Objective To obtain antibiotic-resistant mutants producing metabolites with antitumor activity from wild-type actinomycete strains without antitumor activity. Methods An actinomycete strain L35-1 was used as an initial strain for obtaining antibiotic-resistant mutants, which is a marine-derived wild-type strain without antitumor activity with an inhibition rate of 2.8% at the 1000 μg/ml of high sample concentration on K562 cells. The antibiotic-resistant mutants both from auto-mutagenesis and chemical mutagen-induced mutagenesis were selected by single colony isolation on antibiotic-containing plates according to the method for obtaining drug-resistant mutants in ribosome engineering. The antitumor activity was assayed by the MTT method using K562 cells for the mutants with aqueous acetone extracts of the whole broth of their fermentation.Results A total of 114 neomycin-resistant (ner) and 68 streptomycin-resistant (str) mutants, all from auto-mutagenesis, was obtained on drug-containing plates. Among them, the 7 ner and 3 str mutants appeared to be bioactive with an inhibition rate above 20% at the 100 μg/ml sample concentration on K562 cells. On the other hand, 41 str and 32 ner mutants from DES-induced mutagenesis and 46 ner mutants from NTG-induced mutagenesis were obtained by mutagen-induced mutation coupled with the single colony isolation on antibiotic-containing plates, among which, one str mutant from DES-induced mutagenesis and one ner mutant from NTG-induced mutagenesis were bioactive with an inhibition rate over 20% at the 100 μg/ml sample concentration on K562 cells. Conclusions The present result has revealed that the wild-type actinomycete strains without bioactivity might become a great source initial strains to obtain bioactive mutants by drug-resistant mutation technique.

Adult ; Diagnosis, Differential ; Follow-Up Studies ; Humans ; Knee Joint ; Male ; Mucin-1 ; metabolism ; Neurilemmoma ; metabolism ; pathology ; surgery ; S100 Proteins ; metabolism ; Sarcoma, Synovial ; metabolism ; pathology ; Sweat Glands

This paper is to explore the reasonable applications of automatic exposure control in computed radiography, and to improve the quality of CR images. It is very important to select a suitable KV value in automatic exposure control in computed radiography. At the same time, a suitable ionization chamber, correct density compensation and necesary post-processing should be selected.
Adolescent ; Adult ; Aged ; Automation ; Child ; Child, Preschool ; Female ; Head ; diagnostic imaging ; Humans ; Image Processing, Computer-Assisted ; Infant ; Lumbar Vertebrae ; diagnostic imaging ; Male ; Middle Aged ; Quality Control ; Radiation Dosage ; Radiographic Image Enhancement ; methods ; Radiography, Thoracic ; Tomography, X-Ray Computed ; methods ; standards

OBJECTIVETo prepare a decellularized whole laryngeal scaffold by utilizing a perfusion-decellularized technique, reseed cells on it, and construct decellularized laryngeal muscles.

METHODSPerfusion decellularized larynxes were obtained by common carotid arterious perfusion with detergents. Then they were performed by macroscopic view, histological examination, scanning electron microscopy (SEM) and cartilage viability. Decellularized laryngeal scaffold were then reseeded with inducted mesenchymal stem cells (MSCs). Composites were transferred into greater omentums of rabbits after one day's adherence and harvested after eight weeks. Macroscopic view, histological examination and immunohistochemistry were performed.

RESULTSPerfusion larynxes became transparent after two hours. Histology and SEM indicated that perfusion method showed better decullularized effect. More vintages and collagen fibers but no intact cell or nuclei were retained in the decellularized matrix. Porosity measured by Image pro plus 6.0 was 80.4% +/- 3.2% (x +/- s). Chondrocyte vitality assay indicated chondrocyte vitality rate in the perfusion group was 86.9% +/- 1.5%. After eight weeks, vascularization formed and integrated cartilage frameworks still remained. Histological examination could clearly show the presence of muscle bundles and vessels. Immunohistochemical examination indicated that sarcomeric-alpha actin expressed positively in corresponding areas.

CONCLUSIONSIt is feasible to reseed MSCs into the decellularized laryngeal muscle matrix for constructing tissue-engineered laryngeal muscles. This in vivo maturation into the omentum could be the first step before in situ implantation of the construct.

Animals ; Extracellular Matrix ; Feasibility Studies ; Laryngeal Muscles ; cytology ; physiology ; Larynx, Artificial ; Rabbits ; Regeneration ; Tissue Engineering ; methods ; Tissue Scaffolds

OBJECTIVETo study the effect of testosterone propionate (TP) on the distribution pattern of calcitonin gene-related peptide (CGRP) in two types of motoneuron (Mn) pools in rats.

METHODThe double labeling of cholera toxin B subunit coupled with colloidal gold (CB-Au) retrograde identification combining with immunocytochemistry was mainly used to reveal the distribution pattern of CGRP-like immunoreactivity (CGRP-LI) and its changes in the motoneuron pools labeled by CB-Au.

RESULTTP injected intramuscularly 28 days later significantly decreased CGRP expression in Mn pool innervating extensor digitorum longus (EDL, fast-twitch), comparing with corresponding control and castration group respectively (P < 0.001), while no significant effect on Mn pools innervating soleus (SOL, slow-twitch, P > 0.05) was observed.

CONCLUSIONEDL-Mn pool is more sensitive to testosterone propionate than SOL-Mn pool in regulating CGRP expression.

Animals ; Calcitonin Gene-Related Peptide ; drug effects ; metabolism ; Male ; Motor Neurons ; drug effects ; metabolism ; Muscle Fibers, Fast-Twitch ; cytology ; drug effects ; Muscle Fibers, Slow-Twitch ; cytology ; drug effects ; Rats ; Rats, Wistar ; Testosterone Propionate ; pharmacology

OBJECTIVETo assess the correlations between Survivin and vascular endothelial growth factor (VEGF) in hepatocellular carcinoma (HCC) and their clinical significance.

METHODSThe expressions of Survivin and VEGF in 50 HCC specimens and 20 normal hepatic tissue specimens were detected by immunohistochemistry, and the results were analyzed in relation to the patients' clinicopathologic characteristics.

RESULTSOf the 50 HCC specimens, 32 (64.0%) were positive for Survivin expression, and 34 (68.0%) were positive for VEGF expression. Survivin expression was not detected in normal hepatic tissues, and 2 (10%) of these tissues were positive for VEGF, showing significant difference in Survivin and VEGF expressions between HCC specimens and normal hepatic tissues. Survivin and VEGF expressions were not correlated to the patients' gender, age, tumor size, degree of differentiation and alpha fetoprotein level (P<0.05), but related to the clinical stage and lymph node metastasis of HCC (P<0.05). Correlation analysis indicated a close correlation between the expressions of survivin and VEGF in the HCC specimens (chi 2=6.69, P<0.05).

CONCLUSIONSurvivin and VEGF are over-expressed in HCC tissues, and may theoretically serve as the targets of molecular targeted drugs. Detection of the expressions of Survivin and VEGF in HCC tissues may provide assistance for prognostic evaluation of the patients.

Adult ; Aged ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Inhibitor of Apoptosis Proteins ; Liver Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Microtubule-Associated Proteins ; biosynthesis ; Middle Aged ; Prognosis ; Vascular Endothelial Growth Factor A ; biosynthesis

Aim To study the apoptosis effect of cucurbitacin Ⅱa on non-small cell lung cancer cell lines NCI-H460 and A549 and its underlying mechanism.Methods Cell viability was assessed by CCK-8 assay.The apoptosis effect and cell cycle arrest were detected by Flow cytometry.Western blot was employed to detect the related protein.Results The proliferation of lung cancer cell lines NCI-H460 and A549 was inhibited by CuⅡa, which showed cytotoxic activity with IC50 values of 224.9 nmol·L-1 and 108.3 nmol·L-1 against NCI-H460 and A549 respectively.CuⅡa induced the cells apoptosis and cell cycle arrest at G2/M phase.The results of Western blot showed CuⅡa inhibited the phosphorylation of STAT3 and Cofilin in a dose-dependent manner.Further, CuⅡa inhibited the phosphorylation of Aurora A, in line with the important characteristics of anti-tumor effect of Aurora A kinase inhibitor with blocking cells in the G2/M phase.Conclusion CuⅡa has obvious anti-tumor effect against non-small cell lung cancer, which suggests its value as a lead compound for lung cell carcinoma.

Objective To investigate the health literacy level and health related behavior of high school students in Xuhui District of Shanghai, and to provide basis for health literacy intervention. Methods Using qualitative and quantitative methods, the quantitative part adopted the multi-stage stratified sampling method.A total of 355 aged 15-18 from senior high schools in Xuhui District were selected for face-to-face survey.The qualitative part adopted the form of group interviews, and 20 students from two senior high schools in Xuhui District were selected for interviews. Results The health literacy level of high school students in Xuhui District was 27.32%, which was lower than that of adults in the same period.Among the three dimensions, the level of basic knowledge and concept literacy was the highest, followed by the level of health basic skills literacy, the level of healthy lifestyle and behavior literacy was the lowest; among the six types of health problems, the level of infectious disease prevention literacy and basic medical literacy was at a low level.Statistical analysis of the correct answer rate showed that the answer rate of various questions in the health literacy survey was quite different.Further analysis of the health-related behavioral characteristics of high school students showed that most students had correct health concept and awareness of health change, but they encountered resistance in the process of knowledge transformation into action, which mainly came from academic burden, school education, peer influence and so on.The main channels for high school students to obtain health-related information were classroom, parents, school and various media.They had a high degree of trust in medical professionals and encountered difficulties in seeking health information. Conclusion The health literacy level of senior high school students in Xuhui District of Shanghai needs to be improved.We should explore a health intervention model more suitable for the behavioral characteristics of this group weak links in their health literacy.

OBJECTIVEBuilding a method which can examines virus pathogenic in gastroenteritis excrement specimen.

METHODSChoosing six positive specimens which tested in our laboratory, include adenovirus, calicivirus, rotavirus, bocavirus, astrovirus and enterovirus. Through sequence-independent single primer amplification(SISPA) constructs a gene bank. Looks up the viral gene fragment in gene bank.

RESULTSObtaining corresponding viral acid sequence in six specimens.

CONCLUSIONThis research can examine enterovirus and the virus which cause diarrhea, It make a foundation for further studies the viral cause of disease which the examination not yet discovered at present.

Child, Preschool ; DNA Primers ; genetics ; Diarrhea ; diagnosis ; virology ; Feces ; virology ; Female ; Humans ; Infant ; Male ; Polymerase Chain Reaction ; methods ; Virus Diseases ; diagnosis ; virology ; Viruses ; genetics ; isolation & purification

OBJECTIVETo investigate the effects of sympathetic neurotransmitters and adrenergic receptors on liver fibrosis in murine schistosomiasis.

METHODSMice were infestated with schistosoma by means of pasting cercariae on their abdomens. Thirty mice were randomly divided into a control group and a model group. Hematoxylin eosin and Van Gieson staining were used to view the histopathology of their livers. Immunofluorescence histochemistry and laser scanning confocal fluorescence microscopy were used to measure the a1A and beta2 adrenergic receptors in livers of the two groups of mice. High performance liquid chromatography-electrochemical detector (HPLC-ECD) was used to determine the concentration of norepinephrine (NE) and dopamine (DA) in the plasma of the mice.

RESULTSImmunofluorescence histochemistry showed that a1A and beta2 receptors were present in hepatocytes and hepatic sinusoids of the livers of the mice of the two groups, but there were many more in the livers of the schistosoma infected mice (t=-2.888; t=-6.648) (P<0.05). The results of HPLC-ECD showed that the levels of NE and DA in the model group were higher than those of the control group (t=-3.372; t=-4.428) (P<0.05).

CONCLUSIONSympathetic neurotransmitters and adrenergic receptors may participate in liver fibrogenesis in mice infected with schistosoma.

Animals ; Dopamine ; blood ; Liver ; pathology ; Liver Cirrhosis ; metabolism ; parasitology ; pathology ; Male ; Mice ; Mice, Inbred Strains ; Neurotransmitter Agents ; blood ; Norepinephrine ; blood ; Receptors, Adrenergic ; blood ; Schistosomiasis ; metabolism