Development of a consensus microarray method for identification of some highly pathogenic viruses.

Some highly pathogenic viruses, such as Chikungunya virus, Japanese encephalitis virus, Yellow fever virus, Dengue virus, Hanta virus, SARS-CoV, and H5N1 avian influenza virus can cause severe infectious diseases. However, the consensus method for detecting these viruses has not been well established. A rapid and sensitive microarray approach for detection of these viruses and a panel of specific probes covering nine genera and 16 virus species were designed. 70-mer oligonucleotides were used at the genus level and 50-mer oligonucleotides were at the species level, respectively. To decrease the interference of the host genome in hybridization, the consensus genus primers were designed and used to reverse transcribe only virus genome. The synthesis of the second strand was carried out with a random primer sequence (5'-GTTTCCCAGTAGGTCTCNNNNNNNN-3'). The amplified products were labeled and processed for microarray analyses. This microarray-based method used the highly conserved consensus primers to synthesize specifically the virus cDNA and could identify effectively Chikungunya virus, Japanese encephalitis virus, Yellow fever virus, Dengue virus, Tick borne encephalitis virus, and H5N1 avian influenza virus. Using this method, one unknown virus isolated from pig brain in Shanxi Province, China was identified. This method may have an important potential application for the diagnosis of virus infection.

Mortality and trend analysis of bladder cancer in China from 2004 to 2008 / 上海预防医学

Objective:To analyze bladder cancer mortality in China from 2004 to 2018. Methods:The dataset of bladder cancer mortality from 2004 to 2018， based on 605 national surveillance sites and regularly published by the Chinese Center for Disease Control and Prevention， was collected and age-standardized according to the demographic structure of China in 2000. The crude mortality rate （CMR）， the age-standard mortality rate （ASMR）， and the ratio of ASMRs of rural to urban areas（RR）were calculated to analyze the distributions of the mortality of bladder cancer stratified by age， sex， region （rural/urban areas， eastern/central/western areas） or time. Results:The CMR of bladder cancer in China from 2004 to 2018 was 1.69/105， and the ASMR was 1.09/105. The temporal trend on the CMR of bladder cancer from 2004 to 2018 increased significantly （APC=2.91%，P<0.001）， whereas the trend on the ASMR decreased a little （APC=-1.29，P=0.008）. The temporal trend on the CMR of bladder cancer in the males increased （APC=3.29%，P<0.001）， whereas the trend on the ASMR did not change. The temporal trend on the CMR of bladder cancer in the female increased （APC=2.12%，P<0.001）， whereas the trend on the ASMR decreased （APC=-1.94，P=0.008）. Both the CMR and ASMR of urban areas were higher than those of rural areas （P<0.05）. However， the temporal trend on the ASMR of bladder cancer in the urban decreased significantly （APC=-2.05%，P=0.002）， mainly exhibited in eastern and western urban. The ASMR of eastern urban areas was higher than that of western urban areas. The temporal trend on the ASMR of bladder cancer in the rural did not change （P>0.05）， and no differences in the ASMR were detected between eastern， central and western rural areas. The gaps between rural and urban areas in the eastern （P<0.001） or western （P=0.002） region reduced. Although the temporal trend on the ASMR in urban people over 40 years old decreased significantly， the ASMR of urban people over 60 years old was much higher than that of other age groups in the urban or any age groups in rural areas. Conclusion:The age group over 60 years old in the urban is the major target population for bladder cancer prevention and treatment. Screening， diagnosis and treatment for bladder cancer in rural should be strengthened.

Mortality analysis and time trend of prostate cancer in China from 2004 to 2008 / 上海预防医学

Objective:To obtain the temporal and spatial trends on prostate cancer mortality in China from 2004 to 2018. Methods:The data of prostate cancer mortality was collected from 605 national disease surveillance sites and age-standardized according to the demographic structure of China in 2000. The crude mortality rate （CMR） and age-standardized mortality rate （ASMR）， and the ratio of ASMRs of rural to urban areas（RR）， were calculated to analyze the distributions of the mortality of prostate cancer stratified by age， sex， region （rural/urban areas， eastern/central/western areas） or time. Results:The temporal trend on the CMR of prostate cancer from 2004 to 2018 increased significantly （APC=5.23%，P<0.001）， whereas the trend on the ASMR did not change （APC=0.65%，P=0.336）. The ASMR of urban areas was higher than that of rural areas（P<0.05）. The temporal trend on the ASMR of urban or rural did not change（P>0.05）. The same trend was detected for the RR value （P>0.05）. The ASMR of eastern urban areas was higher than that of the central or the western urban areas. The ASMR of eastern rural areas was higher than that of central and western areas. In the eastern areas， both the temporal trends for the ASMRs of the urban and the rural increased （The urban： APC=1.6%， P=0.015； the rural： APC=1.02%， P=0.013）. However， the RR values for the East， the Center or the West did not change. The ASMR of the people over 60 years old was higher than that of the people under 60 years old. However， the temporal trends on the ASMRs of the group under 60 years old or the group over 60 years did not change （P>0.05）. The ASMR of the people under 60 years old in 2018 was significantly lower than that in 2004（P=0.004）. Conclusion:A significant correlation exists between the death of prostate cancer and age. The mortality of prostate cancer in urban is higher than that in rural. The current screening strategy for prostate cancer has a limited impact on the prognosis of prostate cancer patients in China. Stratified refinement of prostate cancer screening strategies for people aged over 60 years in urban areas and the causal prophylaxis of prostate cancer are priorities for future prostate cancer prevention and control.

Status of pesticide registration and residue analysis for traditional Chinese medicine in China / 中国中药杂志

The present paper outlined pesticide registration status for traditional Chinese medicines (TCMs) and summarized the characteristics of pesticide contamination in different regions of some widely used TCMs by retrieving last 10 years' literatures. At present, the problems of pesticide residues for TCM include less pesticide registrations, widespread high-residue organochlorine pesticides contamination, pesticide abuse, irregular GAP bases and imperfect pesticide limit standards, etc. According to the current situation, we should adopt some control measures to strengthen the quality control of TCMs so as to ensure the safety of TCMs and related products.

Investigation on the endemic foci of new emerged tick-borne encephalitis in Charles Hilary, Xinjiang / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate the recent emerged endemic region of tick-borne encephalitis (TBE) regarding its natural reserves, in Charles Hilary, northern Xinjiang and to isolate and characterize the viral geographic strain.</p><p><b>METHODS</b>Using indirect fluorescent assay to detect tick-borne encephalitis virus (TBEV) specific IgG antibodies from serum of local residents including 2 unconfirmed viral encephalitis patients in 2011 spring-summer. Viruses were isolated from tick samples by inoculating BALB/c mice and BHK-21 cells. For phylogenetic analysis. TBEV NS1 gene fragments were detected by RT-PCR and then subjected to sequence alignment.</p><p><b>RESULTS</b>1760 ticks were captured from the fields to have found that Ixodes persulcatus were dominated among the tick population. Two viral encephalitis patients were diagnosed as TBEV infection. In addition, 35.4% (23/65) local residents were detected positive for presence of TBEV specific-IgG antibodies in serum. After inoculation, morbidity and mortality of BALB/c mice were 72.9% (70/96) and 55.7% (44/79), respectively. TBEV specific-fragments were amplified from brain tissue of dead mice and cells culture supernatant. NS1 sequence alignment showed that the viral isolates were clustered into TBEV far-eastern sub-type, phylogenetically, and were mostly close to the isolates from northeastern China (99%) and Russian strain (98%).</p><p><b>CONCLUSION</b>In this study, a new endemic loci of TBE was firstly described in Charles Hilary natural reserve, northern Xinjiang. TBEV geographic isolates belonged to TBEV far-eastern subtype while Ixodes persulcatus and Dermacentor silvarum played crucial roles for disease transition.</p>

Expression of tick-borne encephalitis virus prM-E protein in insect cells and studies on its antigenicity / 中华实验和临床病毒学杂志

<p><b>BACKGROUND</b>To express the prM-E protein in Sf9 cells, and lay a basis for further study on the function of the viral proteins and development of specific diagnostic reagents.</p><p><b>METHODS</b>The recombinant prM-E protein of tick-borne encephalitis virus was expressed in insect cell Sf9 by RT-PCR amplification of prM-E gene, construction of donor plasmid of Bac-to-Bac baculovirus expression system, homologous recombination of donor plasmid with bacmid DNA at the site of Tn7 and transfection of insect cell Sf9.</p><p><b>RESULTS</b>Recombinant subviral particles, about 30 nm in diameter, consisting of prM-E were observed by electron microscope in the supernatant of infected cells, which indicated that infected cells released virus-like particles (VLPs) into the culture medium. The results of Western-blot and the indirect immunofluorescence assay (IFA) showed that the recombinant proteins retained antigenic and conformational structures similar to those of native virus proteins. Using the recombinant prM-E as antigens to detect samples of patient sera by ELISA and IFA, all of 16 sera from patients with tick-borne encephalitis were positive and all of 6 sera from other patients were negative.</p><p><b>CONCLUSION</b>The prM-E protein expressed in insect cells retains good antigenicity.</p>

An experimental study on the differentiation of bone marrow mesenchymal stem cells into vascular endothelial cells / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the feasibility of differentiation of bone marrow mesenchymal stem cells (MSCs) into vascular endothelial cells and the mechanism of its involvement in wound healing.</p><p><b>METHODS</b>Porcine MSCs were harvested from porcine marrow, and they were isolated and purified by density gradient centrifugation. After being cultured and amplified in vitro, the MSCs were labelled with BrdU (5-bromodeoxy-uridine). Full skin loss wound was created on the back of the mini-swine whose bone marrow was obtained. The labelled MSCs with fibrin glue as the vector were regrafted back to the donor animal wound. The wound tissue specimens were harvested at 2, 4, 6, 8 and 12 post-operation weeks and were immunohistochemically stained by BrdU and factor VIII (FVIII) for comparative study.</p><p><b>RESULTS</b>Most BrdU positive cells aggregated around small blood vessels in the granulation tissue of the wounds. Only individual vascular endothelial cells were BrdU positive. There was FVIII expression in the cytoplasm of BrdU positive cells.</p><p><b>CONCLUSION</b>MSCs were closely correlated with the formation of small blood vessels in granulation tissue during wound healing process. The porcine MSCs possessed the potential to differentiate into vascular entoehelial cells and to participate in wound healing under the micro-enviroment of the wound.</p>

The screening and identification of an unknown virus by DNA microarray / 中华检验医学杂志

Objective In our previous study,we established DNA microarray technology for identification of medical viruses on genus levels and arboviruses on species levels.In this study,we employed these microarrays to determine the pathogen of newly isolated unknown virus in July,2006 from pig brain in Shanxi province.Methods The pathogen isolated from pig brains were inoculated in BHK21 cells.After CPE were observed,the supernatants were collected and RNA was extracted.After reverse transcription and random PCR amplification,the labeled nucleic acids were hybridized with DNA microarrays.Results The hybridization results with medical viruses DNA microarray indicated that the unknown virus belonged to Flavivirus.Combined with epidemiological investigation,we presumed that it might be a kind of arbovirus. Then the labeled specimen were further hybridized with arbovirus DNA microarray and the results confirmed that it was Japanese encephalitis virus(JBV).This coincided with PCR and sequencing analysis.Conclusions The DNA microarray we established previously could be employed to identify unknown viruses.This method provides a new method for determining new viral pathogens.