Objective To study the dynamic changes of transendothelial electrical resistance (TEER) on our in vitro model of brain blood barrier (BBB) made from primary culture of BALB/c mouse brain microvascular endothelial cells (BMVEC), and to explore the relation between TEER and BBB permeability, and search for the best culture condition. Methods BMVEC were isolated from BALB/c mouse and cultured on a transwell insert with special micro-pore. The cells were identified with immunohistochemical methods and electron microscope. TEER over BMVEC was measured after BBB model establishment for determining the 3H-Glucose permeability of BBB in vitro. Results BMVEC cultured in the transwell insert exhibited typical "flagstone" appearance and in a tight monolayer structure under electron microscope. Immunohistochemical detection of ZO-1 protein, a marker antigen of tight junction, showed smooth, continuous and tight junctions between confluent BMVEC. TEER over BMVEC monolayer increased to (346?10) ?/cm2 when the permeability for 3H-Glucose was decreased to the minimum. Conclusion BBB model in vitro made from primary culture of BMVEC in transwell has the basic characteristics of BBB in morphology, electrical resistance and permeability.

Objective To examine the relationship between CCT 5 ,γδ T cell and autoimmune diseases .Methods Recombinant CCT5 protein was cloned , expressed and purified in E.coli.Three peptides of CCT5 protein were used to prepare for anti-CCT5 monoclonal antibodies .Purified CCT5 protein was used to expand γδT cells from pe-ripheral blood mononuclear cells (PBMC).Plasma level of CCT5 in healthy donors, patients with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) were detected by ELISA assays .The correlation analysis between plasma CCT5 concentration and the percentage of different subtypes of γδT cells measured by flow cytometry was made . Results The CCT5 gene was amplified by PCR and the length of the target fragment was 1 750 bp.The expressed 65 ku CCT5 protein was purified and validated by SDS-PAGE.Two paired monoclonal anti-CCT5 antibodies were screened to detect CCT5 protein in plasma.Immobilized recombinant CCT5 protein was able to induce specific sig-nificant amplification of peripheral γδT cells.Correlation analysis of 10 healthy donors indicated significant corre-lation between the plasma CCT 5 concentration and the proportion of Vγ9 and Vδ2 γδ T cells.The plasma CCT5 concentration significantly decreased in autoimmune diseases patients , including RA and SLE .Conclusions These data suggest that CCT 5 could be a novel Vγ9δ2 γδT cell-related factor in autoimmune diseases , which deepen the understanding of Vγ9δ2 γδT cell function in autoimmune diseases .

Objective:To compare the application effect of whole-process seamless nursing and conventional nursing in the perioperative period of ankylosing spondylitis (AS) with kyphosis complicated by cervical spine fracture.Methods:A retrospective cohort study was conducted to analyze the clinical data of 64 patients with AS with kyphosis complicated by cervical spine fracture admitted to Henan Provincial People′s Hospital from April 2017 to December 2022, including 37 males and 27 females, aged 27-73 years [(49.8±14.6)years]. There were 43 patients with vertebral fractures and 21 with intervertebral space fractures. All patients underwent reduction and fixation or correction and fixation. Thirty-two patients admitted from April 2017 to August 2019 received conventional care (conventional nursing group), and 32 patients admitted from September 2019 to December 2022 received whole-process seamless care (seamless nursing group). The two groups were compared concerning the visual analog scale (VAS) before, at 12 hours, 24 hours, 1 month, 3 months after surgery and at the last follow-up, the American Spinal Injury Association (ASIA) score before surgery, at 1 and 3 months after surgery and at the last follow-up, the health survey questionnaire (SF-36) score and patient satisfaction score before surgery and at the last follow-up, as well as the incidence of postoperative complications.Results:All patients were followed up for 6-12 months [(9.8±3.2)months]. There was no statistical difference in preoperative VAS between the two groups ( P>0.05). The values of VAS in the seamless nursing group were (3.9±1.9)points, (4.2±0.7)points, (2.7±0.9)points, (2.6±0.6)points, and (1.7±0.8)points at 12 hours, 24 hours, 1 month, 3 months after surgery and at the last follow-up, respectively, lower than those of the conventional nursing group [(5.7±1.2)points, (5.8±1.1)points, (3.6±1.2)points, (3.2±1.1)points, and (2.4±1.0)points] ( P<0.05 or 0.01). The VAS of the seamless nursing group at 12 hours, 24 hours, 1 month, 3 months after surgery and at the last follow-up was lower than that before surgery (all P<0.05). The VAS at 1 and 3 months after surgery and at the last follow-up was lower than those before and at 12, 24 hours after surgery and the VAS at the last follow-up was lower than those at 1, 3 months after surgery (all P<0.05). The differences among VAS of the conventional nursing group before and at 12, 24 hours after surgery were statistically insignificant (all P>0.05). The VAS of the conventional nursing group at 1 and 3 months after surgery and at the last follow-up was lower than those before surgery and at 12, 24 hours after surgery and the VAS at the last follow-up was lower than those at 1, 3 months after surgery (all P<0.05). There was no statistically significant difference in VAS of the two groups between 12 hours and 24 hours after surgery, and between 1 month and 3 months after surgery (all P>0.05). There was no significant difference in the ASIA scores before surgery between the two groups ( P>0.05). There were significant improvements in ASIA scores in the seamless nursing group at 1, 3 months after surgery and at the last follow-up compared with those of the conventional nursing group ( P<0.05). There were no statistically significant differences in preoperative SF-36 score and patient satisfaction score between the two groups (all P>0.05). At the last follow-up, the SF-36 score and patient satisfaction score of the seamless nursing group were (47.4±6.2)points and (99.5±1.2)points, respectively, which were higher than those of the conventional nursing group [(42.2±7.3)points and (98.1±1.6)points] (all P<0.05). At the last follow-up, the SF-36 score and patient satisfaction score of the seamless nursing group were higher than those before surgery (all P<0.01). The SF-36 score of the conventional nursing group was higher than that before surgery ( P<0.01), but there was no significant difference in patient satisfaction score ( P>0.05). The incidence of postoperative complications in the seamless nursing group was 6.3% (2/32), lower than that of the conventional nursing group [25.0% (8/32)] ( P<0.05). Conclusion:For AS with kyphosis complicated by cervical spine fracture, whole-process seamless nursing is associated with alleviated postoperative pain, improved spinal nervous function, quality of life and degree of satisfaction, and reduced incidence of complications compared with the conventional nursing.

OBJECTIVE：To e stablish the method for simultaneous determination of 10 kinds of active components in Tibetan medicine Siwei jianghuang prescription ，and to optimize its decoction technology. METHODS ：HPLC method was adopted. Using soaking time ，the amount of added water ，decoction time and decoction times as factors ，comprehensive score of the contents of 10 kinds of components and solid extracts rate as response values ，one the basis of single factor test ，Box-Behnken response surface method was used to optimize its decoction technology. RESULTS ：The linear range of gallic acid ，corilagin，magnoflorine， ellagic acid ， hydrochloric jatrorrhizine ， hydrochloride palmatine ， hydrochloride berberine ， bisdemethoxycurcumin， demethoxycurcumin and curcumin were 0.280 6-1.683 6，0.289 6-1.737 6，0.320 8-1.924 8，0.116 0-0.696 0，0.018 9-0.113 5， 0.013 3-0.079 9，0.092 3-0.553 8，0.025 5-0.153 0，0.036 1-0.216 3，0.041 0-0.245 7 µg（all r were 0.999 9），respectively. The limits of quantitation were 0.28，14.48，3.21，11.60，1.89，4.44，0.46，0.26，0.36，0.41 ng，respectively. The limits of detection were 0.11，4.14，1.24，3.32，0.58，1.33，0.13，0.09，0.14，0.12 ng，respectively. RSDs of precision ，stability and reproducibility tests were all lower than 3％. The recoveries were 92.56％-103.69％（RSDs were 0.90％-3.81％，n＝6）. The optimal decoction technology included soaking 60 min，adding 8-fold（mL/g）water，decoction for twice ，lasting for 65 min each time. In 6 validation tests ，comprehensive scores were 3.323 2-3.422 4，and the absolute value of the relative error with the predicted value （3.437 4）was less than 2％.CONCLUSIONS：Established method is simple and repeatable ，and can be used for simultaneous determination of 10 kinds of active components in Siwei jianghuang prescription. Optimized decoction technology is stable and feasible.

AIM: To find specific metabolic markers for women entering peri-menopausal period and patients with menopausal syndrome based on