Objective To explore the most suitable chemostatic condition and a method of cell-counting in reconstituted basement membrane invasion experiment in vitro. Method Reconstituted basement membrane invasion experiment was employed, and conditioned medium, fibronectin (FN), and a combination of conditioned medium and FN were respectively used as chemoattractants was placed in the lower compartment of the chamber. After the filters were stained with hematoxylin and eosin (HE), two kinds of cell-counting methods were employed. One was random cell-counting to count the cells in 5 fields infiltrated in the different areas of the filter membrane under 40?magnification, and the other was whole cell-counting of the infiltrating cells on the whole filter membrane with 4? magnification and image analysis system. Results The number of infiltrating cells in the presence of the chemoattractants was much larger than that of the control group. The number of infiltrating cells of the conditioned medium group was larger than that of FN group. The numbers of infiltrating cells in the combination of conditioned medium and FN groups were significantly larger than those of conditioned medium and FN alone groups. The accuracy of method of random cell-counting under 40? magnification was poorer than that of whole cell-counting with image analysis system. Conclusion Chemoattractants exert strong effect on the tumor cells invasion, and different chemoattractants show different chemotactic abilities. By using image analysis system to count all the infiltrative cells of the whole filter membrane is an objective and accurate cell-counting method.

To develop a blue cap anticoagulant tube blood volume measuring card of to solve the problem of insufficient or excessive blood collection in clinical coagulation specimens.The device was composed of a measuring card,a transparent housing with a base and a tube holder.The measuring card was divided into qualified and unqualified areas,the housing was used to insert the card,the tube holder was used to place blood collection tubes.The device was used by clinical nurses to judge the adequacy of blood collection volume in blue cap anticoagulant tube.After the use of the device,the failure rate of clinical blue cap anticoagulation tube specimens submission was reduced from 6.71‰ to 2.73‰,shortened the time limit for specimen submission.At the same time,the device made the rejection judgment of department specimens more standardized and avoided the acceptance of unqualified specimens caused by subjective judgment errors.The device has simple structure,convenient operation and strong practicability,and has promotion value.

The full length cDNA of silkworm hibadh gene was cloned by RT-PCR and RACE (Rapid amplification of cDNA ends) technique. The hibadh gene and its deduced amino acid sequences were analyzed. The tissue distribution of hibadh gene in 5th instar silkworm larvae was tested by RT-PCR. The expression patterns of hibadh gene in simulated weightless environment were analyzed by real time RT-PCR. The results showed that the full length hibadh cDNA sequence was 1074 bp in lenth, including an open read frame of 969 bp encoding the entire coding region of Hibadh (GenBank accession No. EU719652). The deduced amino acid sequence similarities of hibadh between silkworm and Burkholderia ambifaria, Drosophila melanogaster, Apis mellifera, Xenopus tropicalis, Mus musculus, Homo sapiens were 46%, 43%, 48%, 44%, 45%, 45%, respectively. Signal peptide analysis showed that Hibadh was a secretory protein. There wasn't glycosyl-phosphatidyl inositol anchor site in Hibadh amino acid sequence. Molecular weight and isoelectric point of Hibadh were 34.1 kD and 9.14 respectively. The RT-PCR tests indicated that the hibadh gene expressed in head, silk gland, midgut, cuticle, blood, fat body, tuba malpighii of the 5th instar silkworm larvae. There were different expression patterns of hibadh gene during different silkworm embryo period in simulated weightless environment. Simulated weightlessness resulted in the expression of silkworm hibadh gene up regulated 2.3-fold (P < 0.05), up regulated 4.6-fold (P<0.01), down regulated 7.6-fold (P < 0.01), down regulated 2.6-fold (P < 0.05) during apophysis formation period, inverse period, trachea formation period, and whole embryo period, respectively. There was no significant change of hibadh gene expression during other period of silkworm embryo between simulated weightless and control groups. There were different response patterns to simulated weightless environment between hibadh gene and whole body of silkworm. Gene showed much higher sensitivity compared to whole body in response to environment. This study is useful for the further research on the gravity biological mechanism of hibadh gene.
Alcohol Oxidoreductases ; genetics ; metabolism ; Amino Acid Sequence ; Animals ; Base Sequence ; Bombyx ; enzymology ; genetics ; Cloning, Molecular ; Computer Simulation ; Genes, Insect ; genetics ; Models, Biological ; Molecular Sequence Data ; Weightlessness

Lung is one of the most sensitive target organs of human beings under the shock waves. Due to its serious injury, rapid development and high mortality, blast lung injury has been a widely concerned research topic in the field of military medicine. In the normal physiological state, the body is in a dynamic balance between pro-inflammaton and anti-inflammation, oxidation and anti-oxidation, promoting apoptosis and inhibiting apoptosis. While blast lung injury breaks the balance and causes physiological, biochemical and pathological changes in the body, seriously leading to acute respiratory distress syndrome and multiple organ dysfunction syndrome, and eventually the mortality. So far, the researches on blast lung injury mainly involve damage model, pathogenesis, pathological changes, intervention treatment and so on, which has achieved great research findings. In the review, the authors summarize the progress of molecular mechanism for blast lung injury from the perspective of inflammatory reaction, oxidative stress, apoptosis and so on, which may promote the discovery of new targets for the diagnosis, treatment and rehabilitation intervention of blast lung injury.

Objective In this study,we investigated the active ingredients,targets and molecular mechanisms of Bai zhi based on network pharmacology and mRNA transcriptome sequencing technology for the treatment of microgravity induced bone loss,with a view to providing new therapeutic strategies for microgravity induced bone loss diseases.Methods Investigating the antagonistic effect of the traditional Chinese medicine Bai zhi on microgravity induced bone loss by constructing a hindlimb unloading mice model.18 healthy male mice were randomly divided into Control,HLU and HLU+BZ groups,6 mice in each group,and the effects of Bai zhi on the bone mineral density of the model mice were evaluated after 28 d of continuous gavage.Screening the active ingredients and targets of Bai zhi through TCMSP,Genecards,OMIM,TTD,DisGeNET and other network pharmacology databases.A hindlimb tail suspension unloading animal(HLU)model was established,and the differentially expressed genes(DEGs)responding to mechanical unloading were analyzed using transcriptome sequencing methods,and the targets of Bai zhi active ingredients intersected with the targets of the differentially expressed genes responding to mechanical unloading,so that the core gene targets of Bai zhi for intervening in weightlessness bone loss could be obtained;Construction of Bai zhi component-target protein interaction network(PPI)using String database and Cytoscape software,and enrichment and analysis of key signaling pathways of Bai zhi for treating microgravity induced bone loss using R Studio software.Results Bai zhi effectively restored bone loss in hindlimb tail suspension mice.By quantitative analysis of bone mineral density scanning of hindlimb tail suspension mice,the results showed that Bai zhi significantly restored the loss of bone mineral density in the model mice(P?