Expression of nucleostemin mRNA and protein in the esophageal squamous cell carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the mRNA and protein expression of nucleostemin (NS) in human esophageal squamous cell carcinoma.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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, P < 0.01). 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, P < 0.01). The expression level of both NS protein and mRNA was positively correlated with histological grade, infiltration depth, and lymph node metastasis (P < 0.05), but not with age, gender or pathological type (P > 0.05).</p><p><b>CONCLUSION</b>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.</p>

Investigation on Anaplasma phagocytophilum infection in rodents from forest areas in northeastern China / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate the prevalence of Anaplasma phagocytophilum in rodents from forest areas in northeastern China.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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%.</p><p><b>CONCLUSION</b>A. phagocytophilum infection was presented in rodents from Jilin and Heilongjiang province.</p>

Identification of Amur like virus in Apodemus peninsulae and its molecular characteristics in China / 中华流行病学杂志

<p><b>OBJECTIVE</b>To confirm the existence of Amur-like viruses in Apodemus peninsulae in China, and to understand the molecular characteristics of these viruses.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>

Study on the coinfection status of Borrelia burgdorferi sensu lato and spotted fever group Rickettsia in ticks from Hunchun, Jilin province / 中华流行病学杂志

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>

Investigation on rodents' natural infection of Orientia tsutsugamushi in some areas of Inner Mongolia and Xinjiang, China / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate rodents' natural infection of Orientia tsutsugamushi (Ot) in some areas of Inner Mongolia and Xinjiang, China.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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%.</p><p><b>CONCLUSION</b>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.</p>

Investigation on Borrelia burgdorferi sensu lato in ticks and rodents collected in Da Xing-An Mountains Forest areas of China / 中华流行病学杂志

<p><b>OBJECTIVE</b>To detect and study the types of Borrelia burgdorferi sensu lato in ticks and rodents from Da Xing-An Mountains Forest areas of China.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>

Study on the coinfection of three tick-borne infectious diseases in China using polymerase chain reaction method / 中华流行病学杂志

<p><b>OBJECTIVE</b>To study the existence of Ehrluichiosis, lyme disease and tick-borne spotted fever coinfection in some areas in China.</p><p><b>METHODS</b>Using polymerase chain reaction (PCR), B. burgdorferi sensu lato, spotted fever group (SFG) Rickettsiae and human granulocytic ehrlichia (HGE), Ehrlichia chaffeensis (EC) were detected in ticks and mouse samples collected from Inner Mogolia autonomous region, Heilongjiang province, Beijing and Fujian province.</p><p><b>RESULTS</b>408 Ixodes persulcatus collected from Inner Mogolia autonomous region, HGE and B. burgdorferi sensu lato and SFG Rickettsiae were detected positive, with rates as 6.8%, 7.8%, 45.6%, respectively. 5 (5/408) were coinfection with HGE and B. burgdorferi sensu lato while 1 (1/408) was coinfection with HGE and SFG Rickettsiae. 46 Ixodes persulcatus collected from Helongjiang province were determined positive, with rates as 6.5%, 10.8% and 34.8%, respectively including 1 (1/46) coinfected with HGE and B. burgdorferi sensu lato. 2 of 922 ticks collected from Beijing were detected positive with B. burgdorferi sensu lato. Among 283 groups of Haemaphysalis yeni ticks (3/group) and from 38 rodent samples collected from Ninghua county of Fujian province HCE and B. burgdorferi sensu lato and SFG Rickettsiae were detected. Out of them, 25 groups were positive with EC and the minimal positive rate was 3.8% while 21 rodent samples were positive with EC with a positive rate of 56.4%. 2 ticks and 1 rodent sample were detected positive with EC and spotted fever group.</p><p><b>CONCLUSION</b>Coinfection of HGE and B. burgdorferi sensu lato or spotted fever group Richi did exist in Ixodes persulcatus collected from Inner Mogolia autonomous region and Heilongjiang province. Coinfection of EC and spotted fever group Richi was found in the ticks and rodents collected from Fujian province.</p>

A case-control study on the mxA polymorphisms and susceptibility to severe acute respiratory syndromes / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate the association between the genetic polymorphisms of myxovirus resistance 1 (MxA) gene and susceptibility to severe acute respiratory syndromes (SARS).</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSION</b>mxA promoter-88G/T SNP might be confered to host genetic susceptibility to SARS in Chinese Han population.</p>

A case-control study on the risk factors of severe acute respiratory syndromes among health care workers / 中华流行病学杂志

<p><b>OBJECTIVE</b>To study the factors in relation to severe acute respiratory syndromes (SARS) among health care workers and to develop related protective measures.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>

Study on the correlation between positive rates of SARS RNA in clinical confirmed SARS patients and the appearance of RNA in relation to the development of the disease / 中华流行病学杂志

<p><b>OBJECTIVES</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSION</b>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.</p>

A new type of spotted fever group Rickttsiaes detected in the area of Changbai mountain, Jilin province / 中华流行病学杂志

<p><b>OBJECTIVE</b>In order to find out the current situation of tick-borne spotted fever in the area of Changbai mountain, Jilin province.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>

A case-control study on the vitamin D receptor gene polymorphisms and susceptibility to pulmonary tuberculosis / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate the association between the genetic polymorphisms of VDR gene and susceptibility to pulmonary tuberculosis.</p><p><b>METHODS</b>Case-control study was conducted. PCR-RFLP technique was used to detect the C/T polymorphism in VDR gene. Information on related factors of tuberculosis was collected using a pre-tested questionnaire. Univariate and multivariate logistic analyses were conducted with SPSS software package.</p><p><b>RESULTS</b>A sample of 76 cases and 171 controls was studied. The genotype frequencies of VDR-FF, VDR-Ff and VDR-ff were 38.2%, 44.7%, 17.1% and 52.6%, 40.9%, 6.4% respectively. VDR-ff was significantly overrepresented in case group, the OR (95% CI) was 3.668 (1.483 - 9.071) when comparing with FF genotype. The significant association remained after adjusting BCG immunization and smoking, the OR (95% CI) was 3.036 (1.117 - 8.253).</p><p><b>CONCLUSION</b>The VDR-ff genotype might be associated with the susceptibility to pulmonary tuberculosis in Chinese Han population.</p>

The application of indirect immuno-fluorescence assay in the diagnosis of severe acute respiratory syndrome / 中华流行病学杂志

<p><b>OBJECTIVE</b>To explore the temporal profile of serum antibody against coronavirus in patients with severe acute respiratory syndrome (SARS), and to evaluate the reliability of indirect immuno-fluorescence assay (IFA) in the diagnosis of SARS.</p><p><b>METHODS</b>Clinically confirmed SARS patients, suspected SARS patients, and controls were included in the study. IFA was used to detect the serum antibody against SARS coronavirus. General information about the subjects was collected using a standard questionnaire.</p><p><b>RESULTS</b>The positive rates of specific IgG and IgM against SARS virus within 10 days after onset of the disease were 55.1% and 16.3% respectively and then increased up to 89.8% for IgG and 65.3% for IgM. After 25 days of the onset of the disease, 90.9% patients became positive for both IgG and IgM. Results from chi-square for trend test revealed that the positive rates of both IgG and IgM increased with time (chi(2) for trend = 16.376, P = 0.00005 for IgG; chi(2) for trend = 28.736, P = 0.00000 for IgM). Sensitivity, specificity and agreement value of IFA regarding the diagnosis of SARS were all higher than 90%.</p><p><b>CONCLUSION</b>IFA can be used to assist diagnosis of SARS after 10 days of the onset of disease.</p>

A case-control study on natural-resistance-associated macrophage protein 1 gene polymorphisms and susceptibility to pulmonary tuberculosis / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate association between the natural-resistance-associated macrophage protein 1 (NRAMP1) gene polymorphisms and susceptibility to pulmonary tuberculosis (TB) in Chinese Han population.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>Polymorphisms of D543N and 3'UTR locus in NRAMP1 gene might affect their susceptibility to TB in Chinese Han population.</p>