Objective To establish and verify a time-resolved fluorescence immunochromatographic assay for the detection of S100B protein content based on double-antibody sandwich method,so as to provide a reliable method for the auxiliary diagnosis of neurological disorders and tumors.Methods Time-resolved fluorescent microspheres were used to label mouse anti-human S100B protein IgG2a monoclonal antibody(labeled antibody,clone number 22G7-3) and goat anti-chicken IgY antibody,respectively,and NC membranes were coated with mouse anti-human S100B protein IgG2a monoclonal antibody(coating antibody,clone number 5H2-3) and chicken IgY antibody as detection line(T line) and quality control line(C line),respectively,to prepare fluorescent immunochromatographic test strips.The coating antibody concentration(0.5,1,1.5,2 mg/mL),labeled antibody concentration(5,10,15,20 μg/mL) and detection time(5,8,10,12,15,18,20,22 min) were optimized.Serum and EDTA anticoagulated plasma from the same donor were used as samples to analyze the matrix effects.The linear range,limit of blank,precision,accuracy,anti-interference capability and stability of the method were verified.In addition,the established method and electrochemiluminescence method were used to detect 30 serum samples respectively,and the results were compared.Results The optimal conditions were coating antibody concentration of 2 mg/mL,labeled antibody concentration of 15 μg/mL,and detection time of 12 min.There was a significant matrix effect between serum and EDTA plasma,and plasma T/C was significantly lower than serum(t = 5.075,P < 0.01).Adding 2.5,5,and 10 mmol/L Ca~(2+) could improve the consistency of the two detection results,with R~2 of 0.846 0,0.724 1,and 0.723 9,respectively,while R~2 was 0.154 8 when Ca~(2+) was not added.There was a good linear relationship between S100B antigen concentration and T/C in the range of 0.01-10 ng/mL,and the linear equation was y = 3.857 81 x + 0.483 37,R~2 = 0.997 4.The limit of blank was 0.003 ng/mL.The coefficients of variation(CVs) of precision verification were less than 10%,and the relative deviations of accuracy verification were also less than 10%.Compared with the corresponding control group,bilirubin,hemoglobin,lipids,neuron specific enolase(NSE),glial fibrillary acid protein(GFAP) and ubiquitin carboxyl-terminal hydrolase isozyme L1(UCHL1) exhibited no significant effect on the test results(t = 0.660,0.141,1.691,1.875,0.091 and 0.274,respectively,each P > 0.05).Compared with 0 d,there was no significant difference in the test results of the test strips stored at 37 ℃ for 7,14,21,28,35 and 42 d(t = 0.197,0.451,0.199,1.506,0.074 and 0.768,respectively,each P > 0.05).There was no significant difference in the results of the 30 serum samples detected by the established method and electrochemilumine-scence method(difference range was-0.93-0.88,W =-2.000,P > 0.05).Conclusion The established time-resolved fluorescence immunochromatography method based on the double-antibody sandwich method has good linearity,precision,accuracy,anti-interference ability and stability,which has good correlation with commonly used clinical methods and can be used for the rapid quantitative detection of S100B protein in serum samples.

This paper interprets the content and recommendations of the guidelines on infection prevention and control in long-term care facilities put forward by the World Health Organization （WHO） during the 2019 coronavirus disease （COVID-19） pandemic， and actively explores the key points of nursing and infection prevention and control measures for the long-term care facilities under the background of repeated outbreaks， with the aim of providing care measures and infection prevention and control measures that suit our national conditions to improve the living standards of the elderly and protect them from viral infection amid the recurring pandemic.

A young male diagnosed with severe hemophilia A since childhood, was presented with recurrent joint and urinary bleeding. Annualized bleed rates dropped below five with low dose prophylactic medication.Bleeding in the right knee joint recently aggravated. Due to coexisting HIV infection and advanced hemophilic arthritis, the patient was managed by a multi-disciplinary team(MDT).Total knee arthroplasty was performed by an experienced surgeon using modern prosthesis design and intraoperative navigation technologies.Physical and rehabilitation therapy was provided during the postoperative period, and joint function improved. The MDT managed the young patient with HIV infection and advanced hemophilic arthritis. The patient was diagnosed with osteoporosis thought to have been caused by hemophilia, HIV infection and antiviral drugs; and he received treatment. The treatment of this patient reflects the importance of multidisciplinary cooperation in the management of difficult and rare diseases.

Objective:To analyse the physicochemical properties and structure of major privet pollen allergen Lig v 1 using bioinformatics software and provide a reference for choosing suitable recombinant expression system for Lig v 1 and modifying the allergen Lig v 1 experimentally.Methods:The physicochemical properties were analysed by ProtParam,the signal peptide by SignalP 4.1 Server,the transmembrane helix by TMHMM Server v.2.0,the secondary structure by GOR4,MHCⅡepitopes by NetMHCⅡ2.2 Server,B-cell epitopes by ProteanTM 5.01 and the phylogenetic tree by MEGA 6.Results: Privet major pollen allergen Lig v 1 was stable in Escherichia coli and it doesn′t possess any signal peptide and transmembrane helix.Most secondary structures of Lig v 1 were random coils.Potential region of MHCⅡepitope of Lig v 1 was 30-44.Potential B-cell epitopes possess discontinuous and continuous a mino acid sequences.Lig v 1 and its counterparts from Fraxinus excelsior and Olea europaea were clustered into one group.Conclusion:Escherichia coli is the suitable expression system for recombinant Lig v 1.In silico prediction of the epitopes of Lig v 1 provides a reference for modifying the allergen Lig v 1 experimentally.

Objective To investigate the change of aquaporin 2 (AQP2) mRNA and protein levels in renal collecting duct of SD rats after hypoxin caused by rising of the altitude to 4600 m. Methods Forty male SD rats were randomly divided into 4 groups (24 h, 48 h, 72 h and 1 week group), and 10 rats in Xining city were used as control group. All the 40 SD rats were transported to Kekexili Natural Reservation areas (4600 m) in Qinghai province. Rats of four experimental groups were sacrificed and renal tissue samples were harvested at different time point respectively, the control group rats were treated in Xining city (2260 m) as well. The concentration of plasma antidiuretic hormone (ADH) was measured by radioimmunity method. The expression of AQP2 mRNA and proteins was evaluated by real-time fluorescent quantitative-PCR, Western blot and immunofluorescence assay. Results The concentration of plasma ADH was decreased at 24 h and was only 28.5% of that of control group, reaching the lowest concentration at 48 h [(86.94±6.49) μg/L vs (302.5±310.48) μg/L], then it increased gradually and was similar to the control group at 7 d [(306.46±11.14) μg/L vs (302.53±10.48)μg/L, P＞ 0.05]. There were significant differences of the control group with 24 h, 48 h and 72 h groups, respectively[(302.53± 10.48) μg/L vs (142.46±10.57)μg/L, (86.94±6.49)μg/L, (169.65±11.15) μg/L respectively, P＜0.01]. The change of AQP2 gene expression level was consistent with the change of ADH. It was decreased at the begining when exposure to altitude and it reached its lowest level at 48 h. It was then returned to high level similarly to that of the control group at 7 d (0.09±0.01 vs 0.09± 0.008, P＞0.05 ). There were significant differences of the control group with 24 h, 48 h and 72 h group, respectively (0.09±0.008 vs 0.04±0.005, 0.03±0.002, 0.04±0.003 respectively, P＜0.01 ). Conclusions AQP2 expression in the renal collecting duct of SD rats is altered over the period exposed to altitude. It is decreased in the early hypoxia period, and is increased in later period. This change may be related to the intensity of hypoxia, which is mediated by a potential adaptation mechanisms against hypoxia caused by high altitude.

Objective To investigate the in vivo transduction capability of fusion protein PEP-1-EGFP with mice.Methods Two prokaryotic expression plasmids pET15b-EGFP and pET15b-PEP-1-EGFP were constructed and transformed into E.coli BL21(DE3) to express EGFP and fusion protein PEP-1-EGFP,respectively.The expressed EGFP and PEP-1-EGFP were purified with Ni2+-resin affinity chromatography.Five hundred micrograms of EGFP and PEP-1-EGFP fusion protein were injected into mouse through caudal vein,respectively,the mice were euthanized and perfused with PBS 2 hours after administration.Then,the heart,brain,liver,spleen and kidney were removed and sectioned with a cryostat at 7 ?m for visualization with a inverted fluorescent microscope.ResultsThe brain,heart,liver,spleen and kidney injected with PEP-1-EGFP showed bright and homogenous green fluorescence whereas that with EGFP showed no green fluorescence at all.Conclusion The successful expression and purification of PEP-1-EGFP fusion protein and its efficient transduction into mice in vivo provide a basis for the research on transmembrane delivery of macromolecule drugs mediated by the cell-penetrating peptide,PEP-1.

Aim To explore the effects of heat stress on TNF-?-induced fever and the content of cAMP in rat hypothalamus. Methods ①We examined the peak time of HSP70 expression in hypothalamus after heat stress; ②We studied the change of TNF-?-induced fever and the content of cAMP in hypothalamus after heat stress. Result The maximum expression of HSP70 in hypothalamus was attained at 12 h after heat stress. Heat stress limited TNF-?-induced fever and decreased the content of cAMP in hypothalamus. Conclusion Heat stress can inhibit TNF-?-induced fever in rats, and this effect may be related with the inhibition of cAMP synthesis induced by high expression HSP70 in hypothalamus.

Aim To study the role and mechanism of opioid receptor on IL-1?-induced fever.Methods Rats were dealed with intracerebreventricular administration of naloxone and/or IL-1?.Changes in body temperature were measured.The content of cAMP and HSP70 expression in hypothalamus were detected.Results Naloxone can limit IL-1?-induced fever and decrease the content of cAMP and HSP70 expression in hypothalamus(P