Expression of BmSPI38 tandem multimers in Escherichia coli and its antifungal activity / 生物工程学报

The aim of this study was to prepare tandem multimeric proteins of BmSPI38, a silkworm protease inhibitor, with better structural homogeneity, higher activity and stronger antifungal ability by protein engineering. The tandem multimeric proteins of BmSPI38 were prepared by prokaryotic expression technology. The effects of tandem multimerization on the structural homogeneity, inhibitory activity and antifungal ability of BmSPI38 were explored by in-gel activity staining of protease inhibitor, protease inhibition assays and fungal growth inhibition experiments. Activity staining showed that the tandem expression based on the peptide flexible linker greatly improved the structural homogeneity of BmSPI38 protein. Protease inhibition experiments showed that the tandem trimerization and tetramerization based on the linker improved the inhibitory ability of BmSPI38 to microbial proteases. Conidial germination assays showed that His6-SPI38L-tetramer had stronger inhibition on conidial germination of Beauveria bassiana than that of His6-SPI38-monomer. Fungal growth inhibition assay showed that the inhibitory ability of BmSPI38 against Saccharomyces cerevisiae and Candida albicans could be enhanced by tandem multimerization. The present study successfully achieved the heterologous active expression of the silkworm protease inhibitor BmSPI38 in Escherichia coli, and confirmed that the structural homogeneity and antifungal ability of BmSPI38 could be enhanced by tandem multimerization. This study provides important theoretical basis and new strategies for cultivating antifungal transgenic silkworm. Moreover, it may promote the exogenous production of BmSPI38 and its application in the medical field.

Key active sites of proteases and protease inhibitors: a review / 生物工程学报

Proteases are widely found in organisms participating in the decomposition of proteins to maintain the organisms' normal life activities. Protease inhibitors regulate the activities of target proteases by binding to their active sites, thereby affecting protein metabolism. The key amino acid mutations in proteases and protease inhibitors can affect their physiological functions, stability, catalytic activity, and inhibition specificity. More active, stable, specific, environmentally friendly and cheap proteases and protease inhibitors might be obtained by excavating various natural mutants of proteases and protease inhibitors, analyzing their key active sites by using protein engineering methods. Here, we review the studies on proteases' key active sites and protease inhibitors to deepen the understanding of the active mechanism of proteases and their inhibitors.

Analysis of related factors of time and mortality of severe trauma patients in emergency department based on ISS / 中华急诊医学杂志

Objective:The clinical influencing factors of residence time of patients with multiple trauma in emergency department were evaluated based on ISS, and the correlation between emergency department length of stay and mortality was analyzed.Methods:From October 2013 to November 2016244patients with multiple trauma treated in the emergency department of our hospital were selected to evaluate and record the ISS score of the patients, and to count the general data of the patients, the emergency department length of stay and the outcome of this visit.Results:The study showed that 244 patients were involved, 191 males and 53 females. The emergency emergency department length of stay was (117.25±87.442) min, The data show that 46 cases were clinically converted to death.Conclusions:The higher the ISS score of multiple trauma patients, the longer the emergency department length of stay, the higher the mortality rate of patients with prolonged emergency department length of stay, and the higher the ISS score of multiple trauma patients, the higher the mortality rate.

Study on Relativity of AGEs, Inlfammatory Factor and Growth Factor in Ulcer-curation Process of Diabetic Feet Promoted by Yin-Huang-San / 世界科学技术-中医药现代化

This study was aimed to discuss the external treatment of diabetic foot ulcer with traditional Chinese medicine (TCM) Yin-Huang-San (YHS) and to observe its promoting healing effect. Analysis was made on AGEs correlated with inflammatory factors and growth factors. A total of 60 patients with damp-heat-toxin pattern standard diabetic foot ulcer patients were randomly divided into the treatment group and control group. External application of YHS was given in the treatment group. And external application of metronidazole and glucose injection was given in the control group. The dressing was changed once a day. Blood was drawn 1 d before the treatment, and 7 d, 14 d, 21 d, 28 d after the medication. Improvements of patients’ clinical symptoms were observed. Dynamic monitoring was given on changes of hs-CRP, TNF-α, IL-1, VEGF, EGF, bFGF, PDGF and AGEs. The results showed that the total efficiency of the treatment group was 96.7%; and the total efficiency of the control group was 83.3%. Compared with the control group, external treatment with YHS can obviously improve clinical symptoms of diabetic foot ulcer (P<0.01), reduce AGEs and inflammatory factor, and increase the number of growth factors. AGEs had significant positive correlation with inflammatory factor; it had significant negative correlation with growth factors. It was concluded that YHS was effective in the treatment of diabetic foot ulcer patients with damp-heat-toxin pattern.

Treatment of Arteriosclerosis Obliterans with Jie-Du Tong-Mai Decoction and Influence on ABI, CRP, TCPO2 / 世界科学技术-中医药现代化

This study was aimed to observe the clinical efficacy of Jie-Du Tong-Mai (JDTM) decoction (i.e., detoxi-fication unclogg artery soup) of arteriosclerosis obliterans. A total of 246 arteriosclerosis obliterans cases with the pattern of blood stasis were selected and randomly divided into two groups. In the control group of 120 cases, 0.1 g of aspirin and 20 mg of simvastatin tablets were orally administrated, once a day. In the treatment group of 126 cas-es, JDTM decoction was combined on the basis of oral administration of aspirin and simvastatin tablets. After 8 weeks of treatment, observations were made on improvement of clinical symptoms and changes of ABI, Hs-CRP, TcpO2. The results showed that compared with the control group, clinical symptoms of the treatment group were obvi-ously improved (P< 0.05). And the ABI, CRP, TcpO2 were also obviously increased (P< 0.01). It was concluded that JDTM decoction was effective in the treatment of arteriosclerosis obliterans with the pattern of blood stasis. It al-so indirectly proved the guiding meaning of traditional Chinese medicine that evil causing stasis.

Treatment of lower limb ischemia with combination of traditional Chinese medicine and transplantation of autologous bone marrow mononuclear cells: a report of 23 cases / 中西医结合学报

OBJECTIVE: To evaluate the therapeutic effect of transplantation of autologous bone marrow mononuclear cells combined with traditional Chinese medicine for the treatment of limb ischemia. METHODS: Twenty-three patients with limb ischemia were treated. G-CSF was used to stimulate the bone marrow. The mononuclear cells were separated from the aspirated bone marrow fluid in the stem cell studio. The cell amount was above 1x10(9). The transplantation was performed by the way of intra-muscular multi-injection. Traditional Chinese medicine for replenishing qi to activate blood was prescribed from the first day after operation. The pain, poikilothermia, ulcer or necrosis and ankle/brachial index (ABI) of the ischemic limb were evaluated before and after the treatment. RESULTS: The pain score and poikilothermia score decreased one month after the transplantation, with distinct differences as compared with the scores before the treatment (P<0.05). The ABI increased gradually after the treatment, and one month after the treatment, it was 0.15 higher than that before the treatment. CONCLUSION: Transplantation of autologous bone marrow mononuclear cells combined with traditional Chinese medicine can decrease the symptoms and signs of severe lower limb ischemia effectively, and improve the circulation of the ischemic area.