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Hemorrhagic fever with renal syndrome (HFRS) is a serious public health problem in Shandong Province, China. We conducted an epizootiologic investigation and phylogeographic and phylodynamic analyses to infer the phylogenetic relationships of hantaviruses in space and time, and gain further insights into their evolutionary dynamics in Shandong Province. Our data indicated that the Seoul virus (SEOV) is distributed throughout Shandong, whereas Hantaan virus (HTNV) co-circulates with SEOV in the eastern and southern areas of Shandong. Their distribution showed strong geographic clustering. In addition, our analyses indicated multiple evolutionary paths, long-distance transmission, and demographic expansion events for SEOV in some areas. Selection pressure analyses revealed that negative selection on hantaviruses acted as the principal evolutionary force, whereas a little evidence of positive selection exists. We found that several positively selected sites were located within major functional regions and indicated the importance of these residues for adaptive evolution of hantaviruses.

Hantavirus genome sequences were recovered from lung tissues of Chinese white-bellied rats (Niviventer confucianus) captured in Yunnan province, China. Pairwise comparison of the nucleotide and deduced amino acid sequences of the entire S and partial M and L segments indicated that the newly discovered virus strain, which was designated as strain YN509, was very different from other rodent-borne hantaviruses. Phylogenetic analysis showed that the new strain fit into a clade containing Da Bie Shan virus (DBSV) (also carried by N. confucianus), which is mainly found in Anhui Province in mainland China. Strain YN509 appears to be in a sister taxa of the DBSV group described previously. These data suggest that strain YN509 is a new subtype of DBSV, which appears to be widely distributed in China with a higher genetic diversity than expected.

Many rodent species may be involved in the enzootic maintenance of these agents., A total of 705 rodents from 6 provinces and autonomous regions of mainland People’s Republic of China were tested by PCRs for tick-borne agents (Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, spotted fever group rickettsiae, and Francisella tularensis). Infection rates were 5.5%, 6.7%, 9.1% and 5.0%, respectively. Eighteen (2.6%) rodents of 10 species were positive for 2 or 3 agents. Sequence analysis of PCR products confirmed the presence and genotypes of detected agents. These findings demonstrate that these tick-borne agents cocirculate and that a variety of rodent species may be involved in their enzootic maintenance.

A molecular epidemiological survey was conducted to investigate the presence of pathogenic Borrelia burgdorferi sensu lato (s.l.) species in the forest areas of Zhejiang province, south-east China. A total of 182 ticks of 6 species and 200 rodents of 8 species were collected and individually examined for the presence of B. burgdorferi s.l. DNA by nested PCR targeting the 5S-23S rRNA intergenic spacer. Forty-one ticks of four species, Haemaphysalis concinna, Haemaphysalis longicornis, Rhipicephalus microplus and Haemaphysalis warburconi, were infected with B. burgdorferi s.l., with an overall infection rate of 23 %. Sixteen rodents of four species, Nivivener confucianus, Nivivener coxingi, Apodemus sylvaticus and Rattus losea, were positive for B. burgdorferi s.l., with an overall prevalence of 8 %. MseI RFLP analysis and sequence analysis of the positive PCR products showed that Borrelia spirochaetes in specimens consisted of Borrelia garinii, Borrelia afzelii and Borrelia valaisiana-related group. Forty (98 %) of the B. burgdorferi s.l.-positive ticks were infected with B. garinii and one (2 %) was infected with B. afzelii. Twelve (75 %) of the positive rodents were infected with B. garinii and four (25 %) were infected with the Borrelia spirochaete belonging to B. valaisiana-related group.

Objective To investigate the expression of RECK mRNA and protein in esophageal squamous cell carcinoma (ESCC) and to examine its relationship with the clinicopathologic features.

Objective To compare the differences of mitochondrial DNA (mtDNA) copies among the tissues of esophageal squamous cell carcinoma (ESCC), para-neoplastic tissue and normal mucous membrane of the esophagus, and to study the relationship between the mtDNA and the occurrence and development of esophageal squamous cell carcinoma.

Viral adaptation to the host may be occurring under selective immune pressure., Single nucleotide variations (SNVs) at 5 loci (17564, 21721, 22222, 23823, and 27827) were used to define the molecular epidemiologic characteristics of severe acute respiratory syndrome–associated coronavirus (SARS-CoV) from Beijing patients. Five fragments targeted at the SNV loci were amplified directly from clinical samples by using reverse transcription–polymerase chain reaction (RT-PCR), before sequencing the amplified products. Analyses of 45 sequences obtained from 29 patients showed that the GGCTC motif dominated among samples collected from March to early April 2003; the TGTTT motif predominanted afterwards. The switch from GGCTC to TGTTT was observed among patients belonging to the same cluster, which ruled out the possibility of the coincidental superposition of 2 epidemics running in parallel in Beijing. The Beijing isolates underwent the same change pattern reported from Guangdong Province. The same series of mutations occurring in separate geographic locations and at different times suggests a dominant process of viral adaptation to the host.

A total of 1,345 Ixodes persulcatus ticks collected from northeastern China were investigated for the presence of Anaplasma phagocytophila and Borrelia burgdorferi by a nested polymerase chain reaction (PCR). Sixty-two (4.6%) ticks were positive for A. phagocytophila and 454 (33.8%) were positive for B. burgdorferi. Seven (0.5%) were coinfected with both agents. Sequence analysis of 919-basepair PCR amplicons revealed three types of A. phagocytophila. Type 1 was identical to the published sequences of A. phagocytophilas responsible for human granulocytic ehrlichiosis (HGE). The other two variants differed from the HGE agent sequence at one and four positions, respectively. These findings imply that infection with A. phagocytophila poses a potential health threat to both humans and animals in northeastern China, and that ehrlichiosis should be considered in the differential diagnosis of febrile patients with a history of tick bite, particularly when clinical manifestations are atypical for Lyme disease.

Inspired by the recent discovery of genetically distinct hantaviruses from insectivore species worldwide, we performed a small-scale search for insectivore-borne hantaviruses. In this paper, we report the discovery of a new hantavirus, which was designated the Qian Hu Shan virus (QHSV). This virus was detected in the lung tissues of three stripe-backed shrews (Sorex cylindricauda), which were captured in the Yunnan Province, China. The full-length S genomic segment of the representative QHSV strain YN05-284 was 1661 nucleotides and is predicted to encode a nucleocapsid protein of 429 amino acids that starts at nucleotide position 48. It exhibited the highest similarity with other Sorex-related hantaviruses, with 68.1%-72.8% nucleotide and 71.9%-84.4% amino acid sequence identities. An analysis of a 1430-nucleotide region of the partial M segment exhibited approximately 54.4%-79.5% nucleotide and 43.2%-90.8% amino acid sequence identities to other hantaviruses. A comparison of a 432-nucleotide region of the L segment also showed similar degrees of identity, with 68.9%-78.4% nucleotide and 71.1%-93.8% amino acid sequence identities to other hantaviruses. Phylogenetic analyses using Bayesian methods indicated that QHSV shared the most recent common ancestor with other Sorex-related hantaviruses. The host was identified using a morphological assessment and verified using mitochondrial cytochrome b (mt-Cyt b) gene sequencing. A pair-wise comparison of the 1140-nucleotide mt-Cyt b gene sequence from the host demonstrated that the host was close to S. cylindricauda from Nepal with 94.3% identity. The virus-host association tanglegram, which was constructed using the Dendroscope software, indicated that the QHSV phylogeny and the host phylogeny were approximately matched, which suggests no evidence of host switching for QHSV. Our results contribute to a wider viewpoint regarding the heterogeneity of viruses that infect shrews.

Flaviviruses present a wide range of genetic diversity and exhibit diverse host relationships. Mosquito-borne flaviviruses have recently been isolated and characterized worldwide. Yunnan Province of China is one of the richest areas of species diversity and is the center of multi-species evolution in mainland Asia, which supports the circulation of numerous arthropod-borne viruses (arboviruses). In a screening program of arboviruses, mosquitoes were collected during the mosquito activity season in the Yunnan Province from 2007 to 2010. Eleven flavivirus strains, named Yunnan Culex flaviviruses (YNCxFVs), were obtained from Culex tritaeniorhynchus and Anopheles sinensis specimens. Sequence analyses based on partial nonstructural protein (NS) 5 gene indicated that the YNCxFVs shared 92.8–99.6% nucleotide identity with each other and were similar to the Culex-related flaviviruses. The complete genome of one representative isolate, LSFlaviV-A20-09, was sequenced. The genome was 10,865 nucleotides long and contained a single, long open reading frame (ORF) of 10,080 nucleotides that encoded a 3360-aa polyprotein. This genome was most closely related to the Quang Binh virus (QBV) VN180 strain, an insect-specific flavivirus isolated from Culex mosquitoes in Vietnam, but only had 83.0% nucleotide and 93.8% amino acid identities for the ORF sequence. The genome has approximately 66.3%–68.5% nucleotide sequence and 69.3–73.3% amino acid sequence identities to other Culex flaviviruses, and only has 47.9–57.9% nucleotide sequence and 38.7–55.1% amino acid sequence identities to Coquillettidia-related, Mansonia-related and Aedes-related flaviviruses. Phylogenetic analyses revealed that the LSFlaviV-A20-09 fell into the Culex-related flavivirus clade. Our discoveries provide more information regarding the heterogeneity of viruses that infect mosquitoes.

A total of 372 adult Ixodes persulcatus ticks were collected from vegetation in a forest area of Heilongjiang Province in northeastern China, where Lyme disease is known to be endemic. The ticks were examined for the presence of granulocytic ehrlichiae by heminested PCR with primers derived from the 16S rRNA gene. Of 310 ticks obtained from the Dahe forestry farm, two pools (each containing 5 ticks) were found positive, with a minimum infection rate of 0.6%. Ehrlichial DNA was also detected in one female (1.6%) of 62 ticks collected from the Yulin forestry farm. The overall minimum infection rate of the 372 I. persulcatus adults was 0.8%. The nucleotide sequences of 919-bp PCR products from the three positive tick specimens were identical to each other and very closely related to the members of the Ehrlichia phagocytophila genogroup. This is the first identification of granulocytic ehrlichiae in ticks in Asia and the first report of infection in I. persulcatus anywhere.

We determined the complete nucleotide sequence of a Japanese encephalitis virus (JEV) isolate (designated SH17M-2007) from a pool of Culex tritaeniorhynchus collected in southern China in 2007. The genome consisted of 10,965 nucleotides and included a single open reading frame (10,296 nucleotides) that encodes a 3,432-amino-acid polyprotein. The SH17M-2007 had 97.3 to 98.4% nucleotide identity with two Korean strains (KV1899, K94P05) and two Japanese strains (Ishikawa, JEV/sw/Mie/40/2004), but only 88.8% identity with the Chinese vaccine strain SA14-14-2. Five unique amino acid substitutions including one in the envelope (E) protein (Glu(E-306)-Lys) were found in the SH17M-2007 strain. Phylogenetic relationships based on the full-length nucleotide sequences were similar to those based on the E gene.

To investigate the mRNA and protein expression of nucleostemin (NS) in human esophageal squamous cell carcinoma.The mRNA and protein expression of NS were detected in 31 mucosal atypical hyperplasia specimens, 62 esophageal squamous cell carcinoma specimens and the matched normal esophageal mucosa samples by RT-PCR and immunohistochemistry method, respectively.The positive expression rate of NS protein in normal esophageal mucosa, atypical hyperplasia and esophageal squamous cell carcinoma was 17.7% (11/62), 41.9% (13/31) and 69.4% (43/62), respectively. There was a significant difference among the above three groups (chi2 = 33.676, P0.01). The expression levels of NS mRNA in esophageal squamous cell carcinoma (0.971 +/- 0.121) was significantly higher than that in the atypical hyperplasia (0.913 +/- 0.085) and also in the normal esophageal mucosa (0.866 +/- 0.103; F = 14.829, P0.01). The expression level of both NS protein and mRNA was positively correlated with histological grade, infiltration depth, and lymph node metastasis (P0.05), but not with age, gender or pathological type (P0.05).Our results indicate that nucleostemin mRNA and protein are over-expressed in human esophageal squamous cell carcinoma, and it may be related with its oncogenesis.

By using multilocus sequence analysis, five Borrelia valaisiana -related strains isolated from rodents and ticks in southwestern China were eventually classified as a new genospecies of B. burgdorferi sensu lato rather than B. valaisiana . The finding explained the differences in transmission cycle and phenotype between B. valaisiana strains from Europe and B. valaisiana -related strains from eastern Asia.

To confirm the existence of Amur-like viruses in Apodemus peninsulae in China, and to understand the molecular characteristics of these viruses.Total RNA was extracted from lungs of A. peninsulae captured in Jilin of Northeast China with Trizol reagent. Complete S and partial M segments of Amur virus were amplified and sequenced. Phylogenetic analyses on multiple nucleotide sequences were performed with the Clustal method and DNASTAR software.383 bp cDNA of M segment and 1696 bp of S segment of Amur like virus were recovered from lung tissue of A. peninsulae, named JilinAP06. The full-length of its S gene comprised of 1696 nucleotides with ORF including 1287 nucleotides and encoding a protein which comprised 429 amino acids. The phylogenetic analysis of this sample with other hantaviruses revealed that the complete S and partial M segment sequence of JilinAP06 both were closely related to those Amur viruses such as AP63, AP61, AP1371 and AP1168 found in A. peninsulae from Far East region of Russia and B78 strain, Liu strain and H5 strain, which were all from Chinese patients. The complete S and partial M segment sequence of JilinAP06 had only 81.0% identities with the nucleotide sequences of HV prototype 76-118 strain.Amur-like viruses did exist in A. peninsulae from Northeasern China while A. peninsulae might be the natural reservoir of Amur-like viruses in China and was the important infectious source to HFRS patients which were caused by Amur-like viruses.

To investigate the prevalence of Anaplasma phagocytophilum in rodents from forest areas in northeastern China.PCR amplification, followed by sequence analysis was carried out. The sequences of 16S rRNA and gltA gene fragment amplified from rodent specimens were compared with corresponding part of the sequences deposited in GenBank.A total number of 276 rodents were tested, including 102 in Jilin province, 61 in Helongjiang province and 113 in Inner Mongolia autonomous region. The positive rates were 8.82%, 1.64% and 0.00%, respectively. The infection rate in rodents infected by ticks was 11.30 times higher than that in rodents without ticks (P = 0.002). The S. A. phagocytophilum 16S rRNA sequences from rodents in Jilin and Heilongjiang were identical and differed in 3-5 bases compared with the corresponding parts of A. phagocytophilum from America, Sweden and Japan. Compared with the sequences registered in GenBank, the nucleotide sequence of gltA varied from 87%-97% and its deduced amino acid sequence changed from 84%-99%.A. phagocytophilum infection was presented in rodents from Jilin and Heilongjiang province.

To detect and study the types of Borrelia burgdorferi sensu lato in ticks and rodents from Da Xing-An Mountains Forest areas of China.Nested PCR was performed to amplify 5S-23S rRNA intergenic spacer of B. burgdorferi. Positive products were analysed by restriction fragment length polymorphism (RFLP) and single strand conformation polymorphism (SSCP), specimens showing unique RFLP profile were sequenced and analysed.1336 Ixodes persulcatus, 144 Dermacento silvarum, 144 Haemaphysalis concinna and 145 rodents of 9 species were collected from 16 sections of Da Xing-An Mountains Forest areas of China. Specific fragments were amplified from 293 I. persulcatus and 6 D. silvarum and 5 rodents of 4 species. B. burgdorferi was not detected in H. concinna. Among the positively tested I. persulcatus, 209 contained B. garinii genospecies and 45 contained B.afzelii genospecies based on RFLP. Moreover, B.garinii genospecies consisted of B. garinii 20047 and B. garinii NT29. 17 adult I. persulcatus were simultaneously infected with B. garinii 20047 and B. garinii NT29. Nine adult I. persulcatus were simultaneously infected with B. garinii 20047 and B. afzelii. Four adult I. persulcatus were simultaneously infected with B. garinii 20047 and B. garinii NT29 and B. afzelii. Two D. silvarum were infected with B. garinii 20047, 1 D. silvarum with B. garinii 20047, 2 D. silvarum with B. afzelii. 3 rodents were infected with B. garinii 20047 while 2 rodents were infected with B. garinii NT29. Mixed infection was not found in D. silvarum and rodents. In addition, nine I. persulcatus and one D. silvarum specimens showed unique RFLP pattern. Data from sequential analysis showed that they all belonged to B. garinii. PCR-SSCP profiles of 5S-23S rRNA intergenic spacer of B. burgdorferi in the positive specimens exceeded 36 types; B. garinii 20047 showed 16 types while B. garinii NT29 showing 11 types, B. afzelii showing 9 types. SSCP profiles of the specimens coinfected with multiple B. burgdorferi was relatively complex.The infection of B. burgdorferi was found in the ticks and rodents in Da Xing-An Mountains Forests areas. The infection rate of I. persulcatus was high. B. garinii was predominant genospecies, and the population of B. burgdorferi was heterogeneous in the area. Mixed infections of different B. burgdorferi genospecies in ticks were found. I. persulcatus and Clethrionomys rufocanus were possibly served as major vector and major host for B. burgdorferi, respectively, suggesting that further study is needed to confirm the coinfection in humans and animals in this region.

To investigate rodents' natural infection of Orientia tsutsugamushi (Ot) in some areas of Inner Mongolia and Xinjiang, China.DNAs were extracted from spleens of the captured mice and nested-polymerase chain reaction (nPCR) technique was used to detect the Ot-Sta56 gene. Six positive samples were sequenced and analyzed by Clustal X (5.0) and DNA Club software.A total of 90 rodents were captured in Inner Mongolia, and the overall prevalence of Ot was 6.67%. There was no significant difference in infection rates among the positive rodents species. 20 rodents were captured in Xinjiang, and the prevalence of Ot was 5.00%. The geographical difference in infection rates was not statistically significant between Inner Mongolia and Xinjiang. 9 rodents were captured in farmlands of Inner Mongolia and Xinjiang but there was no positive samples found. 101 rodents were captured in grasslands, and the prevalence of Ot was 6.93%. The Sta56 gene nucleotide sequence homology to Karp strain of N59 (from Microtus maximowiczii), N69 (from Cricetulus barabensis) and X33(from Cricetus cricetus) was 99%. The sequence homology to Taitung-2 strain and TW461 strain of N65 (from C. barabensis) was 94%, and the sequence homology to Taitung-2 strain and TW461 strain of N88(from Apodemus agrarius) was also 94%. The sequence homology to Oishi strain of N90 (from A. agrarius) was 96.00%.Our findings indicated that infections of Ot did exist in rodents captured from Inner Mongolia and Xinjiang. The genotypes of Ot in Inner Mongolia and Xinjiang were quite complex, with some of them belonged to Karp type, and the others belonged to Taitung-2, TW461 and Oishi types which providing evidence for further investigation on the scrub typhus fuci in the two areas.

To understand the coinfection status of Borrelia burgdorferi sensu lato (B.b.s.l) and spotted fever group Rickettsia (SFGR) in Hunchun of Jilin province, China.Polymerase chain reaction (PCR) was used to detect the 5S-23S rRNA intergenic spacer of B. b. s. l and ompA of SFGR in ticks was collected in Hunchun,Jilin province. The amplification products of positive ticks were sequenced, and phylogenetic analysis was conducted by PHYLIP software package.The infection rate of B. b. s. l was 36.0% in Ixodes persulcatus ticks and the SFGR was discovered in I. persulcatus ticks,with an infection rate of 2.0%. The coinfection rate of both agents was 2.0%. In 327 Dermacentor siltarum ticks, the positive rates of B. b. s. l and SFGR were 30.9% and 29.1% respectively. 55 ticks (16.8%) were coinfected with the two pathogens. The sequence analysis of B. b. s. l showed that the B. b. s. l in Jilin area, which were highly homologous, all belonged to B. garinii genotypes. The sequence analysis of SFGR positive products showed that the DNA secquence of the newly detected agent (JL-95) was close to the two previously described rickettsiae which were detected in I. ricinus from Slovakia (called IRS3 and IRS4). Phylogenetic relationships inferred from the comparison of these sequences with those of other genus Rickettsiae indicated that JL-95, IRS3 and IRS4 constituted a new rickettsial genotype and formed a separate cluster among the spotted fever group Rickettsiae.Coinfection of B. b. s. l and SFGR existed in Hunchun, Jilin province. The sequencing of specific fragment confirmed a new SFGR which was different from other rickettsiae known in China.

Tularemia is a disease caused by Francisella tularensis, a gram-negative, facultative intracellular bacterium. F. tularensis is generally believed to be confined to the Northern Hemisphere and has been reported in many American and Eurasian countries, for example, the United States, Mexico, Canada, Japan, the former Soviet Union, Spain, Sweden, and Norway (1). Terrestrial and aquatic mammals such as ground squirrels, rabbits, hares, voles, and water rats are animal reservoirs for transmission of F. tularensis to humans (2). In China, F. tularensis was isolated in Daurian ground squirrels (Spermophilus dauricus) from Tongliao, Inner Mongolia Autonomous Region, in 1957. An outbreak of tularemia caused by contact with infected hares was first reported in Heilongjiang Province in 1959 (3). Thereafter, 6 cases were diagnosed in Qinghai Province in 1965 (4). Epidemiologic investigation identified several natural foci of the disease in Tibet from 1962 to 1972 and in Xinjiang Autonomous Region in 1986 (5,6), where F. tularensis was isolated from patients, Ixodes liberelis and Dermacentor marginatus ticks, and woolly hares (Lepus oiostolos). The latest outbreak occurred in 1986 at a food-processing factory in Shandong Province, where 31 of 36 workers who slaughtered hares became ill (7). Since then, no cases of F. tularensis infection have been reported in either humans or animal hosts. Whether the foci have become quiescent or the disease is underreported is unclear because tularemia is not a reportable disease in China. The objectives of this study were to investigate the presence of F. tularensis in rodents and to determine the subspecies type of the agent in China.

To investigate the association between the genetic polymorphisms of myxovirus resistance 1 (MxA) gene and susceptibility to severe acute respiratory syndromes (SARS).A case-control study was conducted and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect the T/G polymorphism at position-88 in the mxA gene promoter. Information on related factors of SARS was collected using a pre-testing questionnaire. Univariate and multivariate logistic analyses were conducted with SPSS software package.Sixty-six cases and sixty-four controls were selected for the study. Comparing with GG genotype, the proportion of GT genotype were significantly higher in the case group (81.3%) than that in the control group (62.5%)) with an OR (95% CI) of 2.700 (1.208-6.037). Multivariate logistic regression analysis revealed that the significant association remained after factors as wearing masks, protection gowns and eye-protection when contacting with SARS patient etc. were adjusted with an OR (95 % CI) of 2.911 (1.027-8.250).mxA promoter-88G/T SNP might be confered to host genetic susceptibility to SARS in Chinese Han population.

To study the factors in relation to severe acute respiratory syndromes (SARS) among health care workers and to develop related protective measures.Case-control study was applied. A standardized questionnaire was used to collect SARS related information for health care workers who had contacted or treated SARS patients. Univariate analysis was conducted using SPSS 10.0 software package and multivariate logistic regression analysis was conducted using SAS 6.12.Twenty-seven of the 49 factors under study were significantly associated with SARS infection, in which 22 factors were protective, and the other 5 were risk factors. 27 factors were included for multivariate logistic regression analysis. Results showed that six factors as wearing eye glasses, wearing protection gowns, exposure to secrets/mode of contact with SARS patients, types of mask and the working years atc, remained significant association with hospital infection of SARS.SARS infection in heath care workers was related to many factors during the process of diagnoses and/or treatment. It is recommended that adequate masks, eye-protection and protective gowns should be adopted for heath care workers during the process of clinical diagnoses and treatment of SARS patients.

This study investigated the long-term excretion of severe acute respiratory syndrome–associated coronavirus in sputum and stool specimens from 56 infected patients. The median (range) duration of virus excretion in sputa and stools was 21 (14–52) and 27 (16–126) days, respectively. Coexisting illness or conditions were associated with longer viral excretion in stools.

To study the correlation between positive rates of RNA in clinical confirmed severe acute respiratory syndrome (SARS) patients and its appearance in relation to the development of the disease in order to provide scientific basis for early diagnosis, effective prevention and treatment of the disease.One-step reverse transcription-polymerase chain reaction (RT-PCR) was used to amplify the SARS RNA in the clinical specimens from different courses of the disease. The representative amplicons were then sequenced. Chi-square for trend test was performed to study the correlation between positive rates of RT-PCR and at different periods after the onset of the disease.The fragments amplified from the sputum specimens of SARS patients were shown to share 100% homology with the published SARS-associated coronavirus. Of the different clinical specimens, positive rate in the stools appeared to be the highest (21.55%). Chi-square for trend test revealed that the positive rates of stools and sputa of SARS patients decreased with the development of the disease (chi(2) for trend = 12.55 and 16.408, P = 0.0004 and P = 0.000 05 respectively).One-step RT-PCR proved to be an effective method for the detection of SARS-associated coronavirus from clinical specimens. Data as indicated that the positive rates of SARS coronavirus were decreasing in SARS patients along with the disease progression.

In order to find out the current situation of tick-borne spotted fever in the area of Changbai mountain, Jilin province.In this study, a polymerase chain reaction (PCR) method was developed with primers R. rOmpA 190.70p and R. rOmpA 190.701n designed on the basis of rOmpA gene, which is specific for examining spotted fever group Rickttsiaes (SFGR). Six hundred nighty-three ticks were tested and a positive PCR product amplified from D. silvarum specimen (named JL-02) was cloned and sequenced.The SFGR DNA was detected from D. silvarum, Haemaphysalis concinna with the positive rates were 53.81% and 7.41% respectively. Its nucleotide sequence of 587 bp rOmpA and derived amino-acids showed 100.00% similarity with nucleotide sequence of DnS 14 and 99.00% with DnS 28 from the Former Soviet Union according to the result of BLUST and CLUSTAL, which was differential from the DNA sequences of strains previously detected in China.The natural focus of tick-borne spotted fever did exist in the area of Changbai mountain. The DNA sequence of SFGR was similar to that of DnS 14, which was first reported in China.

To investigate association between the natural-resistance-associated macrophage protein 1 (NRAMP1) gene polymorphisms and susceptibility to pulmonary tuberculosis (TB) in Chinese Han population.Hospital-based case-control study design was adopted. Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) technique were used to type three NRAMP1 polymorphisms (INT4, D543N and 3'UTR). Information on related factors of tuberculosis was collected using a pre-tested standard questionnaire. Univariate and multivariate unconditional logistic analyses were conducted using SPSS for window software package. Totally, 110 cases of TB were selected during April 2001 to June 2002, with an average age of (27.7 +/- 12.7) years. Also, 180 cases of healthy control were selected, aged (27.3 +/- 9.2) years in average. Locus of NRAMP1 polymorphism was analysed with univariate method.Univariate analysis demonstrated that the D543N G/A and 3'UTR TGTG+/del genotype occurred more frequently in the cases than in the controls, with crude odds ratios (OR) (95% CI) of 2.22 (1.03 - 4.78) and 1.93 (1.14 - 3.26), respectively. No significant association was observed between TB and INT4 polymorphisms. In multivariate analysis, associations of TB and D543N G/A and 3'UTR TGTG+/del genotypes remained, adjusted for exposure history and bacille Camette-Guérin immunization. Adjusted OR (95% CI) was 3.04 (1.12 - 8.27) and 2.36 (1.20 - 4.64), respectively. Still, no significant association between INT4 polymorphisms and TB was found.Polymorphisms of D543N and 3'UTR locus in NRAMP1 gene might affect their susceptibility to TB in Chinese Han population.

To explore the expression of reversion inducing cysteine-rich protein with Kazal motifs (RECK), vascular endothelial growth factor (VEGF) and endoglin (CD105) protein and its correlation with occurrence, development, invasion and metastasis in esophageal squamous cell carcinoma (ESCC).Streptavidin-peroxidase (SP) immunohisto-chemistry was used to detect expression of RECK and VEGF in 62 cases of ESCC, 31 cases of adjacent atypical hyperplastic epithelium and 62 cases of normal esophageal epithelium. CD105 Mb was used to assess microvessel density (MVD).The expression of RECK was closely correlated with histological grade, infiltrative depth and lymphatic metastasis in ESCC (P0.05). The expression of RECK decreased during cancer development: normal esophageal epithelium (85.5%, 53/62), adjacent atypical hyperplastic epithelium (71.0%, 22/31), and carcinoma (59.7%, 37/62). There was a significant difference among the groups (P0.05). The expression of VEGF protein was closely correlated with infiltrative depth and lymphatic metastasis in ESCC (P0.05). The expression of VEGF protein increased during cancer development: normal esophageal epithelium (29.0%, 18/62), adjacent atypical hyperplastic epithelium (54.8%, 17/31), and carcinoma (67.7%, 42/62). There was a significant difference among the groups (P0.05). MVDCD105 increased in accordance with histological grade, but there was no significant difference (grade I, 36.92 +/- 10.85; grade II, 37.65 +/- 9.50; and grade III, 38.06 +/- 12.19). The MVDCD105 was closely correlated with infiltration and lymphatic metastasis in ESCC (P0.05). The expression of RECK was inversely correlated with the expression of VEGF and CD105.RECK, VEGF and CD105 play important roles in the infiltration, metastasis and carcinogenesis in esophageal carcinoma. Angiogenesis in ESCC may be promoted by over-expression of CD105.

Background Tularemia was reported in China over 50 years ago, however, many epidemical characteristics remain unclear. In the present study, the prevalence of Francisella tularensis in ticks was investigated during an epidemiological surveillance in China and then we measured their genetic diversity by conducting multiple-locus variable- number tandem repeat analysis (MLVA). Results 1670 ticks from 2 endemic areas (Inner Mongolia Autonomous Region and Heilongjiang Province) and 2 non-endemic areas (Jilin and Fujian Provinces) were collected and tested for evidence of tularemia by nested PCR. The prevalence of Francisella tularensis in ticks averaged 1.98%. The positive rates were significantly different among tick species, with Dermacentor silvarum and Ixodes persulatus responsible for all positive numbers. All F. tularensis that were detected in ticks belonged to F. tularensis subsp. holarctica and MLVA disclosed genetic diversity. One subtype was identified in 17 of 33 positive tick samples in three different study areas. Another subtype belonging to F. tularensis subsp. holarctica genotype was described for the first time in the current study. Conclusion The study showed two tick species, D. silvarum and I. persulatus harboring the pathogen of tularemia in natural environment, indicating these two tick species might have a role in tularemia existence in China. MLVA results disclosed the genetic diversity F. tularensis and identified one genotype as the most prevalent among the investigated ticks in China.