ABSTRACT
ObjectiveTo investigate the serological markers associated with posthepatectomy recurrence in patients with hepatocellular carcinoma, and to establish a prognostic model to evaluate whether palliative hepatectomy is suitable for such patients. MethodsA total of 111 patients with hepatocellular carcinoma who underwent hepatectomy in the Affiliated Cancer Hospital of Zhengzhou University from February 2009 to July 2013 and received follow-up were enrolled. Basic clinical data were collected and the patients were divided into recurrence group and non-recurrence group according to whether recurrence was observed during follow-up. The t-test was used for comparison of normally distributed continuous data between two groups and the Wilcoxon rank sum test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test was used for comparison of categorical data between two groups. Survival curves were plotted using the Kaplan-Meier method, and survival differences were analyzed using the log-rank test. A Cox regression analysis was used to perform univariate and multivariate analyses, and the area under the ROC curve (AUC) was used to evaluate prediction efficiency. ResultsThe Kaplan-Meier survival curves showed that the patients with low alpha-fetoprotein (AFP), alkaline phosphatase, gamma-glutamyl transpeptidase (GGT), and fibrinogen and high CXCL13 had a longer median time to recurrence (P＜0.05). AFP (hazard ratio ［HR］［95%CI］=1.69(1.03~2.79), P=0.039), GGT (HR［95%CI］=1.89(1.14~3.14), P=0.014), and CXCL13 (HR［95%CI］=0.54(0.33~0.89), P=0.015) were independent factors associated with posthepatectomy recurrence. The prognostic index PI=0.526×AFP+0.637×GGT-0.616×CXCL13 established based on these factors had an AUC of 0.87, a sensitivity of 93.75%, and a specificity of 63.64% in predicting recurrence within 0-3 months after palliative hepatectomy, with a significant reduction in prediction efficiency for recurrence within 0-6 months (AUC=0.68) or a longer period of time. The recurrence prediction efficiency of this model for palliative hepatectomy was significantly higher than that for radical resection. ConclusionThe prognostic model established based on CXCL13, AFP, and GGT can be used to evaluate the risk of early recurrence after palliative hepatectomy and thus helps clinicians to make diagnosis and treatment decisions based on patients’ benefits.
ABSTRACT
Objective@#To express the NS6 nonstructural protein of human norovirus (NoV) in Escherichia coli, and to detect its enzymatic activity after purification.@*Methods@#Human NoV NS6 gene was cloned into the prokaryotic expression vector pDE1 and then was transformed into E. coli BL21 for expression. NS6 protein was purified by Ni-NTA affinity chromatography and Superdex 200 pg column. The activity of NS6 protein was determined by digestion of fusion protein 15VP1-6P at 37 ℃.@*Results@#Human NoV NS6 protein was stably and highly expressed in E. coli. After purification, the expressed product reached a purity of more than 95%, and the relative molecular weight of NS6 protein was about 23×103 Da. NS6 protein could cleave the fusion protein containing rhinovirus 3C cleavage site.@*Conclusions@#The nonstructural protein NS6 of human NoV was successfully expressed in Escherichia coli, which laid a foundation for further study on the pathogenesis of human NoV.
ABSTRACT
Objective@#To analyze the epidemiological characteristics and pathogen types of viral diarrhea in children under 5 years of age in Shandong province, and provide reference for the prevention and control of viral diarrhea.@*Methods@#A total of 1 017 fecal samples were collected from all children aged 5 years and younger with diarrhea who were admitted to the sentinel hospital of Shandong province from 2012 to 2017 within 3 days. Rotavirus antigen was detected by using an ELISA method . Rotavirus G/P typing was performed by RT-PCR; Norovirus (GI and GII), Sapovirus, and Astrovirus were detected by multiplex RT-PCR, and adenovirus was detected by PCR.@*Results@#In the 1 017 fecal specimens, the overall positive rate was 51.62% (525/1017), and viral nucleic acids were detected in at least 421 samples, and mixed virus infection was found in 104 sampes. The mixed infection accounted for 10.23% (104/1017) of all infections. The positive detection rates of Rotavirus, Calicivirus, Adenovirus and Astrovirus were 34.22% (348/1017), 16.91% (172/1017), 2.56% (26/1017), and 9.64% (98/1017)). The total detection rate of diarrhea virus and the detection rate of Rotavirus were the highest at 12 to 17 months of age, which was 51.72% (105/203) and 20.20% (41/203), respectively. The positive rate of diarrhea in children aged 2 years and younger was 49.36% (502/1017), which was much higher than the positive rate of diarrhea in children over 2 years old (2.26% (23/1017)). The peak of viral diarrhea was found to occur between November and April of the following year. The genotype of rotavirus was dominated by G9 (82.76%), the P genotype was dominated by P[8] (80.46%), and the G/P combination was dominated by G9P[8] (83.87%). Norovirus was the main infection in the Calicivirus (87.21%).@*Conclusions@#From 2012 to 2017, viral diarrhea in children under 5 years of age in Shandong Province was mainly caused by Rotavirus infection, followed by Norovirus. The overall prevalence of viral diarrhea in Shandong was moderate in China, and autumn and winter were the main epidemic season for viral diarrhea.
ABSTRACT
Objective@#The gene engineering technique was used to express the P[6] genotype rotavirus (rotaviruses, RVs) GST-VP8*-Z84 protein from pigs, and the binding characteristics of the protein to oligosaccharide and salivary receptor were studied.@*Methods@#The GST-VP8*-Z84 protein was purified by GST Escherichia coli expression system and affinity chromatography using porcine P[6]. Enzyme-linked immunosorbent assay (ELISA) saliva binding test and oligosaccharide binding test were used to analyze the binding characteristics of the genotype to receptors.@*Results@#Porcine P[6] GST-VP8*-Z84 protein bound well to mucin core 2.@*Conclusions@#The potential receptor of P[6] RV may be the core of mucin, which may provide the experimental basis and theoretical basis for the mechanism of rotavirus and receptor interaction and the development of RV vaccine and highly effective therapeutic drugs.
ABSTRACT
Objective@#During the 2016 winter season, GII.2 norovirus(NoV) suddenly emerged in China. To elucidate its mechanism of epidemic, this study focused on characteristics of binding between the P protein of capsid and histo-blood group antigens (HBGAs).@*Methods@#The research object was GⅡ.2 ZTX strain which had an outbreaks by the end of 2016 in Beijing. Recombinant prokaryotic expression plasmid was constructed, and the expression of virus P protein was determined and purified. The P protein characteristics of binding to HBGAs was studied through saliva and oligosaccharide binding experiments.@*Results@#Soluble P protein was successfully obtained, and combined with type A, B, AB saliva.@*Conclusions@#The result illuminate the combination with new outbreaks of NoV and salivary types, which provided a basis for its pathogenic mechanism and prevention and control measures.
ABSTRACT
Objective@#To investigate the clinical and epidemiological characteristics of Coronavirus HKU1 (Human CoV-HKU1) and NL63 (Human CoV-NL63) in children with acute respiratory tract infection in Nanjing.@*Methods@#From August 2009 to July 2011, 1 286 respiratory samples were collected from the outpatient and hospitalized children in the Children′s Hospital of Nanjing Medical University. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect HCoV-HKU1 and NL63 genes, besides, positive samples were used for common respiratory virus screening. The positive amplification products were cloned, sequenced, homologous and phylogenetic analysis was conducted by molecular biological method .@*Results@#The detection rate of HCoV-HKU1 was 1.1% (14/1 286), the positive sequences shared a 98.2%-100% nucleotide identity with the HCoV-HKU1 strains and mixed infection rate was 92.9%. The main clinical diagnoses were bronchitis, bronchopneumonia and bronchiolitis. The clinical manifestations were cough, fever, wheezing. The detection rate of HCoV-NL63 was 1.5% (19/1 286), the positive sequences shared a 95.6%-100% nucleotide identity with the HCoV-NL63 strains and mixed infection rate were 63.2%. The main clinical diagnosis were acute upper respiratory tract infection, bronchitis, bronchopneumonia. The clinical manifestations were fever, cough, expectoration. No deaths were found in both HCoV-HKU1 and NL63 infections.@*Conclusions@#From August 2009 to July 2011, HCoV-HKU1 and NL63 were detected in children with respiratory tract infection in Nanjing area. HCoV-HKU1 infected cases were lower respiratory tract infection, epidemic in winter and spring, infected cases were mainly under 1 years of age, HCoV-NL63 infected cases including upper respiratory and lower respiratory tract infection, epidemic in the season of summer and autumn. The infected cases were mainly at the age rank from 1 year to 3 years. The clinical manifestations of children infected with coronavirus HKU1 and NL63 are not specificity.
ABSTRACT
Objective@#To evaluate of the specificity of a new norovirus (NoV) detection method of in situ capture real-time quantitative reverse transcription polymerase chain reaction (ISC-RT-qPCR) and to apply the method for the detection of NoVs in fresh strawberry.@*Methods@#A panel of stool samples with different NoV genotypes and various inoculums were used for the experiments.@*Results@#We found that all the tested samples of eight genogroup Ⅱ (GⅡ) NoVs could be detected specifically by ISC-RT-qPCR. Moreover, in contrast to the conventional RT-qPCR method , the situation that the Ct value increased as the inoculum of NoV GⅡ decreased was not shown using ISC-RT-qPCR. When we tested NoVs in strawberry samples by ISC-RT-qPCR, the minimum test limit could reach 1.36 genocopy/10 g of fresh strawberry.@*Conclusions@#ISC-RT-qPCR is an effective and specific technic and it could be applied for the detection of infectious NoVs from stool samples and fresh strawberry samples.
ABSTRACT
Objective@#The VP8* core protein of rotavirus P[6] genotype LL4260 was purified by prokaryotic expression, which is important for further study of protein structure and function.@*Methods@#The P[6] genotype LL4260 strain was obtained by PCR.The recombinant plasmid pET30 a-LL4260VP8*core was inserted into pET30 a vector and transformed into BL21 (DE3) competent cells with the correct recombinant plasmid. The expressed protein is purified by affinity chromatography and molecular sieve chromatography.@*Results@#The pET30 a-LL4260VP8* core region protein is soluble in the supernatant and proteins of approximately 22 kDa are identified by electrophoresis using purified proteins.@*Conclusions@#In this study, LL4260 containing pET30 a-LL4260VP8* core plasmid was successfully constructed and LL4260 strain VP8* protein was expressed.
ABSTRACT
Objective@#To investigate the human bocavirus (HBoV) infection in the children diagnosed with brochiolitis, incomparison with the clinical characteristics of bronchiolitis of HBoV vs. respiratory syncytial virus (RSV) infection alone.@*Methods@#A total of 396 throat swabs were obtained from children diagnosed with bronchiolitis from January 2015 to December 2016 in Pediatric Asthma Center of Gansu Maternal and Child Health Care Hospital, Lanzhou. Specimens were tested for the nucleic acids of HBoV, RSV and other 6 common respiratory viruses by nested-polymerase chain reaction (nested PCR() and reverse transcription-polymerase chain reaction (RT-PCR), and the epidemic status, clinical characteristics of HBoV and RSV were analyzed and compared with each other.@*Results@#The positive rate of virus infection was 53.54%, and HBoV and RSV infections were 9.84% and 24.49% in 396 cases. The numbers of cases of HBoV single infection, RSV single infection, HBoV and RSV mixed infection were 29, 86 and 5. The number of boys infected with HBoV was 27. There was statistical significance in the difference between the genders. The statistical significance also existed in difference of age. The babies at ages ranging from 6 to 12 months had the hifgest positive rate. The first incidence peak of the infection of HBoV was in October 2015. The children with HBoV infection, compared with RSV patients, presented more often with vomiting, diarrhea (P<0.05), while symptoms such as the occurrence of dyspnea, hypoxemia and the hospitalization time were all lower.@*Conclusions@#HBoV is one of the possible causes of brochiolitis in infants, its infection distributed over the whole year. The clinical symptoms, laboratory and radio-graphic changes are similar between the HBoV and RSV infections, the only difference was that the HBoV infection was comparatively mild.
ABSTRACT
0bjective To identify the molecular structure and genome homology characteristics of a GV murine norovirus (MNV) from wild mice.Methods@#Nucleic acids were extracted from collected mouse intestinal specimens and subjected to high-throughput sequencing. The whole genome sequence was obtained by RT-PCR and RACE method .@*Results@#The MNV genome was 7, 832 bp in length, with 76.8%-78.1% nucleotide identity to other selected murine norovirus strains in GenBank. The P2 region, the hypervariable area in viral protein 1 gene shows highest heterology, and the identity was among 60.9%-78.1%, lower than other genes regions. The result demonstrated the higher variability of this newly isolated WNV strain.@*Conclusions@#The wild-type MNV isolated in this study is first reported in China, which enriched the domestic MNV gene pool and provided a reference for understanding the molecular genetic characteristics and evolutionary origin of MNV in China.
ABSTRACT
Objective@#To understand the novel bat-borne hantavirus on Yunnan province.@*Methods@#Eighty-four bats were collected from Puer, Yunnan from July to August in 2016. A hantavirus, named as DodeHV, was identified in Pomona Roundleaf Bat by high-throughput sequencing technology. Nested or Semi-nested reverse transcription-polymerase chain reaction was used to confirm and screen DodeHV, as well as to amplify its genome sequence by specific primers. Then homology and phylogenetic analysis of all three segments were conducted by using software MegAlign and MEGA 6.0.@*Results@#All the positive individuals are Pomona Roundleaf bats by PCR in this study and 4 out of 67 (5.97%) Pomona Roundleaf bats were positive. The complete ORF of DodeHV S Segment, the most sequence of M and the complete sequence of L segment were obtained. It shared the highest homology with the strain XSV-VN1982B4 found on Phu Tho Province, Viet- Nam in 2013, the nucleotide sequence identities of S, M, L segment compared with XSV was 79.0%, 79.2%, 79.9% respectively, and its amino acid sequence identities was 93.4%, 94.8%, 96.6% respectively. Meanwhile, phylogenetic analysis showed that DodeHV was also closely related to strain XSV-VN1982B4.@*Conclusions@#The discovery of DodeHV enriched the virus reservoir of our country which is meaningful to public health. It is indicated that there is potential risk of bat-borne DodeHV at cross-border infection at frontier.
ABSTRACT
Objective@#To explore the receptor binding specificity of VP8* protein of porcine P[19] rotaviruses (RVs) with oligosaccharides.@*Methods@#The porcine P[19] VP8* protein was expressed and purified. The receptor binding specificity of P[19] VP8* was analyzed by oligosaccharide binding and saliva binding assay.@*Results@#The P[19] VP8* protein showed significant binding to mucin cores, especially mucin core 2.@*Conclusions@#Mucin core 2 may be a potential receptor for the porcine P[19] RV, which provides certain basis for the study of virus infection mechanism and RV surveillance.
ABSTRACT
Human norovirus (NoV) is a member of the calicivirus family, can cause nausea, vomiting, abdominal pain and diarrhea as the main clinical symptoms of acute gastroenteritis. Human norovirus infection can be popular in the world, all ages, causing serious burden of disease. Due to the lack of suitable small animal models, there is still a lack of understanding of human immune to norovirus infection and pathogenesis. Murine norovirus (MNV) was originally isolated in immune deficient mice, and causes infection and epidemic in mice. MNV provides an alternative model to study human norovirus infection and the host intestinal immune mechanism. This article will elaborate on two aspects of innate immunity and adaptive immunity.
ABSTRACT
Objective:To study the effect of alum on immune response in mice induced by HBoV1 VP2 VLPs.Methods:BABL/c mice were randomly divided into VLPs experimental group, alum adjuvant experimental group, PBS control group and alum adjuvant control group,the experimental group mice were intramuscular immunization with HBoV1 VP2 VLPs and HBoV1 VP2 VLPs added alum,control group mice were immunization with alum or PBS buffer,then to study the effect of alum on immune response in mice induced by HBoV1 VP2 VLPs by cellular and humoral immune strength.Results: Alum adjuvant decreased cellular immune response induced by HBoV1 VP2 VLPs(P<0.001),enhance the HBoV1 VP2 VLPs immuned serum IgG titer(P<0.05)and IgG activity(P<0.01).Conclusion:Alum adjuvant can enhance humoral immune response induced by HBoV1 VP2 VLPs,but weaken cellular immune response induced by HBoV1 VP2 VLPs,when HBoV1 VP2 VLPs used as a prophylactic vaccine it should add alum adjuvant,when used as a therapeutic vaccine,it should not add alum adjuvant.
ABSTRACT
Objective To understand the epidemiological and clinical characteristics of ＜ 2 years old children hospitalized due to intussusceptions.Methods Clinical and demographic data of ＜2 years old children hospitalized due to intussusception between January 2007 and August 2013 were retrospectively collected in Affiliated Children' s Hospital of Soochow University in Suzhou.The incidence data,age distribution,seasonality and clinical characteristics of hospitalized intussusceptions cases were analyzed.Results A total of 594 intussusception-related hospitalizations were identified during this period in children aged ＜2 years,no death occurred.The crude incidence of hospitalized intussusception was 57.3 per 100 000 in children aged ＜2 years (95％CI:52.8-62.1),and 100.6 per 100 000 in children aged ＜1 year (95％CI:92.1-109.8).The male to female ratio was 1.90 ∶ 1.Up to 85.4％ (507/594) of the cases were aged ＜ 1 year,and 66.2％ (393/594) of the cases were aged 3-8 months.The incidence peaked in age group 5-8 months.The median age of the cases was 6.8 months (QR=4.4),and increased from 6.3 months (QR=4.2) in 2007 to 7.3 months (QR=4.0) in 2013.No obvious seasonality was observed.Main symptoms or signs included vomiting (83.2％,494/594),abdominal mass (81.1％,482/594),and bloody stool (64.5％,383/594).Abdominal ultrasonic testing was the most frequently used diagnostic approach (98.7％,586/594).Up to 86.2％ (512/594) of patients were successfully treated by surgical intervention.The main sites for acute intussusception in children aged ＜2 years were ileo-colic (34.5％,183/530),ileo-ileo (30.8％,163/530) or ileo-ileo-colic (27.9％,148/530).Conclusion The incidence of hospitalized intussusception in children aged ＜2 years was high in Suzhou.It is necessary to establish an active surveillance system to provide baseline data for the evaluation of rotavirus vaccine safety.
ABSTRACT
We studied the epidemiological characteristics of human bocavirus 4 (HBoV4) in children with a- cute gastroenteritis in Lanzhou (China). A total of 331 stool specimens were collected from children aged < 5 years with acute diarrhea at the First Hospital of Lanzhou University between July 2012 and June 2013. Specimens of HBoV were identified by nested polymerase chain reaction assays. Compared with related sequences in GenBank, the HBoV-positive strain isolated in the present study was,quite surprisingly, a rare genotype named HBoV4. This strain was a typical HBoV4,with high levels of nucleotide and amino acid homology to the Thailand strain, JQ267789 (98.9% and 98.7%, respectively), and the USA strain, GQ506568 (97.6% and 97.4%, respectively). This is the first report of HBoV4 as the causative agent for acute gastroenteritis in pediatric patients in China. This strain is one of two genotypes of HBoV that are currently circulating.
Subject(s)
Child, Preschool , Female , Humans , Infant , Male , China , Feces , Virology , Gastroenteritis , Virology , Human bocavirus , Classification , Genetics , Molecular Sequence Data , Parvoviridae Infections , Virology , PhylogenyABSTRACT
Interactions between noroviruses (NoVs) and the receptors of histo-blood group antigens (HB-GAs) affect the infectivity and host susceptibility of NoVs. We elucidated the binding profile of a GII. 12 NoV to HBGAs. First, we synthesized the P domain sequence of the GII. 12 NoV strain Pune (GenBank accession number EU921353). Protein of the P domain was expressed in a prokaryotic system. Formation of the P particle was monitored by gel-filtration chromatography. Antiserum was prepared by immunization of mice with GII. 12 P particles. The binding profile of the GII. 12 NoV Pune strain was determined by binding of the P particle with a panel of saliva samples with various known HBGAs phenotypes. The GII. 12 NoV was bound strongly to saliva samples of subjects with B and AB types and weakly to A, O secretor, and non-secretor saliva samples, suggesting higher affinity with B antigen by GII. 12 NoV. These results were consistent with those determined by a previous crystallography study of GII. 12 NoV. These data suggested that individuals with B and AB blood types may be more susceptible to infection by GII. 12 NoV compared with those with other blood types. Our findings may provide a basis for the prevention and control of an epidemic of GII. 12 NoV.
Subject(s)
Animals , Female , Humans , Mice , Blood Group Antigens , Metabolism , Caliciviridae Infections , Metabolism , Virology , Gastroenteritis , Metabolism , Virology , Genotype , Mice, Inbred BALB C , Norovirus , Genetics , Metabolism , Protein Binding , Receptors, Virus , Metabolism , Viral Proteins , Genetics , MetabolismABSTRACT
Rotavirus is the leading causal agent of severe acute gastroenteritis in children aged <5 years. A specific pharmacologic agent for the treatment of rotavirus-infected children is lacking. In China, only the Luo Tewei oral vaccine (Lanzhou Institute of Biological Products, Shanghai, China), which is produced from Lanzhou lamb rotavirus vaccine (LLR), is available. Studies have hypothesized that the genotype of LLR is G10P[12], To identify the genotype of LLR by reverse transcription-polymerase chain reaction, we showed that the VP7 and VP4 genotypes of LLR were G10 and P[15], respectively, based on sequencing, alignment and phylogenetic analyses. In conclusion, we identified the genotype of rotavirus strain LLR to be G10P[15].
Subject(s)
Humans , China , Genotype , Molecular Sequence Data , Phylogeny , Rotavirus , Chemistry , Classification , Genetics , Rotavirus Infections , Virology , Rotavirus Vaccines , Chemistry , Classification , Genetics , Sequence Homology, Amino Acid , Viral Proteins , Chemistry , GeneticsABSTRACT
MicroRNA (miRNA) are small non-coding molecules of ribonucleic acid. They are about 22 nucleotides in length, single-stranded, and mediate post-translational regulation by the repression or degradation of messenger RNA(mRNA). miRNA play a key part in the proliferation, differentiation and death of cells. Viral infection is one of the most common causes of human disease. Some studies have found that miRNA has a very close relationship with viral infection, which has an effect on viral replication, the immune response and antiviral immunity. Use of miRNA may become the cornerstone of new methods for the diagnosis and treatment of viral infection. This article summarizes the progress of research into miRNA and viral infection.