Adult diarrhea rotavirus in Jilin, China

In May 1986, an outbreak of epidemic acute diarrhea occurred in one city and three counties in Yanbian area. The diarrheal cases were seen in all age groups, the majority of the cases were seen in adult age group. No bacterial pathogens were isolated in 22 fecal samples examined, however rotavirus like particles of 52-68 nm in diameter were found in 50% (11/22) of fecal samples by the immunoelectron microscopy using convalescent sera. Examination by the adult diarrhea rotavirus (ADRV) ELISA kit showed positive reaction in 86% (6/7) of fecal extracts, however in all 22 fecal extracts examination using ELISA kit for detection of group A rotavirus showed negative reaction. The PAGE patterns of viral RNAs similar to those of ADRV were seen in 13 out of 22 fecal extracts. The detection rate by PAGE analysis was 59%. Based on the above data, the etiological agent of the epidemic acute diarrhea, which occurred in Yanbian area, was identified to be ADRV.     

Purification and characterization of adult diarrhea rotavirus: identification of viral structural proteins.

Adult diarrhea rotavirus (ADRV) is a newly identified strain of noncultivable human group B rotavirus that has been epidemic in the People's Republic of China since 1982. We have used sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western (immuno-) blot analysis to examine the viral proteins present in the outer and inner capsids of ADRV and compared these with the proteins of a group A rotavirus, SA11. EDTA treatment of double-shelled virions removed the outer capsid and resulted in the loss of three polypeptides of 64, 61, and 41, kilodaltons (kDa). Endo-beta-N-acetylglucosaminidase H digestion of double-shelled virions identified the 41-kDa polypeptide as a glycoprotein. CaCl2 treatment of single-shelled particles removed the inner capsid and resulted in the loss of one polypeptide with a molecular mass of 47 kDa. The remaining core particle had two major structural proteins of 136 and 113 kDa. All of the proteins visualized on sodium dodecyl sulfate-polyacrylamide gel electrophoresis were antigenic by Western blot analysis when probed with convalescent-phase human and animal antisera. A 47-kDa polypeptide was most abundant and was strongly immunoreactive with human sera, animal sera raised against ADRV and against other group B animal rotaviruses (infectious diarrhea of infant rat virus, bovine and porcine group B rotavirus, and bovine enteric syncytial virus) and a monoclonal antibody prepared against infectious diarrhea of infant rat virus. This 47-kDa inner capsid polypeptide contains a common group B antigen and is similar to the VP6 of the group A rotaviruses. Human convalescent-phase sera also responded to a 41-kDa polypeptide of the outer capsid that seems similar to the VP7 of group A rotavirus. Other polypeptides have been given tentative designations on the basis of similarities to the control preparation of SA11, including a 136-kDa polypeptide designated VP1, a 113-kDa polypeptide designated VP2, 64- and 61-kDa polypeptides designated VP5 and VP5a, and several proteins in the 110- to 72-kDa range that may be VP3, VP4, or related proteins. The lack of cross-reactivity on Western blots between antisera to group A versus group B rotaviruses confirmed that these viruses are antigenically quite distinct.     

Involvement of aquaporins in a mouse model of rotavirus diarrhea

Rotavirus diarrhea is a major worldwide cause of infantile gastroenteritis; however, the mechanism responsible for intestinal fluid loss remains unclear. Water transfer across the intestinal epithelial membrane seems to occur because of aquaporins（AQPs）. Accumulating evidence indicates that alterations in AQPs may play an important role in pathogenesis. Here, we focus on changes in AQPs in a mouse model of rotavirus diarrhea. In the present study, 32 of 35 mice developed diarrhea and mild dehydration within 24 hours after infection with rotavirus strain SA11. Intestinal epithelial cells demonstrated cytoplasmic vacuolation, malaligned villi, and atrophy. AQP1 expression was significantly attenuated in the ileum and colon in comparison with controls; likewise, AQP4 and-8 protein expression were significantly decreased in the colon of rotavirus diarrhea-infected mice. In contrast, AQP3 protein expression was significantly increased in the colon of rotavirus-infected mice in comparison with controls. These results indicate that rotavirus diarrhea is associated with the downregulation of AQP1,-4, and-8 expression. Therefore, AQPs play an important role in rotavirus diarrhea.     

Isolation of an atypical rotavirus causing diarrhea in neonatal ferrets

A rotavirus was isolated from neonatal ferrets (Mustela putorius furo) with diarrhea at a large commercial farm. This virus is classified as an atypical rotavirus, probably belonging to the group C rotaviruses. This classification is based on the lack of the rotavirus group A common antigen and on its distinct dsRNA electropherotype pattern in polyacrylamide gels. The diarrheal disease was reproduced experimentally in neonatal ferrets.     

三种检测婴幼儿腹泻A组轮状病毒的方法学评价

目的对婴幼儿腹泻A组轮状病毒的三种不同检测方法进行评价。方法同时采用胶体金、琼脂糖凝胶电泳、逆转录PCR（RT-PCR）对浙江萧山医院1周320份腹泻患儿的标本进行轮状病毒检测,然后进行方法学比较。结果三种方法的阳性检出率分别为36．3％、48．1％和42．8％,经卡方检验三种方法的检出率之间差异均存在统计学意义（P〈0．05）;以琼脂糖凝胶电泳为检测的最终标准,RT-PCR的敏感性为88．9％,特异性达100％;胶体金法的敏感性71．4％,特异性达96．4％。结论胶体金法检测轮状病毒的敏感性和特异性能够满足临床上对轮状病毒快速筛检的要求;PT-PCR法的敏感性和特异性均最高,既可用于临床检测,又是轮状病毒血清G、基因P分型的重要手段,在对无菌体液和环境样本中轮状病毒的检测具有不可比拟的优势。     

Nasal immunization of mice with virus-like particles protects offspring against rotavirus diarrhea

Rotavirus is the major cause of diarrhea among young infants in both humans and animals. Immune protection of newborns by vaccination is difficult to achieve since there is not enough time to mount an immune response before exposure to the virus. We have designed a vaccination strategy mediating transfer of neutralizing antibodies from the mother to the offspring during pregnancy and/or lactation. Adult female mice were nasally immunized with virus-like particles (VLPs) made of viral proteins VP2 and 6 (VLP2/6) or VP 2, 6, and 7 (VLP2/6/7) derived from the RF rotavirus strain in the presence or absence of cholera toxin. Both vaccines elicited serum and milk antibodies against the respective VPs. Four days after parturition, suckling pups were challenged orally with RF rotavirus. Pups from mothers immunized with VLP2/6/7 but not VLP2/6 were protected against rotavirus diarrhea, indicating that VP7 plays a key role in protection. Protection was mediated by milk rather than serum antibodies, and mucosal adjuvants were not required. In conclusion, VLPs containing VP7 administered nasally to mothers represent a promising vaccine candidate for the protection of suckling newborns against rotavirus-induced diarrhea, even in the absence of a mucosal adjuvant.     

婴幼儿腹泻轮状病毒和腺病毒检测结果分析

目的了解本地区婴幼儿腹泻患者中轮状病毒和腺病毒的感染情况,为临床治疗提供依据。方法收集2013年1月-12月门诊和住院腹泻患儿的粪便,进行轮状病毒和腺病毒抗原检测,并对结果进行统计学分析。结果在检测的2 579例婴幼儿腹泻中轮状病毒阳性为616例,阳性率为23.89%,高发季节为1月、11月和12月,高发年龄组为13-24月婴幼儿,腺病毒阳性共102例,阳性率为3.96%,程散发状况,高发年龄组为7-12月的婴幼儿。结论轮状病毒和腺病毒都能引起婴幼儿腹泻,但是轮状病毒为最主要的病原体,及时检测轮状病毒抗原,为临床治疗和疾病监测提供参考。     

G and P genotypes of rotavirus circulating among children with diarrhea in the Colombian northern coast.

A study on the prevalence of rotavirus G and P genotypes was carried out based on 253 stool specimens obtained from children living in the Colombia northern coast region who were less than 3-years-old and who suffered from acute diarrhea. A previous study had detected the presence of rotavirus A in 90 (36.5%) of the 246 samples tested by enzyme immunoassay (EIA), and these strains were investigated in the present study. Of these, 50 strains yielded an RNA electropherotype, most of which (80.0%) had long profiles and 20.0% of which had short profiles. Genotyping of 84 positive samples indicated that 67.9% of the strains could be typed. G1 (57.9%), was the most predominant VP7 genotype, followed by G3 (21.1%), G9 (15.8%) and G2 (5.3%). Among the VP4 genotypes, P[4] (49.1%) was the most prevalent, followed by P[6] 36.4% and P[8] (14.5%). Neither G4 nor G8 nor P[9] types were detected. The most common G-P combinations were G3 P[4] (8.8%) and G9 P[6] (7.0%), followed by G1 P[4] and G1 P[8] (5.3% each). All G1 P[8] strains showed long RNA profiles, whereas G3 P[4] and G9 P[6] displayed both long and short patterns. Mixed infections involved 21.0% of strains. There was a marked diversity among strains collected, and novel strains, including G9, as well as other atypical combinations of G and P genotypes, such as G9 P[6] and G3 P[4], were found.     

杭州地区腹泻儿童轮状病毒与腺病毒感染调查

目的 分析不同季节及不同年龄段腹泻患儿感染轮状病毒（RV）与腺病毒（Eads）情况,为临床提供早期、快速、准确、可靠的依据,以便及时采取相应的治疗使患儿早日康复。方法 采用杭州艾博医药有限公司提供的轮状病毒（A组）/腺病毒检测试剂盒（乳胶法）,对2012年1月至2015年12月浙江省人民医院门诊和住院就诊的3 368例腹泻儿童粪便标本进行轮状病毒和腺病毒抗原检测。结果 3 368例腹泻儿童粪便中,RV阳性578例占17.16%,Eads阳性106例占3.15%。11月至次年1月为轮状病毒感染的高发月份,1～3岁为儿童感染的高发年龄;腺病毒感染阳性率低,呈散发性。结论 杭州地区近年1～3岁婴幼儿的腹泻主要是由轮状病毒和腺病毒引起,其中轮状病毒感染率明显高于腺病毒,且有明显的季节性;而腺病毒感染呈散发流行,未表现出明显的季节性及年龄分布。     

Antigenemia, RNAemia, and innate immunity in children with acute rotavirus diarrhea

Antigenemia is commonly detected in children with acute rotavirus diarrhea, but the prevalence of viremia has not been clearly defined. We examined antigenemia in plasma and RNAemia in peripheral blood mononuclear cells (PBMC) of children with acute diarrhea by EIA, RT-PCR, and Southern hybridization, using primers and a probe specific to rotavirus NSP4 gene. We detected the presence of rotavirus antigen in 33.3% and almost full-length NSP4 gene in 70.8% of the acute-phase plasma and PBMC, respectively. In contrast, antigenemia and RNAemia were detected in 0% and 4.2% of the convalescent-phase plasma and PBMC, respectively, which were similar to antigenemia (0%) and RNAemia (7.7%) in healthy controls. We demonstrated an increase in the proportions of activated myeloid dendritic cells (mDC) and activated plasmacytoid DC (pDC) in acute-phase PBMC of patients when compared to those in convalescent phase of patients and in PBMC of healthy controls. The activation of mDC peaked on days 2-4 after illness onset, and the activation of acute-phase pDC appeared to correlate with levels of antigenemia. High prevalence of NSP4 gene in acute-phase PBMC indicates possible rotavirus replication in white blood cells, and extraintestinal spread and the activation of DC may have implications for the prevention of rotavirus disease in children.     

Symposium on diarrhea. 6. Infectious diarrhea.

Diarrhea may be primarily infectious in origin. Causes can be conveniently classified according to the etiologic agent, which may be viral, chlamydial, bacterial, protozoal, helminthic or fungal. The most common type of infectious diarrhea in Canada is viral. Bacterial infection, particularly staphylococcal and salmonellal, also is relatively common.     

Symposium on diarrhea. 5. Postoperative diarrhea.

Under normal circumstances many physiologic mechanisms prevent diarrhea. The most important are the normal processes of gastric emptying, digestion and absorption, the functioning of the ileocecal valve and elimination of bacteria from the small bowel. A surgical procedure may disrupt any of these mechanisms or may unmask a previously asymptomatic disorder that now requires appropriate treatment. Treatment of diarrhea related to surgery may be directed toward the diarrhea itself with such agents as tranquilizers and opiates but if these are not soon effective any disrupted functions must be analysed and specific measures taken to correct or compensate for these disruptions. The most useful measures in appropriate cases are the prevention of the dumping syndrome and the administration of pancreatic enzymes, cholestyramine, hydrophilic stool thickeners and broad-spectrum antibiotics. These measures may be ineffective or inadequate and surgical revision may be necessary.     

Symposium on diarrhea. 3. Investigation of chronic diarrhea.

The practical approach to the investigation of diarrhea must be logical and based on anatomic considerations. The site of the underlying disorder may be determined by the clinical picture, and the logic of investigation will be influenced by the history. Important specific investigation in a case of colonic diarrhea include a careful rectal examination, stool inspection, sigmoidoscopy, rectal biopsy and barium enema study. Colonoscopy has been used, but its role has yet to be defined. In a case of small-bowel steatorrhea or diarrhea quantitative chemical estimation of the daily output of stool fat is useful, and to this investigation is added a small-bowel radiograph series and, if the radiographic findings are abnormal, small-bowel biopsy. Other investigations for small-bowel disease may include the breath test with carbon-14-labelled glycocholic acid, the lactose tolerance test, duodenal aspiration for giardiasis, analysis of serum immunoglobulins and, on occasion, isolation of vasoactive intestinal polypeptide hormone (which may aid the diagnosis of functioning tumours of the pancreas or small bowel). Investigations for pancreatic steatorrhea include abdominal radiography, performance of the secretin test and testing of the response to pancreatic replacement therapy. In some patients it may be useful to use endoscopic retrograde cholangiopancreatography to differentiate pancreatic carcinoma and chronic pancreatitis.     

A waterborne outbreak of epidemic diarrhea due to group A rotavirus in Malatya, Turkey

We characterized an outbreak of acute diarrheal disease caused by group A rotavirus that occurred during the Autumn of 2005 in Malatya City, Turkey. A total 9907 patients between 0 to 91 years old (mean age: 25.05?19.67) were included in the epidemic. The patients? data were prospectively collected and statistically analyzed. Microbiologic analyses were performed to determine the etiologic agent. Rapid onset diarrhea (98.36%), abdominal cramps (69%), fever (44.4%) and vomiting (69.6%) were the most common symptoms observed in patients. Rotavirus antigen was detected in 52.7% of the studied patients. RT-PCR analysis led to identification of Group A rotavirus as the causative agent of this epidemic. Simultaneous measurements of the drinking water samples yielded very low chlorine levels; as low as 0 to 0.05 mg/L. The outbreak investigation team indicated possible contamination of a large water depository from a water well, which supplies drinking water to two major districts of the city. Effective chlorination and blockage of the passage between the well and the water depository stopped the outbreak. This outbreak shows the high epidemic potency of rotavirus in large human populations, including all age groups, and underlines the importance of water safety in pipeline systems.     

炎琥宁注射液对小儿轮状病毒腹泻的治疗及对患儿心肌酶的影响

目的探讨炎琥宁注射液对小儿轮状病毒腹泻的疗效及对患儿心肌酶的影响。方法选择2016年1月至2016年12月大兴区妇幼保健院收治的94例小儿轮状病毒腹泻患儿为研究对象,所有患儿按照随机数字表法分为观察组(47例)与对照组(47例)。两组患儿均采用常规治疗,观察组在常规治疗基础上结合炎琥宁注射液治疗。两组患儿疗程均为7d。比较两组患儿的疗效,治疗前后肌酸激酶(CK)、乳酸脱氢酶(LDH)、肌酸激酶心型同工酶(CK-MB)的变化,止泻时间、腹胀消失时间、退热时间,及不良反应发生情况。结果观察组患儿治疗总有效率(93.62%)高于对照组(74.47%),差异有统计学意义(P0.05)。两组患儿治疗后CK、LDH、CK-MB水平较治疗前降低(P0.05),且观察组低于对照组(P0.05)。观察组患儿止泻时间、腹胀消失时间及退热时间均短于对照组(P0.05)。两组患儿不良反应发生率比较差异无统计学意义(P0.05)。结论炎琥宁注射液治疗小儿轮状病毒腹泻效果显著,且可明显改善患儿心肌酶水平,具有一定临床应用价值。