Dengue fever is an acute arbovirus-borne infectious disease caused by dengue virus. Its clinical characteristics include sudden fever， generalized pain， rash， hemorrhage and leukopenia. Severe cases often accompany severe multi-organ damage and even death. The dengue fever epidemic in China has a tendency to spread across the country， and some inland provinces have insufficient experience in early diagnosis and treatment of severe cases. To further standardize the clinical diagnosis and treatment of dengue fever， based on the “Guidelines for the diagnosis and treatment of dengue fever （2014 edition） （2nd edition）”，combined with domestic and international research progress and diagnostic and treatment experience， the“Diagnosis and treatment scheme for dengue fever （2024 edition）” was formulated by the National Health Commission （NHC） and the State Administration of Traditional Chinese Medicine. Based on this version of the diagnosis and treatment scheme， this article interprets the pathogenesis， diagnostic definition and treatment principles， with the aim of improving medical quality， reducing severe illness and mortality rates.
To establish a method for detection of Klebsiella pneumoniae （KP） based on recombinase aided amplification （RAA）-clustered regularly interspaced short palindromic repeats （CRISPR）/CRISPR-associated protein （CRISPR-Cas13a） system.
Methods
CrRNA and RAA primer pairs were designed and screened for the detection of KP gene， while a rapid and accurate method for the detection of KP gene was developed based on RAA-CRISPR-Cas13a technology. The RAA-CRISPR and real time fluorescence quantitative PCR （RT-qPCR） methods were compared by the synthesized KP plasmid. Genomic DNA was extracted from clinical strains of Pseudomonas aeruginosa， Acinetobacter baumannii， Escherichia coli and Streptococcus pneumoniae for RAA-CRISPR detection to evaluate the specificity of this method.Total of 50 clinical samples （including 30 KP culture-positive samples and 20 KP-negative samples） collected from Peking Union Medical College Hospital， Chinese Academy of Medical Sciences， from April 2023 to March 2024 were detected for KP through both RAA-CRISPR and RT-qPCR. The sensitivity， positive agreement rate and negative agreement rate of both methods were compared， respectively. Statistical analysis of data was performed by ANOVA and paired t-tests.
Results
The optimal crRNA and RAA primers for KP detection were selected by KP-positive plasmids， and CRISPR-Cas13a-based KP detection method was established successfully. The detection sensitivity of CRISPR-Cas13a for KP plasmids reached 1 copy/μl， higher than that of RT-qPCR （10 copies/μl）. The specific evaluation results showed that there was no cross-reactivity with nontarget strains， indicating that CRISPR-Cas13a system was specific. Among the sample detection， the sensitivity of RAA-CRISPR and RT-qPCR were 100% （30/30） and 83.3% （25/30）， respectively， using bacterial culture and mass spectrometry technology as the gold standard. The positive concordance rate of both detection were 100% （30/30） and 83.3% （25/30）， and the negative concordance rate was 100%.
Conclusions
A method was established to accurately detect KP gene by RAA amplification technology and CRISPR-Cas13a system. This method can accurately detect KP gene， which can help diagnose KP infection for timely and effective treatment.
To investigate the influence factors for post infectious bronchiolitis obliterans（PIBO） in children with severe pneumonia， and construct and validate a column chart prediction model.
Methods
The medical records of 252 children with severe pneumonia treated in Jianhu Clinical Medical College of Yangzhou University from January 2022 to December 2023 were collected. The children were followed up for 6 months and divided into PIBO group （192 cases） and non-PIBO group （60 cases）according to whether PIBO occurred. The clinical indicators of the two groups were compared to screen the influence factors for PIBO in severe pneumonia children， and independent influence factors were screened by multivariate Logistic regression analysis. A nomogram model was constructed， and the differentiation of the prediction model was evaluated by receiver operating characteristic curve （ROC）. The clinical practicability and calibration degree of the prediction model were evaluated by decision curve and calibration curve.
Results
Among the 252 children with severe pneumonia， a total of 60 cases occurred PIBO， with the incidence rate of 23.81% （60/252）. The age of children in PIBO group ［（18.87 ± 6.72） months］was lower than that of non-PIBO group ［（28.30 ± 9.04） months］（t = 8.648， P ＜ 0.001）； the duration of fever ［（13.77 ±3.75） days］was longer than that of non-PIBO group ［（9.92 ± 3.05） days］（t = 8.057， P ＜ 0.001）； the level of LDH ［（725.78 ± 98.72） U/L］was higher than that of non-PIBO group ［（628.49 ± 88.35） U/L］（t = 7.236， P ＜0.001）； the duration of mechanical ventilation ［（7.10 ± 2.33） days］was longer than that of non-PIBO group［（4.89 ± 0.97） days］（t = 10.541， P ＜ 0.001）， all with significant differences. The results of multivariate binary Logistic regression analysis showed that age （OR = 0.836， 95%CI： 0.773-0.904）， duration of fever （OR =1.548， 95%CI： 1.288-1.861）， LDH （OR = 1.014， 95%CI： 1.008-1.021） and duration of mechanical ventilation （OR =2.060， 95%CI： 1.496-2.836） were all independent influence factors for the occurrence of PIBO in children with severe pneumonia （all P ＜ 0.001）. The area under the ROC curve （AUC） of the column chart prediction model was 0.960 （95%CI： 0.937-0.984）. The calibration curve of the predictive model indicates that the predicted probability of the model was close to the actual probability， and the calibration degree was well.The decision curve DCA of the prediction model was located above the None and All lines， indicating that the model had clinical practicality within this range. The Hosmer Lemeshow Chi-square test showed well fit of the prediction model.
Conclusions
Age， duration of heat， LDH and duration of mechanical ventilation were all independent influence factors for PIBO in children with severe pneumonia. Constructing a column chart prediction model based on these independent influencing factors can provide a practical tool for early identification and intervention of PIBO in children with severe pneumonia.
To explore the dynamic changes of heparin binding protein （HBP） and interleukin-6 （IL-6） levels in children with lobar pneumonia （LP）， and to analyze the correlation between HBP，IL-6 and disease severity.
Methods
The clinical data of 160 children with LP admitted to Suzhou Wujiang Distinct Children's Hospital from January 2022 to August 2023 were analyzed， retrospectively. According to the severity of pneumonia， they were divided into common pneumonia group （93 cases） and severe pneumonia group （67 cases）， and healthy children who underwent physical examination at the same time were selected as healthy control group （60 cases）. The general data and laboratory indicators of children of three groups were analyzed. HBP and IL-6 levels of children in normal pneumonia group and severe pneumonia group were analyzed before treatment and 3 d， 7 d， 14 d after treatment. The effects of gender， age， body mass index（BMI）， duration of fever， duration of hospitalization， lung lobe lesions and pathogen distribution on the levels of ΔHBP and ΔIL-6 were analyzed by hierarchical regression （Forward method）. The correlation between HBP and IL-6 levels between common pneumonia group and severe pneumonia group were analyzed by locally weighted regression （Lowess）. Logistic regression was used to analyze the independent correlation between HBP， IL-6 levels and disease severity. The diagnostic efficacy of HBP， IL-6 alone and in combination for the severity of LP were evaluated by the receiver operating characteristic curves （ROC）. The restricted cubic spline（RCS） model was established to analyze the dose-response relationship with associated strength between HBP level， IL-6 level and disease severity. The clinical practical value of HBP and IL-6 models were evaluated by decision curve analysis.
Results
The levels of white blood cell （WBC）， neutrophil （NEU）， lymphocyte （LYM），platelet （PLT）， neutrophil-lymphocyte ratio （NLR）， erythrocyte sedimentation rate （ESR）， C-reactive protein（CRP）， procalcitonin （PCT）， immunoglobulin A （IgA）， immunoglobulin M （IgM）， immunoglobulin G （IgG），CD4+ T， CD8+ T and CD4+/CD8+ T among children in common pneumonia group， severe pneumonia group and healthy control group were all significantly different （F = 10.899， 68.235， 7.467， 24.068， 41.612， 151.070，283.137， 435.08， 73.047， 68.450， 59.703， 28.519， 32.398， 30.491； all P ＜ 0.001）. The duration of fever， duration of hospitalization， NEU， PLT， NLR， ESR， CRP， PCT， IgA， IgG， IgM， CD4+ T， CD8+ T， CD4+/CD8+ T （t = 7.681，8.628， 3.969， 4.125， 4.474， 5.551， 4.025， 11.996， 7.188， 6.208， 4.005， 3.151， 4.046， 4.463； all P ＜ 0.001）， HBP and IL-6 before treatment as well as 3 d， 7 d and 14 d after treatment in common pneumonia group and severe pneumonia group were all significantly different （HBP： t = 19.684， 17.632， 14.883， 6.72； all P ＜ 0.001. IL-6：t = 11.667， 10.454， 9.444， 18.424； all P ＜ 0.001）. The hierarchical regression model analysis showed that gender，age， body mass index （BMI）， lung lobe disease and pathogen distribution had significant positive effects on ΔHBP and ΔIL-6 （ΔHBP： F = 25.074， 21.935， 17.402， 14.333， 10.577； all P ＜ 0.001. ΔIL-6： F =14.512， 12.249，11.248， 18.218， 20.506； all P ＜ 0.001）. Lowess analysis showed that there was a significant linear positive correlation between HBP and IL-6 of common pneumonia group and severe pneumonia group （r = 0.50， 0.53；both P ＜ 0.001）. Logistic regression analysis showed that after adjusting covariates， HBP and IL-6 were still risk factors for the severity of LP （OR = 1.758， 95%CI： 1.622-1.891， P ＜ 0.001； OR = 1.207， 95%CI： 1.154-1.260，P = 0.001）， and there was an independent correlation with the severity of LP， the trend test of HBP and IL-6 from low to high quintile array was statistically significant （t = 13.002， 6.068； both Ptrend ＜ 0.001）. The sensitivity analysis showed E value were 1.701 and 1.273， respectively. Subgroup analysis showed that there were statistical differences in HBP and IL-6 before treatment as well as 3 d， 7 d and 14 d after treatment between children with Mycoplasma pneumoniae infection and Streptococcus pneumoniae infection in common pneumonia group and severe pneumonia group （all P ＜ 0.001）. ROC curve analysis for diagnosing the severity of LP showed that the combined diagnostic effect of HBP and IL-6 was better， with an AUC of 0.991 （95%CI： 0.980-0.999， P ＜ 0.001），and the sensitivity， specificity and accuracy were 94.03%， 97.85% and 96.25%， which were significantly higher than those of single index diagnosis. RCS model analysis showed that strength association of both HBP and IL-6 levels were associated with the severity of LP in a dose-response relationship （Pfornonlinear = 0.331， 0.544）.The decision curve analysis showed that HBP and IL-6 prediction models had high clinical practical value.
Conclusions
HBP and IL-6 levels are significantly increased in children with LP， and independently correlated with the severity of the disease. They are important biomarkers for evaluating the disease and have important clinical value.
To investigate the effect and interaction of vaginal mucosal immune function and human papilloma virus （HPV）16/18 infection on therapeutic efficacy of aminolevulinic acidphotodynamic therapy （ALA-PDT） in patients with cervical intraepithelial neoplasia （CIN）.
Methods
Total of 124 patients with CIN admitted to Jiangyin Traditional Chinese Medicine Hospital from June 2021 to March 2023 were collected. According to the efficacy of ALA-PDT， the patients were divided into ineffective group （31 cases） and effective group （93 cases）. The general information， vaginal mucosal immune function and HPV16/18 infection status of the two groups were compared. The influencing factors of ineffective ALAPDT in CIN patients were analyzed by multiple Logistic regression analysis， and the interaction between vaginal mucosal immune function and HPV16/18 infection on ALA-PDT efficacy in CIN patients were analyzed by a generalized multi factor dimensionality reduction model.
Results
Age （χ2 = 4.803， P = 0.028）and history of miscarriage （χ2 = 12.949， P ＜ 0.001） between effective group and ineffective group were significantly different. CD3+ T lymphocytes ［（60.05 ± 12.26）% vs. （48.82 ± 9.63）%： t = 4.641， P ＜ 0.001］，CD4+ T lymphocytes ［（39.05 ± 7.05）% vs. （33.10 ± 5.21）%： t = 4.318， P ＜ 0.001］， CD4+/CD8+ T ［（1.38 ± 0.31）vs. （1.09 ± 0.23）： t = 4.783， P ＜ 0.001］， IgA ［（2.21 ± 0.66） g/L vs. （1.61 ± 0.43） g/L： t = 4.731， P ＜ 0.001］，IgG ［（13.68 ± 4.18） g/L vs. （10.35 ± 3.21） g/L： t = 4.051， P ＜ 0.001］， IgM ［（1.26 ± 0.29 g/L） vs. （1.07 ± 0.23） g/L：t = 3.314， P = 0.001］and the proportion of HPV16/18 positivity ［37 （39.78%） vs. 21 （67.74%）： χ2 = 7.299，P = 0.007］of patients in effective group were significantly higher than those of the ineffective group； CD8+ T lymphocytes of patients in effective group ［（27.50 ± 6.08）% vs. （30.36 ± 7.11）%： t = 2.172， P = 0.032］was significantly lower than that of the ineffective group， with significant differences. Logistic regression analysis showed that age （OR = 1.864， 95%CI： 1.100-3.158， P = 0.021）， CD4+/CD8+ T （OR = 0.586， 95%CI： 0.433-0.792， P = 0.001）， IgA （OR = 0.657， 95%CI： 0.497-0.868， P = 0.003） and HPV16/18 （OR = 2.767， 95%CI：1.239-6.180， P = 0.013） were independent influencing factors for ALA-PDT failure in CIN patients. GMDR interaction analysis results showed that CD3+ T lymphocyte， CD4+ T lymphocyte， CD8+ T lymphocyte， CD4+/CD8+ T， IgA and HPV16/18 infection interacted with each other in the clinical efficacy of CIN （test sample accuracy = 0.7342， P ＜ 0.01）.
Conclusions
Age， CD4+/CD8+ T， IgA level and HPV16/18 infection were all independent influencing factors for the efficacy of ALA-PDT in CIN patients. Logistic regression model established based on the above factors could predict the clinical efficacy of ALA-PDT.
To investigate the differences of high mobility group protein B1 （HMGB1）level among children with refractory Mycoplasma pneumoniae pneumonia （RMPP） of different ages and its predictive value for prognosis.
Methods
A retrospective analysis was carried out on 132 children with RMPP admitted to Yuechi County People's Hospital from February 2023 to March 2024. According to the age， children were divided into infant group （≤ 3 years old， 38 cases）， preschool group （3-6 years old， 48 cases）and school-age group （≥ 6 years old， 46 cases）. The proportions of gender， body mass index （BMI）， as well as fever， wheezing， shortness of breath， moist rales， wheezing sounds， expectoration， cough and skin lesions among children of different age groups were compared， respectively. The correlations between HMGB1 and tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， interleukin-8 （IL-8）， immunoglobulin A （IgA），immunoglobulin G （IgG）， immunoglobulin M （IgM）， CD3+ T， CD4+ T， CD8+ T and CD4+/CD8+ T of different ages children were analyzed by multiple linear regression. According to different outcomes， the enrolled children were divided into improved group （101 cases） and deteriorated group （31 cases）. The influencing factors for the prognosis of children with RMPP were screened by multivariate Logistic regression analysis.The synergistic effect of HMGB1 and age on the prognosis of children with RMPP were analyzed by Cox proportional hazards model. The dose-response relationship between HMGB1 levels and the risk of disease deterioration of children with RMPP were analyzed by restricted cubic spline model （RCS）.
Results
Among RMPP children in infant group， preschool group and school-age group， cases with fever （χ2 = 20.696， P ＜0.001）， wheezing （χ2 = 17.109， P ＜ 0.001）， shortness of breath （χ2 = 20.213， P ＜ 0.001）， moist rales （χ2 = 11.305，P = 0.004）， wheezing sounds （χ2 = 19.072， P ＜ 0.001）， expectoration （χ2 = 21.414， P ＜ 0.001）， and levels of ESR （F = 30.461， P ＜ 0.001）， NE （F = 9.848， P ＜ 0.001）， PCT （F = 62.067， P ＜ 0.001）， CRP （F = 12.372，P ＜ 0.001）， HMGB1 （F = 29.395， P ＜ 0.001）， TNF-α （F = 44.713， P ＜ 0.001）， IL-6 （F = 3.206， P = 0.044），IL-8 （F = 22.069， P ＜ 0.001）， IgA （F = 71.892， P ＜ 0.001）， IgG （F = 4.142， P = 0.018）， IgM （F = 17.033，P ＜ 0.001）， CD3+ T cells （F = 22.663， P ＜ 0.001）， CD4+ T cells （F = 10.431， P ＜ 0.001）， CD8+ T cells （F =5.878， P = 0.004）， and CD4+/CD8+ T （F = 13.238， P ＜ 0.001） were all significantly different. Multiple linear regression showed that among RMPP children of the infant group， preschool group and school-age group，CD3+ T， CD4+ T and CD4+/CD8+ T were negatively correlated with the level of HMGB1， while TNF-α， IL-6，IL-8， IgA， IgG， IgM and CD8+ T were positively correlated with the level of HMGB1. Multivariate Logistic regression analysis showed that TNF-α （OR = 1.242， 95%CI： 1.182-2.307， P = 0.048）， IL-6 （OR = 5.766， 95%CI：1.308-10.312， P = 0.035）， IL-8 （OR = 2.445， 95%CI： 1.166-5.156， P = 0.040）， IgA （OR = 2.782， 95%CI： 1.389-9.332， P = 0.009）， IgG （OR = 9.186， 95%CI： 1.224-13.264， P = 0.020）， IgM （OR = 3.354， 95%CI： 1.179-6.196， P =0.037）， CD3+ T （OR = 2.807， 95%CI： 1.193-5.294， P = 0.026）， CD4+/CD8+ T （OR = 1.421， 95%CI： 1.204-5.245，P = 0.042） and HMGB1 （OR = 4.628， 95%CI： 1.318-6.141， P = 0.003） were all independent risk factors influencing the prognosis of children with RMPP. When HMGB1 ＞ 86 ng/ml and age ≥ 6 years coexisted（OR = 2.746， 95%CI： 1.396-4.462， P = 0.013）， the risk of disease deterioration among children with RMPP was higher. RCS model analysis showed that regardless of whether confounding factors were adjusted，there was a non-linear positively correlation between HMGB1 and the deterioration of RMPP in children of different age groups （before adjustment： Pnon-linearity = 0.034， 0.046， 0.069； after adjustment： Pnon-linearity =0.053， 0.067， 0.084）.
Conclusions
HMGB1 level show significant differences among children with RMPP of different ages. Moreover， HMGB1 level was positively correlated with TNF-α， IL-6， IL-8， IgA， IgG， IgM and CD8+ T. On the other hand， HMGB1 level was negatively correlated with CD3+ T， CD4+ T and CD4+/CD8+ T.HMGB1 level was positively correlated with the risk of disease deterioration of children with RMPP and had a certain predictive value for the outcome.
To explore the value of procalcitonin （PCT） level in identification of bacterial bronchitis pathogen types， and investigate the value of guidance of moxifloxacin application.
Methods
Total of 236 patients with bacterial bronchitis admitted to Beijing Gulou Hospital of Traditional Chinese Medicine from November 2018 to December 2023 were selected. Bacterial pathogens types of the patients were detected by fully automatic bacterial identification instrument， and PCT levels of patients in Grampositive group （160 cases） and Gram-negative group （76 cases） were compared. The diagnostic value of PCT level for bacterial bronchial pathogen types identification were analyzed by receiver operating characteristic curve （ROC）. The research subjects were randomly divided into PCT guidance group （Moxifloxacin was given to patients based on PCT level） and control group （Moxifloxacin was given as routine treatment） by a numerical table method， with 118 cases in each group. The disappearance time of symptoms， levels of inflammatory factors and the clinical efficacy of both groups of patients were compared， respectively.
Results
Among the 236 patients with bacterial bronchitis， 160 strains of Gram-positive bacteria were detected， with a detection rate of 67.80%； 76 strains of Gram-negative bacteria were detected， with a detection rate of 32.20%.Compared with the Gram-positive bacterial group ［2.27 （0.72， 6.22） ng/ml］， the PCT level of patients in Gramnegative bacterial group ［7.13 （2.50， 17.65） ng/ml］was significantly increased， with significant difference（U = 6.365， P ＜ 0.001）. The ROC curve analysis of PCT level for pathogen type identification in patients with bacterial bronchitis showed that the Yoden index （0.535） was the highest at a PCT level of 5.42 ng/ml， with corresponding sensitivity and specificity of 0.739 and 0.796， respectively. The area under ROC curve （AUC）was 0.784. The disappearance time of wheezing ［（2.77 ± 0.83） d］， temperature recovery time ［（2.68 ± 0.81） d］， cough disappearance time ［（3.52 ± 1.41） d］and duration of hospitalization ［（5.67 ± 2.46） d］of patients in PCT guidance group were significantly lower than those of the control group ［（4.36 ± 1.32） d， （4.53 ± 1.44） d， （6.27 ± 1.39） d and （9.12 ± 3.52） d］，with significant differences （t = 11.077， 12.163， 15.088， 8.727， all P ＜ 0.001）. After 1 day， 7 days and 14 days of treatment， the cough symptom scores and C-reactive protein （CRP） levels of patients in PCT guidance group were significantly lower than those of the control group； After 7 days and 14 days of treatment， the levels of PCT and white blood cell count （WBC） of patients in PCT guidance group were significantly lower than those of the control group， with significant differences （all P ＜ 0.001）. The total effective rate of patients in PCT guidance group after 14 days of treatment was 93.22% （110/118）， significantly higher than that of control group （77.12%，91/118）， with significant differences （χ2 = 12.110， P = 0.001）.
Conclusions
Serum PCT level can be used as an effective indicator for the pathogenic type identification of bacterial bronchitis. By monitoring serum PCT level，the use of moxifloxacin of patients with bacterial bronchitis can be guided.
To investigate the value of surgical treatment for patients of acquired immune deficiency syndrome （AIDS） complicated with rectal prolapse， and to improve the diagnostic and therapeutic understanding of rectal prolapse.
Methods
The clinical records， diagnosis and treatment of a patient with AIDS complicated with rectal prolapse of grade Ⅲ admitted to Chengdu Anorectal Hospital on November 29th， 2023 were reviewed， retrospectively.
Results
The patient was admitted to hospital presenting with rectal prolapse. Physical examination showed that the anal mass protruded from the anus， in a ring shape，about 10 cm and hierarchical mucosal folds could be seen， which could not be manually reduced. After admission， transanal rectal partial resection and anastomosis and transanal pelvic floor reconstruction were performed under sacral canal anesthesia plus intravenous general anesthesia. The patient's anus regained its normal appearance， structure and function after operation. Three days after the operation， the patient began to defecate， with formed， soft and unobstructed stools. One week after the operation， the wound was basically healed， with no obvious pain， bleeding or edema. Both digital rectal examination and anoscopy showed no abnormalities， and anorectal manometry was normal. There was no recurrence of rectal prolapse during the 1-year follow-up.
Conclusions
Early diagnosis and comprehensive evaluation is important for the patients with AIDS and rectal prolapse. Appropriate surgical procedure is crucial for improving the prognosis and life quality of these patients.
