The coronavirus disease 2019 (COVID-19) is an acute infectious disease caused by the new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, which has led to serious worldwide economic burden. Due to the continuous emergence of variants, vaccines and monoclonal antibodies are only partial effective against infections caused by distinct strains of SARS-CoV-2. Therefore, it is still of great importance to call for the development of broad-spectrum and effective small molecule drugs to combat both current and future outbreaks triggered by SARS-CoV-2. Cathepsin L (CatL) cleaves the spike glycoprotein (S) of SARS-CoV-2, playing an indispensable role in enhancing virus entry into host cells. Therefore CatL is one of the ideal targets for the development of pan-coronavirus inhibitor-based drugs. In this study, a CatL enzyme inhibitor screening model was established based on fluorescein labeled substrate. Two CatL inhibitors IMB 6290 and IMB 8014 with low cytotoxicity were obtained through high-throughput screening, the half inhibition concentrations (IC₅₀) of which were 11.53 ± 0.68 and 1.56 ± 1.10 μmol·L^-1, respectively. SDS-PAGE and cell-cell fusion experiments confirmed that the compounds inhibited the hydrolysis of S protein by CatL in a concentration-dependent manner. Surface plasmon resonance (SPR) detection showed that both compounds exhibited moderate binding affinity with CatL. Molecular docking revealed the binding mode between the compound and the CatL active pocket. The pseudovirus experiment further confirmed the inhibitory effects of IMB 8014 on the S protein mediated entry process. In vitro pharmacokinetic evaluation indicated that the compounds had relatively good drug-likeness properties. Our research suggested that these two compounds have the potential to be further developed as antiviral drugs for COVID-19 treatment.

Traditional Chinese ancient prescriptions have been used for treatment of liver cancer for a long history and the scientific and rational compatibility is a great wealth for modern research and development (R&D) of new drugs. The research and development of new drugs are often accompanied with a large investment, a long cycle and a high risk, especially for the anti-tumor drugs R&D which are facing more risks and lower successful rate. In this research, the regularity of compatibility of drugs was analyzed from 124 anti-hepatoma ancient prescriptions by computer program. The results can offer help to the R&D of anti-hepatoma new drugs and reduce the risk of drug screening. In addition, we surveyed 22 companies in this field from six provinces such as Beijing, Shanghai, Tianjin and so on and obtained 240 risk assessment questionaires. Then we used qualitative analysis method to interpret the greatest impacts for the risks in the process of R&D, production and sales of anti-hepatoma new drugs. The study provides a basis for anti-liver cancer drugs R&D researchers, who can take effective measures to reduce the R&D risks and improve successful rate.
Carcinoma, Hepatocellular ; drug therapy ; history ; China ; Drug Discovery ; history ; Drug Incompatibility ; Drug Prescriptions ; history ; Drugs, Chinese Herbal ; history ; therapeutic use ; History, Ancient ; Humans ; Liver Neoplasms ; drug therapy ; history ; Research ; history

Abstract: Objective　By collecting and sorting the information of varicella cases reported in Liaoning Province from 2006 to 2021, the epidemiological characteristics were analyzed, and the monthly incidence data were predicted, so as to explore the prevention and control strategy of varicella disease in Liaoning Province. Methods　By collecting the characteristic information of varicella cases in Liaoning Province, epidemiological analysis was carried out on the regional, population, and temporal characteristics of varicella incidence. The monthly incidence data of varicella were fitted with Eviews software, seasonal ARIMA model was used for modeling, and models were selected according to SC and AIC. After modeling, the model was used to predict the incidence data in 2022. Results　The incidence rate of varicella in Liaoning Province has increased in recent years. The onset time was "bimodal distribution", with the main peak occurring from November to January of the next year and the secondary peak occurring from May to June. Since 2019, the onset age has shifted backward. From the original 0-<10 age group with the highest incidence rate, it shifted to the 10-<20 age group with the highest incidence rate. From 2006 to 2021, the incidence of varicella mainly concentrated in people aged 0 to <40 years old, and the incidence rate of the population over 40 years old showed a cliff-like decline. The incidence of chickenpox was higher in the central region of Liaoning Province, such as Shenyang, Dalian, Anshan and Panjin, and relatively low in Huludao, Jinzhou, Fuxin and Liaoyang. The distribution of the population was mainly students, followed by kindergartens and scattered children. ARIMA model of monthly incidence data was established by software as ARIMA (1, 0, 1) (1, 1, 1)12. Conclusions　The incidence rate of varicella in Liaoning Province has been rising in recent years. The incidence is obviously seasonal, and the age group of the affected population has moved backward. It is predicted that the incidence will continue to increase in 2022. The prevention and control of varicella should still be the current key work. In order to reduce the population incidence rate, two-dose vaccination strategies should be vigorously promoted the implementation of the, and the inclusion of varicella vaccine in the immunization program should be achieved as soon as possible.

This study is to explore new lead compounds by inhibition of Pin1 for anticancer therapy using temperature sensitive mutants. As Pin1 is conserved from yeast to human, we established a high-throughput screening method for Pin1 inhibitors, which employed yeast assay. This method led to the identification of one potent hits, 8-11. In vitro, 8-11 inhibited purified Pin1 enzyme activity with IC50 of (10.40 +/- 1.68) micromol x L(-1), induced G1 phase arrest and apoptosis, showed inhibitory effects on a series of cancer cell proliferation, reduced Cyclin D1 expression, was defined as reciprocally matched for protein-ligand complex in virtual docking analysis and reduced cell migration ability. In vivo, we could observe reduction of tumor volume after treatment with 8-11 in xenograft mice compared with vehicle DMSO treatment. Altogether, these results provide for the first time the involvement of 8-11 in the anticancer activity against Pin1.
Animals ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cyclin D1 ; metabolism ; Drug Screening Assays, Antitumor ; methods ; G1 Phase ; High-Throughput Screening Assays ; methods ; Humans ; Mice ; NIMA-Interacting Peptidylprolyl Isomerase ; Neoplasms ; pathology ; Peptidylprolyl Isomerase ; antagonists & inhibitors ; Temperature ; Xenograft Model Antitumor Assays ; Yeasts

OBJECTIVETo evaluate the impact of nicotine- and tar-free cigarette smoke extract (fCSE) on the serum testosterone (T) level and erectile function of male rats.

METHODSWe randomized 30 male SD rats to three groups of equal number to receive subcutaneous injection of PBS (1.0 ml / 300 g body weight per day), fCSE (1.0 ml/300 g body weight per day), and reduced glutathione hormone (GSH, 200 mg per kg body weight per day) in addition to fCSE (fCSE + GSH), respectively, all for 8 weeks. Then we evaluated the erectile function of the rats by measuring the maximal intracavernous pressure (MICP), mean arterial pressure (MAP), ICP/MAP ratio, time of stimulation to MICP (Tmax), and cavernosal filling fate (CFR). We determined the serum T level, the activities of superoxide dismutase (SOD) , malondialdehyde (MDA), and nitric oxide synthase (NOS) in the cavernosal tissue, and also observed the morphological changes of the corpus cavernosum.

RESULTSCompared with the controls, the rats of the fCSE group showed obvious decreases in the levels of serum T ([5.37 ± 1.43] vs [3.22 ± 1.11] μg/L), NOS ([2.90 ± 0.27] vs [1.67 ± 0.18] U/mg) , and SOD ([18.41 ± 1.09] vs [13.36 ± 1.18] U/mg prot) and erectile function-related indexes MICP ([85.92 ± 6.36] vs [58.99 ± 10.76] mmHg), MICP/MAP (0.86 ± 0.09 vs [0.56 ± 0.08]), and CFR (2.14 ± 0.44 vs 0.89 ± 0.44), but markedly increased Tmax ([29.90 ± 5.78] vs [42.90 ± 8.56]s), with a positive correlation between the serum T level and CFR (r = 0. 364, P < 0.05). Masson staining revealed a lower ratio of the corpus cavernosum smooth muscle tissue to collagen fiber in the fCSE group (0.27 ± 0.04) than in the control (0.98 ± 0.12). Compared with the fCSE group, the fCSE + GSH group exhibited significantly improved MICP ([58.99 ± 10.76 ] vs [77.95 ± 7.71] mmHg), MICP/MAP (0.56 ± 0.08 vs 0.77 ± 0.09), and CFR (0.89 ± 0.44] vs 1.76 ± 0.42) and shortened Tmax ([42.90 ± 8.56 ] vs [32.10 ± 5.84 ] s). The ratio of the corpus cavernosum smooth muscle tissue to collagen fiber was higher in the fCSE + GSH than in the fCSE group (0.77 ± 0.09 vs 0.27 ± 0.04) but still lower than in the control (0.98 ± 0.12).

CONCLUSIONNicotine- and tar-free cigarette smoke extract reduces the serum T level and erectile function of rats, which is related to oxidative stress. Antioxidant therapy can improve erectile function but has a limited value for morphological protection of the penile tissue.

Animals ; Erectile Dysfunction ; chemically induced ; Male ; Malondialdehyde ; metabolism ; Muscle, Smooth ; pathology ; Nicotine ; Nitric Oxide Synthase ; metabolism ; Penile Erection ; drug effects ; Penis ; pathology ; Rats ; Rats, Sprague-Dawley ; Smoke ; adverse effects ; Superoxide Dismutase ; metabolism ; Tars ; Tobacco ; adverse effects

Animals ; Fatty Liver ; drug therapy ; metabolism ; prevention & control ; Liver ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism ; Taurine ; therapeutic use

OBJECTIVETo observe the effects of diethlhexyl phthalate (DEHP) on lipid peroxidation and the life span in Drosophila melanogaster.

METHODSFed Drosophila with the concentration 0.20% DEHP of exposure after 0, 14, 28 days, the activity of total superoxide dismutase (SOD), CuZn-SOD and the concentration of malondialdehyde were determined. At the same time, the longevity test was carried out to examine the effect of DEHP on the Drosophila's lifespan.

RESULTSThe lifespan of Drosophila was shortened in a dose of DEHP exposed groups. The indexes of mean life span (MLS), 50% lethal time and mean maximum life span in three DEHP-treated groups (concentration of 0.05%, 0.10% and 0.20%) were lower than those of the controlled group respectively (P < 0.01 or P < 0.05). The MLS of both Drosophila sexes were reduced from the control of 64 days and 59 days to the test 60 days-52 days and 54 days-49 days respectively. DEHP decreased the activity of SOD (P < 0.01 or P < 0.05), and lead to a time-dependent relation and an increase in the concentration of malondialdehyde (P < 0.01 or P < 0.05) in the DEHP-exposed Drosophila groups.

CONCLUSIONDEHP might promote the process of lipid peroxidation and shorten the life span in Drosophila melanogaster. It should be one of the reasons in the senescence of Drosophila.

Animals ; Diethylhexyl Phthalate ; pharmacology ; Drosophila melanogaster ; drug effects ; growth & development ; metabolism ; Female ; Lipid Peroxidation ; drug effects ; Longevity ; drug effects ; Male ; Malondialdehyde ; metabolism ; Superoxide Dismutase ; metabolism ; Time Factors

Endoplasmic reticulum (ER) stress occurs in macrophage-rich areas of advanced atherosclerotic lesions and contributes to macrophage apoptosis and subsequent plaque necrosis. The purpose of the present study was to investigate the effects of caveolin-1 (Cav-1) on ER stress-induced apoptosis in cultured macrophages and the underlying mechanisms. RAW264.7 cells were incubated with thapsigargin (TG) to establish ER stress model. And Cav-1 expression was detected by Western blot. After being pretreated with filipin(III), a caveolae inhibitor, RAW264.7 cells were assayed with flow cytometry and confocal laser scanning microscopy to detect cell apoptosis. Moreover, p38 mitogen-activated protein kinase (MAPK) phosphorylation and C/EBP homologous protein (CHOP) expression were detected with Western blot. The results showed that Cav-1 expression was markedly increased at early stage of TG treatment (P < 0.05) and then decreased with prolonged or high dose TG treatments. The increasing of Cav-1 expression induced by TG in RAW264.7 cells was abolished under inhibition of caveolae by filipin(III) (P < 0.05). The effect of TG on apoptosis of RAW264.7 cells was further augmented after pretreatment with filipin(III) (P < 0.05). Western blotting showed that MAPK phosphorylation induced by TG was inhibited by filipin(III) in RAW264.7 cells (P < 0.05), whereas CHOP remained unchanged (P > 0.05). These results suggest that Cav-1 may play a critical role in suppressing ER stress-induced macrophages apoptosis in vitro, and one of the mechanisms may be correlated with the activation of p38 MAPK prosurvival pathway.
Animals ; Apoptosis ; drug effects ; Caveolin 1 ; genetics ; metabolism ; Cell Line ; Endoplasmic Reticulum Stress ; physiology ; Filipin ; pharmacology ; MAP Kinase Signaling System ; Macrophages ; cytology ; drug effects ; Mice ; Thapsigargin ; pharmacology ; Transcription Factor CHOP ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

OBJECTIVETo study the effect of acupuncture on tumor and its mechanism.

METHODSLiver cancer, gastric cancer and hypodermic tumor rat models were made by implantation of replicated Walker-256 cell strain. The 3 model rats were respectively divided into two groups at random, a model control group and an electroacupuncture group. The electroacupuncture groups were treated with electroacupuncture (EA) at "Zusanli" (ST 36), "Hegu" (LI 4) and "San-yinjiao" (SP 6), once each day, 15 min one session, for 15 days. The gross tumor volume and the tumor inhibitory rate, and the levels of humoral immunity index, including serum 1gG, IgM, IgA and C3, C4, and the levels of cellular immunity index, including CD4+, CD8+ and CD4+/CD8+ in the peripheral blood in each group were detected.

RESULTSThe gross tumor volumes in the EA groups were significantly smaller than those in the model control group (P<0.01). The contents of IgG, IgM and C3 in the EA groups increased significantly compared with those in the model control group (P<0.05 or P<0.01). The contents of IgA and C4 in the EA groups did not significantly change (P>0.05). The content of CD4+ and CD4+/CD8+ in the EA groups are significantly higher than those in the model control group (P<0.05 or P<0.01).

CONCLUSIONAcupuncture at "Zusanli" (ST 36), "Hegu" (LI 4) and "Sanyinjiao" (SP 6) can increase immune function and inhibit tumor growth in Walker-256 liver cancer, gastric cancer and hypodermic tumor rats.

Animals ; Carcinoma 256, Walker ; immunology ; pathology ; therapy ; Electroacupuncture ; Male ; Rats ; Rats, Wistar

The purpose of this study was to investigate the influence of electrical stimulation of anterior cingulate cortex (ACC) on spontaneous activity of neurons in thalamic ventrobasal nucleus (VB). Experiments were performed on 12 male Sprague-Dawley rats weighing 250-310 g (4-5 months old). According to Paxinos and Watson's coordinate atlas of the rat, the frontal and parietal cortical areas were exposed by craniotomy, the recording electrodes were then inserted into the VB (P 2.4-4.1 mm, R 2.0-3.5 mm, H 5.2-6.8 mm) and the stimulating electrodes into the ACC (A 1.1-3.0 mm, R 0.0-1.0 mm, H 1.5-2.4 mm). Single-unit activities were recorded extracellularly in the VB by glass micropipettes (impedance 3-8 MOmega) filled with 0.5 mol/L sodium acetate solution containing saturated Fast Green. To study the effects of ACC activation on the spontaneous activities of VB cells, single electrical pulse (0.2 ms duration) was delivered to the ACC by a concentric bipolar stainless steel electrode (0.32 mm outer diameter). An effective ACC stimulation was determined for each VB neuron by gradually increasing the current intensity from 0.1 mA until either a significant change in the spontaneous activity of the VB neuron was observed, or the current intensity reached 0.4 mA. The results showed that ACC stimulation significantly suppressed the spontaneous activities in 12 out of 53 VB neurons (22.6%). (1) After the stimulation was delivered to ACC, the spontaneous activities of different VB neurons were totally suppressed for different time span. (2) There was obvious dose-effect relevance between ACC stimulation intensity and their inhibitory effect. The duration of complete inhibition was prolonged with the increases in the intensity and number of stimulation impulses in ACC. (3) The stimulation in the ACC depressed the spontaneous activities of VB neurons in different forms and this inhibition exhibited an accumulative effect. All these results indicate that the stimulation of ACC exerts an inhibitory influence on the spontaneous activities of VB neurons.
Animals ; Electric Stimulation ; Gyrus Cinguli ; physiology ; Male ; Neurons ; cytology ; Rats ; Rats, Sprague-Dawley ; Thalamic Nuclei ; cytology