Objective:To clone and express panallergen profilin from the pollen of coco(Cocos nucifera Linnaeus).Methods:RT-PCR and RACE methods were applied to clone the full-length panallergen genes from coco pollen and the sequence was analyzed.The specific primers were designed.The ORF of profilin of coco pollen was amplified with RT-PCR and cloned into the expression vector pET 28a.Expression of the recombinant coco pollen profilin was carried out in E.coli BL21(DE3) and the purification of the recombinant protein was performed via affinity chromatography with Ni2+ coupled to sepharose.IgE reactivity to recombinant coco pollen profilin was investigated by immunoblot.Results:The complete sequence of coco pollen profilin was cloned.The sequence was 608 bp and included an open reading frame(396 bp) coding for 131 amino acids.Sequence analysis showed that the deduced protein was an acidic protein with an estimated molecular mass of 14.19 kD and a pI of 4.61.The GeneBank accession number of the clones was EF173598.After overexpressed in E.coli BL21(DE3),the recombinant protein was purified through affinity chromatography with Ni2+ coupled to sepharose.Immunoassay showed that the recombinant allergen has good IgE binding capacity.Conclusion:The profilin of coco pollen is expressed successfully in BL21(DE3),which will be used as a base for further study on coco pollen related allergy.

To synthesize salbutamol immunogen and develop an enzyme immunoassay (ELISA), a new salbutamol immunogen was synthesized using 4-aminobenzoic acid as a linker to connect hapten with carrier protein. An enzyme immunoassay based on the antibody prepared was developed and applied to detect salbutamol residue spiked in swine liver. An unusual coating antigen, clenbuterol-ovalbumin (OVA) conjugate instead of salbutamol-OVA conjugate, was used in the immunoassay and the results were discussed based on the structures of related compounds. The antibodies showed high sensitivity in the heterologous assay when using clenbuterol-OVA as a coating antigen, with an IC50 value of 8.97 ng mL(-1) toward salbutamol. The antibodies prepared showed high cross-reactivity with clenbuterol (107%) and were promising for the simultaneous determination of salbutamol and clenbuterol residues in food and food products. Recovery rates from the salbutamol-spiked swine liver samples were in the range of 70%-99%, while the intra-assay and inter-assay coefficients of variation were <13.3% and <14.3%, respectively. In summary, the antibodies of salbutamol have been successfully prepared. Sensitive and stable analysis for the detection of salbutamol residues in swine liver was obtained based on the competitive ELISA methods developed in this study.

Objective To explore the effect of apolipoprotein E deficiency on astrocyte destruction in the ex vivo spinal cord slice model. Methods Vibratome-cut transverse spinal cord slices from C57BL/6 postnatal day 7 ApoE -/-mice and wild-type mice were cultured on transwell porous supports. After 7 days in the culture , spinal cord slices were exposed to LPS for 5 days. The expression of AQP4, GFAP and Iba1 was detected by immunofluorescence. The concentration of TNF-α and NO were detected by ELISA and method of nitrate reductase, respectively. Results Marked loss of AQP4 and GFAP staining were shown in LPS-affected groups, not in the control group, and the lesion score was higher in ApoE-/-group than WT-group (P<0.05). In addition, the expression of Iba1 and concentration of TNF-α, NO in the LPS-affected groups were more than that of the control group (P < 0.05), and they were higher in the ApoE-/-LPS group than the WT-LPS group (P < 0.05). Conclusions ApoE deficiency exacerbates astrocyte injury, likely through promoting the release of pro-inflammatory mediators from microglia.

Objective To investigate the efficacy of ICG clearance test for hepatic reserve function in patients with hepatic cirrhosis and primary liver carcinoma by pulse dye deusitometry (PDD). Methods Eighty-nine patients with hepatic cirrhosis and forty patients with primary hver carcinoma were enrolled in this study. All patients were classified by Child-Pugh grade. The value of indocyanin green retention at 15 min (ICGRI5) was measured by PDD. Biochemical parameters including CHE, PAB, ALB, PTA and TBA were also examined. Correlation analysis between ICGR15 and those indexes were conducted. Results (1) ICGR15, CHE, TBA, ALB, PTA among Child-Pugh A, B, C in all patients were significantly different (P < 0.05). PAB between Child-Pugh B and A group, C and B group was significantly different (P < 0.05). (2)Increased level of ICGR15 and TBA positively correlated with Child-Pugh scores, while negatively correlated with the level of PTA. The difference of ICGRI5 between Child-Pugh score 5 and score 7, score 9 and score 10 were all significant (P <0.05). (3) The correlation between ICGR15 and other clinical markers is significant. The clinical marker with the best correlation was TBA (r=0.605), was the parameter in most significantly correlated with ICGR15 followed by PTA, CHE, PAB, and ALB (r= -0.598, -0.565, -0.537, -0.424). Conclusion There was good correlation between the clinical markers and the value of ICGR15 which demonstrate superiority to the other indexes. The data also suggested that CHE, TBA, could be used in evaluating liver reserve function.

Objective To observe the damage of high mobility group box 1 (HMGB1) on human umbilical vein endothelial cell (HUVEC) barrier permeability and the protective effect of unfractionated heparin (UFH), and to explore the down-regulated protection effect mechanism of UFH on HMGB1-mediated vascular endothelial cadherin (VE-cadherin) expression. Methods The trypsin-digested HUVEC were subcultured in culture flasks. When the cells were grown to 80%, they were randomly divided into four groups: phosphate buffer (PBS) control group (200 μL PBS), recombinant human high mobility group box 1 (rhHMGB1) treatment group (100 μg/L rhHMGB1), UFH control group (10 kU/L UFH), and UFH pretreatment group (10 kU/L UFH+100 μg/L rhHMGB1). The cells in each group were challenged with different reagent for 24 hours, and the activity of endothelial cells was determined by methyl thiazolyl tetrazolium (MTT) colorimetric assay. The permeability of endothelial cells was measured by Transwell method, and the expression and distribution of VE-cadherin was observed by immunofluorescence. The protein expressions of VE-cadherin and phosphorylated p38 mitogen-activated protein kinase (p-p38MAPK) were determined by Western Blot. Results After treatment with 100 μg/L rhHMGB1 for 24 hours, the activity of endothelial cells was not significantly different from that of the PBS control group (A value: 0.230±0.004 vs. 0.255±0.006, P > 0.05), but the permeability was significantly increased (glucan FD40 fluorescence intensity: 11.05±0.12 vs. 6.34±0.39, P < 0.05). Compared with PBS control group, the fluorescence microscopy showed that the VE-cadherin membrane localization was reduced, the distribution was loose, and there were obvious fissures between cells in rhHMGB1 treatment group, and quantitative analysis showed the protein expression of VE-cadherin was decreased significantly (VE-cadherin/β-actin: 0.16±0.04 vs. 0.31±0.03, P < 0.05), and the expression of p-p38MAPK protein was significantly increased (p-p38MAPK/β-actin: 0.79±0.03 vs. 0.26±0.05, P < 0.05). UFH pretreatment could protect HMGB1-mediated endothelial cell injury, cell permeability was significantly reduced (glucan FD40 fluorescence intensity: 9.11±0.23 vs. 11.05±0.12), fluorescence expression of VE-cadherin was enhanced, membrane localization was significantly increased, quantitative analysis showed that VE-cadherin protein expression was significantly up-regulated (VE-cadherin/β-actin: 0.24±0.02 vs. 0.16±0.04), and p38MAPK phosphorylation level was significantly decreased (p-p38MAPK/β-actin: 0.54±0.05 vs. 0.79±0.03), the difference was statistically significant as compared with rhHMGB1 treatment group (all P < 0.05). There was no significant difference in all parameters between PBS control group and UFH control group. Conclusions UFH can protect the endothelial cell barrier from the HMGB1 by regulating the expression and distribution of VE-cadherin. The mechanism may be related to the inhibition of p38MAPK phosphorylation by UFH.

Objective:To investigate the relations between enlarged perivascular spaces (EPVS) and cognitive impairment in patients with acute mild ischemic stroke.Methods:A total of 234 patients with acute mild ischemic stroke admitted to our hospital from October 2019 to June 2021 were chosen in our study. According to the Montreal Cognitive Assessment (MoCA) scores 7 d after admission, these patients were divided into normal cognitive function group (MoCA scores≥26) and cognitive impairment group (MoCA scores<26). The clinical data and imaging data of patients from the 2 groups were compared. Multivariate Logistic regression analysis was used to determine the independent influencing factors for post-stroke cognitive impairment (PSCI). Spearman correlation analysis was performed to analyze the correlations of severity degrees of EPVS of basal ganglia with MoCA total scores and each cognitive domain scores in patients from cognitive impairment group.Results:Among the 234 patients, 73 (31.2%) had normal cognitive function and 161 (68.8%) had cognitive impairment. As compared with normal cognitive function group, patients from cognitive impairment group had significantly older age, significantly less years of education, statistically higher fasting blood glucose level, significantly higher proportion of patients with moderate and severe basal ganglia EPVS, and significantly higher proportion of patients with white matter lesion (WML) grading 2 and 3 ( P<0.05). Multivariate Logistic regression analysis showed that age, years of education, basal ganglia EPVS and WML grading were independent influencing factors for PSCI ( OR=1.049, 95%CI: 1.007-1.093, P=0.021; OR=0.910, 95%CI: 0.832-0.995, P=0.039; OR=0.760, 95%CI: 1.176-2.637, P=0.006; OR=2.270, 95%CI: 1.219-4.228, P=0.010). Correlation analysis showed that the severity degrees of basal ganglia EPVS were negatively correlated with MoCA scores, and scores of visual space and executive ability scale, attention scale, language scale and delayed recall scale ( P<0.05). Conclusion:Acute mild ischemic stroke patients with older age, less years of education, severer basal ganglia EPVS and higher WML grading trends to have cognitive impairment; basal ganglia EPVS mainly affects the cognitive domains of visual space and executive ability, attention, language, and delayed recall.

This article introduces the health management service specification for a chronic patient population. The concept of this specification comes from the whole process of people-centered and integrated health management of the chronic patient population. It is also based on the World Health Organization′s integrated chronic patient population management model. The completion of the specification has gone through several rounds of expert discussion in related fields in China. The contents include seven key links: suspect selection, clinical diagnosis, health assessment, intervention plan development, intervention plan implementation, follow-up, and health reassessment. It also includes establishing a professional team and developing a support system and mechanism of financing, compensation, and incentives. The specification aims at integrating regional health resources through standardizing service technology and scientific management concepts. It would help to improve the efficiency and quality of health management of chronic patient population in China, thereby reducing the economic burden of chronic diseases for patients and their families, and achieving the goal of improving residents′ health.

OBJECTIVE: To assess the association of polymorphisms of rs3819024 and rs8193037 loci in the promoter region of IL-17A gene with the risk of ischemic stroke (IS) among ethnic Han Chinese from Guangxi. METHODS: The polymorphisms of rs3819024 and rs8193037 loci were detected by a SNaPshot assay and DNA sequencing among 392 IS patients and 443 healthy controls with matched age and gender. RESULTS: The genotypes, dominant model, recessive model, and alleles of rs3819024 polymorphisms showed no significant difference between the two groups, with the P values calculated as 0.150, 0.227, 0.125, 0.594 and 0.202, respectively, and OR (95% CI) as 1.27(0.92-1.74), 1.28(0.86-1.91), 1.27(0.94-1.72), 1.10(0.78-1.54), and 1.13(0.94-1.38), respectively. The genotypes, dominant model, recessive model, and alleles of rs8193037 polymorphisms also showed no significant difference between the two groups, with the P values calculated as 0.722, 0.352, 0.863, 0.345 and 0.969, respectively, and OR (95% CI) as 0.94(0.65-1.35), 2.25(0.41-12.35), 0.97(0.68-1.38), 2.27(0.41-12.48), and 1.01(0.72-1.40), respectively. CONCLUSION Polymorphisms of the rs3819024 and rs8193037 loci of the IL-17A gene are not associated with the susceptibility to IS among ethnic Han Chinese from Guangxi.
Alleles ; Asian Continental Ancestry Group ; Brain Ischemia ; genetics ; Case-Control Studies ; China ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Interleukin-17 ; genetics ; Polymorphism, Single Nucleotide ; Stroke ; genetics

Objective To explore the effects of whole process management pattern on the complication of PICC among cancer patients. Methods From October 2012, we started to build up the whole process management pattern to plan, implement, check and cope with integration of 4 stages key element including environment, safety, quality and patient′s health for PICC patients, and it carried out in the whole hospital. We compared the number of PICC cases, success rate of first PICC insertion and incidence of complication between the time of January to October 2012 and the time of November 2012 to December 2013, and compared the patients′satisfaction of nursing quality. Results After the whole process management pattern used, the total number of PICC climbed from 275 cases to 475 cases and the success rate of first PICC insertion increased from 94. 9% to 98. 9% with statistical difference (χ2 =11. 58,P<0. 01). The incidences of complication declined, of which the rate of mechanical phlebitis was 1. 8%, infection related catheter 2. 1%, and all complications were lower than before the whole process management pattern used (χ2 =5. 99,25. 17, respectively; P <0. 01). The patients′satisfaction increased from 93. 1% to 97. 1% with statistical difference (χ2 =6. 53,P<0. 05). Conclusions The whole process management pattern declines the complications of PICC among cancer patients, and promotes patients safety, releases clinical pressure and enhanced patients′satisfaction and quality of specific nursing.

Objective: In this study, we tested for the presence of four human coronaviruses (HCoVs) in children with respiratory tract disease in Fuzhou, Fujian, China. Methods: Nasopharyngeal aspirates were collected from children with respiratory tract disease from Nov, 2007 to Jan, 2015. A total of 266 clinical samples were tested for HCoVs using reverse-transcription polymerase chain reaction (RT-PCR). The positive products were sequenced and compared with those in GenBank by BLAST. The positive samples were then tested for HCoV-HKU1 and HCoV-NL63 using RT-PCR method . We compared the 440 bp pol gene sequence of the 8 HCoV isolates in Fuzhou, China to other HCoV isolates documented in the GenBank database by using MEGA software version 6.06 and the neighbor-joining method . Results: HCoVs were detected in 8 patients (3.0%) out of the 266 children. Two of 266 (0.38%) were positive for HCoV-HKU1; 1 of 266(0.38%)were positive for HCoV-NL63; 1 of 266 (0.38%) were positive for HCoV-229E; 4 of 266 (1.5%)were positive for HCoV-OC43. All of children who were positive for HCoV had respiratory illness. Two HCoV-HKU1 were found to co-infect with human parainfluenza virus type 3 (HPIV-3). The 8 HCoV strains in our study fell into four clusters. Two strains of HCoV-HKU1 were genotype A. Conclusions HCoV infections were probably associated with upper and lower respiratory illness in children. Additional studies are needed to investigate the potential roles of these HCoVs in diseases.