Effects of neuro-immuno-modulation on healing of wound combined with local radiation injury in rats / 中华创伤杂志(英文版)

<p><b>PURPOSE</b>To investigate effects of neuro-immuno-modulation on wound healing by observing changes of cytokines and hypothalamic-pituitary-adrenal (HPA) axis hormones in acute stress reaction in rats with wound and combined local radiation injury.</p><p><b>METHODS</b>Sixty female Wistar rats (weighting 200 ± 20 g) were randomly divided into normal control group, wound group and combined wound-local radiation (CWR) group (25 Gy local radiation post wound), 20 rats in each group. Contents of IL-1β, IL-6 and IFN-γ and IL-4 in serum were measured and changes of adrenocorticotropic hormone (ACTH) and glucocorticoid (GC) in serum were analyzed by using enzyme-linked immunosorbent assay and radioimmunologic assay, respectively at different time points post wound and radiation.</p><p><b>RESULTS</b>(1) The level of IFN-γ, one of the Th1 cell cytokines increased significantly at 14 d post CWR, which was markedly higher than that in control group and wound group. However, the level of IL-4, IL-1β and IL-6, one of the Th2 cell cytokines, did not show obvious change. (2) Ratio of Th1/Th2 (IFN-γ/IL-4) in wound group and CWR group increased significantly at 7 d after wound and radiation, which suggested that Th1/Th2 balance drifted to Th1 immune response. The ratio of Th1/Th2 in wound group returned to the normal level up to 14 d after the wound and radiation, while the Th1/Th2 ratio in CWR group increased persistently and was much higher than that in control and wound groups. (3) Level of serous ACTH and GC in CWR group increased at 3 d post wound and radiation, and among them, level of GC showed statistically significant increase, which was much higher than that in control and wound groups.</p><p><b>CONCLUSION</b>Level of serous neurohormone GC in rats increased significantly immediately after wound and radiation; while the level of IFN-γ showed significant increase only up to 14 d after wound and radiation, and the Th1/Th2 imbalance sustained till 28 d post wound and radiation. In order to reduce acute damage caused by CWR, organic immune system and nerve system showed up a marked regulate effects simultaneously and mutually. Nonetheless, the excessive stress induced by CWR causes disturbance of immunoregulation, which is one of the key reasons for delayed wound healing in CWR.</p>

Effects and mechanism of CD4+ CD25+ regulatory T cells in mouse experimental colitis treated by CLYSTER No. 1 / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the effects and mechanism of CD4+ CD25+ regulatory T cells (Tregs) in mouse experimental colitis treated by CLYSTER No. 1.</p><p><b>METHOD</b>The mouse model of experimental colitis was established by dinitrochlorobenzene (DNCB)-acetic acid (AA) in mice DNCB and AA. Adult KM mouse were randomly divided into four groups: normal control group, experimental colitis model group, SASP and Chinese medicine therapeutic groups. Proportion of CD4 CD25+ Tregs in peripheral blood (PB) and mesenteric lymph node (MLN) was estimated by flow cytometry at the end of one or two week after treating with SASP and CLYSTER No. 1.</p><p><b>RESULT</b>The model of experimental colitis in mouse was successfully established. Compared with normal control group, the proportion of CD4 CD25 Tregs was markedly decreased in PB and MLN of model control group of experimental colitis. But it was significantly increased in therapeutic groups of SASP and CLYSTER No. 1, and their CD4+ CD25+ Tregs in PB and MLN were much more than the model control group at the end of one or two weeks after treating with SASP and CLYSTER No. 1.</p><p><b>CONCLUSION</b>CD4+ CD25+ Tregs with strong immune suppression could play a central role in the initiation and development of mouse experiment colitis, and the CLYSTER No. 1 might exert its therapeutic effects on UC by the regulation of number and function of CD4+ CD25+ Tregs.</p>

Bridge PCR,An Easy Way for Concatemerizing DNA Tags / 中国生物工程杂志

In MAST (mRNA accessible site tagging),the DNA tags from synthesized library were employed for identifying mRNA accessible sites. A large number of tags were amplified and subcloned for sequencing to verify mRNA binding profiles. A PCR was designed by using one primer which bridges over the tag terminal sequences. In PCR reaction DNA tag fragments were concatemerized by a bridge primer in reaction cycles. The concatemerized tag fragments were subcloned and sequenced. Dozens of the concatemerized sequences contained thousands tags. The PCR was a simple,effective way which for sequencing tags in a high through put manner.

Design and Identification of Antisense Oligonucleotide Targeting to 16S rRNA of E.coli / 中国生物工程杂志

Targeting rRNA of bacteria is a new strategy for antibiotic agent development. The rRNA such as mRNA are naturally self-folded molecules which expose only limited accessible target-sites for binding. These accessible sites are pivotal for designing the effective antisense oligonucleotides, ribozymes, and DNAzymes. MAST, an RNA accessible site screening method, illustrated 6 accessible sites on 16S rRNA by immobilizing 16S rRNA and hybridizing with oligonucleotide library. 5 of the accessible sites were identified valid, and the antisense oligonucleotides targeted to which showed inhibition effectiveness on the proliferation. Among the 5 target sites, one showed the priority of accessibility. Ribozyme designed to this site showed obvious inhibition to the growth when induced expressing in the transfection E.coli.

Screening and cloning of hepatitis C virus non-structural protein 4A interacting protein gene in hepatocytes / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate biological functions of hepatitis C virus (HCV) non-structural protein 4A (NS4A).</p><p><b>METHODS</b>Yeast-two hybrid technique was performed to seek proteins in hepatocytes interacting with HCV NS4A. HCV NS4A bait plasmid was constructed by ligating the NS4A gene with carrier plasmid pGBKT7, then it was transformed into yeast AH109 (alpha type). The transformed yeast cells were amplified and mated with yeast cells Y187 (alpha type) containing liver cDNA library plasmid pACT2 in 2 x YPDA medium. Diploid yeast cells were plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) and synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing X-alpha-gal for selection two times. After extracting plasmid from blue colonies, plasmid DNA was transformed into competent E.coli and analyzed by DNA sequencing and bioinformatics methods.</p><p><b>RESULTS</b>Among twenty-two positive colonies there were eleven positive for metallothionein 2A, three for eukaryotic translation elongation factor 1 alpha 1, two for albumin, two for RNA binding motif protein 21, two for myomesin, one for cytochrome C oxidase II, and one for ATPase.</p><p><b>CONCLUSIONS</b>Genes of HCV NS4A interacting proteins in hepatocytes were successfully cloned and the results pave the way for studying the biological functions of NS4A and associated proteins.</p>

Relationship between expression of Bax and Bcl-2 proteins and apoptosis in radiation compound wound healing of rats / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To study the relationship between the expression of Bax, Bcl-2 proteins, and apoptosis in radiation compound wound healing of rats.</p><p><b>METHODS</b>Apoptosis, Bax and Bcl-2 proteins were estimated by in situ terminal labeling (TUNEL) and immunohistochemical methods.</p><p><b>RESULTS</b>(1) Changes of the apoptosis in wound healing showed three typical characteristics: early occurrence, high frequency and delayed disappearance after radiation to rats when compared with those of simple wound group, which might be an important reason for radiation-induced delayed wound healing. (2) The expression of Bax protein increased evidently with the increment of apoptosis and showed a good corresponding relationship with the apoptotic frequency in the process of wound healing. While the expression of Bcl-2 protein decreased obviously as the apoptosis reached a maximum and showed increasing tendency up to normal level when the apoptosis decreased distinctively.</p><p><b>CONCLUSIONS</b>Bax and Bcl-2 proteins play an important role in the apoptotic regulation of radiation compound wound healing in rats.</p>

Expression of MMP1 and TIMP1 in radiation-combined wound healing and their effects on the healing process and tissue remodeling / 军事医学科学院院刊

Objective:To study the expression of MMP1 and TIMP1 in simple and radiation-combined wound healing and their effects on the healing process and tissue remodeling. Methods: A rat model of radiation-combined wound healing was used. Immunohistochemistry and in situ hybridization were performed which enabled the detection of MMP1 and TIMP1 expression in the healing process. Ultrastructural changes were observed with transmission EM. Results: The wound healing process was impaired and delayed. In rats receiving 25 Gy of gamma ray locally the irradiated wounds healed 6 days later than non-irradiated controls. The following changes in MMP1 and TIMP1 expression were found: (1) In the early inflammatory phase and in the period of granulation tissue formation, MMP1 expression in the newly-formed epidermis of irradiated wounds approximated that in the controls. Later, the epidermal expression of MMP1 in radiation wounds was comparatively increased with the delay of the healing process.On days 3 to 14 after wounding, TIMP1 was weakly positive in the proliferating keratinocytes of control wounds and became negative after epidermal covering, whereas no or only slight epidermal expression was detected in radiation wounds before epidermal covering.(2)MMP1 and TIMP　1 expression in radiation wounds was markedlydecreased in fibroblasts　, endotheliocytes and macrophages as compared with the controls. The expression phase was prolonged due to the delay of the healing process.Conclusions:The reduced expression of MMP1 and TIMP1 in granulation tissue retards such important processes as cell migration, angiogenesis and tissue remodeling, thus retarding the healing process. The expression of MMP1 in the newly-formed epidermis may help the process of reepithelialization,but in the late healing period, overexpression of MMP1 and decreased expression of TIMP1 in the epidermis may hinder the establishment of basal membrane and the formation of granulation tissue, and thus affect the matrix remodeling process.