OBJECTIVE To design and synthesize novel α-naphthylthiol amino acid ester lysine specific demethylase 1(LSD1) inhibitors, evaluate their inhibitory activity with selectivity against LSD1, and explore their binding mechanism through molecular docking and dynamics simulation. METHODS Based on the binding mode of hit compound 3a with LSD1, the α- naphthyl mercapto amino acid ethyl ester small molecule compound were designed by fixing the planar hydrophobic naphthyl ring in the structure, while introducing hydrophilic amino fragment, and they were prepared through a multi-component one-pot cascade reaction. All the compounds were evaluated for their inhibitory activity against LSD1 at concentrations of 5.0 and 1.0 μmol·L–1 using the LSD1 screening platform of research group. The most potent compound was tested for its IC50 value and enzyme selectivity over MAO-A and MAO-B, and its binding mode was investigated through molecular docking and dynamics simulation. RESULTS A total of 13 compounds were obtained, all of which exhibited significant inhibitory effects on LSD1. Among them, nine compounds showed an inhibitory rate of over 50.0% against LSD1 at a concentration of 1.0 μmol·L–1, while compound 3l displaying the best activity with an IC50 value of 0.17 μmol·L–1, 174 times higher than the positive control. It also showed excellent selectivity towards MAO-A and MAO-B. Molecular docking and dynamics simulations indicated that compound 3l inhibited the activity of LSD1 through multiple interactions. CONCLUSION The structures of α-naphthylthiol amino acid ester can serve as lead compounds or active fragments, laying a solid foundation for the subsequent design of LSD1 inhibitors based on structure-oriented drug design.

AIM:The paper is to explore a rapid and simple method for the culture of mouse primary thyroid cells.METHODS:Mouse thyroid cells were isolated by enzyme digestion and cultured with improved medium,and their morphology,characteristics and secretory function were observed within 14 d.RESULTS:In the cultures,the active pri-mary cells were obtained from the thyroid tissue after digestion for 25 min;adherent growth was observed on the 2nd day.And secondary follicles appeared from the 5th to 7th day.Over 95%cells were detected with thyroglobulin.The secretion of total triiodothyronine and total thyroxine maintains over 60%in 7 d.The expression levels of specific genes can still maintain more than 50%in 10 d.CONCLUSION:Mouse thyroid primary cells can be rapidly cultured by this method,and the cells can be used for studying thyroid endocrine secretion within 7 d and studying thyroid genes within 10 d.

Objective:To investigate the impact of Tumor-Infiltrating Lymphocytes (TILs) density in the tumor stroma on the long-term prognosis of hepatocellular carcinoma (HCC) liver transplant patients meeting the Hangzhou criteria.Methods:This study is a retrospective cohort study. The clinical data of 83 patients with hepatocellular carcinoma who met the Hangzhou criteria and underwent allogeneic liver transplantation from January 2018 to December 2023 in the Department of Hepatobiliary Surgery, Beijing Chaoyang Hospital affiliated to Capital Medical University were collected and analyzed. Hematoxylin and Eosin (HE) staining was used to study the density of TILs in the resected liver grafts. Patients were divided into TILs-negative group (TILs<10%, n=31) and TILs-positive group (TILs≥10%, n=52) based on whether the TILs density exceeded 10%. Clinical and pathological characteristics were analyzed, and the significance of alpha-fetoprotein (AFP), TILs density, and microvascular invasion on the prognosis of HCC patients who met the Hangzhou criteria was studied. Measurement data with normal distribution were expressed as mean±standard deviation ( ± s) and compared between groups using t-test. Measurement data with skewed distribution were expressed as M ( Q1, Q3) and compared using rank-sum tests. Categorical data were compared using chi-square test. Kaplan-Meier method was used to study the relationship between various observation indicators and overall survival, and survival curves were plotted. Log-rank test was used to compare the survival rates between groups, and multivariate Cox regression model was used to adjust for the distribution of risk factors between groups. Results:The preoperative AFP level in the TILs-negative group was (15.69±1.21) U/mL, and in the TILs-positive group was (12.17±0.13) U/mL, with a statistically significant difference between the two groups ( P<0.05). In the TILs-negative group, 8 cases had microvascular invasion, and the number of low, moderate, and high differentiation tumors was 8, 23, and 0, respectively. In the TILs-positive group, 3 cases had microvascular invasion, and the number of low, moderate, and high differentiation tumors was 2, 31, and 19, respectively. The results indicated that patients in the TILs-negative group were more likely to have microvascular invasion and poorer tumor differentiation ( P<0.05). All patients were regularly followed up, and the 1-, 2-, and 3-year survival rates in the TILs-negative group and TILs-positive group were 84.0%, 77.6%, 69.8%, and 94.7%, 91.7%, 86.6%, respectively ( P<0.01). Cox proportional hazards model indicated that microvascular invasion ( RR=4.474, 95% CI: 1.172-17.072, P=0.028) and TILs-negative status ( RR=5.081, 95% CI: 1.420-18.184, P=0.012) were independent risk factors for the long-term prognosis of HCC patients who met the Hangzhou criteria. Conclusions:Among HCC patients meeting the Hangzhou criteria, the density of TILs in the tumor stroma is related to AFP levels, tumor differentiation, and the presence of microvascular invasion. TILs-negative status indicates a poorer prognosis for these patients.

Objective:To investigate the decay law of 177Lu-DOTA-TATE in the treatment of neuroendocrine tumors in patients with external radiation and the radiation dose to the surrounding persons. Methods:The radiation dose rate from patients to the surrounding environment after 177Lu-DOTA-TATE treatment was measured using a handheld radiation dosimeter at different times. The decay law of the radiation dose rate with time and distance was analyzed, and the radiation levels at different distances from the patients and the radiation dose of the patients to the surrounding persons were estimated. Results:The external radiation dose rate from the patients decayed exponentially with time rapidly in the early stage and gradually slowly in the later stage. The effective half-life was (1.41±0.25) h (0.9-1.8)h in the rapid decay stage and (40.18±6.0) h (32.7-54.73 h) in the slower decay stage. At 0 h after injection, the normalized mean radiation dose rates at 0.25, 0.5, 1, 2, and 3 m away from the patient were 41.7±6.3, 20.0±3.0, 7.2±3.8, 3.4±0.9, and 1.9±0.4 μSv·h -1·GBq -1, respectively. The radiation doses to doctors, nurses, physicists, and positioning technicians were 2.0, 10.24, 1.08, and 4.05 μSv/patient, respectively. Conclusions:The external radiation dose rate from patients receiving 177Lu-DOTA-TATE treatment rapidly decreases over time and distance. The radiation levels that patients may cause to the surrounding workers are within the allowable range of national standards. After discharge, patients should reduce close and long-term contact with the surrounding persons.

Objective:To explore the strategy of prostate biopsy in patients with prostate specific antigen(PSA)gray zone based on prostate imaging reporting and data system (PI-RADS).Methods:The clinical data of 427 patients who underwent transperineal prostate biopsy in the First Affiliated Hospital of Nanjing Medical University from January 2020 to December 2022 were retrospectively analyzed. The median age was 66 (61, 72) years old. The median PSA was 6.62 (5.46, 8.19) ng/ml. The median PSA density (PSAD) was 0.15 (0.11, 0.21) ng/ml 2. The median prostate volume (PV) was 43.68 (31.12, 56.82) ml. PSA velocity (PSAV) data were available in 65 patients with negative MRI examination(PI-RADS <3), and the median PSAV was 1.40 (0.69, 2.89) ng/(ml· year). Among the patients with positive MRI(PI-RADS≥3), there were 174 patients with only 1 lesion and 83 patients with ≥2 lesions. A total of 170 patients with negative MRI underwent systematic biopsy, and 257 patients with positive MRI underwent systematic combined targeted biopsy. The PI-RADS score, regions of interest(ROI), PSAD, f/tPSA and PSAV were analyzed to explore the biopsy strategy for patients with PSA gray area based on bpMRI imaging. Results:Of the 427 patients included in the study, 194 were positive and 233 were negative. Among the patients with positive biopsy pathology, 140 cases were clinically significant prostate cancer (CsPCa). Among the MRI-negative patients, there were 33 cases with PSAV ≥1.4 ng/(ml·year), and 10 cases of prostate cancer and 6 cases of CsPCa were detected by systematic biopsy.In 32 cases with PSAV <1.4 ng/(ml·year), 3 cases of prostate cancer and 0 case of CsPCa were detected by systematic biopsy. The sensitivity of systematic biopsy for the diagnosis of prostate cancer and CsPCa in patients with PSAV≥1.4 ng/(ml·year) were 76.9% (10/13) and 100.0% (6/6) respectively, the specificity were 55.8% (29/52) and 54.2% (32/59) respectively, the negative predictive value were 90.6% (29/32) and 100.0% (32/32) respectively, and the positive predictive value were 30.3% (10/33) and 18.2% (6/33) respectively. In MRI-positive patients with PI-RADS 3, the prostate cancer detection rates of targeted biopsy combined with systematic biopsy, systematic biopsy and targeted biopsy were 41.7% (45/108), 32.4% (35/108) and 35.2% (38/108), respectively ( P=0.349). The detection rates of CsPCa were 27.8% (30/108), 21.3% (23/108) and 25.0% (27/108), respectively ( P=0.541). In patients with PI-RADS 4-5 and PSAD > 0.15 ng/ml 2, the detection rates of CsPCa in targeted biopsy combined with systematic biopsy, systematic biopsy and targeted biopsy were 67.8% (61/90), 58.9% (53/90) and 67.8% (61/90), respectively ( P=0.354). Conclusions:For MRI-negative patients, all CsPCa could be detected by perineal systematic biopsy when PSAV ≥1.4 ng/(ml·year), and active observation could be performed when PSAV <1.4 ng/(ml·year). For MRI-positive patients, targeted combined systemic biopsy was required when PI-RADS score was 3, and targeted biopsy only could be performed when PI-RADS score ≥4 and PSAD >0.15 ng/ml 2, otherwise targeted combined systemic biopsy was required.

Hypoglycemia can lead to physical and psychological disorders, which are associated with a decrease in quality of life and an increase in mortality risk. Compared to those hospitalized, homebound and community patients have a higher incidence of hypoglycemia and are more vulnerable to the disorders. Hypoglycemia is preventable and treatable, timely intervention can preserve physical well-being of patients and reduce the mortality risk. This article reviews the incidence of hypoglycemia in community diabetic patients, and the knowledge and awareness of patients about hypoglycemia, including the onset inducement, symptoms, hazards, coping and prevention, to provide a reference for the prevention and treatment of hypoglycemia in diabetic patients.

Objective:To explore the pathogenesis of flunarizine-induced parkinsonism from gut-brain axis perspective.Methods:Thirty male C57BL/6 mice were randomly divided into control group and flunarizine group ( n=15). Mice in the control group were given 0.1 mL 50% polyethylene glycol 400+50% saline by gavage once/d for 2 weeks, while mice in the flunarizine group were given 6 mg/mL flunarizine+50% polyethylene glycol 400+50% saline by gavage at a daily dose of 30 mg/kg for 2 weeks. Body mass was recorded 1, 3, 5, 7, 10 and 14 d after drug administration, and motor function was assessed by rotarod test 14 d after drug administration; 16s RNA sequencing was performed in the feces to observe the intestinal flora; intestinal transit function was detected by Evans blue by gavage; and then, the mice were sacrificed and homogenate or frozen sections (brain and intestinal tissues) were prepared; dopamine-ergic neuron expression was detected by Western blotting; RT-qPCR was applied to detect the expressions of inflammatory factors in the substantia nigra, and immunofluorescent staining was used to detect the expressions of ZO-1 and Claudin-5 in the intestinal epithelial tissues. Results:Compared with the control group, the flunarizine group had lower body mass ratio 1, 3, 5, 7, 10 and 14 d after drug administration (ratio to body mass before drug administration). Compared with the control group, the flunarizine group had significantly shortened residence time in rod rotating and lower rotational speed when falling ( P<0.05). Compared with the control group, the flunarizine group had decreased tyrosine hydroxylase protein in the substantia nigra without significant difference ( P>0.05). Compared with the control group, the flunarizine group had significantly increased interleukin-6 and tumor necrosis factor-α in the substantia nigra (1.00±0.00 vs. 2.79±0.83; 1.00±0.00 vs. 3.39±1.37), significantly lower intestinal Evans blue propulsion rate (80.67%±4.51% vs. 50.67%±6.03%), and statistically decreased ZO-1 and Claudin-5 expressions in the colonic epithelial tissues (27.01±1.41 vs. 16.32±2.83; 37.00±2.80 vs. 24.52±2.12, P<0.05). Totally, 576 microorganisms were noted in both control group and flunarizine group, 744 in the control group alone, and 634 in the flunarizine group alone. The intestinal flora β diversity indices in the 2 groups were significantly different based on weighted Unifrac-principle coordinates analysis (PCoA, PCoA1: 39.88%; PCoA2: 30.69%). Compared with the control group, the microbial colony structure of mice in flunarizine group was dominated by phylum thick-walled bacteria and phylum warty microbacteria, and by families Muribaculaceae, Lachnospiraceae and Akkermansiaceae. Compared with the control group, the flunarizine group had significantly decreased relative abundance of Ackermannia spp. and Lactobacillus spp. in the intestinal flora ( P<0.05). Conclusion:Flunarizine may contribute to the pathogenesis of DIP by causing structural disturbances in the intestinal flora and inducing neuroinflammation based on the gut-brain axis.

The global COVID-19 coronavirus pandemic has infected over 109 million people, leading to over 2 million deaths up to date and still lacking of effective drugs for patient treatment. Here, we screened about 1.8 million small molecules against the main protease (M^pro) and papain like protease (PL^pro), two major proteases in severe acute respiratory syndrome-coronavirus 2 genome, and identified 1851M^pro inhibitors and 205 PL^pro inhibitors with low nmol/l activity of the best hits. Among these inhibitors, eight small molecules showed dual inhibition effects on both M^pro and PL^pro, exhibiting potential as better candidates for COVID-19 treatment. The best inhibitors of each protease were tested in antiviral assay, with over 40% of M^pro inhibitors and over 20% of PL^pro inhibitors showing high potency in viral inhibition with low cytotoxicity. The X-ray crystal structure of SARS-CoV-2 M^pro in complex with its potent inhibitor 4a was determined at 1.8 Å resolution. Together with docking assays, our results provide a comprehensive resource for future research on anti-SARS-CoV-2 drug development.
Humans ; Antiviral Agents/chemistry* ; COVID-19 ; COVID-19 Drug Treatment ; High-Throughput Screening Assays ; Molecular Docking Simulation ; Protease Inhibitors/chemistry* ; SARS-CoV-2/enzymology* ; Viral Nonstructural Proteins

Objective To study the effect of self-monitoring of blood glucose information management platform based on the medical consortium on the level of blood glucose control by improving the implementation rate of self-monitoring of blood glucose prescriptions for type 2 diabetes patients in the community.Methods A convenient sampling method was used to select 121 patients with type 2 diabetes from April 2021 to April 2023 who were enrolled in a community of a tertiary hospital medical consortium in Beijing.They were randomly divided into 61 patients in an experimental group and 60 patients in a control group.The control group received routine blood glucose monitoring and guidance education,while the experimental group was managed by a self-monitoring of blood glucose information management platform based on the medical consortium with nurses as the main body.Glycated hemoglobin,glucose test,fasting blood sugar,blood glucose 2 hours after meal,and the completion of self blood glucose monitoring prescriptions were evaluated and analyzed in the first week and the twelfth week of the 2 groups.Results 61 cases in the experimental group and 58 cases in the control group completed the follow-up,and the patients who completed the follow-up were analyzed.There was no statistically significant difference in glycated hemoglobin between the 2 groups at baseline(P=0.117).After 12 weeks,the reduction standard-reaching rate in glycated hemoglobin in the experimental group was significantly different from that in the control group(P=0.014).The number of self-monitoring of fasting blood glucose and the total number of other time periods in experimental group were significantly higher than those in the control group(P＜0.001);the implementation rate of self-monitoring prescription in the experimental group was better than that in the control group(P＜0.001).Conclusion A self blood glucose monitoring information platform was built to manage community type 2 diabetes patients,so that patients can master and implement the personalized blood glucose monitoring prescription formulated by the medical union team with nurses as the main body,and it is conducive to improving the frequency of blood glucose monitoring and the level of blood glucose control of diabetes patients.

For diabetes patients, especially elderly diabetes patients, insulin is widely used as an important treatment to control hyperglycemia.However, since a high percentage of elderly diabetes patients use high doses of insulin, it is common to incur issues such as increased insulin resistance and inappropriate treatment.Exogenous insulin-induced autoantibody production is associated with severe insulin resistance and refractory hyperglycemia and hypoglycemia and is referred to as exogenous insulin antibody syndrome.With hypoglycemia, high insulin levels may be inconsistent with C-peptide levels.Increasing the dose of insulin to control blood glucose may be counterproductive.It is necessary to quickly and accurately make a diagnosis and make personalized adjustments to the glucose-lowering regimen to avoid serious consequences.