Comparison of pediatric and adult antibiotic-associated diarrhea and Clostridium difficile infections

Antibiotic-associated diarrhea(AAD) and Clostridum difficile infections(CDI) have been well studied for adult cases, but not as well in the pediatric population. Whether the disease process or response to treatments differs between pediatric and adult patients is an important clinical concern when following global guidelines based largely on adult patients. A systematic review of the literature using databases Pub Med(June 3, 1978-2015) was conducted to compare AAD and CDI in pediatric and adult populations and determine significant differences and similarities that might impact clinical decisions. In general, pediatric AAD and CDI have a more rapid onset of symptoms, a shorter duration of disease and fewer CDI complications(required surgeries and extended hospitalizations) than in adults. Children experience more community-associated CDI and are associated with smaller outbreaks than adult cases of CDI. The ribotype NAP1/027/BI is more common in adults than children. Children and adults share some similar risk factors, but adults have more complex risk factor profiles associated with more co-morbidities, types of disruptive factors and a wider range of exposures to C. difficile in the healthcare environment. The treatment of pediatric and adult AAD is similar(discontinuing or switching the inciting antibiotic), but other treatment strategies for AAD have not been established. Pediatric CDI responds better to metronidazole, while adult CDI responds better to vancomycin. Recurrent CDI is not commonly reported for children. Prevention for both pediatric and adult AAD and CDI relies upon integrated infection control programs, antibiotic stewardship and may include the use of adjunctive probiotics. Clinical presentation of pediatric AAD and CDI are different than adult AAD and CDI symptoms. These differences should be taken into account when rating severity of disease and prescribing antibiotics.

Efficacy of Lactobacillus rhamnosus GG in treatment of acute pediatric diarrhea: A systematic review with meta-analysis

BACKGROUND Diarrhea is a major infectious cause of childhood morbidity and mortality worldwide.In clinical trials,Lactobacillus rhamnosus GG ATCC 53013(LGG)has been used to treat diarrhea.However,recent randomized controlled trials(RCTs)found no evidence of a beneficial effect of LGG treatment.AIM To evaluate the efficacy of LGG in treating acute diarrhea in children.METHODS The EMBASE,MEDLINE,PubMed,Web of Science databases,and the Cochrane Central Register of Controlled Trials were searched up to April 2019 for metaanalyses and RCTs.The Cochrane Review Manager was used to analyze the relevant data.RESULTS Nineteen RCTs met the inclusion criteria and showed that compared with the control group,LGG administration notably reduced the diarrhea duration[mean difference(MD)-24.02 h,95%confidence interval(CI)(-36.58,-11.45)].More effective results were detected at a high dose≥1010 CFU per day[MD-22.56 h,95%CI(-36.41,-8.72)]vs a lower dose.A similar reduction was found in Asian and European patients[MD-24.42 h,95%CI(-47.01,-1.82);MD-32.02 h,95%CI(-49.26,-14.79),respectively].A reduced duration of diarrhea was confirmed in LGG participants with diarrhea for less than 3 d at enrollment[MD-15.83 h,95%CI(-20.68,-10.98)].High-dose LGG effectively reduced the duration of rotavirus-induced diarrhea[MD-31.05 h,95%CI(-50.31,-11.80)]and the stool number per day[MD-1.08,95%CI(-1.87,-0.28)].CONCLUSION High-dose LGG therapy reduces the duration of diarrhea and the stool number per day.Intervention at the early stage is recommended.Future trials are expected to verify the effectiveness of LGG treatment.

Probiotics for antibiotic-associated diarrhea:Do we have a verdict?

Probiotics use has increased tremendously over the past ten years.This was coupled with a surge of data relating their importance in clinical practice.Antibioticassociated diarrhea,whose frequency has risen recently,was one of the earliest targets with data published more than ten years ago.Unfortunately,available trials suffer from severe discrepancies associated with variability and heterogeneity of several factors.Most published randomized controlled trials and subsequent meta-analyses suggest benefit for probiotics in the prevention of antibiotic-associated diarrhea.The same seems to also apply when the data is examined for Clostridium difficile-associated colitis.However,the largest randomized double-blind placebo-controlled trial to date examining the use of a certain preparation of probiotics in antibiotic-associated diarrhea showed disappointing results,but it was flawed with several drawbacks.The commonest species of probiotics studied across most trials is Lactobacillus;however,other types have also shown similar benefit.Probiotics have enjoyed an impeccable safety reputation.Despite a few reports of severe infections sometimes leading to septicemia,most of the available trials confirm their harmless behavior and show similaradverse events compared to placebo.Since a consensus dictating its use is still lacking,it would be advisable at this point to suggest prophylactic use of probiotics to certain patients at risk for antibiotic-associated diarrhea or to those who suffered previous episodes.

Clinicopathogenic studies of acute diarrhea in children

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Preventing pediatric antibiotic-associated diarrhea and Clostridium difficile infections with probiotics: A metaanalysis

AIM: To assess the efficacy and safety of probiotics for preventing pediatric:(1) antibiotic associated diarrhea and(2) Clostridium difficile(C. difficile) infections.METHODS: On June 3, 2013, we searched Pub Med(1960-2013), EMBASE(1974-2013), Cochrane Database of Systematic Reviews(1990-2013), CINAHL(1981-2013), AMED(1985-2013), and ISI Web of Science(2000-2013). Additionally, we conducted an extensive grey literature search including contact with National Institutes of Health Clinical Trials Registry, abstracts from annual infectious disease and gastroenterology meetings, experts in the field and correspondence with authors. The primary outcomes were the incidence of antibiotic-associated diarrhea(AAD) and C. difficile infections(CDI). Dichotomous outcomes(e.g., incidence of AAD or CDI) were pooled using a random-effects model to calculate the relative risk and corresponding 95% confidence interval(95%CI) and weighted on study quality. To explore possible explanations for heterogeneity, a priori subgroup analysis were conducted on probiotic strain type, daily dose, quality of study and safety of probiotics. The overall quality of the evidence supporting each outcome was assessed using the grading of recommendations, assessment, development and evaluation criteria.RESULTS: A total of 1329 studies were identified with 22 trials(23 treatment arms and 4155 participants) meeting eligibility requirements for our review of prevention of AAD and 5 trials(1211 participants) for the prevention of CDI. Trials in adult populations, trials of uncertain antibiotic exposure or studies which did not provide incidence of AAD were excluded. We found 12 trials testing a single strain of probiotic and 10 trials testing a mixture of probiotic strains. Probiotics(all strains combined) significantly reduced the incidence of pediatric AAD(pooled RR = 0.42, 95%CI: 0.33-0.53) and significantly reduced pediatric CDI(pooled RR = 0.35, 95%CI: 0.13-0.92). Of the two strains with multiple trials, both significantly reduced pediatric AAD: Saccharomyces boulardii lyo(pooled RR = 0.43, 95%CI: 0.32-0.60) and Lactobacillus rhamnosus GG(pooled RR = 0.36, 95%CI: 0.19-0.69). There was no significant effect by type of antibiotic, or by duration or dose of probiotic. No adverse events associated were found in the 22 controlled trials relating to the use of probiotics.CONCLUSION: This meta-analysis found that probiotics significantly prevented pediatric antibiotic associated diarrhea and pediatric CDI, but the efficacy varies significantly by the strain of the probiotic.

A blind, randomized comparison of racecadotril and loperamide for stopping acute diarrhea in adults

AIM: Racecadotril is a specific enkephalinase inhibitor that exhibits intestinal antisecretory activity without affecting intestinal transit. Loperamide is an effective anti-diarrheal agent, but it usually induces constipation. This study is to compare the efficacy, safety, and tolerability of racecadotril versus Ioperamide in the outpatient treatment of acutediarrhea in adults.METHODS: A two-center, randomized, parallel-group,single-blind study was carried out to compare the efficacy,tolerability, and safety of racecadotril (100 mg thrics daily) and Ioperamide (2.0 mg 2 twics daily) in 62 adult patients suffering from acute diarrhea. The main efficacy criterion used was the duration of diarrhea after beginning the treatment (in hours). Other signs and symptoms werealso evaluated.RESULTS: The clinical success rates for these antidiarrheal treatments were 95.7% and 92.0% for racecadotril and Ioperamide respectively. Patients on racecadotril had a median duration of diarrhea of 19.5 h compared with a median of 13 h for patients on Ioperamide.Rapid improvement in anal burn and nausea was found for each drug. However, more patients on Ioperamide suffered from reactive constipation (29.0% vs 12.9%).Itching, another adverse event was notably higher in the racecadotril group (28.6% vs 0%). With regard to other adverse events, the two medications showed similar occurrence rates and similar concomitant medicationusage rates.CONCLUSION: Racecadotril and Ioperamide are rapid,equally effective treatments for acute diarrhea in adults,but Ioperamide treatment is associated with a higher incidence of treatment-related constipation.

Efficacy of stool multiplex polymerase chain reaction assay in adult patients with acute infectious diarrhea

BACKGROUND Recently,stool multiplex polymerase chain reaction(PCR)tests have been developed for identifying diarrhea-causing bacterial pathogens.Furthermore,fecal calprotectin is a well-known effective marker for intestinal mucosal inflammation.AIM To evaluate the efficacy of stool multiplex PCR and fecal calprotectin in acute infectious diarrhea.METHODS Overall,400 patients with acute infectious diarrhea were enrolled from Kangdong Sacred Heart Hospital(January 2016 to December 2018).Multiplex PCR detected 7 enteropathogenic bacteria including Salmonella,Campylobacter,Shigella,Escherichia coli O157:H7,Aeromonas,Vibrio,and Clostridium difficile.We reviewed clinical and laboratory findings using stool multiplex PCR.RESULTS Stool multiplex PCR test detected considerably more bacterial pathogens than stool culture(49.2%vs 5.2%),with Campylobacter as the most common pathogen(54%).Patients with positive stool PCR showed elevated fecal calprotectin expression compared to patients with negative stool PCR(1124.5±816.9 mg/kg vs 609±713.2 mg/kg,P=0.001).C-reactive protein(OR=1.01,95%CI:1.001-1.027,P=0.034)and sigmoidoscopy-detected colitis(OR=4.76,95%CI:1.101-20.551,P=0.037)were independent factors in stool PCR-based detection of bacterial pathogens.Sensitivity and specificity of calprotectin were evaluated to be 70.5%and 60.9%,respectively(adjusted cut-off value=388 mg/kg).CONCLUSION Stool multiplex PCR test has increased sensitivity in detecting pathogens than conventional culture,and it is correlated with calprotectin expression.Stool multiplex PCR and calprotectin may be effective in predicting clinical severity of infectious diarrhea.

Formulation of an Early Warning Infectivity Score System for Adult Patients with Acute Bacterial Diarrhea

The aim of our study was to develop a scoring system to predict whether diarrhea is of a bacterial origin and whether the diarrheal patients constitute a potential source of infection to others. Adults with acute diarrhea （n=424） were enrolled in the study. Logistic regression and standard regression coefficients were used to formulate the Early Warning Infectivity Score System for Adults with Acute Bacterial Diarrhea （EWIS-ABD）. Four risk factors were identified by logistic regression, including body temperature （P〈0.01）, abdominal pain （P〈0.01）, leukocyte count in stool （P〈0.01）, and unclean dietary history （P〈0.01）. EWIS-ABD was thus developed, in which the value 〉5 points was set as an indicator of bacterial diarrhea. The incidence of bacterial diarrhea increased along with the elevated score. EWIS-ABD was more specific for bacterial diarrhea than for viral diarrhea. The accuracy and reliability of EWIS-ABD was high by prospective validation in 478 patients with acute diarrhea.

Frequency of E.coli pathotypes in acute diarrhea of children and its related factorsat Beassat hospital,Sanandaj

Objective:Diarrhea is a leading cause of morbidity and mortality among children in developing countries.The bacterial pathogen most commonly associated with childhood diarrhea is Escherichia coli.A one-year prospective study was carried out in Sanandaj to determine the prevalence and roles of the different E.coli pathotypes in children less than five years of age with acute diarrhea.Methods:Rectal swab were collected prospectively from children with acute diarrhea and transported to the Department of Microbiology,School of Medicine, KUMS,Sanandaj during 2008.The study was approved by the institutional ethics committee.Results:During this study period,rectal swabs were investigated from a total of 466 children 1 to 144 months of age(mean, 29.97 months±S.D) with diarrhea.Among the children,191(41%,191/466) were girls,and 275 (59%,275/466) were boys.The age-specific incidence rates of acute diarrhea among children 13-24 and 1 - 12 months of age were 37.37%(37/99) and 26.26%(26/99),respectively,during the study period.A total of 99 strains of E.coli were detected.EPEC 59(59.59%) and EIEC 22(22.22%),were the most commonly found Escherichia coli strains detected in stools from children.Disk diffusion testing showed E.coli strains resistance to tetracycline(89.89%),chloramphenicol(88.88%),Ampicillin(79.79%),Amoxicillin (75.75%) and Ceficime(75.75%).Among risk factors like age,sex,haemoglubin,fathers and mothers education,food and weight of children only mother's education was significant(P =0.018).Conclusion: In most of the clinical laboratories in Iran,E.coli does not considered as an etiologic agent responsible for diarrhea. Results in this study revealed that E.coli should be considered as an etiologic agent causing acute diarrhea among children.We therefore,recommend the routine isolation and identification of E.coli strains in all the clinical laboratories in Sanandaj.Guidelines for appropriate use of antibiotics in Sanandaj need updating.

Efficacy of Xianglian pill for antibiotic-associated diarrhea: a protocol for systematic review and meta-analysis

Background:Antibiotic-associated diarrhea is a clinical common symptom of antibiotics overuse and occurs in 5%-70%of adults.Xianglian pill has been traditionally considered as an efficient treatment of diarrhea and gastrointestinal diseases for thousands of years.However,no systematic review and meta-analyses have focused on its positive effects.Hence,this protocol for systematic review and meta-analysis was developed to evaluate the effect and clinical safety of Xianglian pill on treating antibiotic-associated diarrhea.Methods:All randomized controlled trials published in Chinese and English and assessed use of Xianglian pill for antibiotic-associated diarrhea will be included.Databases of PubMed,EMBASE,Cochrane Library,China National Knowledge Infrastructure,Chinese Biomedical Literature,Wanfang,and Chinese Science and Technology Periodical Database will be searched for randomized controlled trials from their inception until November 16,2020.Primary outcomes will be the incidence of diarrhea and adverse events,and secondary outcomes will be bowel movements and microbiome characteristics.Two authors will extract data and assess the risk of bias independently.Risk ratio will be used to evaluate the results,and meta-analyses will be conducted using STATA 15.0 software.The review aims to demonstrate the effectiveness of Xianglian pill in the prevention and treatment of antibiotic-associated diarrhea.

Prolonged acute diarrhea in adults: Decades of observation from epidemiological perspective in urban Bangladesh

The study aimed to determine the epidemiological, clinical characteristics, and etiology of adults aged ≥20 years presented with prolonged acute diarrhea (≥7 days). A total of 18,210 adults aged ≥20 years were enrolled in the Diarrheal Disease Surveillance System of icddr,b between 1993-2012 and included into the analysis. Of these, 17,631 (97%) had duration of diarrhea ≤6 days, 418 (2%) diarrhea presented with a duration of 7 - 13 days and rest 161 (1%) had history of diarrhea ≥14 days. A higher proportion of adult individuals who had duration of diarrhea ≥14 days (70% vs. 56%;p male. At least 73% of all patients used oral rehydration solution;but proportion was lower among prolonged acute diarrhea groups. Use of antimicrobials was higher among those with duration 7 - 13 days (81%) and ≥14 days (81%). Diarrhea lasting ≥14 days, 47% were suffering from chronic energy deficiency and 30% had history of smoking. Individuals with diarrhea ≥7 days less frequently presented with vomiting, watery stool, frequency of stool >10 times/24 hours, drowsy or lethargy, fast breathing, some or severe dehydration, received intravenous saline for initial corrections of dehydration and stayed for longer duration in hospital (≥24 hours) but more often presented with abdominal pain. Stool microscopic examination showed less frequent presence of red blood cells (36% vs. 44%;p = 0.043) and fecal leucocytes (50% vs. 59%;p = 0.029) among individuals with diarrhea ≥14 days compared to those with ≤6 days. None was infected with Vibrio cholerae (≥14 days) (3% for 7 - 13 days);however, isolation rate of Aeromonas was higher among adults with duration for ≥14 days (11%). Only 15% with ≥14 days were positive for Shigella contrary to 19% (7 - 13 days) and 56% (≤6 days). Differences in sociodemographic, clinical presentation and etiology varied with duration of diarrhea among adults.

Efficacy of a synbiotic chewable tablet in the prevention of antibiotic-associated diarrhea

Infection by Clostridium difficile, a complication of treatment with antibiotics, causes antibiotic- associated diarrhea (AAD) and can lead to colitis and pseudomembranous colitis. Incidence of C. difficile infection is increasing among the elderly undergoing antibiotics therapy confined to health care facilities, conditions that are expensive to treat, decrease the quality of life and are life threatening. Use of probiotics has been proposed as a method to decrease the incidence of AAD in health care facilities. To examine the efficacy of using probiotics, 120 nursing home residents undergoing antibiotic therapy were provided with a synbiotic tablet containing two probiotics, Saccharomyces boulardii and Bacillus coagulans, and a prebiotic, fructooligosaccharide. Residents were evaluated retrospectively for AAD and C. difficile infection. It was found that 95% of residents treated with antibiotics and taking the synbiotic tablet were free of AAD. More than 97% of the residents did not become infected with C. difficile. No adverse effects were reported. Minor side effects, gastrointestinal upset and nausea, were reported by less than 6% of the residents. The cause of the minor side effects was not known. Only 2.5% of the residents stopped taking the synbiotic tablet because of the gastrointestinal upset. These Results suggest that use of the synbiotic tablet prevents AAD and C. difficile infection in nursing home residents undergoing antibiotic therapy. It is concluded that this synbiotic tablet provides an easy to administer and safe approach to controlling AAD and C. difficile infection in residents in nursing homes.

A scoring system for assessing the severity of acute diarrhea of adult patients

BACKGROUND: Diarrhea is frequently seen in developed and developing countries, and severe diarrhea is characterized by the high risk of death. Thus, it is very important to assess the severity of diarrhea early. We conducted a multi-center study to identify risk factors for the severity of diarrhea in adult patients and formulate an adult diarrhea state score(ADSS) for out-patient clinicians.METHODS: A total of 219 adult patients with acute diarrhea were divided into two groups: 132 patients with mild diarrhea and 87 with severe diarrhea. Logistic regression was used to determine risk factors for the severity of diarrhea. The risk factors were assessed and an ADSS was formulated. Receiver operating characteristic(ROC) analysis was made to evaluate the diagnostic accuracy of ADSS, and the Kappa test was used to confi rm the diagnostic reliability.RESULTS: Five risk factors for evaluating the severity of diarrhea in adults included age(P<0.05), axillary temperature(P<0.01), mean arterial pressure(P<0.01), white blood cell count(WBC; P<0.01), and WBC in stool(P<0.01). The area under the ROC curve for ADSS was 0.958 when the cut off value was 4(a sensitivity of 0.909; a specif icity of 0.874), and the Kappa value was 0.781(P<0.05).CONCLUSION: The risk factors associated with the pathogenic condition of diarrhea were identifi ed, quantifi ed and formulated into an ADSS, which has high diagnostic accuracy and reliability for the early identifi cation of patients with severe acute diarrhea.

Clostridium difficile causing acute renal failure: Case presentation and review

AIM: Clostridium difficile infection is primarily a nosocomial infection but asymptomatic carriers of Clostridium difficile can be found in up to 5% of the general population.Ampicillin, cephalosporins and clindamycin are the antibiotics that are most frequently associated with Clostridium difficile-associated diarrhea or colitis. Little is known about acute renal failure as a consequence of Clostridium difficile-associated diarrhea.METHODS: In this case report, we describe the course of Clostridium difficile-associated diarrhea in an 82-yearold patient developing acute renal failure. Stopping the offending agent and symptomatic therapy brought a rapid improvement of diarrhea and acute renal failure, full recovery was gained 18 d after admission. In a systematic review we looked for links between the two conditions.RESULTS: The link between Clostridium difficilr-associated diarrhea and acute renal failure in our patient was most likely volume depletion. However, in experimental studies a direct influence of Clostridium difficile toxins on renal duct cells could be shown.CONCLUSION: Rapid diagnosis, nonspecific supportive treatment and specific antibiotic treatment, especially in the elderly, may lower excess mortality Clostridium difficile-associated diarrhea and renal failure being possible complications.

Localized pseudomembranous colitis in the cecum and ascending colon mimicking acute appendicitis

A 61-year-old male was admitted to our hospital due to right lower abdominal pain and watery diarrhea for 3 d.Beginning 3 wk before he arrived in our hospital,he took 3 rd-generation cephalosporin(cefixime) for 2 wk due to chronic left ear otitis media.Colonoscopic examination revealed yellowish patches of ulcerations and swelling covered with thick serosanguineous exudate in the cecum and ascending colon.After 7 d of oral metronidazole treatment,his symptoms completely disappeared.We report a case of localized pseudomembranous colitis in the cecum and ascending colon mimicking acute appendicitis associated with cefixime.

Acute hepatitis associated with increased atypical lymphocyte

To the Editor:Hepatitis A and hepatitis E are acute infectious diseases caused by hepatitis A virus(HAV)and hepatitis E virus(HEV),which are mainly transmitted through the fecal-oral route[1].The early clinical symptoms of patients with hepatitis A or E are nonspecific,including fever,chills,abdominal pain,diarrhea,and rash[2].

小儿肠胃康颗粒联合蒙脱石散剂辅助治疗儿童急性细菌性腹泻病的疗效

目的:探讨小儿肠胃康颗粒联合蒙脱石散剂辅助治疗儿童急性细菌性腹泻病的临床疗效。方法:选取2021年3月至2022年2月我院门诊收治的96例急性细菌性腹泻病患儿按简单随机化分组法分为对照组和联合组各48例。两组患儿均给予对症治疗(依据细菌培养及药敏试验结果选择敏感性抗菌药物、调整饮食结构、纠正电解质紊乱及菌群失调等),在此基础上,对照组给予蒙脱石散剂治疗,联合组在对照组治疗基础上给予小儿肠胃康颗粒,疗程为2周。比较两组患儿胃肠道不适症状改善时间,检测治疗前后两组患儿血清胃肠激素及免疫功能变化情况,统计不良反应发生情况。结果:治疗后,联合组治疗总有效率高于对照组(100.00%vs.91.67%),但差异无统计学意义(P>0.05),联合组疗效等级优于对照组(Z=2.691,P<0.05)。治疗后,两组患儿大便次数较同组治疗前减少(P均<0.01),联合组大便次数较对照组更少(t=5.771,P<0.05),且联合组腹泻、脱水、食欲减退、腹痛、腹胀缓解时间及大便性状改变时间均较对照组更短(P均<0.05)。治疗后,联合组胃动素(MOT)、胃泌素(GAS)水平低于对照组,生长抑素(SS)、P物质(SP)水平高于对照组,差异均有统计学意义(P均<0.05)。治疗后,联合组CD3^(+)、CD4^(+)及CD4^(+)/CD8^(+)水平高于对照组(P均<0.05)。两组患儿不良反应发生率比较差异无统计学意义(χ^(2)=0.546,P=0.460)。结论:小儿肠胃康颗粒联合蒙脱石散剂治疗儿童急性细菌性腹泻病整体价值优于单用蒙脱石散剂治疗,可缩短患儿胃肠道不适症状缓解时间,改善胃肠道动力学及免疫功能,安全性较高。

非血缘脐血移植后肠道急性移植物抗宿主病的特点分析

背景:尽管非血缘脐血移植有望成为治愈恶性血液病的重要方法,但移植后肠道急性移植物抗宿主病方面的表现及临床特点仍然需要深入研究。目的:分析非血缘脐血移植后发生肠道急性移植物抗宿主病的临床特点。方法:2016年12月至2020年12月中国科学技术大学附属第一医院血液科造血干细胞移植亚专科668例恶性血液病患者接受非血缘脐血移植治疗,其中138例发生肠道急性移植物抗宿主病,男性76例,女性62例,移植时中位年龄13(1-62)岁。所有患者采取清髓性不含抗人胸腺细胞球蛋白方案进行预处理,以及采用环孢素A联合霉酚酸酯预防移植物抗宿主病。结果与结论:①非血缘脐血移植后肠道急性移植物抗宿主病患者均出现不同程度的腹泻,粪便为黄绿色以及黄褐色水样便或者黏液便,其中53例(38.4%)患者出现血便,82例(57.9%)患者伴有皮肤受累,18例(13.0%)患者继发肠道细菌感染,90例(65.2%)患者合并巨细胞病毒血症;②进一步比较1-2级肠道急性移植物抗宿主病(70例,50.7%)与3-4级肠道急性移植物抗宿主病(68例,49.3%)患者的临床特点,发现3-4级肠道急性移植物抗宿主病患者年龄大于1-2级肠道急性移植物抗宿主病患者(P<0.001),更易合并巨细胞病毒血症(P=0.035),腹泻持续时间更长(P=0.00),住院时间也明显增加(P<0.001),而两组患者在性别、移植前疾病状态、供受者HLA匹配度、疾病诊断、合并皮肤急性移植物抗宿主病、继发肠道感染率等方面无显著差异;③结果表明:非血缘脐血移植后肠道急性移植物抗宿主病的临床特点比较复杂,严重影响患者的预后和生活质量,需要及早识别,精准治疗。

猪急性腹泻综合征冠状病毒S蛋白多克隆抗体的制备及在检测该病毒感染中的应用

为制备猪急性腹泻综合征冠状病毒(SADS-CoV)纤突蛋白(S)的多克隆抗体(PAb),本研究经PCR扩增SADS-Co V S蛋白S1亚基C端结构域(S1-CTD)基因片段(384 bp),并将其克隆至原核表达载体p GEX-6p-1中,构建重组质粒p GEX-6p-1-S1-CTD,经双酶切和测序鉴定正确后,转化大肠杆菌BL21(DE3)感受态细胞,利用IPTG诱导表达,通过western blot鉴定重组S1-CTD蛋白(rS1-CTD)的表达及反应原性。结果显示,r S1-CTD以包涵体的形式表达,在40 ku处出现特异性条带。诱导表达后的r S1-CTD经不同浓度尿素重悬并超声离心,SDS-PAGE检测后切胶纯化,得到纯化的重组蛋白。利用BCA试剂盒测得蛋白的浓度为33μg/m L。将该重组蛋白乳化后经3次免疫新西兰大白兔,并在3免一周后采血,分离血清获得S1-CTD蛋白PAb。将SADS-Co V感染Vero E6细胞24 h后,以获得的兔PAb为一抗,分别采用western blot和间接免疫荧光试验(IFA)检测该PAb的反应原性。Western blot结果显示,在约250 ku处出现特异性条带,而阴性对照组无该条带;IFA结果显示,SADS-Co V感染的细胞中出现绿色荧光,而阴性对照细胞无绿色荧光。将SADS-Co V感染仔猪的回肠组织制备病理切片,以制备的PAb为一抗,通过免疫组织化学(IHC)检测SADS-Co V的抗原。结果显示,该组织切片中出现棕色阳性信号,而阴性对照仔猪回肠组织切片则无该棕色信号。表明该PAb可与感染SADS-Co V的仔猪回肠组织中的相应抗原发生特异性免疫反应。综上所述,本实验制备的S1-CTD蛋白PAb具有良好的反应原性和免疫原性,可以用于western blot、IFA、IHC检测体内外SADS-Co V的感染,为后续SADS-Co V检测方法的建立及S蛋白生物学功能的研究奠定基础。

急性腹泻伴良性惊厥患儿血清NSE、H_2S、UCH-L1水平变化及对预测复发的临床价值分析

目的 探讨急性腹泻伴良性惊厥患儿血清神经元特异性烯醇化酶(NSE)、硫化氢(H_(2)S)、泛素羧基末端水解酶L1(UCH-L1)水平变化及其对疾病复发的预测效能。方法 选择秦皇岛市第一医院2018年8月—2021年8月收治的急性腹泻伴良性惊厥的104例患儿进行前瞻性研究,另选同期于秦皇岛市第一医院行健康体检的30例健康儿童为对照组,所有儿童均取空腹静脉血检测血清NSE、H_(2)S、UCH-L1水平,并对比两组儿童及观察组不同病情患儿的血清NSE、H_(2)S、UCH-L1水平差异;观察组患儿随访2年以观察复发情况,对比有无复发患儿血清NSE、H_(2)S、UCH-L1水平差异,并采用受试者工作特征曲线(ROC)分析急性腹泻伴良性惊厥患儿血清NSE、H_(2)S、UCH-L1水平对复发的预测效能。结果 观察组血清NSE、UCH-L1水平明显高于对照组,H_(2)S水平明显低于对照组(P<0.05);相较于腹泻3~5次/d、惊厥发作1次、单次惊厥持续时间<5 min、惊厥局灶性发作的患儿,腹泻≥6次/d、惊厥发作≥2次、单次惊厥持续时间≥5 min、惊厥全面性发作患儿的血清NSE,UCH-L1水平均明显更高(P<0.05),血清H_(2)S水平明显更低(P<0.05);复发组患儿的血清NSE、UCH-L1水平明显高于未复发组(P<0.05),血清H_(2)S水平明显低于未复发组(P<0.05);ROC曲线分析显示,急性腹泻伴良性惊厥患儿血清NSE、H_(2)S、UCH-L1水平及联合检测均对复发有预测效能(P<0.05),其中单项检测时血清NSE对复发的预测效能较高,曲线下面积(AUC)为0.787,敏感为77.78%,特异度为81.05%;联合检测的预测效能最高,AUC为0.804,敏感性为88.89%,特异度为75.79%。结论 急性腹泻伴良性惊厥患儿的血清NSE、UCH-L1水平明显升高,血清H_(2)S水平降低,且与病情程度存在相关性,可用于预测惊厥复发。