Total Infectomes Characterization of Respiratory Infections in pre-COVID-19 Wuhan, China

At the end of 2019 Wuhan witnessed an outbreak of "atypical pneumonia" that later developed into a global pandemic. Metagenomic sequencing rapidly revealed the causative agent of this outbreak to be a novel coronavirus - SARS-CoV-2. Herein, to provide a snapshot of the pathogens in pneumonia-associated respiratory samples from Wuhan prior to the emergence of SARS-CoV-2, we collected bronchoalveolar lavage fluid samples from 408 patients presenting with pneumonia and acute respiratory infections at the Central Hospital of Wuhan between 2016 and 2017. Unbiased total RNA sequencing was performed to reveal their "total infectome", including viruses, bacteria and fungi. Consequently, we identified 37 pathogen species, comprising 15 RNA viruses, 3 DNA viruses, 16 bacteria and 3 fungi, often at high abundance and including multiple co-infections (12.8%). However, SARS-CoV-2 was not present. These data depict a stable core infectome comprising common respiratory pathogens such as rhinoviruses and influenza viruses, an atypical respiratory virus (EV-D68), and a single case of a sporadic zoonotic pathogen - Chlamydia psittaci. Samples from patients experiencing respiratory disease on average had higher pathogen abundance than healthy controls. Phylogenetic analyses of individual pathogens revealed multiple origins and global transmission histories, highlighting the connectedness of the Wuhan population. This study provides a comprehensive overview of the pathogens associated with acute respiratory infections and pneumonia, which were more diverse and complex than obtained using targeted PCR or qPCR approaches. These data also suggest that SARS-CoV-2 or closely related viruses were absent from Wuhan in 2016-2017.

Extensive Genetic Diversity and Host Range of Rodent-borne Coronaviruses

To better understand the genetic diversity, host association and evolution of coronaviruses (CoVs) in China we analyzed a total of 696 rodents encompassing 16 different species sampled from Zhejiang and Yunnan provinces. Based on the reverse transcriptase PCR-based CoV screening CoVs of fecal samples and subsequent sequence analysis of the RdRp gene, we identified CoVs in diverse rodent species, comprising Apodemus agrarius, Apodemus latronum, Bandicota indica, Eothenomys miletus, E. eleusis, Rattus andamanesis, Rattus norvegicus, and R. tanezumi. Apodemus chevrieri was a particularly rich host, harboring 25 rodent CoVs. Genetic and phylogenetic analysis revealed the presence of three groups of CoVs carried by a range of rodents that were closely related to the Lucheng Rn rat coronavirus (LRNV), China Rattus coronavirus HKU24 (ChRCoV_HKU24) and Longquan Rl rat coronavirus (LRLV) identified previously. One newly identified A. chevrieri-associated virus closely related to LRNV lacked an NS2 gene. This virus had a similar genetic organization to AcCoV-JC34, recently discovered in the same rodent species in Yunnan, suggesting that it represents a new viral subtype. Notably, additional variants of LRNV were identified that contained putative nonstructural NS2b genes located downstream of the NS2 gene that were likely derived from the host genome. Recombination events were also identified in the ORF1a gene of Lijiang-71. In sum, these data reveal the substantial genetic diversity and genomic complexity of rodent-borne CoVs, and greatly extend our knowledge of these major wildlife virus reservoirs.

COVID-19 severity is associated with immunopathology and multi-organ damage

COVID-19 is characterised by dysregulated immune responses, metabolic dysfunction and adverse effects on the function of multiple organs. To understand how host responses contribute to COVID-19 pathophysiology, we used a multi-omics approach to identify molecular markers in peripheral blood and plasma samples that distinguish COVID-19 patients experiencing a range of disease severities. A large number of expressed genes, proteins, metabolites and extracellular RNAs (exRNAs) were identified that exhibited strong associations with various clinical parameters. Multiple sets of tissue-specific proteins and exRNAs varied significantly in both mild and severe patients, indicative of multi-organ damage. The continuous activation of IFN-I signalling and neutrophils, as well as a high level of inflammatory cytokines, were observed in severe disease patients. In contrast, COVID-19 in mild patients was characterised by robust T cell responses. Finally, we show that some of expressed genes, proteins and exRNAs can be used as biomarkers to predict the clinical outcomes of SARS-CoV-2 infection. These data refine our understanding of the pathophysiology and clinical progress of COVID-19 and will help guide future studies in this area.

Complete genome characterisation of a novel coronavirus associated with severe human respiratory disease in Wuhan, China

Emerging and re-emerging infectious diseases, such as SARS, MERS, Zika and highly pathogenic influenza present a major threat to public health1-3. Despite intense research effort, how, when and where novel diseases appear are still the source of considerable uncertainly. A severe respiratory disease was recently reported in the city of Wuhan, Hubei province, China. At the time of writing, at least 62 suspected cases have been reported since the first patient was hospitalized on December 12nd 2019. Epidemiological investigation by the local Center for Disease Control and Prevention (CDC) suggested that the outbreak was associated with a sea food market in Wuhan. We studied seven patients who were workers at the market, and collected bronchoalveolar lavage fluid (BALF) from one patient who exhibited a severe respiratory syndrome including fever, dizziness and cough, and who was admitted to Wuhan Central Hospital on December 26th 2019. Next generation metagenomic RNA sequencing4 identified a novel RNA virus from the family Coronaviridae designed WH-Human-1 coronavirus (WHCV). Phylogenetic analysis of the complete viral genome (29,903 nucleotides) revealed that WHCV was most closely related (89.1% nucleotide similarity similarity) to a group of Severe Acute Respiratory Syndrome (SARS)-like coronaviruses (genus Betacoronavirus, subgenus Sarbecovirus) previously sampled from bats in China and that have a history of genomic recombination. This outbreak highlights the ongoing capacity of viral spill-over from animals to cause severe disease in humans.

Diff erential expression analysis of microRNA gene chip in coronary particles and peripheral blood particles in patients with acute myocardial infarction with thrombus aspiration / 中国介入心脏病学杂志

Objective To investigate the diff erential expression of microRNA in microparticles from coronary blood and peripheral blood in patients with acute myocardial infarction, and to provide clues for further study on the role of myocardial in the pathogenesis of myocardial infarction. Methods Coronary and peripheral blood samples were collected from patients with acute myocardial infarction undergoing thrombus aspiration. Microparticles from coronary and peripheral blood samples were isolated by centrifugation and gene chips were used to sequence the microRNA from the microparticles in the two groups. The diff erences in microRNA expression were identifi ed between two groups and the function of these microRNA were analyzed. Results There were signifi cant diff erences between the microRNA in the microparticles from the coronary blood and peripheral blood in patients with acute myocardial infarction. By constructing expression profi les, 307 diff erentially expressed microRNA were found, with 221 of them were up regulated and 86 of them were down regulated. Conclusion There is signifi cant diff erence between the expression of microRNA in microparticles from the coronary blood and the peripheral blood of patients with acute myocardial infarction forty nine of them are closely related to cardiovascular disease, which can be used as the target of further research.

Synchronous culture of rabbit bone narrow-derived endothelial progenitor cells and smooth muscle progenitor cells / 第二军医大学学报

Objective: To induce synchronous differentiation of rabbit bone marrow-derived mononuclear cells into endothelial progenitor cells (EPCs) and smooth muscle progenitor cells (SPCs), and to study their biological properties and the possibility of them as seed cells for tissue-engineered venous valves. Methods: Gradient density centrifugation was used to obtain bone marrow blood mononuclear cells, which were separately cultured with EGM-2 complete medium containing 5% FBS for differentiation of EPCs and with EBM-2 medium without vascular endothelial growth factor (VEGF) containing 5% FBS and 20 ng/mL platelet-derived growth factor-BB (PDGF-BB) for differentiation of SPCs. The differentiation of EPCs and SPCs was identified by various methods. Results: EPCs were cultured for 10 days and the cells fused into monolayer, showing a "stepping stone" appearance and expressing VEGF receptor-2 (VEGFR-2), von Willebrand factor (vWF) and CD133, but not α-smooth muscle actin (α-SMA); Weibel-Palade bodies were seen within the EPCs cytoplasm under the transmission electron microscope. Biological function tests showed visible EPCs growing on the matrigel in a blood vessel-like form. SPCs were cultured for 14 days and showed the specific features of the vascular smooth muscle growth, namely, the "peak-valley" growth way. SPCs expressed CD34 and α-SMA but not vWF and VEGFR-2. Myofilaments, paralleling with the cell longitudinal axis, were seen under the transmission electron microscope. SPCs could not form vessel-like structures on the matrigel. Conclusion: Mononuclear cells can be obtained through gradient density centrifugation of the bone marrow blood, which can be synchronously induced into EPCs and SPCs, providing economical and easy seed cels for tissue-engineered venous valves.

Usefulness of lumen area parameters determined by intravascular ultrasound to predict functional significance of intermediate coronary artery stenosis / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Coronary artery disease is the leading cause of death in China. Percutaneous coronary intervention is a recent milestone technology for treatment coronary artery disease. However, clinical decision making for patients with intermediate coronary stenosis is still controversial. We designed this study to assess the optimal intravascular ultrasound (IVUS) criteria for predicting functional significance of intermediate coronary lesions.</p><p><b>METHODS</b>We enrolled 141 patients with 165 intermediate coronary lesions located in vessels with a diameter ≥ 2.50 mm. IVUS of intermediate coronary lesions were performed before intervention. Pressure-derived fractional flow reserve (FFR) was measured at maximal hyperemia induced by adenosine infusion. An FFR < 0.80 was considered as abnormal functional significance.</p><p><b>RESULTS</b>For the overall 165 lesions, the mean FFR value was 0.84 ± 0.09. The diameter of the stenosis by visual estimation on angiogram was (59.63 ± 11.29)%. Minimum lumen diameter (MLD), minimum lumen area (MLA) and plaque burden (PB) were (2.00 ± 0.36) mm, (3.88 ± 1.34) mm(2), (67.28 ± 9.89)% respectively by IVUS measurements. An FFR < 0.80 was seen in 43 lesions (30.5%). There was a moderate correlation between IVUS parameters and FFR, including MLD (r = 0.372, P < 0.001), MLA (r = 0.442, P < 0.001) and PB (r = -0.172, P < 0.05). MLA was a predictor for FFR as a continuous variable independent of possible confounding variables (P < 0.05), and MLA and PB, were predictors for FFR < 0.80 as binary variables (P < 0.05). The best cutoff value of MLA to predict FFR < 0.80 was < 3.15 mm(2), with a 73.6% diagnostic accuracy; sensitivity 71.4%, specificity 67.0%, AUC = 0.709, and P < 0.001. The cutoff value of the PB to predict FFR < 0.80 was 65.45%; sensitivity 82.6%, specificity 41.2%, AUC = 0.644, and P < 0.01. If both MLA and PB were taken into account, the negative predictive value and the positive predictive value were 88.7% and 64.8% respectively.</p><p><b>CONCLUSIONS</b>Anatomic measurements of intermediate coronary lesions obtained by IVUS showed a moderate correlation to FFR values. IVUS-derived MLA ≥ 3.15 mm(2) may be useful to exclude FFR < 0.80, but poor specificity limits its applicability for physiological assessment of lesions < 3.15 mm(2). MLA was one of many factors affecting coronary flow hemodynamics. Both MLA and PB should be taken into account when determining functional ischemia.</p>

Genetic characteristics of hantaviruses carried by Microtus maximowixzii in Yakeshi of Inner Mongolia, China / 中华流行病学杂志

Objective To analyze the viral genetic characteristics of hantaviruses carried by Microtus maximowixzii in Yakeshi of Inner Mongolia Autonomous Region and its relationship with Hantaan virus (HTNV) and Seoul virus (SEOV) viruses as well as to identify the natural host of Khabarovsk virus (KHAV).Methods HV specific RNAs were detected by RT-PCR.Complete S and M segment were amplified from the RNA-positive samples.Phylogenetic analysis were performed to estimate the genetic characterization and the relationship with other hantaviruses.Results Fifty two Microtus maximowixzii voles were captured in Yakeshi areas.Of those voles,hanta-viral RNA was tested positive in 5 samples (9.62％).Complete S and M segments sequences were obtained from 5 and 2 lung samples,respectively.The complete S segment was consisted of 1848 to 1861 bp,and the M segment consisted of 3662 bp.These viruses were closely related to each other with 92.5％-96.4％ for the S segment sequences and 88.9％-95.4％ for the M segment sequences.They shared a higher identity with KHAV found previously in Yakeshi and KHAV of Russia.However,they were obviously different from the other hantavirus species.The 5 strains had the consistent secondary structure of nucleocapsid protein (NP) and glycoprotein (GP).When further comparing their secondary structures with those of HTNV and SEOV,our results indicated that there were no obvious differences in NP between KHAV and both HNTV,SEOV but with obvious difference in GP.Based on the S and M segment sequences,phylogenetic analyses revealed that these 5 strains clustered together with KHAV and formed a distinct lineage.Furthermore,all known KHAV strains could be divided into two small branches with a nucleotide divergence more than 5.3％.Conclusion Our research data revealed that KHAV was highly endemic among Microtus maximowixzii in Yakeshi area which supported the notion that Microtus maximowixzii had been the natural host of KHAV in the area.

Association between fractional flow reserve and quantitative coronary angiography parameters in intermediate coronary artery stenosis / 中华心血管病杂志

<p><b>OBJECTIVE</b>To explore the relationship between quantitative coronary angiography (QCA) parameters and fractional flow reserve (FFR) for identifying ideal angiographic parameters predictive of myocardial ischemia.</p><p><b>METHODS</b>The study included 121 lesions with QCA and FFR data from 106 patients [mean age: (63 ± 10) years]. The lesions were grouped into FFR > 0.75 group and FFR ≤ 0.75 group. Assessed parameters by QCA included percentage diameter stenosis, minimum luminal diameter (MLD), percentage area stenosis, minimum luminal area (MLA), reference vessel diameter (RVD) and lesion length (LL). Correlation analysis was used to identify the relationship between QCA parameters and FFR value, and receiver operating characteristic (ROC) curve was used to determine parameters predictive of FFR ≤ 0.75.</p><p><b>RESULTS</b>LL was significantly higher [(14.8 ± 7.9) mm vs. (10.7 ± 5.4) mm, P = 0.024] while MLD [(1.47 ± 0.31) mm vs. (1.82 ± 0.51) mm, P = 0.028], RVD [(2.30 ± 0.50) mm vs. (2.81 ± 0.64) mm, P = 0.036], and MLA [(2.30 ± 1.50) mm(2) vs. (3.60 ± 2.30) mm(2), P = 0.038] were significantly lower in FFR ≤ 0.75 group than in FFR > 0.75 group. LL (r = -0.209, P = 0.040) was negatively correlated with FFR, and MLD (r = 0.414, P = 0.040), RVD (r = 0.303, P = 0.000) and MLA (r = 0.315, P = 0.002) were positively correlated with FFR. ROC analysis showed that MLD ≥ 1.6 mm was the best cut-off value to predict FFR > 0.75 with sensitivity 63%, specificity 82%, and positive predictive value 96%.</p><p><b>CONCLUSIONS</b>QCA derived anatomic parameters of intermediate coronary lesions correlate to FFR value in some extent. MLD ≥ 1.6 mm is the best cut-off value to predict FFR > 0.75 in patients with intermediate coronary lesions.</p>

Predictive value of postprocedural leukocyte count on myocardial perfusion, left ventricular function and clinical outcomes in ST-elevated myocardial infarction after percutaneous coronary intervention / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Baseline white blood cell (WBC) count was correlated with ischemic events occurrence in patients with ST-elevated myocardial infarction (STEMI). However, circulating WBC count is altered after percutaneous coronary intervention (PCI). The aim of this study was to assess the relationship between postprocedural WBC count and clinical outcomes in STEMI patients who underwent PCI.</p><p><b>METHODS</b>A total of 242 consecutive acute STEMI patients who underwent successful primary PCI were enrolled and followed up for two years. WBC counts were measured within 12 hours after PCI. ST-segment resolution (ST-R) and myocardial blush grades (MBG) were evaluated immediately after PCI. Left ventricular ejection fraction (LVEF) was obtained at baseline and 12 - 18 months after PCI.</p><p><b>RESULTS</b>Postprocedural WBC count was an independent inverse predictor of ST-R (OR 0.80, P < 0.0001) and MBG 3 (OR 0.82, P < 0.0001). It was negatively correlated with LVEF (baseline r = -0.22, P = 0.001; 12 - 18 months r = -0.29, P < 0.0001). The best cutoff value of WBC for predicting death was determined to be 13.0 × 10(9)/L. The patients with a postprocedural WBC count above 13.0 × 10(9)/L showed a significantly lower cumulative survival rate (30 days, 82.4% vs. 99.0%, P < 0.0001 and 2 years 75.0% vs. 96.4%, P < 0.0001). Multivariate Cox regression analysis showed that a postprocedural WBC count was a strong independent predictor of 30-day mortality (HR 8.48, P = 0.019) and 2-year mortality (HR 4.93, P = 0.009).</p><p><b>CONCLUSIONS</b>Increased postprocedural WBC count is correlated with myocardial malperfusion and left ventricular dysfunction, and is an independent predictor of poor clinical outcomes in STEMI patients who underwent PCI.</p>

Investigation on the natural infectious status of hantaviruses among small mammals in Longquan city, Zhejiang province / 中华流行病学杂志

Objective To investigate the situation of the natural infection of hantaviruses (HV) in small mammals and to provide evidence for the control and prevention of hemorrhagic fever with renal syndrome (HFRS) in Longquan area,Zhejiang province. Methods Small mammals were captured by night trap, and lung tissue samples were collected and stored in liquid nitrogen. HV antigens were detected by indirect immuno-fluorescence assay (IFA). The partial S genome segment sequences were amplified by RT-PCR. DNAStar program was used for editing and comparing the sequences. Phylogeny was analyzed through PAUP*4.0 software. Results 319 small animals were collected in Longquan, and 9 hantavirus antigen-positive samples were identified. The positive rate of hantavirus in Apodemus agrarius was 4.97% . Phylogenetic tree constructed by partial S segment (620-999 nt) showed that the 9 strains carried by A agrarius from Longquan all belonged to HTNV,and had a closer evolutionary relationship with isolate Z251 from Zhejiang province. Conclusion Our results indicated that the main host was A. agrarius and the infection rate of HTNV was high in Longquan area.

Identification of Haemaphysalis longicomis and Rhipicephalus microplus / 中华流行病学杂志

Objective To establish a method combined morphology and molecular marker for identifying Haemaphysalis longicomis and Rhipicephalus microplus. Methods Ticks were collected from domestic animals and wild environment in epidemic area of Hubei and Henan provinces where cases of fever with thrombocytopenia syndrome were prevalent. We classified the ticks by morphology characteristics before 12S rDNA of ticks were amplified by PCR and subsequently sequenced. Phylogenetic tree was constructed by PAUP4.0. Results The ticks belonged to Haemaphysalis longicomis and Rhipicephalus microplus through observation and analysed by the morphological characteristics of the ticks. 12S rDNA was cloned and sequenced while data confirmed the morphological identification of the results. Conclusion The method based on morphology that combined with molecular marker seemed a good method for the identificaton of ticks.

SIRT1 inhibits IL-1β mRNA transcription in lipopotysaccharide tolerant THP-1 cells / 中华流行病学杂志

Objective To explore the role of silent information regulation 2 homolog 1 (SIRTl) in the regulation of IL-lβ mRNA transcription in lipopolysaccharide(LPS) tolerant THP-1 cells. Methods THP-1 human promonocyte model of endotoxin tolerance that simulates the sepsis leukocyte phenotype was used. Chromatin immunoprecipitation assay (ChIP) and real-timePCR were applied to quantify the binding of SIRTl and histone H3 lys9/H4 lysl6 acetylation to IL-1β promoter. IL-1β mRNA transcription was studied after knocking down the SIRTl. Results Thebinding of SIRTl to IL-1β promoter increased about 5 times in tolerant THP-1 cells (P<0.05) , which was accompanied by the low level of histone H3 lys9/H4 lysl6 acetylation (P<0.05, compared with normal cells). Knocking-down of SIRTl increased the transcription of IL-1β mRNA up to the level of 68% of normal cells (P<0.05) ,which was accompanied by the increase of histone H3 lys9/H4 lysl6 acetylation (P<0.05). However,there was no significant difference of p65 lys310 acetylation between normal and tolerant cells. Conclusion SIRTl inhibited the IL-1 β mRNA transcription in tolerant THP-1 cells but had not related to p65 lys310 acetylation. However, it was related to IL-1 p promoter acetylation.

Genetic analysis of hantaviruses carried by Rattus norvegicus collected from Henan and Neimenggu provinces, China / 中华流行病学杂志

Objective Genetic analysis was performed to infer the relationship between hantaviruses carried by Rattus norvegicus from Henan and Neimenggu provinces and the other known hantavirus and the vaccine strain. Methods Total RNA was extracted from lung tissues with Trizol reagent. The complete M and S segment sequences of strains NM133 and Q12 were amplified by RT-PCR. The purified DNA fragments were directly subjected to sequencing, and then to sequence analysis and phylogenetic analysis. Results The complete S segment sequences of strains NM133 and Q12 were found to be 1770 nt and 1772 nt in length respectively, with one open reading frame encoding 429 amino acids. The complete M segment sequences of both two strains are 3654 nucleotide in length encoding a protein of 1133 amino acids. The two strains shared a high degree of homology with most of known Seoul virus (SEOV) but quite different from Hantaan virus and other hantaviruses. Furthermore, the nucleoprotein and glycoprotein of the two strains had the congruent structure with the vaccine strain Z37. On the S- and M-phylogenetic trees, both strains (NM133 and Q12) were grouped into the first cluster of SEOV, and were more closely related to the strains, such as: Hb8610, R22, HB55, L99, and K24-e7. Conclusion Both strains (NM133 and Q12) belonged to SEOV, and sharing a high degree of homology and similar secondary structure with strains including the vaccine strains Z37, our data suggested that the present vaccine used in China could effectively prevent HFRS caused by SEOV.

Three-year follow-up results of visual inspection with acetic acid/Lugol's iodine (VIA/VILI) used as an alternative screening method for cervical cancer in rural areas / 癌症

<p><b>BACKGROUND AND OBJECTIVE</b>In China, there has been no established national program for cervical cancer prevention, the screening methods and experiences are especially deficient in the rural areas. The aim of this paper is to evaluate the effects of acetic acid/Lugol's iodine (VIA/VILI) used for screening of cervical cancer and pre-cancerous lesions in a rural area of China by analyzing the large-scale population-based screening data from the demonstration site.</p><p><b>METHODS</b>Women aged 30-59 years from Xiangyuan County in Shanxi Province were recruited for cervical cancer screening from 2005 to 2007. VIA/VILI was the primary screening method followed by colposcopy if the VIA/VILI was positive. Cervical lesions were diagnosed by directed biopsy under the colposcopy. The VIA/VILI negative women or cervical intraepithelial neoplasia 1 (CIN1) were re-screened using the same procedure in the next year.</p><p><b>RESULTS</b>In total, 7145 women received the cervical cancer screening, with a participation rate of 74.75%. Their average age was 42.16 years. A total of 1287 women were consecutively screened for three times from 2005 to 2007. The detection rates of CIN2, CIN3 and cervical cancer were 0.70% (9/1287), 1.01% (13/1287) and 0.23% (3/1287) for the first round screening, and were 0.22% (2/976), 0.11% (1/976) and 0% (0/976) for the second round screening, respectively. Only one CIN2 was found in the third round screening. In the years of 2006-2007, 3490 women were screened consecutively twice. The detection rates of CIN2, CIN3 and cervical cancer were 0.26% (9/3490), 0.52% (18/3490) and 0.15% (5/3490) for the first round screening, and 0.40% (14/2943), 0.40% (14/2943) and 0.03% (1/2943) for the second round screening. Likewise, 2 368 women were screened consecutively twice in the years of 2007-2008. The detection rates of CIN2, CIN3 and cervical cancer were 0.55% (13/2368), 0.25% (6/2368) and 0.12% (3/2368) for the first round screening, and 0.42 (10/2040), 0.04% (1/2040) and 0% for the second round screening. The cumulative detection rates for CIN2, CIN3 and cervical cancer were 0.81% (58/7145), 0.74% (53/7145) and 0.17% (12/7145), respectively. And 53.45% (31/58) of CIN2, 68.81% (37/53) of CIN3 and almost all cervical cancers (11/12) were found during the first round screening, except for an early stage cervical cancer (Ia). Only one CIN2 was detected in the third round screening in the same population. The average age of CIN1, CIN2, CIN3 and cervical cancer were 38.65, 40.61, 44.10 and 46.73 years, respectively.</p><p><b>CONCLUSIONS</b>VIA/VILI can be used as an alternative screening method for cervical cancer and high-grade pre-cancerous lesions among the women aged 30-59 years in China's rural areas because of its low cost, easy training for the local health providers, and less depending on facilities. One round screening by VIA/VILI can detect more than a half of CIN2, two-thirds of CIN3 and almost all the cervical cancer in the population, and the detection rates of CIN2/3 can be increased by two consecutive rounds of screening.</p>

Genetic characterization of two Seoul virus strains isolated from patients and rats in Hebei province / 中华流行病学杂志

Objective Complete S and M segments of two Seoul virus (SEOV) strains were obtained to determine their genetic types and characteristics. Methods The complete S and M segments from the isolate Li and lung tissue (sample LF18) were amplified by RT-PCR. Genetic analyses were performed by using DNAStar and PHYLIP program package. Results Their sequences consisted of 1772 nucleotides, and had an open reading frame (ORF, 43 to 1332 nt)encoding a nucleoprotein of 429 amino acids for both two strains. The complete M segment sequences consisted of 3653 nucleotides and had an ORF encoding a GnGc precursor of 1133 amino acids. The GnGc precursor of the two strains had 62 cysteine and 6 N-glycosylation sites. Both two strains shared a high degree of homology with other known SEOV strains including strains L99, Gou3, and vaccine strain Z37, with 87.6% to 99.2% and 83.6% to 97.3% nucleotide identities, respectively. On the S-and M- trees, the two strains LF18 and Li were grouped into the third cluster of SEOV. Conclusion Both LF18 and Li strains belonged to SEOV and shared the congruent genetic characteristics with the vaccine strains Z37. Thus, the bivalent vaccine including the strain Z37 could effectively prevent HFRS which was caused by SEOV in Hebei province.

An epidemiological investigation of the rabies between 1999-2008 in Yancheng city,Jiangsu province / 中华流行病学杂志

Objective To better understand the epidemiology of rabies during the past ten years in Yancheng city,Jiangsu province. Methods Data was collected and analyzed on rabies cases in Yancheng. Density and vaccination rate on Canine,Rate of injured people bit by dogs,and the information of post-exposure prophylaxis were studied. Rabies virus in the dog brains,collected around the epidemic areas of Yancheng,were detected and analyzed. Results A total of 135 human rabies cases occurred from 1999 through 2008,and formed the second epidemic peak since 1958. Of these victims,84% (114) were farmers. In general,the rate of people having dogs were 3%-6% per 100 people,and the injured person-times of 100 dogs were 6.37 per year. Notably,the vaccination rate of dogs was only 20%. Of those people injured by dogs and other animals,77% had received post-exposure treatment,and only 5%-10% had been administered anti-rabies serum. Rabies virus antigen was found in 4 (3.6%) of 111 brain specimens among dogs collected from epidemic areas. Genetic analysis of N and G genes,which were amplified from brain specimens,indicated that these viruses belong to genotype Ⅰ rabies and expressing a close relationship with the Chinese vaccine strain CTN. Conclusion The large number of dogs with low vaccination rate among them,together with the incorrect and low post-exposure treatment in rural areas seemed to be responsible for the outbreak of rabies in Yancheng city.

Cloning and expression of the nucleoprotein gene of Puumala-like virus / 中华流行病学杂志

Objective In order to detect Hokkaido virus(HOKV),a recombinant baculovirus containing the nucleoprotein(NP)gene of HOKV was constructed,and then the NP was expressed in insect cell.Methods The NP gene was cloned into plasmid PCR[R]2.1TA vector and then Was ligated into baculovims donor plasmid pFastBacTM1 after cutting by the restriction enzylne Kpn I and Not I.pFastBacTM1 was subsequently transferred into the One ShortTMTOP10 competent cells and then into DH10BacTM E.coli competent cells.which contained the baculovirus shuttle vector(Bacmid)and the helper plasmid to generate a recombinant bacmid.Results The NP gene was successfully expressed jn St9 insect cell.The expressed recombinant nucleoprotein had been identified in the S19 insect cell by indirect immunofluorescence assay,sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)and Westem blot.The results showed that the recombinant nucleoprotein appeared a molecular weight of 50×103M.and could reacmd with anti-recombinant Puumala virus(PUUV)nucleocapsid monoclonal antibodies and polyclonal antibodies against hantavirus.Conclusion Our results indicated that the recombinant nucleoprotein was SUCCESSfully expressed and having the immunogenicity and reactivity of natural nucleoprotein of HOKV.

Increased postprocedural neutrophil count is an independent predictive factor of poor prognosis in acute ST elevation myocardial infarction patients treated with percutaneous coronary intervention / 中华心血管病杂志

<p><b>OBJECTIVE</b>The aim of the study was to evaluate the prognostic value of the postprocedural neutrophil count in patients with first acute ST elevation myocardial infarction (STEMI) treated with successful primary percutaneous coronary intervention (PCI).</p><p><b>METHODS</b>A total of 226 consecutive STEMI patients underwent successful primary PCI were enrolled. Electrocardiograms were recorded before PCI and 2 hours after PCI. Neutrophil counts were measured within 12 hours after PCI. All patients were followed up for 2 years. Logistic regression analysis was used to evaluate predictive values of postprocedural neutrophil for ST-segment resolution (STR) after PCI and for death, non-fatal myocardial infarction and heart failure at 30 days and 2 years post PCI. Time-to-event analyses were performed using the Kaplan-Meier survival curves in patients with various ranges of postprocedural neutrophil counts.</p><p><b>RESULTS</b>Postprocedural neutrophil count ranged from 2.83x10(9)/L to 18.74x10(9)/L, first quartile, median and fourth quartile were 5.66x10(9)/L, 7.38x10(9)/L and 9.34x10(9)/L respectively. Multivariable logistic analysis showed that when postprocedural neutrophil count increased 1x10(9)/L, the risk of non-STR increased 2.28 fold (OR: 2.28, P=0.009), the risk of death (OR: 1.63, P=0.010) and heart failure (OR: 1.16, P=0.035) at 30 days increased 1.63 and 1.16 folds respectively, and the risk of death (OR: 1.29, P=0.003) and heart failure (OR: 1.20, P=0.007) at 2 years increased 1.29 and 1.20 folds respectively, but the risk of non-fatal myocardial infarction was not affected by postprocedural neutrophil count. Furthermore, the patients with postprocedural neutrophil count>or=9.34x10(9)/L had significant lower 30-day (89.1% vs. 99.1% vs. 98.2%, P=0.010) and 2-year (82.4% vs. 96.1% vs. 96.3%, P=0.003) survival rates compared with the patients with postprocedural neutrophil count from 5.66x10(9)/L to 9.33x10(9)/L and the patients with postprocedural neutrophil count<5.66x10(9)/L (all P<0.05).</p><p><b>CONCLUSION</b>Postprocedural neutrophil count is an independent predictor of short- and long-term death and heart failure in first acute STEMI patients treated with successful primary PCI.</p>

Study on the feasibility of a cell-culture inoculation test in the detection and isolation of street rabies virus / 中华流行病学杂志

Objective Feasibility of using MNA cell-culture inoculation test to detect and isolate the street rabies virus. Methods Using MNA cell-culture inoculation test, fluorescent antibody test (FAT) and sandwich ELISA with double-antibodies to detect 33 specimens of street rabies virus, 20 specimens of negative canine brains and 4 specimens of healthy mice brains. Results 33 specimens of street rabies virus were positive to the cell-culture inoculation test but the others were negative. The concordances of MNA cell-cultured inoculation test with FAT and sandwich ELISA with double-antibodies were both 100%. Conclusion MNA cell-culture inoculation test appeared to be both highly sensitive and specific in detecting the street rabies virus, and could be used in detection and isolation of the virus.