Cryptotanshinone (CPT), a lipid soluble active compound in Salvia miltiorrhiza, has a significant inhibitory effect on multiple malignant tumors, e. g. chronic myeloid leukemia (CML) cells and can effectively enhance imatinib's chemotherapeutic effect. However, its functional molecular mechanism remained unclear. In this experiment, the authors conducted a systematic study on the effect of CPT on the imatinib sensitivity and P-glycoprotein (P-gp) expression in CML cells by using CML cells K562 and imatinib persister K562-R. The MTT assays were performed to determine CPT's impact on the inhibitory effect of imatinib. Annexin V-FITC/PI staining analysis was used to detect the changes in the cell apoptosis rate. The active changes in apoptosis regulatory proteins Caspase-3, Caspase-9 and PARP were determined by Western blot. After the cells were pretreated with the gradient concentration of CPT, the expression of P-gp was analyzed by Western blot and flow cytometry. The changes in intracellular concentrations of imatinib were determined by HPLC analysis. The results indicated that the pretreatment with CPT significantly increased the proliferation inhibiting and apoptosis inducing effects of imatinib on K562 and K562-R cells as well as the degradation product expression of pro-apoptotic proteins Caspase-3, Caspase-9 and PARP, with a significant difference with the control group (P < 0.01). However, CPT showed no impact on the P-gp expression in CML cells and the intracellular concentrations of imatinib. In summary, the findings suggested that CPT enhanced the sensitivity of CML cells to imatinib. Its mechanism is not dependent on the inhibition in P-gp expression and the increase in intracellular drug concentration.
ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; metabolism ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; genetics ; metabolism ; Caspase 9 ; genetics ; metabolism ; Drug Resistance, Neoplasm ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Imatinib Mesylate ; pharmacology ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; drug therapy ; genetics ; metabolism ; physiopathology ; Phenanthrenes ; pharmacology

In order to develop a rapid detection kit for novel avian influenza virus (AIV) subtype H7N9, two sets of specific primers and probes were designed based on the nucleotide sequences of hemagglutinin antigen (HA) and neuraminidase antigen (NA) of novel H7N9 virus (2013) available in GenBank to establish the method of TaqMan probe-based multiplex real-time RT-PCR for rapid detection of AIV subtype H7N9. The primer and probe of HA were for all H7 subtype AIVs, while the primer and probe of NA were only for novel N9 subtype AIVs. The results showed that this method had high sensitivity and specificity. This method was applicable to the testing of positive standard sample with a minimum concentration of 10 copies/microL; it not only distinguished H7 subtype from H1, H3, H5, H6, and H9 subtypes, but also distinguished novel N9 subtype from traditional N9 subtype. A total of 2700 samples from Zhuhai, China were tested by this method, and the results were as expected. For the advantages of sensitivity and specificity, the method holds promise for wide application.
Animals ; Birds ; virology ; Influenza A Virus, H7N9 Subtype ; genetics ; isolation & purification ; physiology ; Influenza in Birds ; prevention & control ; virology ; Real-Time Polymerase Chain Reaction ; methods ; Species Specificity ; Taq Polymerase ; metabolism ; Time Factors

OBJECTIVETo evaluate the inhibitive effect of C-21 steroidal glycosides from the root of Cynanchum auriculatum (CGB) on rat glioma C6 cells.

METHODSC6 cells were treated with CGB for 24, 48,72 h at concentration of 30, 60, 120 mg/L, respectively. MTT assay was used for evaluating cell viability; fluorescence-activated cell sorting analysis after Annexin V/propidium iodide staining or single propidium iodide staining was used to test cell apoptosis and cell cycle.

RESULTSCGB at 30, 60, 120 mg/L concentration-dependently decreased C6 cell viability (P<0.001). CGB at 60 and 120 mg/L induced C6 cell apoptosis and cell cycle arrest. The fraction of G0/G1 cells was increased (P<0.05) and that of S phase cells was decreased (P<0.01).

CONCLUSIONCGB can inhibit the growth of rat glioma C6 cells, and induce apoptosis and G0/G1 cell cycle arrest.

Animals ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cynanchum ; chemistry ; Glioma ; pathology ; Monosaccharides ; pharmacology ; Rats ; Steroids ; pharmacology

In order to study the proliferation inhibition effect of H5N1 subtype avian influenza virus (AIV) with small interfere RNA (siRNA), a total of 4 siRNAs were designed in accordance with the NP and PA genes of H5N1 subtype AIV, the siRNAs were then transfected to chicken embryo fibroblast(CEF), CEF was infected with H5N1 subtype AIV after 6 hrs. Virus titer of cell supernatant was tested at 16-56hrs post infection, and pathological changes of the cells was observed; mRNA levels of NP, PA, HA and p13-actin gene were tested at 36hrs post infection. The results showed that these 4 siRNAs could inhibit the prolif-eration of H5N1 subtype AIV in CEF in varying degrees, and one siRNA targeting PA was best per-formed. The experimental results also showed that the inhibition effect was decreased with the time prolonged. This research provides a basis for further studying RNAi on AIV prevention and control.
Actins ; genetics ; Animals ; Chick Embryo ; DNA Primers ; genetics ; Fibroblasts ; virology ; Hemagglutination ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Hemagglutinins ; genetics ; Humans ; Influenza A Virus, H5N1 Subtype ; genetics ; growth & development ; physiology ; RNA Interference ; RNA Replicase ; genetics ; RNA, Small Interfering ; chemical synthesis ; genetics ; RNA-Binding Proteins ; genetics ; Real-Time Polymerase Chain Reaction ; Specific Pathogen-Free Organisms ; Transfection ; Viral Core Proteins ; genetics ; Viral Proteins ; genetics ; Virus Replication

AIMTo explore the effects of calcitonin-gene-related peptide (CGRP) on LPS-induced MMP-9 secretion by alveolar macrophages (AM) in vitro.

METHODSThe supernatant of LPS-induced Wistar rat AM from different intervention groups were collected to measure the activity by gelatin zymography.

RESULTS(Only secreting a small amount of MMP-9 with unstimulated AM, LPS stimulated MMP-9 production in a concentration-dependent manner (p < 0.01). (2) The activity of MMP-9 in CGRP intervention groups at different levels were significantly lower than those in non-intervention group (p < 0.01). (3) The inhibiting effects of CGRP were diminished by H-7 and W-7, an antagonist of protein kinase C (PKC) and calmodulin (CaM) (p < 0.05).

CONCLUSIONThese data suggested that CGRP involved in the MMP-9 secretion by AM, partly, via PKC and CaM pathway.

Animals ; Cells, Cultured ; Female ; Lipopolysaccharides ; adverse effects ; Macrophages, Alveolar ; secretion ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Rats ; Rats, Wistar ; Receptors, Calcitonin Gene-Related Peptide ; metabolism

OBJECTIVETo study the current situation and the cause of schistosomiasis resurgence in order to provide reference for formulation of control strategy.

METHODSData in 1999 - 2003 and baseline data in some areas were collected and analyzed retrospectively.

RESULTSResurgence was seen in 6.15% (16/260) of the areas and one farm where transmission of schistosomiasis had been interrupted and 33.33% (21/64) of the areas already under control. Snails appeared to have been rebounded only in six counties (farm) while in thirty two counties that rebound was seen in both snails and disease prevalence. Tendency of increase in the total numbers of patients, acute patients and cattle with schistosomiasis, areas with snails were seen from 1999 to 2003.

CONCLUSIONSEnvironmental, ecological, societal factors such as flood, acequia, lack of expenditure and lack of incentives at work etc. contributed to the resurgence of epidemics in those areas that criteria had been reached. Surveillance and supervision on the sources of infection and snail diffusion, especially in the areas where the transmission of schistosomiasis had already been under control.

Animals ; Cattle ; China ; epidemiology ; Communicable Diseases, Emerging ; epidemiology ; prevention & control ; Disasters ; Disease Reservoirs ; Ecology ; Humans ; Prevalence ; Retrospective Studies ; Risk Factors ; Schistosomiasis japonica ; drug therapy ; epidemiology ; prevention & control ; Snails ; parasitology

OBJECTIVETo analyze the change of tendency on schistosomiasis epidemics in China in the last 5 years.

METHODSData on schistosomiasis epidemics in the history and particularly in the last 5 years were collected. Tendency and the re-emerging status after 1998 were analyzed.

RESULTSData in 2003 showed that in 42%, 40% and 53% of the provinces, counties and townships with epidemics, the transmission of the disease has been interrupted or controlled. The number of estimated patients of schistosomiasis and areas with snails were also reduced by 92.74% and 73.56%, in 2003. The annual estimated number of chronic cases was around 800 000 and 31 321.5 hectare of snail infested areas were newly identified in recent 5 years. Among 20 national villages under longitudinal surveillance, 30%, 70% and 35% of the villages were presented a tendency of increase in the rates of human infection, bovine infection and Oncomelania snails infection, respectively. A total of 38 counties from 7 provinces have re-emerged in schistosomiasis transmission after those counties having reached criteria of transmission under control or interrupted. In 6 non-endemic counties, snails were presented, and 16 marshlands in Xan river were found with appearance of acute cases of schistosomiasis. More snail infested areas were found in Shanghai, Zhejiang, and Fujian. Both snail infested areas and newly infected cases were occurred in urban areas along the Yangtze River.

CONCLUSIONThe tendency of increase was presented in focal areas along the Yangtze River, due to changes of environmental, ecological, societal and economic status, as well as on the forces of control.

Animals ; China ; epidemiology ; Disease Reservoirs ; Humans ; Praziquantel ; therapeutic use ; Prevalence ; Schistosomiasis japonica ; drug therapy ; epidemiology ; prevention & control ; Snails ; parasitology

Carcinoma, Hepatocellular ; drug therapy ; Chemotherapy, Adjuvant ; Hepatic Artery ; Humans ; Liver Neoplasms ; drug therapy ; Postoperative Period ; Randomized Controlled Trials as Topic ; Treatment Outcome

OBJECTIVEThe severe acute respiratory syndrome (SARS) is a highly contagious infection caused by a newly discovered strain of coronavirus (SARS-CoV). During the outbreak of SARS in the first half of 2003, children appeared to be less susceptible to the SARS coronavirus and pediatric patients presented with a less aggressive clinical course than adult patients did, demonstrating the traits which were rarely observed in other viral contagious disease. The present study aimed to preliminarily examine the presence of serum specific antibodies against severe acute respiratory syndrome (SARS)-associated coronavirus virus (SARS-CoV) in pediatric SARS patients and explore the possibility of subclinical infection in children/adults through close association with SARS cases.

METHODS(1) Clinicians and nurses visited families and collected general and epidemiological information about the subjects using a standard questionnaire and took serum specimens. (2) Specific antibodies against SARS-CoV were assayed with two methods, indirect immunofluorescence assay (IFA) for detecting IgG antibodies and enzyme linked immunosorbent assay (ELISA) for mixed antibodies. Serum specimens tested included those from 21 clinically confirmed pediatric SARS cases (aged from 8 months to 14 years, 11 male and 10 female) and their 23 parents who had close contact with the children, 36 adult patients in convalescence stage of SARS, 24 children (aged 1.5 to 14 years) and other 34 adults who had close contact with infected adults.

RESULTS(1) The positive rates of specific IgG and mixed antibodies against SARS-CoV were 38% (8/21) and 33% (7/21) in pediatric cases; whereas the rates were 75% (27/36) and 69% (25/36) in adult patients. (2) The proportion of the patients who had close contact to SARS patients was 7/8 among the antibody-positive group vs. 1/13 for the antibody-negative group (P < 0.05). (3) The IgG antibody emerged in one of 24 children, whose mother, a nurse, had suffered from SARS (4%). (4) Among 23 parents of children with SARS, one was positive for IgG and the mixed antibodies, whose grandson and husband suffered from SARS; The IgG antibody and the mixed antibodies were also positive in another adult who had close contact with adult SARS cases (3%).

CONCLUSIONS(1) SARS-CoV infection was confirmed by serological methods in 38.1% of clinically diagnosed pediatric SARS cases, which leads to the assumption that correct diagnosis of pediatric SARS requires more accurate and efficient ways, for example, screening for antigen or gene of SARS-CoV. (2) The proportion of the patients who had close contact to SARS patients among antibody-positive cases was higher than that in antibody-negative cases. (3) It is possible that subclinical SARS CoV infection exists in children and adults, although the rate of occurrence is low. The data of the present study did not confirm that SARS had subclinical infection among adults who had close contact to pediatric SARS cases.

Adolescent ; Adult ; Antibodies, Viral ; blood ; immunology ; Child ; Child, Preschool ; Enzyme-Linked Immunosorbent Assay ; Female ; Fluorescent Antibody Technique, Indirect ; Humans ; Immunoglobulin G ; blood ; immunology ; Infant ; Male ; SARS Virus ; immunology ; Severe Acute Respiratory Syndrome ; blood ; epidemiology ; immunology

Objective To explore the mechanism and protective effect of ulinastatin on vascular endothelial cells ( VEC) in sepsis rats.Methods Forty SD male rats were randomly divided into four groups:sham oper-ation group, model group and low and high dose test groups ( ulinastatin, 10 ×104 ,20 ×104 U? kg -1 ) .The sepsis rat models were established with cecal ligation and puncture ( CLP).The level of tumor nuclear fac-tor-α( TNF-α) , interleukin 1β( IL-1β) and IL-6 in serum was assayed by enzyme linked immunosorbent assay method.The level of ni-trogen ( NO) and inducible nitric oxide synthase ( iNOS) was determina-ted by kits.VEC apoptosis was detected by flow cytometry.The activity of cysteinyl aspartate specific proteinase 3 ( Caspase 3) was measured by spectrophotometry.The expression of B-cell lymphoma-2 ( Bcl-2 ) , Bcl-2 associated X protein ( Bax ) and the activation of nuclear factor kappa B ( NF -κB ) signal pathway were assayed by Western blot. Results Compared with sham operation group, the level of TNF-α, IL-1β, IL-6, NO, iNOS was higher, the number of apoptotic VEC was higher, the activity of Caspase 3 and the expression of Bax was higher, the expression of Bcl-2 was lower, the phosphorylation of NF -κB p65 and IκBαwas higher in the model group, the difference was significant ( all P<0.01) .Compared with model group, low and high dose test groups could inhibit the change, the difference was significant (P<0.05).The ratio of Caspase 3 activity was (1.00 ±0.21,3.70 ±0.25,1.83 ±0.14,1.53 ±0.13). Conclusion These results suggested ulinastatin inhibited the injury of VEC in sepsis rats via resisting to inflammation and VEC apoptosis, which might be associated with NF-κB signal pathway.