA by gavage for 6 weeks,respectively.Body weight and fasting blood glucose of the mice were recorded every week.After the treatments,the kidneys of the mice were collected for measurement kidney/body weight ratio,examination of glomerular size with PAS staining,and detection of α-SMA and PCNA expressions using Western blotting and immunohistochemistry.SV40 MES 13 cells cultured in normal glucose(5.6 mmol/L)and high glucose(30 mmol/L)were treated with 24.4 mmol/L mannitol and 5 mmol/L 3-MA for 24 h,respectively,and the changes in cell viability and PCNA expression were examined using CCK8 assay and Western blotting.Bioinformatics analysis of the intersecting gene targets of diabetic kidney disease(DKD)and 3-MA was performed,and the results were verified by Western blotting both in vivo and in vitro.Results In the diabetic mice,treatment with 3-MA produced a short-term hypoglycemic effect,reduced the kidney/body weight ratio and glomerular hypertrophy,and decreased the expressions of α-SMA and PCNA in the renal cortex.In the in vitro study,3-MA significantly lowered the viability and reduced PCNA expression in SV40 MES 13 cells exposed to high glucose.The results of bioinformatic analysis identified AKT1 as the key gene in the therapeutic mechanism of 3-MA for DKD.Western blotting confirmed that 3-MA inhibited the phosphorylation of AKT and S6 in both the renal cortex of diabetic mice and high glucose-treated SV40 MES 13 cells.Conclusion 3-MA suppresses mesangial cell proliferation and alleviates early diabetic renal injury in mice possibly by inhibiting AKT signaling.

Objective To investigate the mechanism mediating the regulatory effect of lncRNA MAGI2-AS3 on cisplatin(DDP)resistance in non-small cell lung cancer(NSCLC).Methods MAGI2-AS3 and miR-1269a expression levels were detected by qRT-PCR in DDP-sensitive lung cancer cell lines(A549 and H1299)and their resistant counterparts(A549/DDP and H1299/DDP).In A549 and H1299 cells with MAGI2-AS3 silencing and A549/DDP and H1299/DDP cells overexpressing MAGI2-AS3,the effects of 20 μmol/L DDP on cell viability and apoptosis were examined with CCK-8 assay,colony formation assay,flow cytometry and Western blotting,and the changes in epithelial-mesenchymal transition(EMT)were assessed with wound healing and Transwell assays.The interaction between MAGI2-AS3,miR-1269a and PTEN was predicted using GEPIA,StarBase and miRDB and verified with luciferase reporter gene assay and radioimmunoprecipitation(RIP)assay.A miR-1269a mimic and pcDNA3.1-PTEN plasmid were used to perform the rescue assay.Results MAGI2-AS3 expression was significantly downregulated in lung cancer tissues(P<0.05)in association with a poor prognosis(P<0.05).In the two DDP-resistant lung cancer cell lines,MAGI2-AS3 expression was significantly lowered as compared with the sensitive cells.Silencing MAGI2-AS3 significantly enhanced cell viability and promoted EMT of A549 and H1299 cells irrespective of DDP treatment,and also decreased DDP-induced apoptosis of the cells.In A549/DDP and H1299/DDP cells,MAGI2-AS3 overexpression strongly repressed cell viability and EMT irrespective of DDP treatment and promoted DDP-induced cell apoptosis.Luciferase reporter gene and RIP assays confirmed the binding of MAGI2-AS3 with miR-1269a and the binding of miR-1269a with 3'-UTR domain of PTEN.The rescue assay demonstrated that MAGI2-AS3 acted as a sponge for miR-1269a to promote PTEN expression and downregulate AKT phosphorylation,thus inhibiting EMT and promoting DDP-induced apoptosis of A549/DDP cells.Conclusion MAGI2-AS3 enhances DDP sensitivity of NSCLC by targeted regulation of the miR-1269a/PTEN/AKT signaling axis.

Bladder cancer ranks 12th in the statistical spectrum of death from malignant tumor pa-tients,its incidence rate is high,the patients population is showing a trend of youthfulness,moreover it is easy to develop metastasis and recurrence.In recent years,the immunotherapy has been gradually promoted in the patients with advanced bladder cancer who cannot receive the cisplatin chemotherapy or who are resistant to chemotherapy,in particular,the immune checkpoint inhibitors(ICIs)represented by programmed cell cleath-1(PD-1)/programmed cell cleath-ligand 1(PD-L1)are dominant,although ICIs represented by PD-1/PD-L1 has achieved good efficacy in immunotherapy.However,due to the increase in immune escape events,only a-bout 30％of patients benefit from immunotherapy.Therefore,there is an urgent need to develop the treatment regimen for the patients with bladder cancer immune escape.Recently,the tumor microenvironment(TME)has become a research hotspot,especially the immunosuppressive cells in TME.There are 5 types of cells with immunosuppressive function in TME:tumor-associated macrophages(TAMs),regulatory T cells(Tregs),bone marrow-derived suppressor cells(MDSCs),tumor-associated central granulocytes(MDSCs)and tumor-associated fibroblasts(CAFs),which play an important role in tumor immune escape.This paper elaborated the composition and function of TAMs in TME and prospects the tumor promoting mechanism of bladder cancer TAMs and targeted treatment of bladder cancer TAMs.

Objective To explore the mechanism of 3-methyladenine(3-MA)for alleviating early diabetic renal injury.Methods Mouse models of streptozotocin(STZ)-induced diabetes mellitus were randomized into model group and 3-MA treatment group for daily treatments with normal saline and 10 mg/kg 3-MA by gavage for 6 weeks,respectively.Body weight and fasting blood glucose of the mice were recorded every week.After the treatments,the kidneys of the mice were collected for measurement kidney/body weight ratio,examination of glomerular size with PAS staining,and detection of α-SMA and PCNA expressions using Western blotting and immunohistochemistry.SV40 MES 13 cells cultured in normal glucose(5.6 mmol/L)and high glucose(30 mmol/L)were treated with 24.4 mmol/L mannitol and 5 mmol/L 3-MA for 24 h,respectively,and the changes in cell viability and PCNA expression were examined using CCK8 assay and Western blotting.Bioinformatics analysis of the intersecting gene targets of diabetic kidney disease(DKD)and 3-MA was performed,and the results were verified by Western blotting both in vivo and in vitro.Results In the diabetic mice,treatment with 3-MA produced a short-term hypoglycemic effect,reduced the kidney/body weight ratio and glomerular hypertrophy,and decreased the expressions of α-SMA and PCNA in the renal cortex.In the in vitro study,3-MA significantly lowered the viability and reduced PCNA expression in SV40 MES 13 cells exposed to high glucose.The results of bioinformatic analysis identified AKT1 as the key gene in the therapeutic mechanism of 3-MA for DKD.Western blotting confirmed that 3-MA inhibited the phosphorylation of AKT and S6 in both the renal cortex of diabetic mice and high glucose-treated SV40 MES 13 cells.Conclusion 3-MA suppresses mesangial cell proliferation and alleviates early diabetic renal injury in mice possibly by inhibiting AKT signaling.

Objective To investigate the mechanism mediating the regulatory effect of lncRNA MAGI2-AS3 on cisplatin(DDP)resistance in non-small cell lung cancer(NSCLC).Methods MAGI2-AS3 and miR-1269a expression levels were detected by qRT-PCR in DDP-sensitive lung cancer cell lines(A549 and H1299)and their resistant counterparts(A549/DDP and H1299/DDP).In A549 and H1299 cells with MAGI2-AS3 silencing and A549/DDP and H1299/DDP cells overexpressing MAGI2-AS3,the effects of 20 μmol/L DDP on cell viability and apoptosis were examined with CCK-8 assay,colony formation assay,flow cytometry and Western blotting,and the changes in epithelial-mesenchymal transition(EMT)were assessed with wound healing and Transwell assays.The interaction between MAGI2-AS3,miR-1269a and PTEN was predicted using GEPIA,StarBase and miRDB and verified with luciferase reporter gene assay and radioimmunoprecipitation(RIP)assay.A miR-1269a mimic and pcDNA3.1-PTEN plasmid were used to perform the rescue assay.Results MAGI2-AS3 expression was significantly downregulated in lung cancer tissues(P<0.05)in association with a poor prognosis(P<0.05).In the two DDP-resistant lung cancer cell lines,MAGI2-AS3 expression was significantly lowered as compared with the sensitive cells.Silencing MAGI2-AS3 significantly enhanced cell viability and promoted EMT of A549 and H1299 cells irrespective of DDP treatment,and also decreased DDP-induced apoptosis of the cells.In A549/DDP and H1299/DDP cells,MAGI2-AS3 overexpression strongly repressed cell viability and EMT irrespective of DDP treatment and promoted DDP-induced cell apoptosis.Luciferase reporter gene and RIP assays confirmed the binding of MAGI2-AS3 with miR-1269a and the binding of miR-1269a with 3'-UTR domain of PTEN.The rescue assay demonstrated that MAGI2-AS3 acted as a sponge for miR-1269a to promote PTEN expression and downregulate AKT phosphorylation,thus inhibiting EMT and promoting DDP-induced apoptosis of A549/DDP cells.Conclusion MAGI2-AS3 enhances DDP sensitivity of NSCLC by targeted regulation of the miR-1269a/PTEN/AKT signaling axis.

Objective:To construct a deep learning-based artificial intelligence endoscopic ultrasound (EUS) bile duct scanning substation system to assist endoscopists in learning multi-station imaging and improve their operation skills.Methods:A total of 522 EUS videos in Renmin Hospital of Wuhan University and Wuhan Union Hospital from May 2016 to October 2020 were collected, and images were captured from these videos, including 3 000 white light images and 31 003 EUS images from Renmin Hospital of Wuhan University, and 799 EUS images from Wuhan Union Hospital. The pictures were divided into training set and test set in the EUS bile duct scanning system. The system included filtering model of white light gastroscopy images (model 1), distinguishing model of standard station images and non-standard station images (model 2) and substation model of EUS bile duct scanning standard images (model 3), which were used to classify the standard images into liver window, stomach window, duodenal bulb window, and duodenal descending window. Then 110 pictures were randomly selected from the test set for a man-machine competition to compare the accuracy of multi-station imaging by experts, advanced endoscopists and the artificial intelligence model.Results:The accuracies of model 1 and model 2 were 100.00% (1 200/1 200) and 93.36% (2 938/3 147) respectively. Those of model 3 on the internal validation dataset in each classification were 97.23% (1 687/1 735) in liver window, 96.89% (1 681/1 735) in stomach window, 98.73% (1 713/1 735) in duodenal bulb window, and 97.18% (1 686/1 735) in duodenal descending window. And those on the external validation dataset were 89.61% (716/799) in liver window, 92.74% (741/799) in stomach window, 90.11% (720/799) in duodenal bulb window, and 92.24% (737/799) in duodenal descending window. In the man-machine competition, the accuracy of the substation model was 89.09% (98/110), which was higher than that of senior endoscopists [85.45% (94/110), 74.55% (82/110), and 85.45% (94/110)] and close to the level of experts [92.73% (102/110) and 90.00% (99/110)].Conclusion:The deep learning-based EUS bile duct scanning system constructed in the current study can assist endoscopists to perform standard multi-station scanning in real time more accurately and improve the completeness and quality of EUS.

Objective:To investigate the clinical efficacy of different doses of atorvastatin in the treatment of chronic heart failure.Methods:A total of 100 patients with chronic heart failure who received treatment in the Third People's Hospital of Bengbu Medical College from June 2019 to June 2020 were included in this study. They were randomly divided into a control group (conventional treatment + atorvastatin calcium tablet 10 mg, n = 49) and an observation group (conventional treatment + atorvastatin calcium tablet 20 mg, n = 51). Before and after 12 weeks of treatment, serum levels of N-terminal pro-brain natriuretic peptide (NT-proBNP) and inflammatory cytokines such as high-sensitivity C-reactive protein (hs-CRP), interleukin-6 (IL-6), interleukin-10 (IL-10) as well as ventricular remodeling (echocardiographic index) were compared between the two groups. Results:After treatment, serum level of NT-proBNP was (506.56 ± 62.37) pg/mL and (660.85 ± 74.55) pg/mL, serum level of hs-CRP was (14.74 ± 2.69) mg/L and (23.31 ± 3.45) mg/L, serum level of IL-6 was (36.77 ± 4.78) ng/L and (43.12 ± 5.22) ng/L, serum level of IL-10 was (12.36 ± 2.63) ng /L and (15.39 ± 3.56) ng/L, left ventricular end diastolic diameter was (37.74 ± 6.83) mm and (42.36 ± 7.73) mm, left ventricular ejection fraction was (45.78 ± 3.31)% and (42.63 ± 2.56)%, and the ratio of early (E wave) to late (A wave) diastolic velocities (E/A) was (1.44 ± 0.06) and (1.21 ± 0.03), respectively in the observation and control groups. There were significant differences in serum levels of NT-proBNP, hs-CRP, IL-6 and IL-10 as well as LVEDD and LVEF between the observation and control groups ( t = 3.73, 3.07, 3.58, 3.68, 2.99, 3.12 and 2.98, all P < 0.05). Conclusion:Atorvastatin is beneficial to the prognosis of patients with chronic heart failure. The therapeutic efficacy of 20 mg atorvastatin per day is better than that of 10 mg atorvastatin per day.

Objective:To explore the clinical manifestations of 4 pedigrees with transthyretin related familial amyloid polyneuropathy (TTR-FAP).Methods:The clinical data were collected and analyzed from 4 pedigrees with TTR-AFP, admitted to our hospital from July 2017 to May 2019; 20 patients and 2 asymptomatic carriers of the TTR mutation gene were included. In particular, the detailed data of the 4 probands affected with TTR-FAP came from the 4 different pedigrees were collected. Results:In these 20 patients, the age of onset ranged from 30 to 65; the first symptoms of diarrhea, constipation, alternating episodes of constipation and diarrhea were found; there were damaged peripheral nerve and inexplicable weight loss; cardiomyopathy was noted in 9 patients; orthostatic hypotension was noted in 9 patients, sexual dysfunction in 5, abnormal urination in 6, and blurred vision or corestenoma in 3. TTR mutation gene was confirmed in 7 patients and pathological diagnosis was found in 3 patients. Diflunisal was used in one patient and tafamidis was used 2 patients. Twelve died and 8 patients survived among 20 patients with disease progression. All the 4 probands were male, with an average age of 49.3 years; all patients had different degrees of sensorimotor peripheral neuropathy, autonomic neuropathy and cardiomyopathy; electrophysiological examination suggested length dependent sensory motor peripheral neuropathy of the extremities, with axonal damage as the evidence; and cardiac hypertrophy was noted in echocardiography. The sural nerve biopsy of the 3 probands showed positive Congo red staining. Medical whole exon sequencing indicated that 2 probands had pathogenic mutations (TTR-E74K and TTR-A140S), and 1 proband had likely pathogenic mutation (TTR-S70R). Two asymptomatic carriers of the TTR gene mutation remained normal condition. Conclusion:The clinical manifestations of TTR-FAP include progressive sensorimotor and autonomic neuropathy, and multi-system disorders, such as combining with gastrointestinal problems, hypertrophic myocardium, inexplicable weight loss and blurred vision or corestenoma, which might be important reminders for diagnosis of TTR-FAP.

Objective: To investigate the clinical features, diagnosis and treatment of febrile infection-related epilepsy syndrome (FIRES). Methods: The clinical data of 12 children with FIRES admitted to Xiangya Hospital of Central South University from 2015 to 2018 were retrospectively analyzed. The basic information, clinical manifestations, electroencephalogram, imaging examination, treatment and prognosis were analyzed. Results: Of the 12 patients, 7 were male and 5 were female. The age of onset was (7.0±3.7)years (1.3 year to 13 years). The average hospitalization time (34-86 days, median 52 days). Twelve patients were healthy before the disease, and had fever before convulsion. The interval between fever and seizure was (3.5±1.7)days (1-7 days). The status epilepticus and consciousness deficit were the main clinical manifestations. The electrogram of 8 patients showed status epilepticus when admitted. 12 patients had disturbance of consciousness; the acute episodes were focal seizures (100%, 12/12) and generalized tonic-clonic seizures (41.7%, 5/12). All patients used 3-5 antiepileptic drugs (median 4), all treated with hormones and gamma globulin. 4 patients with ketogenic diet (KD) were treated within 2 weeks of onset, and the average duration from onset to electroencephalogram (EEG) improvement was (19.2±5.0)days. In 8 patients who did not use KD within 2 weeks of onset, the average duration from onset to EEG improvement was (29.9±9.6)days. Conclusions FIRES is more common in normal children with school age. The main manifestation is refractory status epilepticus in the days after acute fever, focal episodes of seizures, anti-epileptic drug resistance. Early initiation of KD produces a favorable prognosis.