This study aimed to amplify major genome segments (VP7, VP4, VP6, VP2 and NSP2-5) of porcine G3 group A rotavirus (GARV) LLZ212 isolated in our laboratory, determine their genotypes, and explore the evolutionary relationships between G3 GARV strains isolated from humans and pigs in Lulong County, Hebei Province, China. Major genome segments of seven GARV strains were amplified by reverse transcription-polymerase chain reaction (RT-PCR) and the segments were sequenced. The genome segments of seven GARV strains were determined by the online RotaC genotyping tool (RotaC v2.0). The reference sequences of each GARV genome segment were downloaded from GenBank. Homology and phylogenetic evolutionary analyses were conducted using the MEGA 5.0 and DNAStar software packages. LLZ212 isolated from pigs in Lulong had the following genotype: G3-P[8]-I5-C1-N1-T1-E1-H1. All human GARV strains had the following genotype: G3-P[8]-I1-C1-N1-T1-E1-H1. The VP7, VP4, NSP4 and NSP5 genes of the LLZ212 strain had the highest nucleotide identities with the human GARV E885, CMH014/07, Wa and RMC321 strains, respectively, and these clustered together in a sublineage. The VP6, NSP4 and NSP5 genes of the LLZ212 strain shared the highest nucleotide identities with the porcine GARV PRG921 strain, while VP2 associated most closely with porcine GARV OSU strain, and these also clustered in a sublineage. A rare porcine G3-P[8]-I5-C1-N1-T1-E1-H1 GARV strain was identified, which may represent a reassortment between porcine and human viruses. In conclusion, the VP7, VP4, NSP4 and NSP5 genes of LLZ212 share high levels of sequence identity with human GARV, while VP2, VP6, NSP2 and NSP3 cluster with porcine GARV.
Animals ; Capsid Proteins ; genetics ; Child, Preschool ; China ; epidemiology ; Evolution, Molecular ; Genotype ; Humans ; Infant ; Male ; Molecular Sequence Data ; Phylogeny ; Rotavirus ; classification ; genetics ; isolation & purification ; Rotavirus Infections ; epidemiology ; veterinary ; virology ; Swine ; Swine Diseases ; epidemiology ; virology ; Viral Nonstructural Proteins ; genetics

Rotaviruses, which are recognized as one of the major etiological agents among infants and young children with diarrhea, consist of three concentric layers of protein capsid with the enclosed double-stranded RNA genome. Rotaviruses infect host cells mainly by identifying the specific receptors on cell surfaces and binding to them. Therefore, receptors are important factors for viruses infecting cells. So far, there have been many receptors found to be involved in rotavirus infection, including sialic acid, integrin, Toll-like receptor, and blood group antigen. This article provides an overview of receptors involved in rotavirus infection.
Animals ; Humans ; Receptors, Virus ; genetics ; metabolism ; Rotavirus ; genetics ; physiology ; Rotavirus Infections ; genetics ; metabolism ; virology

This study aimed to study the epidemiological and clinical characteristics of human bocavirus 1-4 (HBoV1-4) in children with acute diarrhea in Lanzhou and to investigate the association between HBoV and acute gastroenteritis. A total of 331 stool samples were collected from children aged under 5 years with acute diarrhea at the Department of Pediatrics, the First Hospital, Lanzhou University, between July 2012 and June 2013. Nested PCR was used to screen for HBoV and a general PCR was employed to screen other common diarrhea viruses. We found human bocavirus 1, 2, 3 and 4 in 26, 15, 7 and 1 cases, respectively. There was no specific seasonal distribution of HBoV, with infections occurring throughout the year. HBoV was mostly found in children aged between 7 and 12 months, with a mean age of 11.04 months (+/- 6.92 months), and 93.88% of affected children were aged under 2 years. Overall, 71.3% of mixed infections were mixed and the majority of other infections were caused by rotavirus. There was no statistical difference in the incidence of fever and vomiting associated with HBoV infection. A rare virus strain, HBoV4 (LZFB086), was identified, which showed highest levels of nucleotide sequence identity (99.0%) with a single Thai HBoV strain (JQ267789). No case of HBoV2B was found. In conclusion, HBoV1 was a major etiological pathogen of HBoV in pediatric cases in Lanzhou. HBoV4 was detected in feces for the first time in China. The rate of mixed infections was high and rotavirus was dominant. The data presented suggests that HBoV is not a major causative agent of gastroenteritis.
China ; epidemiology ; Diarrhea ; epidemiology ; virology ; Feces ; virology ; Human bocavirus ; classification ; genetics ; isolation & purification ; Humans ; Infant ; Molecular Sequence Data ; Parvoviridae Infections ; epidemiology ; virology ; Phylogeny ; Seasons

Abstract:This study aims to investigate the genetic characteristics of group A rotavirus (GARV) G9P[8] strains from infantile diarrhea samples in Hebei Lulong region from 2009 to 2011. We randomly selected five GARV G9P[8] strains in Hebei Lulong region from 2009 to 2011, amplified the 11 gene fragments of GARVs by RT-PCR, and analyz their full-genome sequences by homology and phylogenetic analysis with DNAStar and MEGA. The nucleotide homology between strains LL11131077 and LL11131083 in 2011 was significantly higher than hat etween them and the other three strains in 2009 and 2010. The G9P[8] GARVs circulating in Hebei Lulong region from 2009 to 2011 elenged to the same genotype as the prevalent G9P[8] GARVs in other parts of the world. However,the two strains in 2011, compared with those in 2009 and 2010, were located in a different sub-branch of the phylogenetic tree and had amino acid mutations at many sites.
China ; Feces ; virology ; Genome, Viral ; Genotype ; Humans ; Molecular Sequence Data ; Phylogeny ; Rotavirus ; classification ; genetics ; isolation & purification ; Rotavirus Infections ; virology ; Viral Proteins ; genetics

OBJECTIVETo establish an animal model of lung injury in SD rats using intratracheal instillation of unknown polymer and to provide the base for exploring the molecular mechanism of lung tissue injury induced by occupational exposure.

METHODSOne hundred forty SD rats were randomly divided into seven groups, including the control group 1 which was exposed to normal solution, the control group 2 which was not exposed to any one and five treatment groups which were exposed to 1 ml unknown polymer (0.5 ml for each lung) at the doses of 40, 30, 20, 10 and 5 mg/ml, respectively by intratracheal instillation. The rats were sacrificed on the 1st, 3rd, 7th, 10th, 14th, 21th and 28th day after exposure, then the lung tissues were examined pathologically and the blood bio-chemical analysis was conducted.

RESULTSThe results of blood biochemical analysis indicated that ALT and AST levels in rats exposed to 30 and 40 mg/ml unknown polymer were significantly higher than those in control groups. Intratracheal instillation of unknown polymer can causes PLF in experimental animals on the 14th days after exposure. The results of pathological examination exhibited that the lung tissue injury in rats exposed to unknown polymer for 14 days or more was found and the dose-effect relationship was observed.

CONCLUSIONAn animal model of lung injury in SD rats induced by unknown polymer with intratracheal instillation was established successfully. The results of pathological examination showed that the types of rat lung injury were similar to the clinical lung injury after exposure to unknown polymer, which provided a base for studying the mechanism of lung injury caused by occupational exposure to unknown polymer.

Animals ; Disease Models, Animal ; Female ; Lung ; drug effects ; pathology ; Lung Injury ; chemically induced ; Male ; Polymers ; toxicity ; Rats ; Rats, Sprague-Dawley

Objective · To explore the prognostic factors for postoperative radiotherapy for patients with locally advanced esophageal squamous cell carcinoma (ESCC) and the effects of lymph nodes metastasis status on the survival of patients and failure modes of treatment. Methods · Data of 121 patients with locally advanced ESCC who underwent radical resection and postoperative radiotherapy from 2006 to 2013 were collected. The overall survival (OS) and disease-free survival (DFS) were calculated using Kaplan-Meier. Univariate analysis and multivariate analysis were performed to investigate prognostic factors with the log-rank test and the Cox regression model. The differences in OS and recurrence patterns between patients with different lymph node metastasis status were compared. Results · The median DFS of all patients was 22.57 months and median OS was 32.90 months.Multivariate analysis showed that KPS score, length of lesion, and positive lymph nodes ratio (pLNR) were independent prognostic factors for DFS and OS. For patients with positive lymph nodes, the median DFS of patients with pLNR ≤ 0.15 and pLNR>0.15 were 33.43 and 19.20 months (P=0.04).Patients without nodal skip metastasis (NSM) had better median OS than patients with NSM, but the difference was not statistically significant. OS was significantly worse in patients with pLNR>0.15 and NSM than in other patients with positive lymph nodes (median OS of 14.33 vs 32.50 months, P=0.02).pLNR had a better prognostic value for OS than the number of positive lymph nodes (AUC=0.673, P=0.04). Analysis of the failure patterns showed that more distant metastases were observed in patients with pLNR>0.15, while more local and regional recurrences were observed in patients with pLNR ≤ 0. 15. Conclusion · The status of lymph nodes metastasis is associated with the prognosis of postoperative radiotherapy for patients with locally advanced ESCC. pLNR has a better prognostic value for OS for patients with positive lymph nodes. The recurrence pattern varies in patients with different pLNR after postoperative radiotherapy. The patients with high pLNR and NSM have poor prognosis.

Objective To explore the feasibility and clinical outcome of laparoscopy-assisted gastrectomy with D2 dissection by comparing the efficacy of open surgery on radical gastrectomy with D2 dissection for gastric cancer patients. Methods The patients with gastric cancer from October 2008 to August 2013 were divided into two groups according to the different surgical methods. Among them, 175 patients underwent laparoscopic surgery (laparoscopy-assisted surgery group, Group L), and 163 patients underwent laparotomy surgery (open surgery group, Group O). The number of lymph nodes dissected, postoperative recovery, complications, mortality and survival rate of the two groups were compared and analyzed. Results There was no significant difference in the number of lymph nodes resect between the two groups [Group L and Group O: (26.3 ± 13.9) vs (26.8 ± 10.2), t = -0.40, P = 0.684]. Compared with open surgery, the laparoscopy-assisted surgery showed significantly less intraoperative blood loss and quicker recovery of gastrointestinal function. The postoperative hospitalization time of laparoscopic group was less than that of laparotomy group, the difference was statistically significant (P < 0.05). No significant difference was found in 3-year survival rate between the two groups (Group L vs Group O: 92.00％ vs 92.63％, P = 0.262). Conclusions Compared to open surgery, laparoscopic gastrectomy is safer and has quicker postoperative recovery. There is no statistical difference in the number of resect lymph nodes between lapamscopic and open gastrectomy.

The United Nations Environment Program estimates that approximately 20% of agricultural land and 50% of cropland in the world is salt-stressed. The gene NHX (Na+/H+ exchanger) encodes functional protein that catalyzes the countertransport of Na+ and H+ across membranes and may play an important role in plant salt tolerance. To clone the NHX from the wild plant Populus euphratica collected in Tarim basin and Xinjiang Wujiaqu district into a T-vector, designed primer was used to amplify 1kb NHX cDNA fragment with RT-PCR. Total RNA was extracted from Populus euphratica tissue (plant tissue was collected from Tarim basin and Xinjiang Wujiaqu district and stored in liquid nitrogen) according to the Plant RNA Mini Kits of Omega. First cDNAs were synthesized from 1 microg total RNA of Populus euphratica seedling. A pair of primers were used to perform RT-PCR. The amplified DNA fragment was purified and cloned into pMD18-T vector. However, 1kb and 2.3kb fragment were obtained from Tarim basin and Xinjiang Wujiaqu district and named as PtNHX and PwNHX, respectively. Sequence analysis reveals that the cloned PtNHX fragment of Populus euphratica contains partial NHX coding region with 98%, 86%, 84% and 80% identity comparing with Atriplex gemelini, Suaeda maritima, Arabidopsis thaliana and Oryza sativa, respectively. This analysis suggests that NHX gene would be highly conserved in terms of evolution in plant; and it also suggests that the NHX gene of Populus euphratica also would have the similarity with that of Arabidopsis. It may be of great importance in improvement of the plant salt tolerance and breed of crop. At the same time, sequence analysis shows that PwNHX gene includes a coding region about 1350bp with 99% identity comparing with transposon Tn10 IS10-left transposase of Shigella flexneri. On the one hand, the NHX gene may lose its function because it was inserted a fragment in coding region. On the other hand, its product may play a important role in salt tolerance. Populus grow in saline soil. It speculates that it may have other salt tolerance mechanism in Populus. The transposon can be used as transposon tagging to clone other genes and it will help us to understand farther the salt tolerance mechanism.
Amino Acid Sequence ; DNA Transposable Elements ; genetics ; DNA, Complementary ; Genetic Vectors ; genetics ; Molecular Sequence Data ; Plant Proteins ; chemistry ; genetics ; Populus ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Alignment ; Sequence Homology, Amino Acid ; Shigella flexneri ; genetics

OBJECTIVETo explore the relationship between Chinese medicine (CM) constitutive susceptibility and syndrome diversity in diabetic nephropathy (DN).

METHODSEpidemiologic investigation on constitution adopting the "Constitution in Chinese Medicine Questionnaire" (CCMQ), and survey on syndrome type by CM syndrome scale (preliminary) were carried out in 180 DN patients. Cluster analysis on symptom items was used to determine the syndrome type, and canonical correlation analysis was used to analyze the relationship between patients' constitution and syndrome.

RESULTSBaseline levels in all enrolled patients were not different statistically. Cluster analysis showed 8 syndromes existed in DN patients, namely: I, qi-yin deficiency with qi-stagnancy type; II, yin-yang deficiency with heat-water-blood stasis type; III, qi-yin deficiency with dampness-heat type; IV, yin-yang deficiency with blood-stasis and heat type; V, qi-yin deficiency with stagnant heat type; VI, yin-yang deficiency with inner dampness-heat stagnancy type; VII, yin deficiency with heat stagnancy type; and VIII, Kidney (Shen)-Spleen (Pi) deficiency with stagnant heat type. Correlation analysis on the 8 syndromes and the 9 constitutions showed statistical significant correlations between syndrome III and dampness-heat constitution (P=0.0001); syndrome IV and blood-stasis constitution (P=0.0001); and syndrome VII and yin-deficiency constitution (P=0.0180).

CONCLUSIONCertain relationship revealed between CM constitutions and syndrome types; constitution decides the disease genesis, its syndrome type and prognosis, as well as the change of syndromes.

Aged ; Body Constitution ; Cluster Analysis ; Diabetic Nephropathies ; therapy ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Syndrome

AIMTo modify the structure of dehydroepiandrosterone (DHEA).

METHODSUsing hairy root cultures of Anisodus tanguticus to perform biotransformation of DHEA, using chromatographic and spectral techniques to isolate and identify the products.

RESULTS(1) The MS medium without plant hormone was suitable for the growth of the hairy root. (2) DHEA was converted into five products: androst-4-ene-3, 17-dione (I); 6alpha-hydroxyandrost-4-ene-3, 17-dione (II); 6alpha, 17beta-dihydroxyandrost-4-ene-3-one (III); androst-4-ene-3, 6, 17-trione (IV) and 17beta-hydroxyandrost-4-ene-3-one (V).

CONCLUSIONIt is the first time to use hairy root cultures of Anisodus tanguticus for the biotransformation of DHEA and five DHEA-related compounds were obtained.

Androstenedione ; chemistry ; isolation & purification ; Androstenes ; chemistry ; isolation & purification ; Biotransformation ; Culture Media ; Dehydroepiandrosterone ; chemistry ; metabolism ; Molecular Structure ; Plant Roots ; metabolism ; Plants, Medicinal ; metabolism ; Solanaceae ; metabolism ; Tissue Culture Techniques