Design, synthesis, and activity of 5-cyano-2-thiacetyl aromatic pyrimidinone as DENV NS5 RdRp inhibitors / 药学学报

To screen novel anti-dengue virus (DENV) NS5 RdRp enzyme inhibitors, a series of 5-cyano-2-thiacetoaryl pyrimidinone compounds were designed and synthesized by molecular hybridization method with HCV NS5B RdRp inhibitor 3jc and ZIKV NS5 RdRp inhibitor 4w as lead compounds. The anti-DENV activity of these compounds was evaluated by MTT assay and plaque assay and five compounds showed anti-DENV activity. The most active compound 7a'k showed better anti-DENV activity than that of the positive control ribavirin (EC50 = 7.86 μmol·L-1 vs EC50 = 18.07 μmol·L-1), and the other four compounds showed almost the same anti-DENV activity as ribavirin. Finally, the prediction and simulation of the binding mode through molecular provided new ideas for the further development of this new DENV NS5 RdRp inhibitor.

Trends in the asthma burden among children and adolescents in China from 1990-2019 / 中国学校卫生

Objective@#To explore trends in the asthma burden among Chinese children and adolescents 1-19 years old during 1990-2019.@*Methods@#Based on data from the 2019 Global Burden of Disease Study, joinpoint regression was used to analyze the dynamic changes in the gender and age specific asthma burden, and the asthma burden in China was compared with countries that have different socio demographic indices(SDI). In addition, trends in asthma burden attributed to different risk factors were also investigated.@*Results@#The asthma burden decreased slightly from 1990 to 1996 [annual percent change (APC)=-1.7%], then rapidly decreased from 1996 to 2005 (APC=-5.7%). The age standardized disability adjusted life years (DALYs) rate decreased from 158.55/100 000 to 88.59/100 000 in patients 1-19 years of age. From 2005 to 2017, the DALYs rate for asthma increased slowly, then rapidly. In 2017, the DALYs rate peaked at 176.18/100 000, then decreased to 126.79/100 000 in 2019. The burden of asthma in boys was higher than girls, and the DALYs rate for asthma in the group 5-9 years of age was higher than the remaining age groups. Furthermore, the age standardized DALYs rate for asthma among Chinese children and adolescents was relatively low among countries with a different SDI. In addition, the DALYs rate attributed to high body mass index increased in all age groups in China. Specifically, the average APC (AAPC) was 2.9% in group 1-4 years of age and the AAPC was 4.2% in the remaining age groups. The DALYs rate attributed to occupational asthmagens in the group 15-19 years of age decreased from 1990 to 2019 and the AAPC was -2.5%.@*Conclusion@#The asthma burden was relatively low among Chinese children and adolescents, and there were gender and age differences. The gender and age specific DALYs rate for asthma had a tendency to decrease, increase, then decrease. More attention should be paid to boys and the group 5-9 years of age, and strengthen the intervention of obesity and occupational asthmagens.

Research progress on the methods of observation and analysis of dental bonding interfaces / 口腔疾病防治

@#Dental bonding technology and materials have been used widely in dentistry because of their excellent properties. The development of novel bonding technology and materials is constantly being performed to improve the effect of dental bonding restorations. Observation and analysis of the dental bonding interface is one of the most important methods for laboratory evaluation of bonding efficiency. This paper aims to review the methods of observation and analysis of dental bonding interfaces to provide a reference for the selection of evaluation methods in dental bonding research. The features of 6 methods, including scanning electron microscopy (SEM), transmission electron microscopy (TEM), confocal laser scanning microscopy (CLSM), Raman spectroscopy (RS), optical coherence tomography (OCT) and atomic force microscopy (AFM), were described and summarized. Among these methods, SEM and TEM are used most often in the analysis of fine structures; CLSM and OCT are used for the acquisition of characteristic image signals, such as microleakage and exogenous and endogenous fluorescence; and RS and AFM can test chemical composition and mechanical properties.

Ciclesonide inhibits Zika virus replication in vitro / 药学学报

Zika virus (ZIKV) is an emerging mosquito-borne virus that is associated with severe congenital brain malformations in the fetus and Guillain-Barré syndrome in adults. However, there are currently no drugs or preventive vaccines approved for ZIKV infection. Here, ciclesonide has been found significantly against ZIKV activity by plaque and cytotoxicity assays in vitro, and its 50% effective concentration (EC50) to ZIKV SZ01 and MR766 are (0.40 ± 0.22) and (1.59 ± 1.08) μmol·L-1, respectively. Its 50% cytotoxic concentration (CC50) to Vero cells are (64.70 ± 7.33) μmol·L-1; Virus yield reduction and Western blot assays showed that ciclesonide can inhibit replication of ZIKV. In addition, ciclesonide can also inhibit the replication of ZIKV in A549 cells; the results of time of drug addition analysis indicated that ciclesonide mainly acts on the ZIKV RNA synthesis stage. Ciclesonide can also inhibit the internalization of ZIKV. These results indicated that ciclesonide is a potential drug against ZIKV.

Distribution characteristics of asthma in preschool children and the protective effect of breastfeeding / 中国学校卫生

Objective@#To clarify the distribution of asthma in preschool children aged 3-6 years old and the protective effect of breastfeeding on asthma in three provinces in the middle and lower reaches of the Yangtze river in China.@*Methods@#The parents of 27 200 preschoolers chosen from kindergartens in 11 cities of Hubei, Jiangsu and Anhui Province were investigated with children’s asthma and breastfeeding during October to November in 2017.@*Results@#The asthma detection rate for preschoolers was 4.0%,5.0% for boys and 2.9% for girls. The asthma detection rate for children aged 3,4,5,and 6 years old was 3.2%,3.7%,4.4%,and 4.9%. 3 448 (12.7%) and 7 630 (28.1%) preschoolers were never breastfed or mainly breastfed. The rate of exclusive breastfeeding was 50.0% for 1 to 5 months and only 10.2% over 6 months. Meanwhile,there was a significant protective effect on asthma (OR=0.88,95%CI=0.77-0.89) by exclusive breastfeeding for 1-5 months,which was still statistically significant after adjusting for confounding factors. After adjusting for confounding factors,exclusive breastfeeding over 6 month have significant effected on preschool children aged 4 years old without allergic conjunctivitis history (OR=0.63,95%CI=0.40-0.99). Furthermore,exclusive breastfeeding over 6 month had important effect on preschool children aged 4 years old with eczema history (OR=0.52,95%CI=0.27-0.99).@*Conclusion@#Exclusive breastfeeding has protective effect on asthma in preschool children. Exclusive breastfeeding over 6 months even has better protective effect on asthma for the 4-years-old children without the history of allergic conjunctivitis and eczema.

Epidemiological characteristics of 107 pediatric partients with COVID-19 during the initial phase of the outbreak / 中国学校卫生

Objective@#To describe the epidemiological characteristics and clinical manifestations of the 2019 coronavirus disease (COVID-19) in pediatric patients, and to provide data support and decision-making basis for the prevention and control of COVID-19.@*Methods@#Cases of children aged 0-17 years reported by provincial health commissions in Anhui, Shandong, Zhejiang and Henan provinces were collected to analyze their spatial, temporal, and demographic distribution.@*Results@#By 24:00 on February 6, 2020, a total of 107 pediatric patients had been reported in the four provinces, accounting for 3.8% (107/2 808) of the total cases reported in the four provinces during the same period. Anhui, Shandong, Zhejiang and Henan provinces had 25, 25, 28 and 29 cases, respectively. Cases ranged in age from 5 days after birth to 17 years, with a median age of 8 years. Boys accounted for 58.9%. Totally 38 cases had a history of sojourn in Wuhan or Hubei, 63 cases had a history of exposure to confirmed cases, and 6 cases with unknown exposure history. A group of 52 family clustering were found in 107 cases. All cases presented mild symptoms, no serious and no death.@*Conclusion@#Children were also susceptible to the COVID-19. Before February 2, the index pediatric cases were mainly the first generation cases, and after February 3, these pediatric cases were mainly the secondary-generation cases and those who had close contact with confirmed cases. The monitoring of children with secondgeneration cases and close contact with COVID-19 cases were valued.

Inhibitory effect of PI3Kδ inhibitor idelalisib on proliferation of human myeloid leukemia cells and the reversal effect on drug resistance to adriamycin / 中南大学学报(医学版)

OBJECTIVES@#To investigate the effect of adriamycin (ADM), idelalisib or ADM and their combination on cell proliferation and intracellular concentration of ADM, and to explore the reversal effect of idelalisib on drug resistance to ADM.@*METHODS@#The K562 and K562/ADM cells were respectively treated with ADM and idelalisib at different concentrations. The 50% inhibitory concentration (IC@*RESULTS@#The cell survival rates were significantly decreased in a dose-dependent manner when the cells were treated with different doses of ADM (0.001-10.000 mg/L ). The IC@*CONCLUSIONS@#Idelalisib exerts effect on inhibition of the proliferation in myeloid leukemia K562 and K562/ADM cells, which may partially reverse the drug resistance of K562/ADM cells to ADM. The mechanisms for the effect of idelalisib may be related to increasing the accumulation of ADM and inducing the cell apoptosis in the K562 and K562/ADM cells.

Noscapine inhibits migration of colon cancer SW480 cells by down-regulating Wnt3a/β-catenin signaling pathway and cadherin 17 / 中国肿瘤生物治疗杂志

@# Objective: To explore the effects of noscapine (Nos) on the expression of cadherin 17 (CDH17) in colon cancer SW480 cells and the mechanism of Nos on cell migration. Methods: SW480 cells were divided into the control group, empty vector (si-EV) group, CDH17 interference (si-CDH17) group, Nos treatment group, and CDH17 interference+Nos treatment (si-CDH17+Nos) group. Small interfering RNA (siRNA) was used to knockdown CDH17, and the selected concentration of Nos was (55.30±2.21) µg/ml (IC50). The mRNA expression of CDH17 was detected by qPCR; the apoptosis and migration abilities of SW480 cells were observed by Hoechst33258 staining and Transwell assay; the contents of VEGF, MMP2 and MMP9 in SW480 cells were measured by ELISA, and the protein expressions of CDH17, Wnt3a and β-catenin were determined by WB. Results: Compared with the control group, mRNA and protein expressions of CDH17 obviously decreased, cell apoptosis and migration significantly reduced, while the contents of VEGF, MMP2 and MMP9 as well as the protein expressions of Wnt3a and β-catenin significantly decreased in Nos treatment group, siCDH17 group and si-CDH17+Nos treatment group (all P<0.01).The effect of si-CDH17+Nos treatment was more significant than that of si-CDH17 (P<0.01). Conclusion: Nos induces apoptosis and inhibits the migration of human colon cancer SW480 cells, which may be related to the down-regulation of CDH17 expression and inhibition of the Wnt3a/β-catenin signaling pathway.

The study of azvudine activity against dengue viruses in vitro / 药学学报

In this study, azvudine (FNC), hydrochloride salt of azvudine (FNC-HCl) and triphosphate azovudine (FNC-TP) were tested against DENV-Ⅱ recombinant virus (DENV-Ⅱ Luc+). The inhibitory activity of FNC, FNC-HCl and FNC-TP on DENVs were detected by plaque assay. The effect on the expression of DENV-Ⅱ envelope protein E was detected by Western blot; the inhibitory of DENV-Ⅱ viral RNA by compounds was detected by real-time quantitative PCR. MTT assay was used to determine the cytotoxicity of the three compounds on Vero cells. The results showed that FNC, FNC-HCl and FNC-TP inhibited the viral replication by inhibition of renilla luciferase activity of DENV-Ⅱ Luc+. The 50% effective concentration (EC50) of FNC, FNC-HCl and FNC-TP in the inhibition of DENVs replication were from 0.54-25.42 μmol·L-1, while that of ribavirin was 40.78 ±1.02 μmol·L-1 as the positive control. Western blot and real time quantitative PCR results showed that FNC, FNC-HCl and FNC-TP significantly inhibited the expression of DENV-Ⅱ E protein, and the replication of DENV-Ⅱ viral RNA. The 50% cytotoxic concentrations of FNC, FNC-HCl and FNC-TP were all greater than 3 000.00 μmol·L-1. The results suggest that in vitro anti-DENVs activities of FNC, FNC-HCl and FNC-TP are superior to ribavirin, which are expected to become new candidates of anti-DENV drugs.

The combined anti-HIV-1 effect of chloroquine and antiretroviral drugs in vitro / 药学学报

The study is aimed to evaluate the anti-HIV-1 effect of chloroquine in combination with antihuman immunodeficiency virus (HIV) drugs, and inhibition of plasmacytoid dendritic cells (pDC) activation and type I interferon (IFN-I) production by Toll-like receptor 7 (TLR7) agonist stimulation. We investigated the anti-HIV-1ⅢB, HIV-1KM018 activity of chloroquine and chloroquine combined with rategrivir (RAL), enfuvirtide (T-20), indinavir (IDV) and efavirenz (EFV) in vitro by luciferase activity assay system and ELISA method for p24 antigen. We measured the effect of chloroquine on the activation of pDC in combination with RAL and IDV, respectively. Quantitative PCR was used to evaluate the activity of chloroquine in combination with RAL and IDV in the upregulation of interferon (IFN)-α and IFN-β. Chloroquine showed less cytotoxicity to C8166, TZM-bl and PBMC cells, and the 50% cytotoxic concentration values were 85.02 ±0.28, 73.67 ±5.10 and 91.84 ±4.10 μmol·L-1, respectively. The anti-HIV-1ⅢB activity of chloroquine combination with RAL, T-20, IDV and EFV were moderate in synergy, strong in synergy, additive and moderate antagonism, respectively. The anti-HIV-1KM018 activity of chloroquine in combination with RAL, IDV were moderate synergy, minor synergy. There was no significant difference between the chloroquine monotherapy and chloroquine combined with RAL, IDV in the down-regulation of pDC activation and IFN-α, IFN-β expression levels. We have found that chloroquine combined with different anti-HIV drugs represent different degrees of synergism, antagonism or additive anti-HIV-1 effect. Chloroquine in combination with RAL and IDV did not have influence on the inhibitory effect of chloroquine on pDC activation and type I interferon secretion induced by TLR7 agonist. The results suggest that chloroquine may be used to enhance the therapeutic activities of anti-HIV medicines.

Differential Diagnosis of Solitary Pulmonary Inflammatory Lesions and Peripheral Lung Cancers with Contrast-enhanced Computed Tomography

OBJECTIVES: To clarify differences between solitary pulmonary inflammatory lesions and peripheral lung cancers with contrast-enhanced computed tomography. METHODS: In total, 64 and 132 patients with solitary pulmonary inflammatory masses/nodules and peripheral lung cancers, respectively, were enrolled in this study. Their computed tomographic findings were summarized and compared retrospectively. RESULTS: Compared with the peripheral lung cancers, the inflammatory lesions were located closer to the pleura (p<0.0001). The majority of the inflammatory lesions were patchy and oval-shaped (82.8%), whereas most of the tumors were lobulated (82.6%). Almost all the inflammatory cases were unclear (93.8%), whereas most of the tumors had spiculated margins (72.7%). Computed tomography values were significantly higher for the inflammatory lesions than for the cancers (p<0.0001). More than half of the inflammatory lesions had defined necrosis (59.3%). Furthermore, 49.2% of the cancers enhanced inhomogeneously, but only 24.6% had ill-defined necrosis or cavities. The peripheral zones of 98.4% of the inflammatory lesions and 72.7% of the tumors were unclear, with peripheral scattered patches (92.2%) and beam-shaped opacity (66.7%) being the most common findings, respectively. Adjacent pleural thickening was more frequent for the inflammatory lesions than the cancers (95.3% vs. 21.1%, p<0.0001), whereas pleural indentation was found in 67.4% of the subjects with cancer. In addition, hilar (p=0.034) and mediastinal (p=0.003) lymphadenopathy were more commonly detected in the cancers than in the inflammatory cases. CONCLUSIONS: Contrast-enhanced computed tomography findings for pulmonary inflammatory lesions and peripheral lung cancers were significantly different in many aspects. Developing a comprehensive understanding of these differences is helpful for directing their management.

The anti-HIV-1 activities of benzophenones non-nucleoside reverse transcriptase inhibitors in vitro / 药学学报

To evaluate the anti-HIV-1 activities of 5 benzophenones non-nucleoside reverse transcriptase inhibitors (NNRTIs) such as DY1203, DY1204, DY1119, DY1208 and DY1209 in vitro, the cytotoxicity of 5 compounds were tested on C8166, MT-4, H9 and PBMC with the MTT assay. The anti-HIV-1 activities of compounds were evaluated on laboratory-adapted strain, drug-resistant strains and primary isolated strains by p24 antigen expression ELISA. The inhibition of HIV-1 recombinant reverse transcriptase activity was assessed by ELISA assay. Among 5 compounds, DY1203 and DY1204 showed low cytotoxicities with CC50 greater than 200 μg·mL-1. DY1119, DY1208 and DY1209 showed strong anti-HIV-1 activities against HIV-1IIIB, HIV-174V, HIV-1RF/V82F/184V, HIV-1NL4-3 gp41(36G) N42S, HIV-1KM018, HIV-1TC-1 and HIV-1Wan. However, NNRTIs drug-resistant strain HIV-1A17 showed different resistance to these compounds. The 5 compounds proved active against HIV-1 recombinant reverse transcriptase. DY1208 is expected to become a new lead compound for its high therapeutic index. The results can provide new information for HIV-1 drug research and promote the development of new HIV-1 drugs.

Wikstroelide M potently inhibits HIV replication by targeting reverse transcriptase and integrase nuclear translocation / 中国天然药物

AIM@#To evaluate the anti-HIV activity and mechanism of action of wikstroelide M, a daphnane diterpene from Daphne acutiloba Rehder (Thymelaeaceae).@*METHODS@#The anti-HIV activities of wikstroelide M against different HIV strains were evaluated by cytopathic effect assay and p24 quantification assay with ELISA. The inhibitory effect of wikstroelide M on HIV reverse transcription was analyzed by real-time PCR and ELISA. The effect of wikstroelide M on HIV-1 integrase nuclear translocation was observed with a cell-based imaging assay. The effect of wikstroelide M on LEDGF/p75-IN interaction was assayed by molecular docking.@*RESULTS@#Wikstroelide M potently inhibited different HIV-1 strains, including HIV-1IIIB, HIV-1A17, and HIV-19495, induced a cytopathic effect, with EC50 values ranging from 3.81 to 15.65 ng·mL⁻¹. Wikstroelide M also had high inhibitory activities against HIV-2ROD and HIV-2CBL-20-induced cytopathic effects with EC50 values of 18.88 and 31.90 ng·mL⁻¹. The inhibitory activities of wikstroelide M on the three HIV-1 strains were further confirmed by p24 quantification assay, with EC50 values ranging from 15.16 to 35.57 ng·mL⁻¹. Wikstroelide M also potently inhibited HIV-1IIIB induced cytolysis in MT-4 cells, with an EC50 value of 9.60 ng·mL⁻¹. The mechanistic assay showed that wikstroelide M targeted HIV-1 reverse transcriptase and nuclear translocation of integrase through disrupting the interaction between integrase and LEDGF/p75.@*CONCLUSION@#Wikstroelide M may be a potent HIV-1 and HIV-2 inhibitor, the mechanisms of action may include inhibition of reverse trascriptase activity and inhibition of integrase nuclear translocation through disrupting the interaction between integrase and LEDGF/p75.

Identifying risk factors of immune reconstitution inflammatory syndrome in AIDS patients receiving highly active anti-retroviral therapy

Immune reconstitution inflammation syndrome typically occurs within days after patients undergo highly active anti-retroviral therapy and is a big hurdle for effective treatment of AIDS patients. In this study, we monitored immune reconstitution inflammation syndrome occurrence in 238 AIDS patients treated with highly active anti-retroviral therapy. Among them, immune reconstitution inflammation syndrome occurred in 47 cases (19.7%). Immune reconstitution inflammation syndrome patients had significantly higher rate of opportunistic infection (p < 0.001) and persistently lower CD4+ cell count (p < 0.001) compared to the non-immune reconstitution inflammation syndrome patients. In contrast, no significant differences in HIV RNA loads were observed between the immune reconstitution inflammation syndrome group and non-immune reconstitution inflammation syndrome group. These data suggest that a history of opportunistic infection and CD4+ cell counts at baseline may function as risk factors for immune reconstitution inflammation syndrome occurrence in AIDS patients as well as potential prognostic markers. These findings will improve the management of AIDS with highly active anti-retroviral therapy.

Effect of Trigonella foenum-graecum 4-hydroxyisoleucine on high-glucose induced insulin resistance in 3T3-L1 adipocytes of mice / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effect of 4-hydroxyisoleucine (4-HIL), an active component of Trigonella Foenum-graecum L. on high glucose induced insulin resistance (IR) in 3T3-L1 adipocytes, and to explore underlying molecular mechanisms.</p><p><b>METHODS</b>3T3-L1 adipocytes were treated with 25 mmol/L glucose and 0.6 nmol/L insulin to induce IR. They were intervened by different concentrations of 4-HIL (at 5, 10, and 20 micromol/L). [3H]-Deoxy-D-glucose up-taking method was used to detect the glucose uptake. The mRNA expression of cellular tumor necrosis factor-alpha (TNF-alpha) was detected by polymerase chain reaction (PCR). The content of TNF-alpha in the culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA). Palmitic acid (PA) acted as the control.</p><p><b>RESULTS</b>After intervened by 25 mmol/L glucose and 0.6 nmol/L insulin for 18 h, the insulin-stimulated glucose transportation in 3T3-L1 adipocytes was inhibited by 63%. The mRNA expression of cellular TNF-alpha in adipocytes significantly increased, when compared with that in normal adipocytes (P < 0.05). The level of TNF-alpha secreted in the culture supernatant was increased by 70 pg/mL (P < 0.05). Similar changes occurred in the PA group. After exposure to 4-HIL (5, 10, or 20 micromol/L) for 24 h, the glucose transportation was increased by 35%, 50%, and 60%, respectively. PCR results showed that along with increasing 4-HIL concentrations, the mRNA expression of cellular TNF-alpha showed a decreasing trend, showing statistical difference when compared with the model group and the PA group (P < 0.05). Compared with the model group, the TNF-alpha level in the supernatant was respectively reduced by 10 pg/mL, 18 pg/mL, and 39 pg/mL after intervention (P < 0.05).</p><p><b>CONCLUSION</b>4-HIL could remarkably improve high glucose-induced IR in 3T3-L1 adipocytes. Meanwhile, 4-HIL could inhibit the secretion of TNF-alpha.</p>