Kashin-Beck disease(KBD)is a chronic,endemic,degenerative osteoarthropathic condition that is predominantly found in China's Mainland.KBD is a progressive disease,and the signs and symptoms become more severe in a...Kashin-Beck disease(KBD)is a chronic,endemic,degenerative osteoarthropathic condition that is predominantly found in China's Mainland.KBD is a progressive disease,and the signs and symptoms become more severe in adulthood owing to the continuous development of lesions in the joint cartilage[1].Although childhood KBD has largely been controlled,adult KBD is an important health concern in rural regions.Unfortunately,in the last century,many adult KBD patients still remain during serious epidemics.展开更多
Hepatic echinococcosis, also called echinococcosis, is a health-threatening disease commonly found in pasture, and belongs to parasitic zoonoses. The purpose of this study was to investigate the prevalence and the ris...Hepatic echinococcosis, also called echinococcosis, is a health-threatening disease commonly found in pasture, and belongs to parasitic zoonoses. The purpose of this study was to investigate the prevalence and the risk factors of echinococcosis in Qinghai province in order to provide fundamental data for prevention and control of echinococcosis in Qinghai province. A total of 23 445 people from 21 counties were enrolled in this study by multi-stage stratified random sampling. Echinococcosis was diagnosed by using B-mode ultrasonography and serological tests. The results showed that the prevalence of echinococcosis was 4.47%（95%CI： 4.21%–4.73%） and serum positive rate（seroprevalence） was 15.47%（95%CI： 14.92%–16.02%） in 2010. The distribution of echinococcosis differed in age, sex, ethnicity, occupation and regions in Qinghai（P0.05）. GLMM analysis revealed that gender（female vs. male）, ethnicity（Tibetan vs. other ethnicities）, profession（herders vs. other professions） and region（autonomous prefectures vs. cities） were significant risk factors for echinococcosis（P0.05）. It was concluded that the prevalence of echinococcosis in 2010 was about 4% in Qinghai province, and the distribution of echinococcosis in Qinghai was associated with age, sex, ethnicity and profession.展开更多
Kashin-Beck disease（KBD） is an endemic degenerative osteoarthropathy of uncertain etiology. The aim of our study was to identify changes in C-telopeptide of type Ⅱ collagen（CTX-Ⅱ）, pyridinoline（PYD）, and deoxy...Kashin-Beck disease（KBD） is an endemic degenerative osteoarthropathy of uncertain etiology. The aim of our study was to identify changes in C-telopeptide of type Ⅱ collagen（CTX-Ⅱ）, pyridinoline（PYD）, and deoxypyridinoline（DPD） among KBD patients. 54 KBD patients and 78 healthy controls were included this study. Urinary samples were collected and measured by ELISA. The median quantities of PYD, CTX-Ⅱ, and DPD of KBD patients were 1107.73 ng/μmol.cre, 695.11 ng/μmol.cre, and 1342.34 pml/μmol.cre, while the median quantities of healthy controls were 805.59 ng/μmol.cre, 546.47 ng/μmol.cre, and 718.15 pml/μmol.cre, respectively. The differences between KBD patients and healthy controls were statistically significant（Z = 4.405, 3.653, and 3.724; P 〈 0.001）. The higher levels of PYD, CTX-Ⅱ, and DPD detected in KBD patients indicate that they could be used as biomarkers of KBD.展开更多
Objective:To identify full length cDNA sequence of lactate dehydrogenase(LDH) from adult Echinococcus granulosus(E.granulosus) and to predict the structure and function of its encoding protein using bioinformatics met...Objective:To identify full length cDNA sequence of lactate dehydrogenase(LDH) from adult Echinococcus granulosus(E.granulosus) and to predict the structure and function of its encoding protein using bioinformatics methods.Methods:With the help of NCBI,EMBI, Expasy and other online sites,the open reading frame(ORF),conserved domain,physical and chemical parameters,signal peptide,epitope,topological structures of the protein sequences were predicted and a homology tertiary structure model was created:Vector NT1 software was used for sequence alignment,phylogenetic tree construction and tertiary structure prediction. Results:The target sequence was 1 233 bp length with a 996 bp biggest ORF encoding 331 amino acids protein with typical L-LDH conserved domain.It was confirmed as full length cDNA of LDH from E.granulosus and named as EgLDH(GenBank accession number:HM748917).The predicted molecular weight and isoelectric point of the deduced protein were 3 5516.2Da and 6.32 respectively.Compared with LDHs from Taenia solium,Taenia saginata asiatica,Spirometra erinaceieuropaei.Schistosoma japonicum,Clonorchis sinensis and human,it showed similarity of 86% ,85% ,55% ,58% ,58% and 53% ,respectively.EgLDH contained 3 putative transmembrane regions and 4 major epitopes(54aa-59aa.81aa-87aa,97aa-102aa,307aa-313aa),the latter were significant different from the corresponding regions of human LDH.In addition,some NAD and substrate binding sites located on epitopes 54aa-59aa and 97aa-102aa,respectively.Tertiary structure prediction showed that 3 key catalytic residues 105R,165D and 192H forming a catalytic center near the epitope 97aa-102aa,most NAD and substrate binding sites located around the center.Conclusions:The full length cDNA sequences of EgLDH were identified.It encoded a putative transmembrane protein which might be an ideal target molecule for vaccine and drugs.展开更多
Objective LcrV is an important component for the development of a subunit vaccine against plague.To reduce immunosuppressive activity of LcrV,a recombinant LcrV variant lacking amino acids 271 to 326 （rV270） was pre...Objective LcrV is an important component for the development of a subunit vaccine against plague.To reduce immunosuppressive activity of LcrV,a recombinant LcrV variant lacking amino acids 271 to 326 （rV270） was prepared by different methods in this study.Methods A new strategy that produced non-tagged or authentic rV270 protein was designed by insertion of rV270-thrombin-hexahistidine fusion gene into the vector pET24a,or by insertion of hexahistidine-enterokinase-rV270 or hexahistitine-factor Xa-rV270 fusion gene into the vector pET32a.After Co2＋ affinity chromatography,a purification strategy was developed by cleavage of His tag on column,following Sephacryl S-200HR column filtration chromatography.Results Removal of His tag by thrombin,enterokinase and factor Xa displayed a yield of 99.5%,32.4% and 15.3%,respectively.Following Sephacryl S-200HR column filtration chromatography,above 97% purity of rV270 protein was obtained.Purified rV270 that was adsorbed to 25% （v/v） Al（OH）3 adjuvant in phosphate-buffered saline （PBS） induced very high titers of antibody to rV270 in BALB/c mice and protected them （100% survival） against subcutaneous challenge with 106 CFU of Y.pestis virulent strain 141.Conclusion The completely authentic rV270 protein can be prepared by using enterokinase or factor Xa,but they exhibited extremely low cleavage activity to the corresponding recognition site.Thrombin cleavage is an efficient strategy to prepare non-tagged rV270 protein and can be easily operated in a large scale due to its relatively low cost and high cleavage efficacy.The recombinant rV270 can be used as a key component to develop a subunit vaccine of plague.展开更多
Objective To explore potential serum biomarkers of children with Kashin-Beck Disease(KBD)and the metabolic pathways to which the biomarkers belong.Methods A two-stage metabolomic study was employed.The discovery cohor...Objective To explore potential serum biomarkers of children with Kashin-Beck Disease(KBD)and the metabolic pathways to which the biomarkers belong.Methods A two-stage metabolomic study was employed.The discovery cohort included 56 patients,51 internal controls,and 50 external controls.The metabolites were determined by HPLC-(Q-TOF)-MS and confirmed by Human Metabolome Databases(HMDB)and Metlin databases.MetaboAnalyst 3.0 and the Kyoto Encyclopedia of Genes and Genomes(KEGG)database were used to analyze the metabolic pathways of the candidate metabolites.The use of HPLC-(Q-TRAP)-MS enabled quantitative detection of the target metabolites which were chosen using the discovery study and verified in another independent verification cohort of 31 patients,41 internal controls,and 50 external controls.Results Eight candidate metabolites were identified out in the discovery study,namely kynurenic acid,N-α-acetylarginine,6-hydroxymelatonin,sphinganine,ceramide,sphingosine-1 P,spermidine,and glycine.These metabolites exist in sphingolipid,glutathione,and tryptophan metabolic pathways.In the second-stage study,five candidate metabolites were validated,including kynurenic acid,N-α-acetylarginine,sphinganine,spermidine,and sphingosine-1 P.Except for spermidine,all substances exhibited low expression in the case group compared with the external control group,and the difference in levels of sphinganine,spermidine,and sphingosine-1 P was statistically significant.Conclusion The direction of change of levels of sphinganine,spermidine,and sphingosine-1 P in the two-stage study cohorts was completely consistent,and the differences were statistically significant.Therefore,these substances can be used as potential biomarkers of KBD.Furthermore,these results raise the possibility that sphingolipid metabolic pathways may be closely related to KBD.展开更多
Objective To investigate reciprocal regulation between Fur and two RyhB homologs in Yersinia pestis(Y.pestis),as well as the roles of RyhBs in biofilm formation.Methods Regulatory relationships were assessed by a comb...Objective To investigate reciprocal regulation between Fur and two RyhB homologs in Yersinia pestis(Y.pestis),as well as the roles of RyhBs in biofilm formation.Methods Regulatory relationships were assessed by a combination of colony morphology assay,primer extension,electrophoretic mobility shift assay and DNase I footprinting.Results Fur bound to the promoter-proximal DNA regions of ryhB1 and ryhB2 to repress their transcription,while both RyhB1 and RyhB2 repressed the expression of Fur at the post-transcriptional level.In addition,both RyhB1 and RyhB2 positively regulated Y.pestis biofilm exopolysaccharide(EPS)production and the expression of hmsHFRS and hmsT.Conclusion Fur and the two RyhB homologs exert negative reciprocal regulation,and RyhB homologs have a positive regulatory effect on biofilm formation in Y.pestis.展开更多
Brucellosis is one of the most widespread zoonotic diseases, with the most frequent complication being osteoarticular changes. The aim of this study was to assess the changes of C-terminal telopeptide of type II colla...Brucellosis is one of the most widespread zoonotic diseases, with the most frequent complication being osteoarticular changes. The aim of this study was to assess the changes of C-terminal telopeptide of type II collagen （CTX-II） in patients infected with brucellosis. A total of 84 brucellosis patients and 43 volunteers were selected and divided into brucellosis vs. control groups. Serum samples were subjected to serological tests for brucellosis,展开更多
Kaschin-Beck disease (KBD) is an endemic and chronic osteoarthropathy characterized by pathological aspects including chondrocyte degeneration, necrosis, progressive loss of articular cartilage, and secondary degenera...Kaschin-Beck disease (KBD) is an endemic and chronic osteoarthropathy characterized by pathological aspects including chondrocyte degeneration, necrosis, progressive loss of articular cartilage, and secondary degenerative osteoarthrosis of epiphyseal cartilage, epiphyseal plate cartilage, and articular cartilage, during puberty[1]. The main clinical symptoms are limb joint pain, thickening, deformation, limited movement, muscle atrophy, and in case of more severely affected patients, short fingers (toes), short limbs, and even short stature[1].展开更多
Fluorosis is a serious health problem,and an endemic disease in certain areas in China,India,and some countries in Africa[1].There are three types of fluorosis which includes drinking-water type fluorosis,coal-burning...Fluorosis is a serious health problem,and an endemic disease in certain areas in China,India,and some countries in Africa[1].There are three types of fluorosis which includes drinking-water type fluorosis,coal-burning pollution type fluorosis,and brick tea-type fluorosis.Coal-burning pollution type fluorosis and brick tea-type fluorosis are found only in China's Mainland mainland.展开更多
Plague,caused by Yersinia pestis,is a natural focus infectious disease.In China,plague is classified as category A,with the highest risk and hazard among the infectious diseases.Qinghai used to be considered as one of...Plague,caused by Yersinia pestis,is a natural focus infectious disease.In China,plague is classified as category A,with the highest risk and hazard among the infectious diseases.Qinghai used to be considered as one of the most serious areas of plague in China.In recent years,thank to the measures in eight aspects summarized as the“Qinghai model”which were adopted to prevent and control the human plague in Qinghai,Qinghai has not experienced any plague case reported for eight years.In early 2020,coronavirus disease 2019(COVID-19)outbroke in China.The Qinghai model on plague was employed to deal with the COVID-19 emergency in Qinghai Province.The Qinghai Center for Disease Control and Prevention(Qinghai CDC)and hospitals,along with the departments of public security,animal husbandry and other departments,quickly tracked and treated the patients with severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)and started surveillance programs on close contacts timely.At present,the cure rate of patients has reached 100%,and close contacts have been effectively quarantined and tested to avoid the spread of COVID-19.The findings from the study suggest that the prevention and control measures undertaken in Qinghai Province might be effective in dealing with the category A infectious diseases such as COVID-19 and other diseases.展开更多
Background:As part of an ongoing program that aims to use early detection and timely treatment to improve the control of echinococcosis,especially in younger age groups,we undertook a series of active surveys among Qi...Background:As part of an ongoing program that aims to use early detection and timely treatment to improve the control of echinococcosis,especially in younger age groups,we undertook a series of active surveys among Qinghai-Tibetan children in the Qinghai Province of Northwestern China in 2011 and 2012.The significant outcomes that resulted from this study emphasize the need to draw attention to echinococcosis,both alveolar echinococcosis(AE)and cystic echinococcosis(CE),so that policy development is promoted and suitable avenues for control are identified in the highly endemic areas on the Tibetan Plateau.Methods:A total of 19629 primary school students,aged 6-18 years,with a dominant Tibetan background underwent abdominal ultrasound examination,and 86.4%of the compliant students donated 2-5 ml of venous blood for serological tests.All the abnormal ultrasound results were recorded.If identified as echinococcosis,the disease lesion was assessed according to the WHO-Informal Working Group on Echinococcosis(WHO-IWGE)classification for AE and CE.Among the surveyed students,the prevalence by school was compared among geo-locations,sex and age groups.The clinical image presentations were analyzed according to lesion number,size,the location in the liver and the classification stage.Statistical significance was set at P-value<0.05 for comparisons among groups.Results:A total of 341 students(1.7%)were identified by ultrasound as having either CE(119,0.6%)or AE(222,1.1%).The highest prevalence rates of childhood AE cases occurred in the Tehetu(12.1%)and Moba(11.8%)townships in Dari County.There was a high seropositive rate(37.0%)and a heterogeneous distribution of cases,with a prevalence ranged from 0 to 12.1%for AE and 0-2.9%for CE.Moreover,the seropositive rate ranged from 0.7-45.1%across different schools.Conclusions:The high prevalence of echinococcosis in Qinghai-Tibetan primary school students reflects a lack of knowledge about Echinococcus spp.transmission.The combination of systematic education for children and regularly performed anthelmintic treatment for dogs could achieve the goal of sustainable hydatidosis control.展开更多
The present study is to determine the suitable protoscolices （PSCs） density for long-time culturing in vitro. The PSCs were divided into eight groups with different densities and the viability tests were carried out...The present study is to determine the suitable protoscolices （PSCs） density for long-time culturing in vitro. The PSCs were divided into eight groups with different densities and the viability tests were carried out with 0.1% methylene blue staining. Then the infection ability of cultured PSCs was assessed by the mean cyst weight of mice inoculated intraperitoneally with PSCs after 8 months post-infection.展开更多
Background:The Qinghai-Tibet Plateau(QTP)of China is an extensive pastoral and semi-pastoral area,and because of poverty and bad hygiene conditions,Brucella is highly prevalent in this region.In order to adequately pr...Background:The Qinghai-Tibet Plateau(QTP)of China is an extensive pastoral and semi-pastoral area,and because of poverty and bad hygiene conditions,Brucella is highly prevalent in this region.In order to adequately prevent this disease in the QTP region it is important to determine the identity of Brucella species that caused the infection.Methods:A total of 65 Brucella isolates were obtained from human,livestock and wild animals in Qinghai,a Chinese province in east of the QTP.Two molecular typing methods,MLVA(multi-locus variable-number tandem-repeat analysis)and MLST(multi locus sequence typing)were used to identify the species and genotypes of these isolates.Findings:Both MLVA and MLST typing methods classified the 65 isolates into three species,B.melitensis,B.abortus and B.suis,which included 60,4 and 1 isolates respectively.The MLVA method uniquely detected 34(Bm01~Bm34),3(Ba01~Ba03),and 1(Bs01)MLVA-16 genotypes for B.melitensis,B.abortus and B.suis,respectively.However,none of these genotypes exactly matched any of the genotypes in the Brucella2012 MLVA database.The MLST method identified five known ST types:ST7 and ST8(B.melitensis),ST2 and ST5(B.abortus),and ST14(B.suis).We also detected a strain with a mutant type(3-2-3-2-?-5-3-8-2)of ST8(3-2-3-2-1-5-3-8-2).Extensive genotype-sharing events could be observed among isolates from different host species.Conclusions:There were at least three Brucella(B.melitensis,B.abortus and B.suis)species in Qinghai,of which B.melitensis was the predominant species in the area examined.The Brucella population in Qinghai was very different from other regions of the world,possibly owing to the unique geographical characteristics such as extremely high altitude in QTP.There were extensive genotype-sharing events between isolates obtained from humans and other animals.Yaks,sheep and blue sheep were important zoonotic reservoirs of brucellosis causing species found in humans.展开更多
Background:Echinococcus multilocularis causes alveolar echinococcosis(AE)and is widely prevalent in Qinghai Province,China,where a number of different species have been identified as hosts.However,limited information ...Background:Echinococcus multilocularis causes alveolar echinococcosis(AE)and is widely prevalent in Qinghai Province,China,where a number of different species have been identified as hosts.However,limited information is available on the Qinghai vole(Lasiopodomys fuscus),which is hyper endemic to Qinghai Province and may represent a potential intermediate host of E.multilocularis.Thus,L.fuscus could contribute to the endemicity of AE in the area.Methods:Fifty Qinghai voles were captured from Jigzhi County in Qinghai Province for the clinical identification of E.multilocularis infection via anatomical examination.Hydatid fluid was collected from vesicles of the livers in suspected voles and subjected to a microscopic examination and PCR assay based on the barcoding gene of cox 1.PCR-amplified segments were sequenced for a phylogenetic analysis.E.multilocularis-infected Qinghai voles were morphologically identified and subjected to a phylogenetic analysis to confirm their identities.Results:Seventeen of the 50 Qinghai voles had E.multilocularis-infection-like vesicles in their livers.Eleven out of the 17 Qinghai voles presented E.multilocularis infection,which was detected by PCR and sequencing.The phylogenetic analysis showed that all 11 positive samples belonged to the E.multilocularis Asian genotype.A morphological identification and phylogenetic analysis of the E.multilocularis-infected Qinghai voles confirmed that all captured animals were L.fuscus.Conclusions:L.fuscus can be infected with E.multilocularis and plays a potential role in the life cycle and epidemiology of E.multilocularis in the Qinghai-Tibetan Plateau of China.展开更多
The molecular pathogenesis of T-2 toxin-induced cartilage destruction has not been fully unraveled yet. The aim of this study was to detect changes in serum metabolites in a rat anomaly model with articular cartilage ...The molecular pathogenesis of T-2 toxin-induced cartilage destruction has not been fully unraveled yet. The aim of this study was to detect changes in serum metabolites in a rat anomaly model with articular cartilage destruction. Thirty healthy male Wistar rats were fed a diet containing T-2 toxin （300 ng/kg chow） for 3 months. Histopathological changes in femorotibial cartilage were characterized in terms of chondrocyte degeneration/necrosis and superficial cartilage defect, and the endogenous metabolite profile of serum was determined by UPLC/Q-TOF MS. Treated rats showed extensive areas of chondrocyte necrosis and superficial cartilage defect in the articular cartilage. In addition, 8 metabolites were found to change significantly in these rats compared to the control group, including lyso PE （18：0/0：0）, lyso PC（14：0）, lyso PC[18：4 （6Z,9Z,12Z,15Z）], lyso PC[（16：1（9Z）], lyso PC（16：0）, L-valine, hippuric acid, and asparaginyl-glycine. These 8 metabolites associated with cartilage injury are mainly involved in phospholipid and amino acid metabolic pathways.展开更多
Cestode larvae spend one phase of their two-phase life cycle in the viscera of rodents, but cases of cestodes infecting subterranean rodents have only been rarely observed. To experimentally gain some insight into thi...Cestode larvae spend one phase of their two-phase life cycle in the viscera of rodents, but cases of cestodes infecting subterranean rodents have only been rarely observed. To experimentally gain some insight into this phenomenon, we captured approximately 300 plateau zokors(Eospalax baileyi), a typical subterranean rodent inhabiting the Qinghai-Tibet Plateau, and examined their livers for the presence of cysts. Totally, we collected five cysts, and using a mitochondrial gene(cox1) and two nuclear genes(pepck and pold) as genetic markers, we were able to analyze the taxonomy of the cysts. Both the maximum likelihood and Bayesian methods showed that the cysts share a monophyly with Taenia mustelae, while Kimura 2-parameter distances and number of different sites between our sequences and T. mustelae were far less than those found between the examined sequences and other Taeniidae species. These results, alongside supporting paraffin section histology, imply that the cysts found in plateau zokors can be regarded as larvae of T. mustelae, illustrating that zokors are a newly discovered intermediate host record of this parasite.展开更多
Background:The prevalence of human brucellosis in Qinghai Province of China has been increasing rapidly,with confirmed cases distributed across 31 counties.However,the epidemiology of brucellosis transmission has not ...Background:The prevalence of human brucellosis in Qinghai Province of China has been increasing rapidly,with confirmed cases distributed across 31 counties.However,the epidemiology of brucellosis transmission has not been fully elucidated.To characterize the infecting strains isolated from humans,multiple-locus variable-number tandem repeats analysis(MLVA)and whole-genome single-nucleotide polymorphism(SNP)-based approaches were employed.Methods:Strains were isolated from two males blood cultures that were confirmed Brucella m elitensis positive following biotyping and MLVA.Genomic DNA was extracted from these two strains,and whole-genome sequencing was performed.Next,SNP-based phylogenetic analysis was performed to compare the two strains to 94 B.melitensis strains(complete genome and draft genome)retrieved from online databases.Results:The two Brucella isolates were identified as B.melitensis biovar 3(QH2019001 and QH2019005)following conventional biotyping and were found to have differences in their variable number tandem repeats(VNTRs)using MLVA-16.Phylogenetic examination assigned the 96 strains to five genotype groups,with QH2019001 and QH2019005 assigned to the same group,but different subgroups.Moreover,the QH2019005 strain was assigned to a new subgenotype,llj,within genotype II.These findings were then combined to determine the geographic origin of the two Brucella strains.Conclusions:Utilizing a whole-genome SNP-based approach enabled differences between the two B.m elitensis strains to be more clearly resolved,and facilitated the elucidation of their different evolutionary histories.This approach also revealed that QH2019005 is a member of a new subgenotype(Hj)with an ancient origin in the eastern Mediterranean Sea.展开更多
Objective:This study aims to explore the association between the density of Himalayan marmot (Marmota himalayana) and climatic factors such as temperature, precipitation, humidity, vapour pressure, sunshine percentage...Objective:This study aims to explore the association between the density of Himalayan marmot (Marmota himalayana) and climatic factors such as temperature, precipitation, humidity, vapour pressure, sunshine percentage, wind velocity, which are closely associated with global climate change, and to provide a reference for plague prevention and control. Methods: We conducted a regression analysis to find the possible climate factors associated with the density of Himalaya marmot, and analyzed the response characters of Himalayan marmot to climate change.Results: Dailyprecipitation days(>=0.1 mm) and sunshine percentage were significantly associated with thedensityofHimalayan marmot(p<0.01). Conclusion: Climate change was associated with the risk of plague. This phenomenon is valuable for Himalayan marmot and plague prevention. More studies are needed to understand the impact of climate change on Himalayan marmot and plague.展开更多
Bacterial pathogens have evolved various mechanisms to modulate host immune responses for successful infection. In this study, RNA- sequencing technology was used to analyze the responses of human monocytes THP1 to Ye...Bacterial pathogens have evolved various mechanisms to modulate host immune responses for successful infection. In this study, RNA- sequencing technology was used to analyze the responses of human monocytes THP1 to Yersinia pestis infection. Over 6000 genes were differentially expressed over the 12 h infection. Kinetic responses of pathogen recognition receptor signaling pathways, apoptosis, antigen processing, and presentation pathway and coagulation system were analyzed in detail. Among them, RIG-I-like receptor （RLR） signaling pathway, which was established for antiviral defense, was significantly affected. Mice lacking MAVS, the adaptor of the RLR signaling pathway, were less sensitive to infection and exhibited lower IFN-13 production, higher Thl-type cytokines IFN-γ and IL-12 production, and lower Th2-type cytokines IL-4 and IL-13 production in the serum compared with wild-type mice. Moreover, infection of pathogenic bacteria other than E pestis also altered the expression of the RLR pathway, suggesting that the response of RLR pathway to bacterial infection is a universal mechanism.展开更多
基金supported by basic research projects of Qinghai province[2017-ZJ-770]projects of Qinghai province health and family planning commission[2017-wjzdx-38]+1 种基金the 2020"Kunlun TalentsHigh-tech Innovation and Entrepreneurial Talents"project to cultivate leading talents of Qinghai province。
文摘Kashin-Beck disease(KBD)is a chronic,endemic,degenerative osteoarthropathic condition that is predominantly found in China's Mainland.KBD is a progressive disease,and the signs and symptoms become more severe in adulthood owing to the continuous development of lesions in the joint cartilage[1].Although childhood KBD has largely been controlled,adult KBD is an important health concern in rural regions.Unfortunately,in the last century,many adult KBD patients still remain during serious epidemics.
文摘Hepatic echinococcosis, also called echinococcosis, is a health-threatening disease commonly found in pasture, and belongs to parasitic zoonoses. The purpose of this study was to investigate the prevalence and the risk factors of echinococcosis in Qinghai province in order to provide fundamental data for prevention and control of echinococcosis in Qinghai province. A total of 23 445 people from 21 counties were enrolled in this study by multi-stage stratified random sampling. Echinococcosis was diagnosed by using B-mode ultrasonography and serological tests. The results showed that the prevalence of echinococcosis was 4.47%（95%CI： 4.21%–4.73%） and serum positive rate（seroprevalence） was 15.47%（95%CI： 14.92%–16.02%） in 2010. The distribution of echinococcosis differed in age, sex, ethnicity, occupation and regions in Qinghai（P0.05）. GLMM analysis revealed that gender（female vs. male）, ethnicity（Tibetan vs. other ethnicities）, profession（herders vs. other professions） and region（autonomous prefectures vs. cities） were significant risk factors for echinococcosis（P0.05）. It was concluded that the prevalence of echinococcosis in 2010 was about 4% in Qinghai province, and the distribution of echinococcosis in Qinghai was associated with age, sex, ethnicity and profession.
基金supported by the Basic Research Projects of Science and Technology in Qinghai Province(2017-ZJ-770)
文摘Kashin-Beck disease（KBD） is an endemic degenerative osteoarthropathy of uncertain etiology. The aim of our study was to identify changes in C-telopeptide of type Ⅱ collagen（CTX-Ⅱ）, pyridinoline（PYD）, and deoxypyridinoline（DPD） among KBD patients. 54 KBD patients and 78 healthy controls were included this study. Urinary samples were collected and measured by ELISA. The median quantities of PYD, CTX-Ⅱ, and DPD of KBD patients were 1107.73 ng/μmol.cre, 695.11 ng/μmol.cre, and 1342.34 pml/μmol.cre, while the median quantities of healthy controls were 805.59 ng/μmol.cre, 546.47 ng/μmol.cre, and 718.15 pml/μmol.cre, respectively. The differences between KBD patients and healthy controls were statistically significant（Z = 4.405, 3.653, and 3.724; P 〈 0.001）. The higher levels of PYD, CTX-Ⅱ, and DPD detected in KBD patients indicate that they could be used as biomarkers of KBD.
基金Supported by National Nature Science Foundation of China(No:30860070)
文摘Objective:To identify full length cDNA sequence of lactate dehydrogenase(LDH) from adult Echinococcus granulosus(E.granulosus) and to predict the structure and function of its encoding protein using bioinformatics methods.Methods:With the help of NCBI,EMBI, Expasy and other online sites,the open reading frame(ORF),conserved domain,physical and chemical parameters,signal peptide,epitope,topological structures of the protein sequences were predicted and a homology tertiary structure model was created:Vector NT1 software was used for sequence alignment,phylogenetic tree construction and tertiary structure prediction. Results:The target sequence was 1 233 bp length with a 996 bp biggest ORF encoding 331 amino acids protein with typical L-LDH conserved domain.It was confirmed as full length cDNA of LDH from E.granulosus and named as EgLDH(GenBank accession number:HM748917).The predicted molecular weight and isoelectric point of the deduced protein were 3 5516.2Da and 6.32 respectively.Compared with LDHs from Taenia solium,Taenia saginata asiatica,Spirometra erinaceieuropaei.Schistosoma japonicum,Clonorchis sinensis and human,it showed similarity of 86% ,85% ,55% ,58% ,58% and 53% ,respectively.EgLDH contained 3 putative transmembrane regions and 4 major epitopes(54aa-59aa.81aa-87aa,97aa-102aa,307aa-313aa),the latter were significant different from the corresponding regions of human LDH.In addition,some NAD and substrate binding sites located on epitopes 54aa-59aa and 97aa-102aa,respectively.Tertiary structure prediction showed that 3 key catalytic residues 105R,165D and 192H forming a catalytic center near the epitope 97aa-102aa,most NAD and substrate binding sites located around the center.Conclusions:The full length cDNA sequences of EgLDH were identified.It encoded a putative transmembrane protein which might be an ideal target molecule for vaccine and drugs.
基金supported by the National Key Program for Infectious Diseases of China (2009ZX10004-4001)
文摘Objective LcrV is an important component for the development of a subunit vaccine against plague.To reduce immunosuppressive activity of LcrV,a recombinant LcrV variant lacking amino acids 271 to 326 （rV270） was prepared by different methods in this study.Methods A new strategy that produced non-tagged or authentic rV270 protein was designed by insertion of rV270-thrombin-hexahistidine fusion gene into the vector pET24a,or by insertion of hexahistidine-enterokinase-rV270 or hexahistitine-factor Xa-rV270 fusion gene into the vector pET32a.After Co2＋ affinity chromatography,a purification strategy was developed by cleavage of His tag on column,following Sephacryl S-200HR column filtration chromatography.Results Removal of His tag by thrombin,enterokinase and factor Xa displayed a yield of 99.5%,32.4% and 15.3%,respectively.Following Sephacryl S-200HR column filtration chromatography,above 97% purity of rV270 protein was obtained.Purified rV270 that was adsorbed to 25% （v/v） Al（OH）3 adjuvant in phosphate-buffered saline （PBS） induced very high titers of antibody to rV270 in BALB/c mice and protected them （100% survival） against subcutaneous challenge with 106 CFU of Y.pestis virulent strain 141.Conclusion The completely authentic rV270 protein can be prepared by using enterokinase or factor Xa,but they exhibited extremely low cleavage activity to the corresponding recognition site.Thrombin cleavage is an efficient strategy to prepare non-tagged rV270 protein and can be easily operated in a large scale due to its relatively low cost and high cleavage efficacy.The recombinant rV270 can be used as a key component to develop a subunit vaccine of plague.
基金supported by the National Natural Science Foundation[NO.81372937]。
文摘Objective To explore potential serum biomarkers of children with Kashin-Beck Disease(KBD)and the metabolic pathways to which the biomarkers belong.Methods A two-stage metabolomic study was employed.The discovery cohort included 56 patients,51 internal controls,and 50 external controls.The metabolites were determined by HPLC-(Q-TOF)-MS and confirmed by Human Metabolome Databases(HMDB)and Metlin databases.MetaboAnalyst 3.0 and the Kyoto Encyclopedia of Genes and Genomes(KEGG)database were used to analyze the metabolic pathways of the candidate metabolites.The use of HPLC-(Q-TRAP)-MS enabled quantitative detection of the target metabolites which were chosen using the discovery study and verified in another independent verification cohort of 31 patients,41 internal controls,and 50 external controls.Results Eight candidate metabolites were identified out in the discovery study,namely kynurenic acid,N-α-acetylarginine,6-hydroxymelatonin,sphinganine,ceramide,sphingosine-1 P,spermidine,and glycine.These metabolites exist in sphingolipid,glutathione,and tryptophan metabolic pathways.In the second-stage study,five candidate metabolites were validated,including kynurenic acid,N-α-acetylarginine,sphinganine,spermidine,and sphingosine-1 P.Except for spermidine,all substances exhibited low expression in the case group compared with the external control group,and the difference in levels of sphinganine,spermidine,and sphingosine-1 P was statistically significant.Conclusion The direction of change of levels of sphinganine,spermidine,and sphingosine-1 P in the two-stage study cohorts was completely consistent,and the differences were statistically significant.Therefore,these substances can be used as potential biomarkers of KBD.Furthermore,these results raise the possibility that sphingolipid metabolic pathways may be closely related to KBD.
基金supported by the Basic Application Research Project of Science and Technology Department of Qinghai province[2020-ZJ-788]the Key Laboratory of National Health Commission on Plague Control and Prevention[2019PT310004]+1 种基金the Key Laboratory for Plague Prevention and Control of Qinghai Province[p2020-ZJ-Y23]the Qinghai Province High-end Innovative Talents Thousand Talents Program(2019)。
文摘Objective To investigate reciprocal regulation between Fur and two RyhB homologs in Yersinia pestis(Y.pestis),as well as the roles of RyhBs in biofilm formation.Methods Regulatory relationships were assessed by a combination of colony morphology assay,primer extension,electrophoretic mobility shift assay and DNase I footprinting.Results Fur bound to the promoter-proximal DNA regions of ryhB1 and ryhB2 to repress their transcription,while both RyhB1 and RyhB2 repressed the expression of Fur at the post-transcriptional level.In addition,both RyhB1 and RyhB2 positively regulated Y.pestis biofilm exopolysaccharide(EPS)production and the expression of hmsHFRS and hmsT.Conclusion Fur and the two RyhB homologs exert negative reciprocal regulation,and RyhB homologs have a positive regulatory effect on biofilm formation in Y.pestis.
基金supported by the National Natural Scientific Foundation of China(30972556)
文摘Brucellosis is one of the most widespread zoonotic diseases, with the most frequent complication being osteoarticular changes. The aim of this study was to assess the changes of C-terminal telopeptide of type II collagen （CTX-II） in patients infected with brucellosis. A total of 84 brucellosis patients and 43 volunteers were selected and divided into brucellosis vs. control groups. Serum samples were subjected to serological tests for brucellosis,
基金supported by the National Natural Science Foundation of China [Grant number:81372937]
文摘Kaschin-Beck disease (KBD) is an endemic and chronic osteoarthropathy characterized by pathological aspects including chondrocyte degeneration, necrosis, progressive loss of articular cartilage, and secondary degenerative osteoarthrosis of epiphyseal cartilage, epiphyseal plate cartilage, and articular cartilage, during puberty[1]. The main clinical symptoms are limb joint pain, thickening, deformation, limited movement, muscle atrophy, and in case of more severely affected patients, short fingers (toes), short limbs, and even short stature[1].
基金supported by the basic research projects of Qinghai province[2017-ZJ-770]。
文摘Fluorosis is a serious health problem,and an endemic disease in certain areas in China,India,and some countries in Africa[1].There are three types of fluorosis which includes drinking-water type fluorosis,coal-burning pollution type fluorosis,and brick tea-type fluorosis.Coal-burning pollution type fluorosis and brick tea-type fluorosis are found only in China's Mainland mainland.
基金funded by the Key Laboratory of National Health Com-mission Project(2019PT310004)the Key Laboratory for Plague Pre-vention and Control of Qinghai Province(2020-ZJ-Y23).
文摘Plague,caused by Yersinia pestis,is a natural focus infectious disease.In China,plague is classified as category A,with the highest risk and hazard among the infectious diseases.Qinghai used to be considered as one of the most serious areas of plague in China.In recent years,thank to the measures in eight aspects summarized as the“Qinghai model”which were adopted to prevent and control the human plague in Qinghai,Qinghai has not experienced any plague case reported for eight years.In early 2020,coronavirus disease 2019(COVID-19)outbroke in China.The Qinghai model on plague was employed to deal with the COVID-19 emergency in Qinghai Province.The Qinghai Center for Disease Control and Prevention(Qinghai CDC)and hospitals,along with the departments of public security,animal husbandry and other departments,quickly tracked and treated the patients with severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)and started surveillance programs on close contacts timely.At present,the cure rate of patients has reached 100%,and close contacts have been effectively quarantined and tested to avoid the spread of COVID-19.The findings from the study suggest that the prevention and control measures undertaken in Qinghai Province might be effective in dealing with the category A infectious diseases such as COVID-19 and other diseases.
基金Our research was supported by the Natural Science Foundation of Qinghai,China for this project(No.2015-HZ-809).
文摘Background:As part of an ongoing program that aims to use early detection and timely treatment to improve the control of echinococcosis,especially in younger age groups,we undertook a series of active surveys among Qinghai-Tibetan children in the Qinghai Province of Northwestern China in 2011 and 2012.The significant outcomes that resulted from this study emphasize the need to draw attention to echinococcosis,both alveolar echinococcosis(AE)and cystic echinococcosis(CE),so that policy development is promoted and suitable avenues for control are identified in the highly endemic areas on the Tibetan Plateau.Methods:A total of 19629 primary school students,aged 6-18 years,with a dominant Tibetan background underwent abdominal ultrasound examination,and 86.4%of the compliant students donated 2-5 ml of venous blood for serological tests.All the abnormal ultrasound results were recorded.If identified as echinococcosis,the disease lesion was assessed according to the WHO-Informal Working Group on Echinococcosis(WHO-IWGE)classification for AE and CE.Among the surveyed students,the prevalence by school was compared among geo-locations,sex and age groups.The clinical image presentations were analyzed according to lesion number,size,the location in the liver and the classification stage.Statistical significance was set at P-value<0.05 for comparisons among groups.Results:A total of 341 students(1.7%)were identified by ultrasound as having either CE(119,0.6%)or AE(222,1.1%).The highest prevalence rates of childhood AE cases occurred in the Tehetu(12.1%)and Moba(11.8%)townships in Dari County.There was a high seropositive rate(37.0%)and a heterogeneous distribution of cases,with a prevalence ranged from 0 to 12.1%for AE and 0-2.9%for CE.Moreover,the seropositive rate ranged from 0.7-45.1%across different schools.Conclusions:The high prevalence of echinococcosis in Qinghai-Tibetan primary school students reflects a lack of knowledge about Echinococcus spp.transmission.The combination of systematic education for children and regularly performed anthelmintic treatment for dogs could achieve the goal of sustainable hydatidosis control.
基金supported by International Collaboration on Drug and Diagnostics Innovation of Tropical Diseases in China (International S&T Cooperation 2010DFB73280)Technology Development Research for Science Research Institute (2011EG150312)the Youth Project of Shanghai Municipal Health Bureau (87)
文摘The present study is to determine the suitable protoscolices （PSCs） density for long-time culturing in vitro. The PSCs were divided into eight groups with different densities and the viability tests were carried out with 0.1% methylene blue staining. Then the infection ability of cultured PSCs was assessed by the mean cyst weight of mice inoculated intraperitoneally with PSCs after 8 months post-infection.
基金supported by the Science and Technology Plan of Qinghai Province(NO.2013-Z-749)the Youth Innovation Promotion Association of Chinese Academy of Sciences(NO.2015352).
文摘Background:The Qinghai-Tibet Plateau(QTP)of China is an extensive pastoral and semi-pastoral area,and because of poverty and bad hygiene conditions,Brucella is highly prevalent in this region.In order to adequately prevent this disease in the QTP region it is important to determine the identity of Brucella species that caused the infection.Methods:A total of 65 Brucella isolates were obtained from human,livestock and wild animals in Qinghai,a Chinese province in east of the QTP.Two molecular typing methods,MLVA(multi-locus variable-number tandem-repeat analysis)and MLST(multi locus sequence typing)were used to identify the species and genotypes of these isolates.Findings:Both MLVA and MLST typing methods classified the 65 isolates into three species,B.melitensis,B.abortus and B.suis,which included 60,4 and 1 isolates respectively.The MLVA method uniquely detected 34(Bm01~Bm34),3(Ba01~Ba03),and 1(Bs01)MLVA-16 genotypes for B.melitensis,B.abortus and B.suis,respectively.However,none of these genotypes exactly matched any of the genotypes in the Brucella2012 MLVA database.The MLST method identified five known ST types:ST7 and ST8(B.melitensis),ST2 and ST5(B.abortus),and ST14(B.suis).We also detected a strain with a mutant type(3-2-3-2-?-5-3-8-2)of ST8(3-2-3-2-1-5-3-8-2).Extensive genotype-sharing events could be observed among isolates from different host species.Conclusions:There were at least three Brucella(B.melitensis,B.abortus and B.suis)species in Qinghai,of which B.melitensis was the predominant species in the area examined.The Brucella population in Qinghai was very different from other regions of the world,possibly owing to the unique geographical characteristics such as extremely high altitude in QTP.There were extensive genotype-sharing events between isolates obtained from humans and other animals.Yaks,sheep and blue sheep were important zoonotic reservoirs of brucellosis causing species found in humans.
基金Our research was supported by the“Basic Research Project of the Qinghai Science and Technology Department”(2016-ZJ-791)the“International Cooperation Project of the Qinghai Science and Technology Department”(2015-HZ-809).
文摘Background:Echinococcus multilocularis causes alveolar echinococcosis(AE)and is widely prevalent in Qinghai Province,China,where a number of different species have been identified as hosts.However,limited information is available on the Qinghai vole(Lasiopodomys fuscus),which is hyper endemic to Qinghai Province and may represent a potential intermediate host of E.multilocularis.Thus,L.fuscus could contribute to the endemicity of AE in the area.Methods:Fifty Qinghai voles were captured from Jigzhi County in Qinghai Province for the clinical identification of E.multilocularis infection via anatomical examination.Hydatid fluid was collected from vesicles of the livers in suspected voles and subjected to a microscopic examination and PCR assay based on the barcoding gene of cox 1.PCR-amplified segments were sequenced for a phylogenetic analysis.E.multilocularis-infected Qinghai voles were morphologically identified and subjected to a phylogenetic analysis to confirm their identities.Results:Seventeen of the 50 Qinghai voles had E.multilocularis-infection-like vesicles in their livers.Eleven out of the 17 Qinghai voles presented E.multilocularis infection,which was detected by PCR and sequencing.The phylogenetic analysis showed that all 11 positive samples belonged to the E.multilocularis Asian genotype.A morphological identification and phylogenetic analysis of the E.multilocularis-infected Qinghai voles confirmed that all captured animals were L.fuscus.Conclusions:L.fuscus can be infected with E.multilocularis and plays a potential role in the life cycle and epidemiology of E.multilocularis in the Qinghai-Tibetan Plateau of China.
基金financially supported by the National Natural Science Foundation of China[No.81372937]
文摘The molecular pathogenesis of T-2 toxin-induced cartilage destruction has not been fully unraveled yet. The aim of this study was to detect changes in serum metabolites in a rat anomaly model with articular cartilage destruction. Thirty healthy male Wistar rats were fed a diet containing T-2 toxin （300 ng/kg chow） for 3 months. Histopathological changes in femorotibial cartilage were characterized in terms of chondrocyte degeneration/necrosis and superficial cartilage defect, and the endogenous metabolite profile of serum was determined by UPLC/Q-TOF MS. Treated rats showed extensive areas of chondrocyte necrosis and superficial cartilage defect in the articular cartilage. In addition, 8 metabolites were found to change significantly in these rats compared to the control group, including lyso PE （18：0/0：0）, lyso PC（14：0）, lyso PC[18：4 （6Z,9Z,12Z,15Z）], lyso PC[（16：1（9Z）], lyso PC（16：0）, L-valine, hippuric acid, and asparaginyl-glycine. These 8 metabolites associated with cartilage injury are mainly involved in phospholipid and amino acid metabolic pathways.
基金supported by the West Light Foundation of the Chinese Academy of Sciences and the Chinese Academy of Sciences President Scholarship(to G.Lin)
文摘Cestode larvae spend one phase of their two-phase life cycle in the viscera of rodents, but cases of cestodes infecting subterranean rodents have only been rarely observed. To experimentally gain some insight into this phenomenon, we captured approximately 300 plateau zokors(Eospalax baileyi), a typical subterranean rodent inhabiting the Qinghai-Tibet Plateau, and examined their livers for the presence of cysts. Totally, we collected five cysts, and using a mitochondrial gene(cox1) and two nuclear genes(pepck and pold) as genetic markers, we were able to analyze the taxonomy of the cysts. Both the maximum likelihood and Bayesian methods showed that the cysts share a monophyly with Taenia mustelae, while Kimura 2-parameter distances and number of different sites between our sequences and T. mustelae were far less than those found between the examined sequences and other Taeniidae species. These results, alongside supporting paraffin section histology, imply that the cysts found in plateau zokors can be regarded as larvae of T. mustelae, illustrating that zokors are a newly discovered intermediate host record of this parasite.
基金supported by National Key R&D Program of China(Grant No.2020YFA0907101)Major Infectious Diseases such as AIDS and Viral Hepatitis Prevention and Control Technology Major Projects(2018ZX10201002)the National Natural Scientific Foundation of China(81860588).
文摘Background:The prevalence of human brucellosis in Qinghai Province of China has been increasing rapidly,with confirmed cases distributed across 31 counties.However,the epidemiology of brucellosis transmission has not been fully elucidated.To characterize the infecting strains isolated from humans,multiple-locus variable-number tandem repeats analysis(MLVA)and whole-genome single-nucleotide polymorphism(SNP)-based approaches were employed.Methods:Strains were isolated from two males blood cultures that were confirmed Brucella m elitensis positive following biotyping and MLVA.Genomic DNA was extracted from these two strains,and whole-genome sequencing was performed.Next,SNP-based phylogenetic analysis was performed to compare the two strains to 94 B.melitensis strains(complete genome and draft genome)retrieved from online databases.Results:The two Brucella isolates were identified as B.melitensis biovar 3(QH2019001 and QH2019005)following conventional biotyping and were found to have differences in their variable number tandem repeats(VNTRs)using MLVA-16.Phylogenetic examination assigned the 96 strains to five genotype groups,with QH2019001 and QH2019005 assigned to the same group,but different subgroups.Moreover,the QH2019005 strain was assigned to a new subgenotype,llj,within genotype II.These findings were then combined to determine the geographic origin of the two Brucella strains.Conclusions:Utilizing a whole-genome SNP-based approach enabled differences between the two B.m elitensis strains to be more clearly resolved,and facilitated the elucidation of their different evolutionary histories.This approach also revealed that QH2019005 is a member of a new subgenotype(Hj)with an ancient origin in the eastern Mediterranean Sea.
基金The Basic Application Project of Qinghai Provincegrant number:2013-Z-747
文摘Objective:This study aims to explore the association between the density of Himalayan marmot (Marmota himalayana) and climatic factors such as temperature, precipitation, humidity, vapour pressure, sunshine percentage, wind velocity, which are closely associated with global climate change, and to provide a reference for plague prevention and control. Methods: We conducted a regression analysis to find the possible climate factors associated with the density of Himalaya marmot, and analyzed the response characters of Himalayan marmot to climate change.Results: Dailyprecipitation days(>=0.1 mm) and sunshine percentage were significantly associated with thedensityofHimalayan marmot(p<0.01). Conclusion: Climate change was associated with the risk of plague. This phenomenon is valuable for Himalayan marmot and plague prevention. More studies are needed to understand the impact of climate change on Himalayan marmot and plague.
基金supported by the National Basic Research Program of China(Nos.2012CB518704 and 2013CB910804)the National Natural Science Foundation of China(No.31170122)the Basic Research Programs of Science and Technology Department Foundation of QingHai Province(No.2013-Z-748)
文摘Bacterial pathogens have evolved various mechanisms to modulate host immune responses for successful infection. In this study, RNA- sequencing technology was used to analyze the responses of human monocytes THP1 to Yersinia pestis infection. Over 6000 genes were differentially expressed over the 12 h infection. Kinetic responses of pathogen recognition receptor signaling pathways, apoptosis, antigen processing, and presentation pathway and coagulation system were analyzed in detail. Among them, RIG-I-like receptor （RLR） signaling pathway, which was established for antiviral defense, was significantly affected. Mice lacking MAVS, the adaptor of the RLR signaling pathway, were less sensitive to infection and exhibited lower IFN-13 production, higher Thl-type cytokines IFN-γ and IL-12 production, and lower Th2-type cytokines IL-4 and IL-13 production in the serum compared with wild-type mice. Moreover, infection of pathogenic bacteria other than E pestis also altered the expression of the RLR pathway, suggesting that the response of RLR pathway to bacterial infection is a universal mechanism.
