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.

Objective To investigate the ameliorative effects and potential mechanism of Lithocarpus litseifoliu on renal function and inflammation in mice with hyperuricemic nephropathy(HN).Methods The HN model was established by the combined administration of adenine and potassium oxyzate.The mice were randomly divided into normal control group,model control group,benzbromarone group,and high,medium and low dose groups of Lithocarpus litseifoliu.Different drugs were given to the mice,and their body mass was recorded once a week.The levels of uric acid(UA),creatinine(Cr),urine protein(UP),blood urea nitrogen(BUN)and urine urea nitrogen(UUN)as well as the levels of tumor necrosis factor α(TNF-α)and interleukin 6(IL-6)in serum or urine of each group were collected and measured on the 21st day.Hematoxylin-eosin(HE)staining was performed to observe kidney tissue injury in mice;real-time PCR(RT-PCR)was performed to determine ATP-binding cassette subfamily G member 2(ABCG2),urate transporter protein(URAT1),glucose transporter protein 9(GLUT9),and cytosolic factor NF-κB p50(κB p50)in kidney tissues.Results Compared with the normal control group,the body mass of mice in the model control group was significantly lower after the second weeks of modeling(P＜0.05),and the levels of UA,Cr,UP,BUN,UUN,TNF-α,IL-6 contents and GLUT9 mRNA and κB p50 mRNA expression contents of kidney tissues were significantly higher(P＜0.01,P＜0.05).Compared with the model control group,the levels of Cr,UP,BUN and UUN contents and renal tissue nuclear cytokine κB p50 mRNA expression were significantly lower in the high,medium and low dose groups of Lithocarpus litseifoliu(P＜0.01,P＜0.05).The UA levels were significantly lower in the high dose group of Lithocarpus litseifoliu(P＜0.05),and renal ABCG2 mRNA expression was significantly higher in the medium dose group(P＜0.01).The renal URAT1 mRNA expression was significantly decreased in the low dose group(P＜0.01).Conclusion Lithocarpus litseifoliu has shown ameliorative effects on HN mice,and the mechanism may be related to the modulation of renal uric acid transporters,improvement of renal function and anti-inflammation effects.

Objective:To investigate whether octyl ester derivative of ginsenoside Rh2(Rh2-O)can induce immunogenic cell death of Huh-7 hepatocellular carcinoma cells and possible mechanism.Methods:Huh-7 cells were cultured in vitro,and divided into control group,Rh2-O group,positive control group(mitoxantrone treatment).Viability and apoptosis of cells were detected by CCK-8 and flow cytometry,respectively.Concentrations of high mobility family protein 1(HMGB1)and adenosine triphosphate(ATP)in supernatant were detected by ELISA and chemiluminescence assay,respectively.Membrane eversion of calreticulin(CRT)was detected by immunofluorescence assay.ROS level in cells was detected by fluorescence probe DCFH-DA,and expressions of proteins associated with endoplasmic reticulum stress signaling pathway were detected by Western blot.Results:Rh2-O treatment significantly reduced cell viability,promoted apoptosis,induced secretion of HMGB1,ATP,membrane eversion of CRT,increased accumulation of ROS in cells,and enhanced expressions of endoplasmic reticulum stress-related proteins PERK,eIF2α,p-eIF2α(all P<0.05).After addition of endoplasmic reticulum stress inhibitor 4-phenylbutyric acid(4-PBA),membrane eversion of CRT induced by Rh2-O was significantly inhibited(P<0.05).Conclusion:Rh2-O can induce immunogenic cell death in hepatocellular carcinoma cells,whose mechanism may be associated with activation of endoplasmic reticulum stress and promotion of CRT membrane eversion.

Pulmonary fibrosis is a chronic， progressive and irreversible interstitial lung disease. At present， there is no specific drug for the treatment of pulmonary fibrosis， and many TCM monomers have potential therapeutic value for pulmonary fibrosis， among which flavonoids are the main representative. For example， total flavones of Astragalus memeranaceus and scutellarin can reduce inflammatory cell infiltration， lung injury and extracellular matrix （ECM） deposition by interfering with transforming growth factor-β1/drosophila MAD protein signaling pathway. Total flavonoids of Oxytropis falcata Bunge and salidroside can inhibit lung inflammation by mediating JAK/signal transduction and transcriptional activator signaling pathway， and prevent the epithelial interstitial transition （EMT） process. Quercetin and Ginkgo biloba leaf extract can reduce the apoptosis of macrophages by inhibiting the nuclear factor-κB signaling pathway and play an anti-pulmonary fibrosis role. Urushetin and proanthocyanidins can promote the morphological recovery of myofibroblasts and reduce ECM deposition through the phosphatidylinositol 3-kinase/protein kinase B/mammalian target protein of rapamycin signaling pathway. Naringin and luteolin can inhibit scorch death of macrophage and inflammation response， and improve lung function and lung tissue injury through NOD-like receptor heat protein domain related protein 3 signaling pathway. The ethanol extract of Phyllanthus emblica and calycosin can improve the inflammatory injury and fibrosis of lung tissue by activating the signaling pathway of nuclear transcription factor erythro2-related factor 2/antioxidant response element. Isogliquiritin can inhibit the phenotypic transformation of epithelial cells and reverse EMT progression by inhibiting extracellular signal-regulating kinase signaling pathway. In the future， scholars should consider developing appropriate drug carriers to improve their bioavailability and further study drug targets and pathways， to provide evidence for the development of traditional Chinese medicine monomers of flavonoids into clinical practice.

Objective:To explore the clinical efficacy of ultrasound-indicated cervical cerclage in twin pregnancies.Methods:This retrospective cohort study included 96 asymptomatic twin pregnancies with cervix length (CL)≤2.5 cm at 16-27 +6 weeks indicated by transvaginal ultrasound from January 2013 to May 2022 in Fujian Maternity and Child Health Hospital. They received either cervical cerclage (cerclage group, n=45) or conservative treatment (conservative group, n=51). The subgroup analysis was further performed after stratifying the subjects into the CL≤1.0 cm subgroup, the CL>1.0-≤1.5 cm subgroup, and the CL>1.5 cm subgroup. Two independent samples t-test, Mann-Whitney U test, and Chi-square test were used to compare the differences in the pregnancy and perinatal outcomes between different groups. Multivariate logistic regression was used to analyze the impact of cervical cerclage on pregnancy outcomes of women with different CLs. Results:The average gestational age at delivery and the prolonged gestation were significantly greater in the cerclage group than those in the conservative group [35.9 (34.9-37.0) weeks vs 34.9 (29.1-36.1) weeks; 10.7 (9.6-13.1) weeks vs 8.7 (4.8-11.0) weeks, Z=－2.59 and -3.63, both P<0.05]. The incidences of preterm birth before 34 weeks and 28 weeks, chorioamnionitis, and preterm premature rupture of membrane were lower in the cerclage group than those in the conservative group [17.8% (8/45) vs 45.1% (23/51), χ2=8.16; 2.2% (1/45) vs 15.7% (8/51), Fisher' exact test; 8.9% (4/45) vs 25.5% (13/51), χ2=4.52; 15.6% (7/45) vs 33.3% (17/51), χ2=4.03; all P<0.05]. There were no statistically significant differences in the proportion of postpartum hemorrhage and live births between the two groups (both P>0.05). In the women with CL≤1.0 cm, the cerclage group had a greater gestational age at delivery [36.0 (34.8-37.3) weeks vs 34.9 (28.6-35.4) weeks, Z=－2.61, P=0.009], greater prolonged gestation [12.3 (9.7-13.9) weeks vs 7.3 (3.4-9.1) weeks, Z=－3.34, P=0.001], higher birthweight [2 430.0 (2 173.8-2 646.3) g vs 1 900.0 (1 372.5-2 435.0) g, Z=－3.06, P=0.002], lower incidence of adverse neonatal outcomes [16.7% (6/36) vs 36.7% (11/30), χ2=4.22, P=0.048] compared with the conservative group. Multivariate Logistic regression analysis showed that cerclage reduced the risks of preterm birth before 34 and 32 weeks [ aOR (95% CI)=0.083 (0.009-0.790) and 0.092 (0.009-0.939), both P<0.05]. However, cerclage did not reduce the risk of preterm birth in women with CL>1.5 cm or CL>1.0-≤1.5 cm (both P>0.05). Conclusion:Ultrasound- indicated cervical cerclage can reduce the risk of preterm birth before 34 and 32 weeks, prolong pregnancy, and improve perinatal outcomes in asymptomatic twin pregnancies with CL≤1.0 cm.

ObjectiveTo observe the effects of Fuzitang （FZT） on the proliferation of MH7A cells， the human rheumatoid arthritis synovial fibroblasts， and the expression of miR-155 and explore its anti-rheumatoid arthritis mechanism. MethodMH7A cells were cultured in vitro and divided into a blank group， high- （25 g·L^-1） and low-dose （12.5 g·L^-1） FZT groups， and a positive drug group （hydroxychloroquine， 0.006 25 g·L^-1）. The cell proliferation was detected by cell counting kit-8（CCK-8） method， and the change in the MH7A cell cycle was detected by flow cytometry. The mRNA expression of miR-155 and its downstream genes， including SH2 domain-containing inositol 5-phosphatase-1（SHIP-1）， protein kinase B 3（Akt3）， and mammalian target of rapamycin（mTOR）， was detected by Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）， and the protein expression of phosphatidylinositol 3-kinase （PI3K）， Akt3， and mTOR was detected by Western blot. ResultFZT in vitro in a concentration of 6.25 g·L^-1 above could inhibit the proliferation of MH7A cells in the significant dose- and time-effect manner. Compared with the blank group， the FZT groups showed increased proportions of cells in the G₂/M phase （P<0.05）， and the high-dose FZT group showed a decreased proportion of cells in the G₀/G₁ phase （P<0.05）. The arresting effect of FZT on the cell cycle was in a significant dose-effect manner. Compared with the blank group， the FZT groups showed down-regulated miR-155 and mTOR mRNA expression （P<0.05）， and the high-dose FZT group showed up-regulated SHIP1 mRNA expression and down-regulated Akt3 mRNA expression （P<0.05）. Compared with the blank group， the FZT groups showed reduced protein expression of PI3K， Akt3， and mTOR （P<0.05）. ConclusionFZT can significantly inhibit the proliferation of MH7A cells， and the mechanism is related to the promotion of the expression of SHIP-1 and down-regulation of the gene expression of the PI3K/Akt3/mTOR signaling pathway by down-regulating the expression of miR-155.

To explore the effects and molecular mechanism of icariin on the vascular function of mice with type 1 diabetes induced by alloxan, type 1 diabetic mice model was established by intraperitoneal injection with 200 mg/kg alloxan.After oral administration with icariin (60, 120 mg/kg) daily for 2 weeks, blood glucose, body weight, food intake and water intake were detected.To evaluate the impact of icariin on the function of isolated vascular ring contraction and relaxation, thoracic aortas of mice were removed and the Ach-induced vascular ring relaxation, Phe-induced vascular ring contraction, SNP-induced vascular ring relaxation and KCl-induced vascular ring contraction response were detected.To further confirm the mechanism of icariin to improve vascular function, human umbilical vein endothelial cells (HUVECs) were induced by high glucose (HG) in vitro.Western blot was used to detect the effect of icariin on eNOS, p-eNOS, p38 MAPK and p-p38 MAPK expressions in HG-induced human umbilical vein endothelial cells (HUVECs).The results indicated that icariin significantly ameliorated the weight loss and dampened the increase in water intake of the diabetic mice.Meanwhile, icariin had a certain ameliorative effect on blood glucose and food intake without significant difference.The results of isolated thoracic aortas vascular rings contraction and vasodilation function indicated that icariin significantly improved Phe-induced vascular contraction and Ach‐induced vascular relaxation.Meanwhile, icariin had a certain ameliorative effect on KCl-induced vascular contraction response without significant difference.However, no significant change was observed on endothelium‐independent vascular rings relaxation response induced by SNP after treatment with icariin.Results of Western blot showed that icariin inhibited the expression of p-p38 MAPK and induced expression of p-eNOS in the high glucose-induced HUVECs cell model.Therefore, icariin may attenuate alloxan-induced type 1 diabetic mice vascular diastolic function by inhibiting expression of p-p38 MAPK and inducing expression of p-eNOS.

Humans ; Pulmonary Disease, Chronic Obstructive ; Smoke/adverse effects* ; Tobacco ; Tobacco Smoke Pollution

Objective:To evaluate the auxiliary diagnosis value of bacterial culture, polymerase chain reaction (PCR) and serum anti-pertussis toxin immunoglobulin G (AntiPT-IgG) level detection in suspected pertussis.Methods:A total of 110 suspected cases of pertussis treated in the Department of Pediatrics of Wuhu No.1 People′s Hospital from June 2018 to May 2019 were recruited for the study.The nasopharyngeal swabs of all cases were collected for Bordetella pertussis culture and specific nucleic acid PCR detection.Serum samples of 78 cases were collected for the detection of AntiPT-IgG level by enzyme linked immunosorbent assays.Results:The positive rates of bacterial culture group and PCR group were 21.8% and 30.0%, respectively, with no statistically significant difference ( χ2=1.198, P>0.05). The culture positive rate of cases with the duration of cough<2 weeks was 32.1%, which was signi-ficantly higher than that of cases with the duration of cough about 2-4 weeks (14.3%) or >4 weeks (9.1%) ( χ2=6.522, P<0.05). The PCR positive rate of cases with the duration of cough <2 weeks was 39.6%, which was also significantly higher than that of cases with the duration of cough about 2-4 weeks (25.7%) or > 4 weeks (13.6%) ( χ2=6.126, P<0.05). The mean value for serum AntiPT-IgG level of 78 cases was (75.727±78.454) IU/mL, the median AntiPT-IgG levels of cases with the duration of cough<2 weeks and about 2-4 weeks were 5.909 IU/mL and 20.948 IU/mL, respectively, and the positive rates were 14.7% and 38.1%, respectively.The AntiPT-IgG level of cases with the duration of cough> 4 weeks and that at convalescent stage were (79.281±68.254) IU/mL and (107.242±75.750) IU/mL, and the positive rates were 39.1% and 57.1%, respectively. Conclusions:In the vaccine era, the results of pathogenic and serological tests should be combined to assist the clinical diagnosis of pertussis.The positive rate of bacterial culture and specific nucleic acid pathogen detection in children with cough duration less than 2 weeks is high, and the serological diagnosis is more effective after the duration of cough is over 4 weeks.

Objective:To investigate the incidence of asthma and the characteristics of acute attacks in children aged 3-14 in different administrative areas of Wuxi, and to analyze the correlation of asthma with the outdoor environmental factors.Methods:A total of 10 175 children aged 3-14 were randomly selected from 5 different administrative regions in Wuxi City, Jiangsu Province for childhood asthma epidemiological survey from January to December in 2017.Results:The complete questionnaires were collected from 8 318 children with a response rate of 81.70%.There were 6 734 valid questionnaires.Asthma occurred to 259 children, and the prevalence rate was 3.84%.The prevalence of asthma in male and female children was 4.89% (163 cases) and 2.82% (96 cases), respectively.The ratio of male patients to female patients was 1.70∶1.00, and the difference was significant ( χ2=19.72, P<0.01). Children were most prone to suffer from asthma attacks at the turn of seasons (87/259 cases, 33.59%). The frequency of asthma was the lowest from June to August (9/259 cases, 3.47%). Among 244 cases attacked by acute asthma, the prevailing time period of acute attacks was irregular in 88 patients (36.07%), bedtime in 64 cases (26.23%). In different regions of the city, the lowest prevalence rate of asthma was found in Binhu District (50/1 830 cases, 2.73%), the highest prevalence rate of asthma was found in Xinwu District (71/1 502 cases, 4.72%), the difference between the highest and lowest prevalence rates of asthma was significant ( χ2=13.19, P<0.05). In terms of the air quality in different admi-nistrative regions of Wuxi, the nitrogen dioxide (NO 2) concentration, the carbon monoxide (CO) concentration, PM 10 and PM 2.5 in Binhu District were lower than other 4 regions.In different seasons in Wuxi city, the sulfur dioxide(SO 2) concentration, the NO 2 concentration, the CO concentration, PM 10 and PM 2.5 from June to August were significantly lower than those in other months.The prevalence of asthma in children in different administrative districts of Wuxi was weakly positively correlated with the SO 2 concentration, the NO 2 concentration and PM 10 ( r=0.10, P<0.01; r=0.22, P<0.01; r=0.06, P<0.01, respectively). The prevalence of asthma was weakly negatively associated with the ozone(O 3) concentration ( r=-0.06, P<0.01). Acute asthma attacks were weakly positively correlated with the SO 2 concentration ( r=0.22, P<0.01), the NO 2 concentration ( r=0.28, P<0.01), the CO concentration ( r=0.23, P<0.01), PM 10 ( r=0.18, P<0.01) and PM 2.5 ( r=0.18, P<0.01), and weakly negatively correlated with the O 3 concentration ( r=-0.40, P<0.01). Conclusions:The prevalence of asthma in Wuxi is higher in boys than in girls, and the frequency of asthma attacks is related to the season.The SO 2, NO 2, CO, PM 10, PM 2.5 and O 3 concentration affect the prevalence and acute attacks of asthma.