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.

OBJECTIVETo evaluate the effect of adenovirus-mediated double suicide gene (CD/TK) for selective killing of breast cancer cells.

METHODSVascular endothelial growth factor (VEGF)-expressing MCF-7 cells and normal human mammary epithelial cells that did not express VEGF were infected with the adenovirus containing VEGFP-CD/TK-GFP genes. CD/TK gene expression in the infected cells was detected by RT-PCR. After treatment of the infected cells with GCV and/or 5-FC, the cell growth status was evaluated using MTT assay, and the cell cycle changes were detected with flow cytometry. In nude mice bearing human breast cancer, the recombinant adenovirus vector was injected directly into the tumor followed by intraperitoneal injection of the prodrugs GCV and/or 5-FC, and the subsequent tumor growth was observed.

RESULTSThe recombinant adenovirus achieved similar infection rates in MCF-7 and human mammary epithelial cells, and the rates increased gradually with the multiplicity of infection (MOI) of the virus. RT-PCR demonstrated the presence of CD/TK gene product in infected MCF-7 cells, but not in the infected mammary epithelial cells. The infected MCF-7 cells, but not the mammary epithelial cells, were highly sensitive to the pro-drugs. The CD/TK fusion gene system showed significantly greater efficiency than either of the single suicide gene in killing the target cells (P<0.01). At the MOI of 100, treatment of the infected cells with the pro-drugs resulted in increased cell percentage in G(0)-G(1) phase and decreased percentage in S phase. In nude mice bearing MCF-7 cell-derived subcutaneous tumor, treatment with the double suicide gene system significantly inhibited the tumor growth, showing much stronger effect than either of the single suicide gene (P<0.01).

CONCLUSIONThe adenovirus-mediated CD/TK double suicide gene driven by VEGF promoter combined with GCV and 5-FC treatment can be an effective therapy against experimental breast cancer, and produces much greater efficacy than the single suicide gene CD/TK combined with GCV or 5-FC.

Adenoviridae ; genetics ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Cytosine Deaminase ; genetics ; metabolism ; Female ; Flow Cytometry ; Flucytosine ; pharmacology ; Ganciclovir ; pharmacology ; Genes, Transgenic, Suicide ; genetics ; Genetic Therapy ; methods ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Recombinant Fusion Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Thymidine Kinase ; genetics ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo study the selective cytotoxic effect of lentivirus-mediated double suicide gene (CD/TK) against human gastric carcinoma cells SGC-7901 in vitro.

METHODSSGC-7901 cells were infected with FGW-KDRP-CD/TK vector and the infection efficiency was observed under a fluorescence microscope. The morphological changes of the infected cells were observed by Giemsa staining. Flow cytometry (FCM) was employed for cell cycle analysis, and the expression of CD/TK was detected by RT-PCR. The infected cells were then treated with the prodrugs ganciclovir (GCV) and/or 5-fluorocytosine (5-FC) at different concentrations, and the cytotoxic effects were evaluated using MTT method.

RESULTSThe infection efficiency of the lentiviral vector in SGC-7901 cells increased with the titer of the virus, which produced no significant effect on the cancer cell morphology in vitro or on the percentages of G0-G1, G2-M and S phase cells (P>0.05). RT-PCR demonstrated the expression of CD/TK gene in SGC-7901 cells infected by FGW-KDRP-CD/TK. The infected cells were highly sensitive to the prodrugs with a dose-dependent cytotoxic effect within a specific concentration range of the drugs, whereas the non-infected cells were not sensitive to the prodrugs. Combined use of the two prodrugs produced an obviously stronger inhibitory effect than either of the them (P<0.05). When combined, GCV and 5-FC at the concentration of 0.1+40, 1+80, 10+160, and 100+320 mg/L demonstrated a synergetic effect with a CDI<1.

CONCLUSIONLentivirus-mediated CD/TK fusion gene system can selectively kill gastric cancer cells, and the two prodrugs show a synergistic cytotoxic effect.

Adenocarcinoma ; genetics ; pathology ; Cell Line, Tumor ; Cytosine Deaminase ; biosynthesis ; genetics ; Cytotoxins ; pharmacology ; Genes, Transgenic, Suicide ; genetics ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; Lentivirus ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; Stomach Neoplasms ; genetics ; pathology ; Thymidine Kinase ; biosynthesis ; genetics ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; metabolism

OBJECTIVETo find a good way to diagnose VD, value the effect of Yishen Yangnao capsule on VD and try to find some rules of changes in Chinese medicine syndromes.

METHODPatients were randomly divided into treating group and western medicine comparison group. It's the phase III clinical research of Rishen Yangnao capsule curing VD, judging the validity and security of it, using dukexi slice as comparison drug. Some of the patients did the examination of P300.

RESULTThe total validity of Yishen Yangnao capsule is 56.3% (contract team is 60.0%). The improve rate of ADL is 0.1069% (contract team is 0.1134%). The scores of Chinese medicine syndrome descend.

CONCLUSIONYishen Yangnao capsule has the same effect as dukexi slice in curing VD at the side of intelligence situation and life ability.

Activities of Daily Living ; Aged ; Almitrine ; therapeutic use ; Capsules ; Dementia, Vascular ; drug therapy ; physiopathology ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; therapeutic use ; Event-Related Potentials, P300 ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Neuroprotective Agents ; isolation & purification ; therapeutic use ; Phytotherapy ; Treatment Outcome ; Yohimbine ; therapeutic use

OBJECTIVETo measure the degree of satisfaction on various environmental and health components and to discuss the impact of Olympic Games among the residents so as to make relative policy suggestions.

METHODSIn 2006, permanent residents over 15 years old lived in the Asian Games Village Community (where the 29th Olympic Games to be held) were selected to conduct a household's survey, while 1610 valid questionnaires were collected. The questionnaire included demographic information, degrees of satisfaction on various health-related environmental components, living condition and on Olympic events.

RESULTSThe top 4 aspects with the highest satisfaction rates were "overall rates of satisfaction on current life" "green space", "housing conditions" and "water quality", which were 50.43%, 48.59%, 38.95%, 37.08%, respectively. Residents' satisfaction on "impact of hosting the Olympic Games on China's international image", "China's economic development level", "living conditions" and "personal life" were 65.53%, 56.09%, 47.27%, 46.40%, respectively. Data from partial correlation analysis showed that the total scores of satisfaction on environment and life had positive correlation with the total scores of Olympic satisfaction (P < 0.05). The satisfaction degree on Olympic event through factor analysis showed that 10 entries of the Olympic impact could be reflected by two factors--the influence of image to the nation and impact on personal income. Logistic regression showed that the impact of Olympic Games on personal income, the impact of Olympic Games on the image of the nation and standard of living, gender, education level were independent influencing factors of the total scores of environment and life satisfaction (P < 0.05).

CONCLUSIONOther than "green space", most of health-related environment components of Beijing had low degree of satisfaction among inhabitants from the 'typical' communities in Beijing. However, residents had a higher degree of satisfaction on the impact of the Olympic Games to the country's image, the country's economic development level, the environment and personal standard of living.

Adult ; China ; Female ; Humans ; Life Style ; Male ; Middle Aged ; Personal Satisfaction ; Quality of Life ; psychology ; Social Environment ; Sports ; Surveys and Questionnaires

OBJECTIVETo study the selective killing effects of adenovirus (Ad)-mediated double suicide gene system driven by KDR promoter (KDR-CdglyTK) on the human hepatic carcinoma cells and human umbilical vein endothelial cells (HUVECs).

METHODSKDR-expressing BEL-7402 and HUVECs and HepG2 cells that did not express KDR were infected by KDR-CdglyTK, and the infection efficiency and the expression of CdglyTK in the cells was detected by RT-PCR. The infected cells were treated with the the prodrugs 5-FC and GCV at different concentrations, and the cell-killing effects and bystander effects were evaluated by MTT method.

RESULTSAt the multiplicity of infection (MOI) of 100, the recombinant AdKDR-CDglyTK showed similar infection efficiency in the 3 cell lines. RT-PCR demonstrated CDglyTK expression in the recombinant adenovirus and the 3 infected cell lines. BEL-7402 and HUVECs infected by the KDR-CdglyTK, but not the HepG2 cells, were highly sensitive to the prodrugs (P<0.001). Bystander effects of the double suicide gene system were observed in the coculture of the infected and non-infected BEL-7402 and HUVECs.

CONCLUSIONThe double suicide gene system driven by KDR promoter has specific killing effect on KDR-expressing hepatocellular carcinoma cells and HUVECs.

Adenoviridae ; genetics ; Apoptosis ; genetics ; Cells, Cultured ; Cytosine Deaminase ; genetics ; metabolism ; Endothelial Cells ; cytology ; Genes, Transgenic, Suicide ; genetics ; Genetic Therapy ; Genetic Vectors ; Humans ; Liver Neoplasms ; pathology ; Promoter Regions, Genetic ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Thymidine Kinase ; genetics ; metabolism ; Tumor Cells, Cultured ; Umbilical Veins ; cytology ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; metabolism

The effects of cephradinum and ceftazidime on the metabolism of Escherichia coli (E. coli) DH5alpha was determined by microcalorimetry. The microbial activity was recorded as power-time curves through an ampoule method with a TAM Air Isothermal Microcalorimeter at 37 degrees C. The parameters such as the growth rate constant (k), inhibitory ratio (I), the maximum power output (Pm) and the time (tm) corresponding to the maximum power output were calculated. The results show that the ceftazidime has a better inhibitory effect on E. coli DH5alpha than cephradinum.
Anti-Bacterial Agents ; administration & dosage ; pharmacology ; Calorimetry ; methods ; Ceftazidime ; administration & dosage ; pharmacology ; Cephradine ; administration & dosage ; pharmacology ; Escherichia coli ; drug effects ; growth & development ; metabolism ; Microbial Sensitivity Tests

A rice pse(t) (premature senescence, tentatively) mutant line, was isolated from 4,500 independent T-DNA inserted transgenic lines. The symptoms of premature senescence appeared more severely than those of the control plants (Zhonghua 11, japonica) at the last development stage. To characterize the mutant and provide basic information on the candidate genes by mapping to a physical region of 220-kb, experiments were carried out in two phytotrons under controlled temperature of 24 degrees C and 28 degrees C, respectively. The content of chlorophyll, soluble protein and MDA (malondialdehyde), net photosynthesis, the antioxidant enzyme activities of SOD (superoxide dismuase) (EC 1.15.1.1) and POD (peroxidase) (EC 1.11.1.7) and the peptidase activities of leaves were measured from top to bottom according to the leaf positions at the flowering stage. Compared with the control plant, the mutant showed the following characteristics: (1) Higher net photosynthesis rate (P(n)) appeared in the 1st and 2nd leaves, contents of chlorophyll and soluble protein were also higher in the 1st leaf; (2) The activities of SOD, POD and peptidase were higher according to the leaf position from top to bottom; (3) The symptom of premature senescence was accelerated in the mutant at 28 degrees C treatment. The MDA content and the SOD and POD activities between the 24 degrees C and 28 degrees C treatment mutants were not significantly different. Content of chlorophyll and soluble protein of leaves mutant decreased rapidly at 28 degrees C treatment. The results show that pse(t) is sensitive to high temperature. The probable function of PSE(T) is discussed.
Aging ; physiology ; Antioxidants ; metabolism ; Apoptosis ; physiology ; Mutation ; Oryza ; classification ; genetics ; growth & development ; Photosynthesis ; physiology ; Plant Leaves ; classification ; genetics ; growth & development ; Plant Proteins ; genetics ; metabolism

OBJECTIVETo analyze the clinical evaluation of Parkinson's disease (PD) patients receiving integrated Chinese and Western medicine therapy.

METHODSOne hundred and twenty patients were enrolled and randomly allocated to a control group or treatment group. Patients in the two groups received placebo and Bushen Huoxue Granule (, BHG), respectively. Both groups received baseline levodopa and benserazide (Madopar). The effects of treatment were assessed monthly during the 9-month treatment. Means of evaluation included Unified PD Rating Scale (UPDRS) scores (II and III), sleep scale score, 10 m turn back test (getting up time, 10 m×2 times, and turning time), timing motor test (TMT)-left and TMT-right, which were treated as the dependent variables; and age, sex, duration of PD, Hoehn and Yahr (H-Y) stage and Madopar dosage of admitted PD patients were as the independent variables. Multiple linear regression was used to analyze these factors.

RESULTSH-Y stage significantly affected UPDRS II score, UPDRS III score, and getting up time (P<0.01). Madopar dosage and H-Y stage significantly affected the 10 m×2 times (P<0.05 or <0.01). Madopar dosage significantly affected the sleep scale score (P<0.05). There were also significant correlations between age and TMT-left or TMT-right (P<0.01), and duration of PD and TMT-right (P<0.05).

CONCLUSIONSThe six assessed means of clinical evaluation (including UPDRS II and UPDRS III scores, sleep scale score, getting up time, 10 m×2 times, and turning time) are sensitive indexes in all PD patients. H-Y stage and Madopar dosage are the major factors influencing means of clinical assessment of PD treatment.

Aged ; Aged, 80 and over ; Female ; Humans ; Integrative Medicine ; Male ; Medicine, Chinese Traditional ; adverse effects ; Middle Aged ; Parkinson Disease ; physiopathology ; therapy ; Sleep ; Time Factors ; Treatment Outcome

OBJECTIVETo investigate the changes of antisperm antibodies (AsAb), sexual hormones, and inhibin B (INH B) in patients before and after testicular torsion, as well as the effects of these factors on testicular function and reproduction.

METHODSTen patients with single acute testicular torsion (left side 9 and right side 1), aged 16-45 years (19.6 on average), disease course of 3-6 days (averaging 4.7 days), underwent surgical removal of the damaged testis. Before and after the operation, serum AsAb (IgG, IgM, IgA) and INH B were measured by ELISA, and serum follicle-stimulating hormone (FSH), luteotropic hormone (LH), and testosterone (T) determined by chemoluminescence autoanalyzer.

RESULTSAfter the operation, the AsAb levels rose significantly and remained high for at least 26 weeks. The level of INH B was the lowest in the 3rd week and restored to normal in the 12th week, with significant difference between preoperation and the 3rd or the 6th week after the operation. The levels of LH and INH B in the 26th week were elevated significantly compared with the 6th.

CONCLUSIONTesticular injury induced the elevation of AsAb, which would last a very long time. The change of INH B was closely related with the injury of the testis, which reflected the degree of testicular injury and functional restoration of the patients after the operation. Our study showed that AsAb and INH B can be used as useful tools for monitoring testicular function and reproduction.

Adolescent ; Adult ; Autoantibodies ; blood ; Follicle Stimulating Hormone ; blood ; Humans ; Inhibins ; blood ; Luteinizing Hormone ; blood ; Male ; Middle Aged ; Orchiectomy ; Spermatic Cord Torsion ; immunology ; physiopathology ; surgery ; Spermatozoa ; immunology ; Testis ; physiopathology ; Testosterone ; blood