Analysis Of Prognostic Factors In Patients With Postoperative Intracranial Infection And Screening Of CircRNAs In Infected Cerebrospinal Fluid

Objective:In the clinical part of this study,we collect the clinical cases of patients with intracranial infection following neurosurgery,and study the clinical factors related to the prognosis of patients with central nervous system infection after neurosurgery,for the purpose of guiding the.clinical work and improving the prognosis of patients with infection.In the laboratory part of this study,we use gene chip technology and qRT-PCR technology to analyze the cerebrospinal fluid?CSF?of patients diagnosed with postoperative intracranial infection,to study the circRNAs of significant differential expression in the infected CSF,and use bioinformatics to analyze its possible mechanism,for the purpose of using it as the biomarker of intracranial infection for laboratory diagnosis of cerebrospinal fluid,and improving the prognosis of patients.Methods:The clinical part of this study was a retrospective study;including 113 cases of patients diagnosed with central nervous system infection after neurosurgery.The multiple clinically relevant factors in the cases were analyzed by univariate analysis,among them,the continuous variables were analyzed by two independent samples t-test or two independent samples rank sum test,the categorical variables were analyzed by chi-square test or Fisher's exact test.Finally,the significantly different factors were analyzed by multi-factor logistic regression.The laboratory part of this research included 33 patients diagnosed with central nervous system infection after neurosurgery,and collected cerebrospinal fluid in their infectious and non-infective states.The first step was to collect cerebrospinal fluid samples from 3 patients,the collected cerebrospinal fluid of infected and non-infected states were analyzed by circRNA array,and circRNAs with significantly different expression in the infected cerebrospinal fluid were screened out,in which 14 circRNAs were selected for validation.In the second step,the qRT-PCR technology was used to validate the 14 candidate circRNAs with the samples analyzed in the circRNA array,and 3 circRNAs were selected for the expanded sample validation.In the third step,the qRT-PCR technology was used to analyze cerebrospinal fluid samples collected from 30 patients to validate significantly different expression of the three circRNA as biomarkers for intracranial infection following neurosurgery,and finally bioinformatics was used to analyze its possible mechanism.Results:The clinical part of this study included age,primary disease,infection-related surgery time?days?,infection-related surgery times,emergency surgery,drainage time,antibiotic treatment time?days?,cerebrospinal fluid leucocytes?×106/L?,CSF erythrocytes?X106/L?,the ratio between erythrocytes and leucocytes of CSF,CSF mononuclear cell percentage?%?,CSF multinucleated cell percentage?%?,the glucose content in the CSF?mmol/l?,CSF protein?mg/1?,CSF chlorine?mmol/l?,duration of infection?days?and other factors for univariate analysis;The analysis results showed emergency surgery,the number of pre-infection operations,the drainage of tube placement before infection,the number of antibiotics used,the number of antibiotics used?days?,the number of leucocytes in the cerebrospinal fluid?106/L?,the glucose content in the cerebrospinal fluid?mmol/l?,and CSF proteins?mg/l?,bacterial cultured?categorized?,number of bacterial cultured,and the duration of infection?P<0.05?have significant differences.Influential factors that are significant in univariate analysis are included in logistic analysis,and conclusions are drawn.Tube placement time before infection?b:-0.113,OR:0.893,95%CI:0.805-0.991,P:0.033?,number of therapeutic antibiotics?b:-1.470,OR:0.230,95%CI:0.072-0.738,P:0.013?,the number of leukocytes in the cerebrospinal fluid?b:-0.016,OR:0.984,95%CI:0.970-0.998,P:0.027?are all risk factors for cure,and the duration of application of antibiotics?days??b:0.176,OR:1.193,95%CI:1.063-1.339,P:0.003?is a protective factor for cure.In the laboratory part of this study,the first step,circRNA array found 214 circRNAs with significantly different expression in infected cerebrospinal fluid,of which 72 circular RNAs were up-regulated and 142 were down-regulated expression.Through data analysis,select the one.14 circRNAs with a large fold difference and P value meeting the requirements for significant differences were selected for the validation.The circRNAs of upregulation is hsa_circRNA₄02768,hsa_circRNA₄02632,hsa_circRNA₀61260,hsa_circRNA₀48410,hsa_circRNA₀48148,hsa_circRNA₄02482,the circRNAs of downregulation is hsa_circRNA₀55243,hsa_circRNA₁01900,hsa_circRNA₄05481,hsa_circRN A₀07059,hsa_circRNA₁01366,hsa_circRNA₁02213,hsa_circRNA₀08636,hsa_circRNA₀09012.In the second step,the qRT-PCR technology was used to validate the 14 candidate circRNAs with the samples used in the circRNA array.3 circRNAs with a large fold change and a small p value were selected for validation.The up-regulated circRNA was hsa_circRNA₄02632,the down-regulated circRNAs was hsa_circRNA₀08636 and hsa_circRNA₄05481.The qRT-PCR technology was used to validate the 3 candidate circRNAs with the expanded samples collected from 30 patients diagnosed with central nervous system infection after neurosurgery.The up-regulated circRNA is hsa_circRNA₄02632 with a fold change of 2.065,and the P value is much less than 0.05.The hsa_circRNA₀08636 is down-regulated whose fold change is 0.416 and the P value is much less than 0.05,the hsa_circRNA₄05481 expression is down-regulated,the fold change is 0.452,and the P value is much less than 0.05.The functions of these three circRNAs are unclear.Bioinformatics analysis had been performed.FoxO and AMPK signaling pathways are the most important pathways for hsa_circRNA₀08636.Hsa_circRNA₀08636 may be related to oxidative stress and inflammatory responses,have broad application prospects in the diagnosis of intracranial infection.hsa_circRNA₄02632 can be enriched in the insulin resistance pathway,and hsa_circRNA₄05481 can be enriched in endometrial cancer,the Fc-epsilon-RI signaling pathway and the TGF-beta signaling pathway,both of which have the potential to diagnose intracranial infection.Conclusions:The clinical part of this study analyzed 113 cases of patients with intracranial infection after neurosurgery and concluded that the influencing factors related to the prognosis of patients with intracranial infection after neurosurgery were:Tube placement time,Number of therapeutic antibiotics,The number of leukocytes in the cerebrospinal fluid are all risk factors for cure,and the duration of application of antibiotics?days?is a protective factor for cure.The laboratory part of this study analyzed cerebrospinal fluid samples from 33 patients with intracranial infection after neurosurgery.The up-regulated circRNA was hsa_circRNA₄02632,the down-regulated circRNA were hsa_circRNA₀08636 and hsa_circRNA₄05481,all of which had significant expression differences and had potential as biomarkers for laboratory diagnosis of intracranial infection after clinical neurosurgery by some possible mechanism according to the bioinformatics analysis.