We designed and synthesized eighteen lycorine derivatives with five different structural types, and evaluated their antiviral activities on a HCoV-OC43-infected H460 cell model. Structure-activity relationships suggested that the introduction of appropriate substituents on the 6N atom of lycorine was beneficial to activity. Compound 6a gave a good activity with the half effective concentration (EC₅₀) and selectivity index (SI) values of 2.36 μmol·L^-1 and 16.52, respectively. Surface plasmon resonance (SPR) result indicated that 6a might target the non-structural protein 12 (NSP12) subunit in RNA-dependent RNA polymerase (RdRp) of SARS-CoV-2 with the dissociation constant (K_D) value of 1.36 μmol·L^-1. Molecular docking indicated that 6a might act on nidovirus RdRp-associated nucleotidyltransferase (NiRAN) catalytic center of NSP12, distinct from the mechanism of nucleoside-like drugs such as remdesivir. This study provides scientific data for the development of lycorine derivatives into a new class of anti-SARS-CoV-2 small molecule inhibitors.

In this study, we designed and synthesized 12 novel aloperine derivatives with different core structures. Among them, compound 3 with a ten-membered ring core was obtained through a special ring expansion reaction after γ-H Huffman elimination of quaternary ammonium salt, and the structure was verified by X-single crystal diffraction. Furthermore, their antiviral activity against human β-coronavirus HCoV-OC43 was evaluated by CCK-8 assay. Quaternary ammonium salt 2a and 3 had a good inhibitory effect against HCoV-OC43, and 2a had the highest anti-HCoV-OC43 activity with an EC₅₀ values of 3.77 μmol·L^-1 and a SI value of over 53.1. Schrӧdinger molecular docking results showed that both 2a and 3 might display their anti-HCoV-OC43 activity by directly acting on host TMPRSS2 and SR-B1. The results expanded the structural types of endocyclic aloperine and the function against coronavirus, and provided useful scientific data for the development of pharmaceutical applications of these compounds.

Aim To investigate the protective effect of bicyclol on tumor necrosis factor-α/D-galactosamine (TNF-α/D-GalN) induced liver injury and its possible mechanisms. Methods Male C57 mice were given 50, 100, and 200 mg · kg

The level of intracellular keratin 8(KRT-8) is associated with liver diseases, whose expression is increased in hepatitis C virus (HCV)-infected patients with hepatocarcinoma and in cultural cells infected with HCV. However, it is not clear whether KRT-8 will impact HCV replication. In this paper, the HCV replication was analyzed in response to high expression and silence of KRT-8. The inhibitory activities against wild-type and mutant HCV were also analyzed by silence of KRT-8 or combined with known anti-HCV drug telaprevir. Results showed that the protein level of KRT-8 was increased in proportion with the HCV replication. The high expression was found to facilitate HCV replication, while the silence of KRT-8 was able to inhibit HCV replication and enhanced the anti-HCV activity of telaprevir. It also inhibited A156T and D168V mutant HCV, which are resistant to protease inhibitors. These results suggest that KRT-8 is a co-factor for HCV replication. Down-regulation of KRT-8 can inhibit wild type and mutant HCV replication to enhance the anti-HCV activity of known anti-HCV drugs. Therefore, KRT-8 may be a new target in the development of anti-HCV agents.

APOBEC3 is a class of cytidine deaminase, which is considered as a new member of the innate immune system, and APOBEC3G belongs to this family. The research about APOBEC3G is a new direction of innate immune defense mechanism against virus. APOBEC3G has the restrictive activity on many viral replications, which deaminates dC to dU in the viral genome and then induces extensive hypermutation. APOBEC3G can also interrupt viral replication at several phases such as reverse transcription, replication, nucleocapsid and so on by non-deamination mechanisms. However, virus can encode viral proteins to counteract the restriction activity of APOBEC3G. Elucidation of the antagonistic interaction between APOBEC3G and the virus will be contributed to development of new antiviral drugs in the future.
APOBEC-3G Deaminase ; Animals ; Cytidine Deaminase ; genetics ; metabolism ; DNA Replication ; Deamination ; HIV-1 ; physiology ; Hepacivirus ; genetics ; physiology ; Hepatitis B virus ; genetics ; physiology ; Humans ; Paramyxoviridae ; genetics ; physiology ; Retroviridae ; physiology ; Virus Replication ; vif Gene Products, Human Immunodeficiency Virus ; metabolism

OBJECTIVETo evaluate the efficacy and optimal re-implantation time of two-stage revision for management of periprosthetic infection following hip arthroplasty.

METHODSWe retrospectively analyzed the clinical data of 15 patients (15 hip joints) undergoing two-stage ipsilateral total hip arthroplasty (THA) revision from January, 2006 to January, 2010. In the first stage, after surgical debridement and thorough removal of all the implants, a self-made Vancomycin-loaded cement spacer was implanted. The second stage operation was performed 3-6 months later for debridement and removal of the antibiotic-loaded spacer, followed by re-implantation of Vancomycin-loaded bone cement prosthesis in 9 cases and cementless prosthesis in 6 cases. The patients were followed up for 9-46 months (mean 25 months) after the operation.

RESULTSNo reinfection or prosthesis loosening/displacement was found in these cases after the operation. The Harris score increased from 40.3 before the operation to 54.0 after the first-stage operation, and to 88.2 at the last follow-up.

CONCLUSIONTwo-stage revision is effective for treatment of periprosthetic infection following hip arthroplasty, and 3-6 months can be the optimal interval between the two the first-stage and second-stage operation for re-implantation.

Adult ; Aged ; Arthroplasty, Replacement, Hip ; methods ; Female ; Hip Prosthesis ; Humans ; Male ; Middle Aged ; Prosthesis-Related Infections ; surgery ; Reoperation ; Retrospective Studies ; Treatment Outcome

Plant active components characterized of many different structures and activities on multiple targets, have made them to be the important sources of inhibitors on HIV-1. For finding leading compounds with new structure against HIV-1, three key HIV-1 replicative enzymes (reverse transcriptase, protease and integrase) were used as screening models. The in vitro activities of 45 plant derived components isolated from Schisandraceae, Rutaceae and Ranunculaceae were reported. Within twelve triterpene components isolated, eight compounds were found to inhibit HIV-1 protease, in these eight active compounds, kadsuranic acid A (7) and nigranoic acid (8), inhibited both HIV-1 protease and integrase; Among fifteen lignans, meso-dihydroguaiaretic acid (15) and kadsurarin (16) were active on HIV-1 reverse transcriptase, and 4, 4-di(4-hydroxy-3-methoxyphenly)-2, 3-dimethylbutanol (13) active on HIV-1 integrase. All of the six alkaloids, seven flavones, and five others compounds were not active or only with low activities against HIV-1 replicative enzymes. Further studies of the triterpene components showing strong inhibitory activities on HIV-1 were warranted.
Alkaloids ; chemistry ; isolation & purification ; pharmacology ; Anti-HIV Agents ; chemistry ; isolation & purification ; pharmacology ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Flavones ; chemistry ; isolation & purification ; pharmacology ; Guaiacol ; analogs & derivatives ; chemistry ; isolation & purification ; pharmacology ; HIV Integrase ; drug effects ; HIV Protease ; drug effects ; HIV Reverse Transcriptase ; antagonists & inhibitors ; Lignans ; chemistry ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; Ranunculaceae ; chemistry ; Rutaceae ; chemistry ; Schisandraceae ; chemistry ; Triterpenes ; chemistry ; isolation & purification ; pharmacology

For obtaining new structural compounds with unique resistance profiles or novel mechanisms of action on HIV-1 from natural products, anti-HIV-1 drug screening models were used in vitro. Norcantharidin (NCTD), a derivative from cantharidin, was found to have inhibitory activities on HIV-1(IIIB) p24 antigen in lymphocyte lines MT-4, CEM and H9. It inhibited HIV-1 strain 018a (sensitive to zidovudine) from replicating with EC50 (50% effective concentration) of 14.9 micromol L(-1) and also inhibited HIV-1 strain 018c (resistant to zidovudine) from replicating with EC50 of 20.2 micromol L(-1) in primary lymphocytes peripheral blood mononuclear cells (PBMC). Norcantharidin showed synergistic activity with zidovudine on HIV-1(IIIB) in MT-4 cells, the combination index was less than 0.3. But, it was not active on HIV-1 integrase, reverse transcriptase or protease in vitro. As the structure of norcantharidin is unique and different from that of all clinic drugs approved, it would be possible to obtain new and effective compounds against HIV-1 with low toxicities after modification of norcantharidin.
Anti-HIV Agents ; pharmacology ; Bridged Bicyclo Compounds, Heterocyclic ; pharmacology ; Cell Line ; Drug Resistance, Viral ; Drug Synergism ; HIV Core Protein p24 ; metabolism ; HIV Integrase ; metabolism ; HIV-1 ; metabolism ; Humans ; Leukocytes, Mononuclear ; cytology ; virology ; Peptide Hydrolases ; metabolism ; RNA-Directed DNA Polymerase ; metabolism ; T-Lymphocytes ; cytology ; virology ; Virus Replication ; Zidovudine ; pharmacology

OBJECTIVETo explore and evaluate the activities of composite extract from Salvia Yunnanensis and in cell cultures (DS-MEF) for inhibition of human immuno-deficiency virus type 1 (HIV-1) in vitro and in cell cultures.

METHODSThe inhibitory activity of DS-MEF on HIV-1 reverse transcriptase (RT), protease (PR) and integrase (IN) were detected in vitro with radionuclide 3H incorporation, fluorescence assay and enzyme-linked immunosorbent assay respectively. The human T-lymphocyte MT-4 cell line, human T-lymphocyte H 9 cell line chronically infected with HIV-1 IIIB, and the fresh peripheral blood mononuclear cell (PBMC) of healthy persons as well as the laboratory passed HIV-1 IIIB and the clinically isolated HIV-1 AZT sensitive 018a or resistant 018c infected cell cultures were used for evaluating the cytotoxicities and inhibitory activities of DS-MEF on HIV-1 P 24 antigen. The acute toxicities of DS-MEF on KM mice were determined by gastric gavages and intraperitoneal injections with various dosages.

RESULTSThe IC50 of DS-MEF for inhibiting HIV-1 IN, RT and PR were 2.59 +/- 0.50 mg/L, 27.39 +/- 11.18 mg/L and 9.38 +/- 2.45 mg/L respectively. In MT-4 cell cultures infected with HIV-1 III, TC50 were 13.19 +/- 6.07 mg/L, IC50 and SI of anti-HIV-1 activity were 0.224 +/- 0.163 mg/L and 58.7; in chronically infected H 9 cell cultures, TC50 were 18.11 +/- 9.84 mg/L, IC50 on HIV-1 P 24 antigen and SI were l7.230 +/- 21.114 mg/L and 1.1 respectively; TC50 in HIV-1 infected PBMC cultures were 288.70 +/- 0.08 mg/L; IC50 on AZT sensitive HIV-1 018a: 26.42 +/- 11.16 mg/L, and SI: 10.9; On AZT resistant HIV-1 018c, IC50: 27.87 +/-5.35 mg/L, and SI: 10.4. Moreover, DS-MEF showed synergistic effect with AZT or nevirapine (NVP) on HIV-1 IIIB in MT-4 cell cultures, the respective combination index was 0.78 or 0.67. DS-MEF showed no acute toxicity in KM mice with the dosage up to 20 g/kg via gastrogavage, and the 50% lethal dose (LD50) via intraperitoneal injection was 1.18 g/kg.

CONCLUSIONDS-MEF is a promising anti- HIV-1 agent with low toxicity in mice and possesses multi-targets and effective activities.

Animals ; Anti-HIV Agents ; pharmacology ; therapeutic use ; Cell Line ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; HIV Infections ; drug therapy ; virology ; HIV-1 ; drug effects ; enzymology ; physiology ; Humans ; Leukocytes, Mononuclear ; virology ; Male ; Mice ; Mice, Inbred BALB C ; Salvia ; chemistry ; T-Lymphocytes ; virology ; Viral Proteins ; antagonists & inhibitors ; Virus Replication ; drug effects

OBJECTIVETo select the optimal pregnancy time window of embryonic pig skin precursor tissue for xenotransplantation and study its ability in wound repair.

METHODSSkin precursor tissues were obtained from pig fetus of fetal age of 35, 42, 56, 70 days, and were minced into microskin and transplanted to dorsal wounds of BALB/c nude mice, then they were covered with residual skin after plastic surgery of patients or adult pig skin (white). The characteristics of growth and development were observed after transplantation. Pathological examination was performed on 6 and 12 post operation weeks respectively to observe the tissue structure and tumorigenicity.

RESULTSSkin precursor tissues from fetal pig survived and developed after transplantation, and the microskin fused. New tissue area from skin precursor tissues with fetal age of 42 days was (47 +/- 6) mm2, which was higher than that of 35 days (18 +/- 8 mm2), 56 days (31 +/- 12 mm2), 70 days (20 +/- 8 mm2, P < 0.05). The skin precursor developed into "intact skin" with hair, sebaceous glands and sweat glands, and melanocytes were also detected in epidermis. The newly-grown skin tissue included epidermal and dermal layer, and obvious dermal papillae. Teratoma was not found after transplantation in skin precursor tissue with fetal age of 56, 70 days.

CONCLUSIONFetal pig skin precursor tissue with fetal age of 56 days can be used to repair wound as xenotransplantation.

Animals ; Fetal Tissue Transplantation ; Fetus ; Gestational Age ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Skin Transplantation ; Swine ; Transplantation, Heterologous ; Wound Healing