1.Analysis of factors associated with spread through air spaces(STAS) of small adenocarcinomas(≤2 cm) in peripheral stage ⅠA lungs and modeling of nomograms
Jing FENG ; Wei SHAO ; Xiayin CAO ; Jia LIU ; Jialei MING ; Ya’nan ZHANG ; Jianbing YIN ; Jin CHEN ; Honggang KE ; Lei CUI
Chinese Journal of Thoracic and Cardiovascular Surgery 2024;40(3):129-136
		                        		
		                        			
		                        			Objective:To investigate the relationship between spread through air spaces(STAS) of peripheral stage ⅠA small adenocarcinoma of the lung(≤2 cm) and related factors such as clinical and CT morphological features, and to construct a nomogram model.Methods:Relevant clinical, pathological and imaging data of patients who underwent lung surgery and were diagnosed as peripheral stage ⅠA small lung adenocarcinoma by postoperative pathology in the Affiliated Hospital of Nantong University from 2017 to 2022 were collected, of which cases that met the inclusion criteria from 2017 to 2021 served as the training group, and those that met the inclusion criteria in 2022 served as the validation group. The independent risk factors for the occurrence of STAS in peripheral stage ⅠA lung small adenocarcinoma were investigated by using univariate analysis and multifactorial logistic regression analysis, based on which a nomogram prediction model was constructed, and the subjects were analyzed by using the receiver operating characteristic curve( ROC), correction model, etc. were used to evaluate the model. Results:A total of 430 patients who met the criteria were included, including 351 patients in the training group(109 STAS-positive and 242 STAS-negative) and 79 patients in the validation group(23 STAS-positive and 56 STAS-negative). Univariate analysis showed that the patients in the two groups showed a significant difference in age(>58 years old), gender, smoking history, tumor location(subpleural, non-subpleural), pleural pull, nodule type, nodule maximal diameter, solid component maximal diameter, consolidation tumor ratio(CTR), lobulation sign, burr sign, bronchial truncation sign, vascular sign(includes thickening and distortion of blood vessels in/around the nodes), satellite lesions, and ground-glass band sign were statistically significant( P<0.05). The results of multifactorial logistic regression analysis showed that CTR( OR=4.98, P<0.001), lobulation sign( OR=4.07, P=0.013), burr sign( OR=3.66, P<0.001), and satellite lesions( OR=3.56, P=0.009) were the independent risk factors for the occurrence of STAS. Applying the above factors to construct the nomogram model and validate the model, the results showed that the ROC curve was plotted by the nomogram prediction model, and the area under the ROC curve( AUC) of the training set was 0.840(sensitivity 0.835, specificity 0.734), and the validation set had an AUC value of 0.852(sensitivity 0.786, specificity 0.783), and the training set and validation set calibration curves have good overlap with the ideal curve. Conclusion:CTR, lobular sign, burr sign, and satellite lesions are independent risk factors for STAS, and the nomogram model constructed in this study has good predictive value.
		                        		
		                        		
		                        		
		                        	
2.Distribution and Drug Resistance of Pathogens Causing Bloodstream Infection in Patients with Hematological Malignancies
Ming YANG ; Huai-Xin GENG ; Jin-Ge TAI ; Hai-Lian SHAO ; Jing-Wen CHEN ; Ke DONG
Journal of Experimental Hematology 2024;32(2):583-587
		                        		
		                        			
		                        			Objective:To investigate distribution and drug resistance of pathogens of bloodstream infection in patients with hematological malignancies,in order to provide reference for clinical infection control and treatment.Methods:The clinical information of blood culture patients in the hematology department of our hospital from January 2016 to December 2021 was reviewed.They were divided into transplantation group and non-transplantation group according to whether they had undergone hematopoietic stem cell transplantation.The types of pathogens and their drug resistance were analyzed.Results:Two hundred and ninety-nine positive strains of pathogenic bacteria were detected.In the transplantation group,Gram-negative bacteria accounted for 68.5%(50/73),Gram-positive bacteria accounted for 6.8%(5/73),and fungi accounted for 24.7%(18/73).The resistance rate of Escherichia coli to the third-generation cephalosporins was 77.8%,and 11.5%to carbapenems.The resistance rate of Klebsiella pneumoniae to the third-generation cephalosporins was 50.0%,and 56.2%to carbapenems.In the non-transplantation group,Gram-negative bacteria accounted for 64.1%(145/226),Gram-positive bacteria accounted for 31.0%(70/226),and fungi accounted for 4.9%(11/226).Gram-positive bacteria were mainly Enterococcus faecium(6.6%,15/226)and Coagulase-negative Staphylococci(6.2%,14/226).The fungi were all Candida tropicalis.The resistance rate of Escherichia coli to the third-generation cephalosporins was 63.8%,and 10.3%to carbapenems.The resistance rate of Klebsiella pneumoniae to the third-generation cephalosporins was 46.3%,and 26.8%to carbapenems.Conclusion:The types of pathogenic bacteria in bloodstream infection in patients with hematological malignancies are varied.Gram-negative bacteria is the main pathogenic bacteria.The resistance of pathogenic bacteria to antibiotics is severe.Antibiotics should be used scientifically and reasonably according to the detection and resistance of pathogenic bacteria.
		                        		
		                        		
		                        		
		                        	
3.The Preservation of HBV,HCV,HIV Viral Nucleic Acids in Plasma by Dry Spot Method and the Duration of Preservation
Jing ZHANG ; Chun-Yan SHAO ; Ling-Ling REN ; Ling-Ling GAO ; Ming-Hui WANG ; Qun LUO
Journal of Experimental Hematology 2024;32(6):1869-1874
		                        		
		                        			
		                        			Objective:To establish a method for preserving viral nucleic acids in plasma using a blood collection card based on the dry spot method,to predict the duration of nucleic acid preservation by establishing the Arrhenius equation,and to demonstrate the feasibility of this preservation method for the re-testing of nucleic acids in blood samples retained by blood banks.Methods:Plasma samples positive for HBV,HCV,and HIV nucleic acids were prepared into preservation cards in the form of dry plasma spots for storage.The prepared preservation cards were placed under accelerated storage conditions at 37,45,50,and 55 ℃.The preservation cards were periodically retrieved from each temperature condition for nucleic acid extraction,and the nucleic acid samples were purified for subsequent PCR testing,with the recorded CT values.An Arrhenius equation model was established between the expiration time and the storage temperature,thereby predicting the validity period of nucleic acid preservation in blood collection cards under specified storage temperature conditions.Results:For the plasma samples positive for HBV,HCV,and HIV nucleic acids preserved using the dry spot method,the regression equations for the duration with temperature were as follows:y=-11546x+31.74 for HBV,y=-12949x+36.88 for HCV,and y=-12204x+34.48 for HIV,with the correlation coefficient r greater than 0.98 for all.It was predicted that at a storage temperature of 4 ℃,the preservation periods for HBV,HCV,and HIV viral nucleic acids using the dry spot method would be 20792 days,19289 days,and 14285 days,respectively.At a storage temperature of 20 ℃,the preservation periods would be 2135 days 1502 days,and 1289 days,respectively.Conclusion:The nucleic acids of the three common viral pathogens in blood samples,when preserved using the dry spot method,conform to a first-order reaction pattern in the accelerated degradation experiment.The relationship between the rate of nucleic acid degradation and the absolute temperature of storage is consistent with the Arrhenius equation.Based on the calculations using this equation,the stability and validity period of plasma nucleic acid samples preserved using the dry spot method can reach a minimum of 3 .5 years under storage conditions not exceeding 20 ℃,which essentially meets the requirements for the preservation period of blood samples retained by blood banks.
		                        		
		                        		
		                        		
		                        	
4.Molecular study of a case with variant of RHCE*ce allele in haplotype dce resulting in weakened e antigen
Yongkui KONG ; Hecai YANG ; Ming SHAO ; Yinghui CHEN-LI ; Wanjin ZHANG ; Xiaoyan ZHANG ; Jing WANG ; Xianping LYU ; Qiankun YANG
Chinese Journal of Medical Genetics 2024;41(9):1039-1044
		                        		
		                        			
		                        			Objective:To explore the RH genotype for a female with RhD(-) blood type and its molecular basis. Methods:A 26-year-old female who had attended the outpatient clinic of the First Affiliated Hospital of Zhengzhou University in August 2019 was selected as the study subject. Peripheral blood samples were collected from the proband and her parents for Rh phenotyping with gel card method. PCR-sequence-based typing (PCR-SBT) and DNA sequencing were used to determine the RHD zygosity and RH genotype of the proband and her parents. Homology modeling of Rh proteins was performed with bioinformatic software, and protein structural alterations caused by the variant was simulated by molecular dynamics. This study was approved by the Medical Ethics Committee of the First Affiliated Hospital of Zhengzhou University (Ethics No. 2023-KY-0870-003). Results:Serological tests showed that the proband and her father both had weakened e antigen of the Rh phenotype. PCR-SBT and DNA sequencing showed that the genotypes of the proband and her parents were dce/ dCE, dce/ DcE and dCE/ DcE, respectively. And the genotypes of the RHD and RHCE of the proband were RHD*01N.01/ RHD*01N.16, RHCE*01.01/RHCE*04, respectively. Protein simulation and molecular dynamics analysis revealed that the ce_16C variant resulted from RHCE* ce (c.48G>C) may alter the structure of intracellular and extracellular loops, mainly affecting the mobility of extracellular loops 2, 6 and intracellular loops 3, 4. Conclusion:Variant of the RHCE* ce allele c. 48G>C probably underlay the weakened e antigen in this proband.
		                        		
		                        		
		                        		
		                        	
5.Development of a national health standard:Guideline for pediatric transfusion
Rong HUANG ; Qingnan HE ; Mingyan HEI ; Minghua YANG ; Xiaofan ZHU ; Jun LU ; Xiaojun XU ; Tian-Ming YUAN ; Rong ZHANG ; Xu WANG ; Jinping LIU ; Jing WANG ; Zhili SHAO ; Mingyi ZHAO ; Jia GUO ; Xiny-In WU ; Jiarui CHEN ; Qirong CHEN ; Rong GUI ; Yongjian GUO
Chinese Journal of Blood Transfusion 2024;37(7):839-844
		                        		
		                        			
		                        			Children and adults differ significantly in physiology,biochemistry and immune function,which leads to sig-nificant differences in blood transfusion strategies between children and adults.To guide the clinical transfusion practice of pediatric patients and improve the prognosis of children,the National Health Commission organized the formulation and re-lease of the health industry standard Guideline for Pediatric Transfusion(WS/T 795-2022).This paper will briefly introduce some concepts that help understand of the Standard and the preparation process of the Standard,and explain and interpret the preparation of the"scope","general provisions"and"factors to consider"of the Standard,hoping to contribute to the understanding and implementation of the Standard.
		                        		
		                        		
		                        		
		                        	
6.Recognition of abnormal changes in echocardiographic videos by an artificial intelligence assisted diagnosis model based on 3D CNN.
Kai Kai SHEN ; Xi Jun ZHANG ; Ren Jie SHAO ; Ming Chang ZHAO ; Jian Jun CHEN ; Jian Jun YUAN ; Jing Ge ZHAO ; Hao Hui ZHU
Chinese Journal of Cardiology 2023;51(7):750-758
		                        		
		                        			
		                        			Objective: To investigate the diagnostic efficiency and clinical application value of an artificial intelligence-assisted diagnosis model based on a three-dimensional convolutional neural network (3D CNN) on echocardiographic videos of patients with hypertensive heart disease, chronic renal failure (CRF) and hypothyroidism with cardiac involvement. Methods: This study is a retrospective study. The patients with hypertensive heart disease, CRF and hypothyroidism with cardiac involvement, who admitted in Henan Provincial People's Hospital from April 2019 to October 2021, were enrolled. Patients were divided into hypertension group, CRF group, and hypothyroidism group. Additionally, a simple random sampling method was used to select control healthy individuals, who underwent physical examination at the same period. The echocardiographic video data of enrolled participants were analyzed. The video data in each group was divided into a training set and an independent testing set in a ratio of 5 to 1. The temporal and spatial characteristics of videos were extracted using an inflated 3D convolutional network (I3D). The artificial intelligence assisted diagnosis model was trained and tested. There was no case overlapped between the training and validation sets. A model was established according to cases or videos based on video data from 3 different views (single apical four chamber (A4C) view, single parasternal left ventricular long-axis (PLAX) view and all views). The statistical analysis of diagnostic performance was completed to calculate sensitivity, specificity and area under the ROC curve (AUC). The time required for the artificial intelligence and ultrasound physicians to process cases was compared. Results: A total of 730 subjects aged (41.9±12.7) years were enrolled, including 362 males (49.6%), and 17 703 videos were collected. There were 212 cases in the hypertensive group, 210 cases in the CRF group, 105 cases in the hypothyroidism group, and 203 cases in the normal control group. The diagnostic performance of the model predicted by cases based on single PLAX view and all views data was excellent: (1) in the hypertensive group, the sensitivity, specificity and AUC of models based on all views data were 97%, 89% and 0.93, respectively, while those of models based on a single PLAX view were 94%, 95%, and 0.94, respectively; (2) in the CRF group, the sensitivity, specificity and AUC of models based on all views data were 97%, 95% and 0.96, respectively, while those of models based on a single PLAX view were 97%, 89%, and 0.93, respectively; (3) in the hypothyroidism group, the sensitivity, specificity and AUC of models based on all views data were 64%, 100% and 0.82, respectively, while those of models based on a single PLAX view were 82%, 89%, and 0.86, respectively. The time required for the 3D CNN model to measure and analyze the echocardiographic videos of each subject was significantly shorter than that for the ultrasound physicians ((23.96±6.65)s vs. (958.25±266.17)s, P<0.001). Conclusions: The artificial intelligence assisted diagnosis model based on 3D CNN can extract the dynamic temporal and spatial characteristics of echocardiographic videos jointly, and quickly and efficiently identify hypertensive heart disease and cardiac changes caused by CRF and hypothyroidism.
		                        		
		                        		
		                        		
		                        			Male
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Artificial Intelligence
		                        			;
		                        		
		                        			Retrospective Studies
		                        			;
		                        		
		                        			Echocardiography/methods*
		                        			;
		                        		
		                        			Heart Diseases
		                        			;
		                        		
		                        			Hypertension
		                        			;
		                        		
		                        			Hypothyroidism
		                        			
		                        		
		                        	
7.Expert consensus on the prevention and treatment of adverse reactions in subcutaneous immunotherapy(2023, Chongqing).
Yu Cheng YANG ; Yang SHEN ; Xiang Dong WANG ; Yan JIANG ; Qian Hui QIU ; Jian LI ; Shao Qing YU ; Xia KE ; Feng LIU ; Yuan Teng XU ; Hong Fei LOU ; Hong Tian WANG ; Guo Dong YU ; Rui XU ; Juan MENG ; Cui Da MENG ; Na SUN ; Jian Jun CHEN ; Ming ZENG ; Zhi Hai XIE ; Yue Qi SUN ; Jun TANG ; Ke Qing ZHAO ; Wei Tian ZHANG ; Zhao Hui SHI ; Cheng Li XU ; Yan Li YANG ; Mei Ping LU ; Hui Ping YE ; Xin WEI ; Bin SUN ; Yun Fang AN ; Ya Nan SUN ; Yu Rong GU ; Tian Hong ZHANG ; Luo BA ; Qin Tai YANG ; Jing YE ; Yu XU ; Hua Bin LI
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2023;58(7):643-656
8.Discussion on "sugar needle - comfortable acupuncture and moxibustion".
Tao HUANG ; Xiang-Hong JING ; Neng-Gui XU ; Qiang WU ; Yong-Ming LI ; Can-Hui LI ; Ye-Meng CHEN ; Wei-Dong LU ; Yi-Fan YANG ; Tian-Jun WANG ; Shao-Bai WANG
Chinese Acupuncture & Moxibustion 2023;43(3):341-344
		                        		
		                        			
		                        			The cases of feeling comfort during acupuncture and moxibustion treatment in literature were summarized and its biological basis was explored. A simple classification of comfort was made, and the importance of obtaining comfort in acupuncture treatment was pointed out. Considering the pursuit of less pain and harmlessness in modern clinical treatment, sugar needle should be advocated and popularized in current clinical practice of acupuncture and moxibustion.
		                        		
		                        		
		                        		
		                        			Sugars
		                        			;
		                        		
		                        			Moxibustion
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		                        			Acupuncture Therapy
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		                        			Emotions
		                        			;
		                        		
		                        			Needles
		                        			
		                        		
		                        	
9.Mechanism of n-butanol alcohol extract of Baitouweng Decoction in treatment of vulvovaginal candidiasis based on negative regulation of NLRP3 inflammasome via PKCδ/NLRC4/IL-1Ra axis.
Kai-Fan HU ; Ling MO ; Hao ZHANG ; Dan XIA ; Gao-Xiang SHI ; Da-Qiang WU ; Tian-Ming WANG ; Jing SHAO ; Chang-Zhong WANG
China Journal of Chinese Materia Medica 2023;48(6):1578-1588
		                        		
		                        			
		                        			This study aimed to explore the mechanism of n-butanol alcohol extract of Baitouweng Decoction(BAEB) in the treatment of vulvovaginal candidiasis(VVC) in mice based on the negative regulation of NLRP3 inflammasome via PKCδ/NLRC4/IL-1Ra axis. In the experiment, female C57BL/6 mice were divided randomly into the following six groups: a blank control group, a VVC model group, high-, medium-, and low-dose BAEB groups(80, 40, and 20 mg·kg~(-1)), and a fluconazole group(20 mg·kg~(-1)). The VVC model was induced in mice except for those in the blank control group by the estrogen dependence method. After modeling, no treatment was carried out in the blank control group. The mice in the high-, medium-, and low-dose BAEB groups were treated with BAEB at 80, 40, and 20 mg·kg~(-1), respectively, and those in the fluconazole group were treated with fluconazole at 20 mg·kg~(-1). The mice in the VVC model group received the same volume of normal saline. The general state and body weight of mice in each group were observed every day, and the morphological changes of Candida albicans in the vaginal lavage of mice were examined by Gram staining. The fungal load in the vaginal lavage of mice was detected by microdilution assay. After the mice were killed, the degree of neutrophil infiltration in the vaginal lavage was detected by Papanicolaou staining. The content of inflammatory cytokines interleukin(IL)-1β, IL-18, and lactate dehydrogenase(LDH) in the vaginal lavage was tested by enzyme-linked immunosorbent assay(ELISA), and vaginal histopathology was analyzed by hematoxylin-eosin(HE) staining. The expression and distribution of NLRP3, PKCδ, pNLRC4, and IL-1Ra in vaginal tissues were measured by immunohistochemistry(IHC), and the expression and distribution of pNLRC4 and IL-1Ra in vaginal tissues were detected by immunofluorescence(IF). The protein expression of NLRP3, PKCδ, pNLRC4, and IL-1Ra was detected by Western blot(WB), and the mRNA expression of NLRP3, PKCδ, pNLRC4, and IL-1Ra was detected by qRT-PCR. The results showed that compared with the blank control group, the VVC model group showed redness, edema, and white secretions in the vagina. Compared with the VVC model group, the BAEB groups showed improved general state of VVC mice. As revealed by Gram staining, Papanicolaou staining, microdilution assay, and HE staining, compared with the blank control group, the VVC model group showed a large number of hyphae, neutrophils infiltration, and increased fungal load in the vaginal lavage, destroyed vaginal mucosa, and infiltration of a large number of inflammatory cells. BAEB could reduce the transformation of C. albicans from yeast to hyphae. High-dose BAEB could significantly reduce neutrophil infiltration and fungal load. Low-and medium-dose BAEB could reduce the da-mage to the vaginal tissue, while high-dose BAEB could restore the damaged vaginal tissues to normal levels. ELISA results showed that the content of inflammatory cytokines IL-1β, IL-18, and LDH in the VVC model group significantly increased compared with that in the blank control group, and the content of IL-1β, IL-18 and LDH in the medium-and high-dose BAEB groups was significantly reduced compared with that in the VVC model group. WB and qRT-PCR results showed that compared with the blank control group, the VVC model group showed reduced protein and mRNA expression of PKCδ, pNLRC4, and IL-1Ra in vaginal tissues of mice and increased protein and mRNA expression of NLRP3. Compared with the VVC model group, the medium-and high-dose BAEB groups showed up-regulated protein and mRNA expression of PKCδ, pNLRC4, and IL-1Ra in vaginal tissues and inhibited protein and mRNA expression of NLRP3 in vaginal tissues. This study indicated that the therapeutic effect of BAEB on VVC mice was presumably related to the negative regulation of NLRP3 inflammasome by promoting PKCδ/NLRC4/IL-1Ra axis.
		                        		
		                        		
		                        		
		                        			Female
		                        			;
		                        		
		                        			Animals
		                        			;
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Mice
		                        			;
		                        		
		                        			Candidiasis, Vulvovaginal/drug therapy*
		                        			;
		                        		
		                        			Inflammasomes/genetics*
		                        			;
		                        		
		                        			Interleukin-18
		                        			;
		                        		
		                        			NLR Family, Pyrin Domain-Containing 3 Protein/genetics*
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		                        			1-Butanol/pharmacology*
		                        			;
		                        		
		                        			Fluconazole/therapeutic use*
		                        			;
		                        		
		                        			Interleukin 1 Receptor Antagonist Protein/therapeutic use*
		                        			;
		                        		
		                        			Mice, Inbred C57BL
		                        			;
		                        		
		                        			Candida albicans
		                        			;
		                        		
		                        			Cytokines
		                        			;
		                        		
		                        			Drugs, Chinese Herbal/pharmacology*
		                        			;
		                        		
		                        			Ethanol
		                        			;
		                        		
		                        			RNA, Messenger
		                        			;
		                        		
		                        			Calcium-Binding Proteins/therapeutic use*
		                        			
		                        		
		                        	
10.Analysis of transmission dynamics and effectiveness of control of local epidemics caused by the Omicron BA.2 and BA.5.2 COVID-19 strains in Fujian Province
Wen-Jing YE ; Sheng-Gen WU ; Mei-Rong ZHAN ; Zheng-Qiang HUANG ; Shao-Jian CAI ; Wu CHEN ; Jian-Ming OU ; Jie-Feng HUANG ; Tian-Mu CHEN ; Yan-Qin DENG ; Kui-Cheng ZHENG
Chinese Journal of Zoonoses 2023;39(11):1065-1071
		                        		
		                        			
		                        			This study evaluated the scientific nature and effectiveness of iterative optimization of prevention and control measures for local outbreaks caused by the BA.2 and BA.5.2 COVID-19 strains in Fujian Province in 2022,to provide a scientif-ic basis for responding to future new or recurrent respiratory infectious diseases.According to the theory of infectious disease dynamics,relevant information regarding the local epidemic situation caused by the BA.2 sub-type Omicron virus strain in March 2022 and BA.5.2 sub-type Omicron virus strain in October 2022 in Fujian Province was collected.The susceptible exposed infectious removed(SEIAR)model of COVID-19 infection with a latent period and asymptomatic infected persons was used to analyze the transmission dynam-ics of two local epidemic situations,and evaluate the preven-tion and control effects.The incubation period of the BA.2 epidemic was 3 days(1~9 days),the intergenerational inter-val was 3 days(1~5 days),and the initial Rt was 3.0(95%CI:2.7~3.3).The incubation period of the BA.5.2 epidemic was 2 days(1~6 days),the intergenerational interval was 1 day(0~2 days),and the initial R,was 1.9(95%CI:1.7~2.1).The fittingresults for the BA.2 and BA.5.2 epidemics were good,and no statistical difference was observed between the predic-ted and actual numbers of cases(x2BA.2=31.53,x2BA.5.2=27.88,P>0.05).If an emergency response had not been initiated,the BA.2 epidemic would have continued to spread andpeak on April 7th,with an estimated 638 035 cases.The BA.5.2 epidemic would have rapidly spread,reaching a peak on November 14th,with an estimated 685 940 cases.If one incubation period were detected early,the scale of the BA.2 epidemic would have decreased by 25.73%;if two incubation periods were detected early,the scale would have decreased by 79.56%,and if one incubation period had been delayed,the scale would have expanded by 13.72%.If one incubation period had been detected early in the BA.5.2 epidemic,the scale would have decreased by 35.04%;if two incubation periods had been detected early,the scale would have decreased by 92.47%;and if one incubation period had been delayed,the scale would have increased by 19.75%.The guiding ideology,and the prevention and control measures for handling two local epidemics were optimized and iterated.Our study indicated that implementing the"four early"measures ef-fectively decreased the scale of the epidemic,and earlier detection was associated with more significant control effects.This study provides valuable information for the prevention and control of new or recurrent respiratory infectious diseases.
		                        		
		                        		
		                        		
		                        	
            
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