AIM: To construct the prokaryotic expression plasmid of His-tagged human high mobility group box 1 fusion protein (hHMGB1) and to express the fusion protein in E. coli for the affinity purification. METHODS: The cDNA coding region of HMGB1 was amplified by PCR from pGEX4T-HMGB1 and cloned into a modified pET14b vector following the routine procedure. After identification by enzyme digestion, PCR and sequencing, the plasmid was transformed into BL21 (DE_3) competent cells, and the His-HMGB1 fusion protein was induced for expression with isopropyl-beta-D-thiogalactopyranoside (IPTG), and further purified by Ni-NTA affinity chromatography. The protein was filtered for sterilization and used to stimulate human umbilical vein endothelial cells (HUVECs). 24 hours later, the cultured supernatant of HUVECs was collected for the detection of cytokines/chemokines with LiquiChip system. RESULTS: The His-HMGB1 fusion protein expression plasmid was identified by enzyme digestion and sequencing. The purified His-tagged fusion protein was analyzed by SDS-PAGE and Western blotting with specific anti-His antibody. It was found that the production of IL-8 from HUVECs was highly induced in a dose-dependent manner by HMGB1. CONCLUSION: The His-tagged HMGB1 fusion protein expression plasmid was successfully constructed, and purified. Recombinant HMGB1 protein has a high bioactivity on the induction of cytokines in HUVECs, which may significantly facilitate the future study of HMGB1 biological functions.

Ventilators are currently the most commonly used auxiliary mechanical ventilation equipment in clinical practice, and play an important role in the treatment of hypoxemia. Ventilator-associated pneumonia (VAP) is a special type of pulmonary parenchymal inflammation of nosocomial infection in patients with mechanical ventilation, which leads the increase of mortality and affects the prognosis of patients. The non-standard management of ventilator pipeline joints and the aerosol formed by bacterial condensate splashed from ventilator pipeline pollute the air, resulting in cross infection, which are the important reasons for VAP. The existing ventilator pipeline joint cap can achieve the effect of preventing pollution, but the clinical application compliance is not high. Based on the above factors, teachers, students and medical staff of the School of Public Health and Management, Guangxi University of Chinese Medicine and other units have designed a device for preventing pollution and air pollution of the ventilator pipeline joint, and obtained the national utility model patent of China (ZL 2020 2 1361981.X). The device is composed of a model lung, a mask body, a suspension part and a beam mouth part, etc. The use method is simple, and can be applied to the pollution avoidance of invasive ventilator and non-invasive ventilator pipeline joints, to reduce the occurrence of VAP in patients and the occupational exposure of medical staff.

Objective To investigate the effects of hypothyroidism and drug intervention on the cognitive function of rat offspring through the establishment of a rat model of experimental hypothyroidism in pregnancy.Methods A to-tal of 20 SD female rats were randomly divided into control group(CON group)and hypothyroid group(PTU group).The hypothyroid model was established by propylthiouracil(PTU),the thyroid hormone levels of female mice were detected by electrochemiluminescence immunoassay,and the differences between the two groups were compared.After successful modeling,the male mice were mated in cages,and the hypothyroid group was randomly divided into no intervention group(group Ⅰ),first trimester intervention group(group Ⅱ)and second and third tri-mester intervention group(group Ⅲ).The Morris water maze(MWM)experiment was used to test the learning and memory ability of the rats.The morphological structure of hippocampal neurons,the expression of nucleoprotein(NeuN)and synapse-associated protein(SYN)in mature neurons were observed by hematoxylin-eosin staining(HE),Nissl staining and immunohistochemistry.Results ① Compared with the CON group,the female mice in the PTU group had a significant increase in TSH and a significant decrease in FT4(P＜0.05).② In the positio-ning navigation test,the evasion latency of the pups in each group was gradually shortened.On the 5th day,the in-cubation period of group Ⅰ was significantly longer than that of groups CON,Ⅱ,Ⅲ(P＜0.05).There was little change between groups Ⅱ and Ⅲ and CON groups(P＞0.05).③ The residence time of group Ⅱ was significantly different from that in group Ⅰ during the space exploration stage(P＜0.05).There was a significant difference be-tween the number of pups crossing the platform in group Ⅰ and groups CON,Ⅱ and Ⅲ(P＜0.05).④ There was no significant difference between HE staining and Nissl staining in hippocampal tissues of rats in each group.How-ever,compared with the CON group,the average absorbance of NeuN and SYN proteins in the hippocampus of mice in groups Ⅰ,Ⅱ and Ⅲ was significantly reduced(P＜0.05).Conclusion Hypothyroidism will have adverse effects on the cognitive function and hippocampal neuron development of pregnant rats,and the effects of interven-tion on the cognitive function of offspring at different stages of pregnancy are different,the earlier the intervention,the smaller the damage to cognitive function.

OBJECTIVE To optimize the extraction technology of volatile components from Wuyao decoction. METHODS On the basis of single factor investigation ，the extraction technology of volatile components from Wuyao decoction was optimized and validated by Box-Behnken design-response surface technology using the contents of bomyl acetate ，cyperotundone，α-cyperone， ligustilide and dehydrocostuslactone ， extraction rate of volatile oil as indexes ， with extraction time ， soaking time and liquid-material ratio as factors. On this basis ，the extraction state of the decoction was quantified. RESULTS The optimal extraction technology was as followed ：the ratio of liquid -material was 13∶1（mL/g），soaking time was 0.5 h，and the extraction time was 6 h in the boiling state. The comprehensive scores of the three validation experiments were 0.948 7，0.948 4 and 0.948 6 respectively （RSD＝0.02％，n＝3），and the deviation from the predicted value （0.947 9）was no more than 1％. The boiling state of the decoction in 180 ℃ oil bath was taken as the sudden boiling state. CONCLUSIONS The optimized extraction technology is stable and feasible.

OBJECTIVE To optimize the cladding process of volatile oil saturated and aqueous solution method of classic prescription Sanhua decoction. METHODS β-Cyclodextrin was used as the wrapping material, through Plackett-Burman combined with Box-Behnken design tests, the optimal packaging process of Sanhua decoction’s total volatile oil was screened. The clathrate was verified by microscopy, thin layer chromatography, and ultraviolet spectrophotometry, and the change of volatile composition before and after the envelope was determined by headspace-gas chromatography-mass spectrometry. RESULTS The best coating process of the total volatile oil of the classic prescription Sanhua decoction was as follows: speed 2 000 r·min-1, wrapping 1 h, feed-liquid ratio 1∶10, baling temperature 30 ℃, material ratio 1∶9. The comprehensive scores of the three validation experiments were 0.981 0, 0.999 9 and 0.963 4, and the average value was 0.981 4, which was 2.87% different from the predicted value of the model. CONCLUSION The saturated aqueous solution method is simple and stable, and the volatile components are basically unchanged before and after the encapsulation, which provides a reference for the development of Sanhua decoction and the research on the volatile oil in the classic recipe.