Objective To detect 10 RNA viruses including Alphavirus in Togoviridae, Flavivirus in Flaviridae, Hantavirus and Nairovirus in Bunyavirudae and SARS-CoV in Coronavirudae by using genechip technique. Methods The universal PCR primers of Alphavirus and Flavivirus and the PCR primers specific for HFRSV in Hantavirus and XJHFV in Nairovirus were designed by DNAStar software. PCR primers specific for SARS-CoV were adopted from WHO website. In addition, all the PCR primers specific for each virus were designed inside the regions of universal primers. These specific primers were utilized for amplification of cDNA probes. The concentration of probes, the hybridization temperature and duration, the formulation of hybridization solution and the washing conditions were optimized. Results The specific hybridization signals could be obtained when the concentration of probes was 0.3?g/?l. Good hybridization signals could be obtained for all the 10 RNA viruses when the hybridization solution contained 20% formamide, and the hybridization reaction was conducted at 60℃ for 1.5 hours. Two or four pathogens could be detected simultaneously when the target nuclear acids were amplified by multiplex PCR. Conclusion The results showed that the virus pathogens could be detected by genechip technique, and the key step was to design suitable primers and probes.

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Coronaviruses (CoVs) are generally associated with respiratory and enteric infections and have long been recognized as important pathogens of livestock and companion animals. Mouse hepatitis virus (MHV) is a widely studied model system for Coronavirus replication and pathogenesis. In this study, we created a MHV-A59 temperature sensitive (ts) mutant Wu"-ts18(cd) using the recombinant vaccinia reverse genetics system. Virus replication assay in 17C1-1 cells showed the plaque phenotype and replication characterization of constructed Wu"-ts18(cd) were indistinguishable from the reported ts mutant Wu"-ts 18. Then we cultured the ts mutant Wu"-ts 18(cd) at non-permissive temperature 39.5℃, which "forced" the ts recombinant virus to use second-site mutation to revert from a ts to a non-ts phenotype. Sequence analysis showed most of the revertants had the same single amino acid mutation at Nsp16 position 43. The single amino acid mutation at Nsp16 position 76 or position 130 could also revert the ts mutant Wu"-ts 18 (cd) to non-ts phenotype, an additional independent mutation in Nsp13 position 115 played an important role on plaque size. The results provided us with genetic information on the functional determinants of Nsp16. This allowed us to build up a more reasonable model of CoVs replication-transcription complex.

OBJECTIVETo investigate the Western Area Surge (WAS) program in the Ebola outbreak of Sierra Leone, and to analyze its implementing effect.

METHODSThe subject of this study was 3,813 laboratory confirmed Ebola hemorrhagic fever (EHF) cases reported in Sierra Leone from November 19, 2014 through January 27, 2015, a period before and after the implementation of the WAS program. To analyze and make conclusions according to the working experience of China Mobile Laboratory Reponses Team in the fight of Ebola outbreak, using WHO published EHF case definition to make diagnosis and compare the number of bed numbers, confirmed EHF cases, samples tested, and positive rates before and after implementation of WAS program.

RESULTSFrom the implementation of WAS program on 17th December 2014 to half a month later, the total numbers of Ebola holding and treatment centers increased from 640 to 960, six additional laboratories were established. On January, 2015, another two laboratories from America and The Netherlands were established. The numbers of samples tested one month before and after WAS program were 7,891 and 9,783, respectively, with an increase of 24.0 percent, while the positive rate of Ebola virus decreased from 22.2% (1,752/7,891) to 11.0% (1,077/9,783). The positive rate of blood samples decreased from 39.6% (248/626) in the month before WAS program to 27.4% (131/478) (χ2=17.93, P<0.001) in the mother after WAS program, the positive rate of blood samples 22.7% (103/454) to 10% (62/609) (χ2=31.03, P<0.001), accordingly. After 3 weeks of WAS program, in addition to Western Area, another four hotspots in Sierra Leone had also reported a significant decrease of the numbers of confirmed EVD cases. Forty-two days after implementation of WAS program, the daily number of laboratory confirmed EHF cases decreased from 63 to 10.

CONCLUSIONWAS program played a vital role in controlling the EHF outbreak rapidly in Sierra Leone. It could also provide guidance for the control similar large infectious diseases outbreak in the future.

Objective:To compare the efficacy and side effects between simultaneous and sequential integrated boost intensity-modulated radiotherapy after operation for high-grade glioma.Methods:We retrospectively analyzed 142 patients with high-grade glioma who underwent postoperative radiotherapy from January 2010 to December 2017. All patients were divided into the simultaneous and sequential integrated boost intensity-modulated radiotherapy groups. Concurrent temozolomide chemotherapy was delivered during radiotherapy in two groups. The follow-up outcomes were statistically compared between two groups.Results:For the whole group, the median overall survival (OS) was 24 months, the median progression-free survival (PFS) was 17 months, and the median disease-free survival (DFS) was 25 months. In the simultaneous and sequential integrated boost intensity-modulated radiotherapy groups, the median OS were 27.2 and 21.0 months ( P=0.950), the median PFS were 21.2 and 15.0 months ( P=0.21), and the median DFS were 28.0 and 18.0 months ( P=0.171), and the disease control rates were 92.86% and 85.17%( P=0.541), respectively. There was no statistical difference in OS, PFS, DFS, short-term efficacy and side effects between two groups. However, the conformity index in the simultaneous integrated boost intensity-modulated radiotherapy group was better than that in the sequential integrated boost intensity-modulated radiotherapy group ( P=0.032). Conclusions:Postoperative simultaneous and sequential integrated boost intensity-modulated radiotherapy yield no statistical differences in the survival, short-term efficacy and side effects in the treatment of high-grade glioma. However, the conformity index in the simultaneous integrated boost intensity-modulated radiotherapy group is significantly better, which can be recommended for postoperative radiotherapy of high-grade glioma.

This study aimed to evaluate the therapeutic potential of inhibiting protein arginine methyltransferase 5(PRMT5)in cisplatin-induced hearing loss.The effects of PRMT5 inhibition on cisplatin-induced auditory injury were determined using immunohistochemistry,apoptosis assays,and auditory brainstem response.The mechanism of PRMT5 inhibition on hair cell survival was assessed using RNA-seq and Cleavage Under Targets and Tagment-quantitative polymerase chain reaction(CUT&Tag-qPCR)analyses in the HEI-OC1 cell line.Pharmacological inhibition of PRMT5 significantly alleviated cisplatin-induced damage to hair cells and spiral ganglion neurons in the cochlea and decreased apoptosis by protecting mitochondrial function and preventing the accumulation of reactive oxygen species.CUT&Tag-qPCR analysis demonstrated that inhibition of PRMT5 in HEI-OC1 cells reduced the accumulation of H4R3me2s/H3R8me2s marks at the promoter region of the Pik3ca gene,thus activating the expression of Pik3ca.These findings suggest that PRMT5 inhibitors have strong potential as agents against cisplatin-induced ototoxicity and can lay the foundation for further research on treatment strategies of hearing loss.

Objective: To investigate the clinical effect of lithium disilicate glass ceramic cantilever resin-bonded fixed partial dentures (CRBFPDs) on single anterior tooth loss to provide a reference for the selection of restoration methods for single anterior tooth loss. Methods: This study was reviewed and approved by the Ethics Committee, and informed consent was obtained from the patients. Forty-two patients with less than two anterior teeth with monomaxillary loss were included in this study. After 6 months, 1 year, 2 years, and 3 years, the aesthetic and functional effects of the restorations and the periodontal health status were evaluated, and the visual analog scale (VAS) was used to assess patient satisfaction. Results: During the observation period, the connector fractured in one case within 3 months. One case had debonded within 2 years. The aesthetic restoration effect of all lithium disilicate glass ceramic CRBFPDs was categorized as Class A. The periodontal health was good, there was no clinical absorption in the soft and hard tissues of the abutment or subbridge, periodontal status according to the evaluation indices was classified as class A, and the total satisfaction rate of the patient was 100%. Conclusion For single anterior tooth loss patients, lithium disilicate glass ceramic cantilever resin-bonded fixed partial denture can achieve the restoration effect of less invasion, better adhesion, aesthetics, comfort and good biocompatibility. With high patient satisfaction, it can be considered an ideal restoration method for replacing a single anterior tooth.

We report a complete genomic sequence of rare isolates (minor genotype) of the SARS-CoV from SARS patients in Guangdong, China, where the first few cases emerged. The most striking discovery from the isolate is an extra 29-nucleotide sequence located at the nucleotide positions between 27,863 and 27,864 (referred to the complete sequence of BJ01) within an overlapped region composed of BGI-PUP5 (BGI-postulated uncharacterized protein 5) and BGI-PUP6 upstream of the N (nucleocapsid) protein. The discovery of this minor genotype, GD-Ins29, suggests a significant genetic event and differentiates it from the previously reported genotype, the dominant form among all sequenced SARS-CoV isolates. A 17-nt segment of this extra sequence is identical to a segment of the same size in two human mRNA sequences that may interfere with viral replication and transcription in the cytosol of the infected cells. It provides a new avenue for the exploration of the virus-host interaction in viral evolution, host pathogenesis, and vaccine development.
Base Sequence ; China ; Cluster Analysis ; Gene Components ; Genetic Variation ; Genome, Viral ; Genotype ; Molecular Sequence Data ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus ; genetics ; Sequence Analysis, DNA ; Severe Acute Respiratory Syndrome ; genetics

Cisplatin-related ototoxicity is a critical side effect of chemotherapy and can lead to irreversible hearing loss. This study aimed to assess the potential effect of the DNA methyltransferase (DNMT) inhibitor RG108 on cisplatin-induced ototoxicity. Immunohistochemistry, apoptosis assay, and auditory brainstem response (ABR) were employed to determine the impacts of RG108 on cisplatin-induced injury in murine hair cells (HCs) and spiral ganglion neurons (SGNs). Rhodamine 123 and TMRM were utilized for mitochondrial membrane potential (MMP) assessment. Reactive oxygen species (ROS) amounts were evaluated by Cellrox green and Mitosox-red probes. Mitochondrial respiratory function evaluation was performed by determining oxygen consumption rates (OCRs). The results showed that RG108 can markedly reduce cisplatin induced damage in HCs and SGNs, and alleviate apoptotic rate by protecting mitochondrial function through preventing ROS accumulation. Furthermore, RG108 upregulated BCL-2 and downregulated APAF1, BAX, and BAD in HEI-OC1 cells, and triggered the PI3K/AKT pathway. Decreased expression of low-density lipoprotein receptor-related protein 1 (LRP1) and high methylation of the LRP1 promoter were observed after cisplatin treatment. RG108 treatment can increase LRP1 expression and decrease LRP1 promoter methylation. In conclusion, RG108 might represent a new potential agent for preventing hearing loss induced by cisplatin via activating the LRP1-PI3K/AKT pathway.