Objective Currently, there is no commercially available quantitative detection kit for the main Felis domestic allergen (Fel d 1) in China. To establish a rapid detection method for Fel d 1, this study aims to prepare monoclonal antibodies against Fel d 1 protein. Methods The codon preference of Escherichia coli was utilized to optimize and synthesize the Fel d 1 gene. The prokaryotic expression plasmid pET-28a-Fel d 1 was constructed and used to express and purify the recombinant Fel d 1 protein. Subsequently, the recombinant protein was immunized into BALB/c mice and monoclonal antibodies (mAbs) were prepared by the hybridoma technique. An indirect ELISA was established using the recombinant Fel d 1 as the coating antigen, and hybridoma cell lines were screened for positive clones. The specificity and antigenic epitopes of the mAbs were confirmed by Western blot analysis. Finally, the selected hybridoma cells were injected into the peritoneal cavities of BALB/c mice for large-scale monoclonal antibody production. Results The recombinant plasmid pET-28a-Fel d 1 was successfully constructed, and soluble Fel d 1 protein was obtained after optimizing the expression conditions. Western blot and antibody titer assays confirmed the successful isolation of two hybridoma cell lines, 7D11 and 5H4, which stably secreted mAbs specific to Fel d 1. Antibody characterization revealed that the 5H4 mAb was of the IgG2a subtype and could recognize the amino acid region 105-163 of Fel d 1, while the 7D11 mAb was the IgG1 subtype and could recognize the amino acid region 1-59. Conclusion The high-purity recombinant Fel d 1 protein produced in this study provides a promising alternative for clinical immunotherapy of cat allergies. Furthermore, the monoclonal antibody prepared in this experiment lays a material foundation for the in-depth study of the biological function of Fel d 1 and the development of ELISA detection.
Animals ; Antibodies, Monoclonal/biosynthesis* ; Mice, Inbred BALB C ; Cats ; Mice ; Allergens/genetics* ; Glycoproteins/genetics* ; Enzyme-Linked Immunosorbent Assay ; Hybridomas/immunology* ; Recombinant Proteins/genetics* ; Female ; Antibody Specificity

Objective:To study the risk factors for sepsis with acute pancreatitis (AP) in the intensive care unit, Nomogram models were constructed and evaluatedand.Methods:Clinical data of 306 AP patients were admitted to the First Department of Critical Care Medicine. The Second Affiliated Hospital of Anhui Medical University from October 2013 to October 2023 were included, they were divided into sepsis group and non-sepsis group according to whether sepsis occurred. Baseline data and laboratory test results of the two groups were compared. The study subjects were randomly(random number) divided into training set and verification set, and the optimal variables were selected by LASSO regression. Then multivariate Logistic regression analysis was performed to construct a Nomogram risk prediction model. Then the prediction efficiency of the model was evaluated.Results:A total of 306 AP patients were included, including 127 patients with sepsis and 179 patients without sepsis. According to the ratio of 7:3, 215 cases were randomly (random number) divided into the training set and 91 cases were verified. Multivariate Logistic regression analysis showed that BMI, APACHEⅡ score, platelet, IL-6, total bilirubin and complications were independent risk factors for sepsis in patients with AP. According to the above risk factors, a Nomogram prediction model was established. The area under ROC curve of the model in training set and verified set were 0.913 (95% CI 0.875-0.951) and 0.901 (95% CI 0.837-0.965) respectively. The calibration curve shows that the predicted probability is basically consistent with the actual probability. Decision curve analysis shows that it has good clinical net benefit in a large threshold range. Conclusions:The independent risk factors of AP associated with sepsis in ICU are BMI, APACHEⅡscore, platelet, IL-6, total bilirubin and complications. The Nomogram risk predictive model based on these factors has good predictive power.

Aim To investigate the effect of 96 h paradoxical sleep deprivation(PSD) on learning & memory ability of rats and the concomitant gene expression change in the rat hippocampus.Methods PSD model was established by Modified Multiple Platform Method(MMPM).Learning and memory ability of the PSD model was assessed through hexagonal maze compared with the control, and the gene expression in hippocampus of each group was detected by gene chip technology.After that, GO and KEGG enrichment analysis were applied to determine the changes of the gene expression in PSD model compared with the control.Results PSD affected the memory consolidation process in rats.The gene chips showed that 100 genes expressed differently in the hippocampus of PSD model compared with the control, and among them, 25 genes were up-regulated and 75 genes down-regulated.The up-regulated genes mainly involved in cell growth and differentiation, learning and memory, inflammation, oxidative stress, and biological processes such as synaptic transmission, while the down-regulated genes mainly involved in immune response, central nervous system development, synaptic transmission, metabolism, signal transduction, neurotransmitter secretion, cell apoptosis, and so on.KEGG analysis showed that genes differentially expressed mainly involved in cGMP-PKG, metabolism, NF-kappa B and Pathways in cancer, and so on.Conclusion In rats, PSD leads to memory deficit, as well as a variety of gene expression changes in hippocampus of the PSD model.

Objective Cerebral microbleeds (CMBs) are important indicators of cerebral small vessel disease .However, it is still unclear whether endothelial dysfunction is involved in CMBs .The aim of this study is to investigate the correlation between CMBs and soluble E-selectin (sE-selectin) in patients with acute ischemic stroke . Methods Based on the results of MRI (3.0 T) susceptibility weighted imaging , we divided patients with first acute ischemic stroke into a CMBs group ( n=63 ) and a non-CMBs group (n=63), and recruited another 45 volunteers with normal MRI findings as controls .We collected and conducted comparative a-nalysis on the demographic data , biochemical variables ( including the sE-selectin level ) , vascular risk factors , and the number of CMBs of the patients . Results Ordinal logistic regression analysis showed a significantly positive correlation between sE -selectin and the number of CMBs (OR=1.062, 95%CI:1.023-1.103, P=0.002), higher systolic blood pressure associated with more CMBs (OR=1.014, 95%CI:1.002-1.025, P=0.021). Conclusion Serum sE-selectin is significantly positively correlated with and can be used as a biological marker for the severity of CMBs .

Objective To investigate the differences in the epidemiological characteristics of patients in pre-hospital medical care before and after “5 · 12” Wenchuan earthhquake in Deyang city.Methods Comparison was made between the epidemiological features of cases treated in Deyang emergency medical center from the January 1,2007 through the December 31,2007 before the earthquake (the pre-earthquake group) and those treated from the January 1,2009 through the December 31,2009 after the earthquake ( the post-earthquake group) with the criteria of international classification of diseases (ICD-10).Results ( 1 ) There were 11 325 patients medically cared before earthquake and 16 265 patients after disaster.The distribution of sex,age,the top five causes,the seasonal and diurnal variations of emergency call,radius of service circle of pre-hospital medical care cases were no significantly different between the two groups ( P ＞ 0.05 ).( 2 ) The time required for scheduling ambulance and time consumed for the transportation of patients in the post-earthquake group were less than those in the pre-earthquake group ( P ＜ 0.05 ).Conclusions The efficiency of medical care done by 120 Emergency Command Center in Deyang city has been improved significantly in ease of without change in epidemiological features of patients after reconstruction of disaster area.

Transcranial Doppler (TCD) is a non-invasive,convenient and inexpensive method for cerebral hemodynamic assessment widely used in clinical practice.Monitoring of cerebrovascular reserve capacity with TCD can provide relevant information for clinical decision making in the treatment of carotid stenosis.During carotid angioplasty and stenting,TCD can be used to evaluate effect of stenting on hemodynamics,and predict major complications,particularly postoperative hyperperfusion.

Objective To investigate the enlarged perivascular space (EPVS) and its clinical significance in patients with cerebral small vessel disease (CSVD).Methods One hundred seventy-four patients with CSVD and 86 patients without CSVD admitted to Jinling Hospital,Clinical School of Nanjing University School of Medicine from October 2011 to February 2012 were recruited.All patients underwent cranial MRI examination (including diffusion-weighted imaging and fluid attenuated inversion recovery sequences).The numbers of EPVS and anatomic distribution in all the subjects of both groups were analyzed.The receiver operator characteristic (ROC) curve was used to investigate its diagnostic critical value of anatomic distribution.Results Multivariate logistic regression analysis showed that EPVS in basal ganglia region (odds ratio [OR] 1.491,95％ confidence interval [CI] 1.165-1.909; P =0.002) and EPVS in centrum semiovale (OR 1.279,95％ CI 1.022-1.601;P=0.032) were independently associated with CSVD.EPVS in the basal ganglia region and the centrum semiovale in patients with CSVD was significantly more than that in patients with non-CSVD (all P ＜0.001).Its corresponding diagnosis cut-off points of CSVD were 4 and 6 respectively.The area under the ROC curve and the diagnostic sensitivity and specificity were 0.859,72.4％,93.0％ and 0.808,65.5％,95.3％,respectively.Conclusions EPVS contributes to the diagnosis of CSVD.When using EPVS to diagnose CSVD,the anatomical sites need to be distinguished and establish appropriate diagnostic critical value.

Objective To observe the expression of IL-6 mRNA and study the mechanism of acupuncture in focal cerebral ischemia.Methods In situ hybridization was used to detect the expression of IL-6 mRNA in the sham operation group, control group and acupuncture group 2 h, 6 h, 12 h, 24 h after reperfusion in the brain of middle cerebral artery occlusion rats, meanwhile and also observe ultrastructure in cortex of ischemic brain tissue 24 h after reperfusion.Results The sham operation group showed a basic expression of IL-6 mRNA in cortex and striate body. The cells expressing IL-6 mRNA were increased 2 h after ischemic reperfusion in cortex and striate body and peaked 24 h after reperfusion in control group and acupuncture group. The cells expressing IL-6 mRNA in cortex of ischemic area were stronger in acupuncture group than in control group at every research time point besides 2 h after ischemic reperfusion. The cells expressing IL-6 mRNA striate body of ischemic area were stronger in acupuncture group than in control group at every research time point. Acupuncture can lighten the degeneration of cell nucleolus and necrosis of cell, resume the integrity of the nucleolus, improve the function of mitochondrial and endoplasmic, and have significant difference with control group. Conclusion Acupuncture can increase the cells expressing of IL-6 mRNA.