Objective To establish a simple and sensitive detection method of Sendai virus ( SeV ) by reverse transcription loop-mediated isothermal amplification ( RT-LAMP) technique. Methods According to the published Gen-Bank sequences (DQ219803. 1), six pairs of primers were designed targeting the conserved region of SeV. The amplifica-tion products were detected with a LAMP real-time Turbidimeter. (LA-302). Through optimizing the LAMP primers and re-action conditions, a rapid and specific detection method of SeV was established. Meanwhile, the amplified products were colored by fluorescence detection reagent after completion of the reaction, so that the amplification could be visualized and detected by naked eyes. Then, methodological evaluation of the RT-LAMP was tested. Results The method of RT-LAMP showed a highly efficient amplification for SeV viral target gene which was performed at 63℃ for 60 min with the LAMP re-al-time Turbidimeter (LA-302). The detection limit was 2. 1 TCID50, 100 times higher than that of RT-PCR, and no cross-reaction with other RNA and DNA viruses of mice was observed. The results of SeV LAMP reaction was visualized and the tube could be directly observed by naked eyes with the addition of fluorescence detection reagent. The results were consist-ent with the results detected by real-time tubidimeter. 92 clinical samples were detected byRT- LAMP, RT-PCR and indi- rect ELISA, and the coincidence rate was 100%. Conclusions This established SeV RT-LAMP detection method is fast, specific, highly sensitive,easy to perform under simple conditions, and is suitable for rapid detection of Sendai viirus.

BACKGROUNDThe expressive level of glucose-regulated protein 78 (GRP78) is elevated and correlated with resistance of chemotherapy drugs in breast cancer cell. However, little is known about the relationship between its expression and drug resistance in non-small cell lung cancer (NSCLC). The aim of this study was to explore the relationship between drug resistance and the expression of GRP78 in NSCLC.

METHODSDrug sensitivity test was used to detect the resistance to 8 chemotherapy drugs in 52 NSCLC fresh surgical samples by methylthiazoletrazolium (MTT), and expression of GRP78 was detected by immunohistochemistry method. Spearman correlation assay was used to investigate the correlation between the GRP78 expression and drug resistance.

RESULTSThe resistance rates to paclitaxel (PTX), adriamycin (ADM), carboplatin (CBP), topotecan (TPT), navelbine (NVB), vincristine (VCR), cisplatin (DDP) and etoposide (VP-16) of the 52 samples were 42.31%, 57.69%, 63.46%, 65.38%, 67.31%, 73.08%, 78.85%, 90.38%, respectively. Fourteen cases showed the complete resistance to the total 8 chemotherapy drugs. Furthermore, the expression of GRP78 was stronger in poorly differentiated cancer as compared with the well and moderately differentiated cancer (P < 0.05), so as in stage II and III cancer than in stage I cancer (P < 0.05). Spearman correlation assay showed that there was a correlation between the chemotherapeutics resistance to ADM, VP-16, VCR, TPT and the expression of GRP78 in NSCLC (P < 0.05).

CONCLUSIONSIt is feasible to detect the drug sensitivity to chemotherapy for tumor cells by MTT method. The results of chemosensitivity assay in vitro are indicative of clinical drug administration in NSCLC. The detection of GRP78 isalso indicative of the resistance to chemotherapy drugs and the differentiation and the clinical stage in NSCLC.

Objective:To explore the death risk factors of extremely severe burn patients, establish a death risk nomogram predicting model, and investigate the predictive value for death risk of extremely severe burn patients.Methods:The medical records of 231 extremely severe burn patients (190 males and 41 females, aged 18-60 years) who were admitted to the Institute of Burn Research of the First Affiliated Hospital of Army Medical University from January 2010 to October 2018 and met the inclusion criteria were analyzed retrospectively. According to the final outcome, the patients were divided into survival group of 173 patients and death group of 58 patients. The sex, age, severity of inhalation injury, total burn area, full-thickness burn area, burn index, rehydration coefficient and urine volume coefficient of the first and second 24 h after injury, the first base excess, shock index, and hematocrit (HCT) after admission, whether to have pre-hospital fluid infusion, use of ventilator, and use of continuous renal replacement therapy (CRRT), and abbreviated burn severity index (ABSI ) and Baux score on admission of patients in the two groups were recorded or calculated. According to the use of ventilator, the patients were divided into with ventilator group of 131 patients and without ventilator group of 100 patients, and the death, total burn surface area, burn index, incidence and severity of inhalation injury were recorded. According to the use of CRRT, the patients were divided into with CRRT group of 59 patients and without CRRT group of 172 patients, and the death, total burn surface area, and burn index were recorded. Data were statistically analyzed with t test, chi-square test, and Mann-Whitney U test to screen the death related factors of patients. The indexes with statistically significant differences between survival group and death group were included in the multivariate logistic regression analysis to screen the independent death risk factors of patients, and the death risk nomogram predicting model was constructed based on the results.The Bootstrap method was used to validate the death risk nomogram predicting model internally. The predictive value of the nomogram model for predicting death risk of patients was detected by drawing calibration graph and calculating concordance index, and the death risk scores of 231 patients were acquired according to the death risk nomogram model. The receiver′s operating characteristic (ROC) curve was drawn, and the optimal threshold and the sensitivity and specificity of optimal threshold in the ROC curve and the area under the curve were calculated. Results:(1) There were statistically significant differences in burn index, ABSI on admission, severity of inhalation injury, total burn area, full-thickness burn area, rehydration coefficient at the first 24 h after injury, use of ventilator, use of CRRT, and Baux score on admission of patients between the two groups ( Z=-7.696, -7.031, χ2=18.304, 63.065, 23.300, 13.073, 34.240, 59.586, t=-7.536, P<0.01). (2) There were statistically significant differences in death, incidence and severity of inhalation injury, total burn area, and burn index of patients between with ventilator group and without ventilator group ( χ2=34.240, 17.394, 25.479, Z=-6.557, -7.049, P<0.01). (3) There were statistically significant differences in death, total burn area, and burn index of patients between with CRRT group and without CRRT group ( χ2=62.982, Z= -47.421, -6.678, P<0.01). (4) The use of ventilator, use of CRRT, and burn index were independent risk factors for the death of extremely severe burn patients (odds ratio=3.277, 5.587, 1.067, 95% confidence interval=1.073-10.008, 2.384-13.093, 1.038-1.096, P<0.05 or P<0.01). (5) The initial concordance index of nomogram predicting model was 0.90 and the corrected concordance index was 0.89. The concordance indexes before and after correction were higher and similar, which showed that the nomogram had good concordance and predictive effect. The optimum threshold of ROC curve was 0.23, the sensitivity and specificity of optimum threshold were 86.0% and 80.0%, respectively, and the area under ROC curve was 0.90 (95% confidence interval=0.86-0.94, P<0.01). Conclusions:Severe burns and damage and/or failure of organ are the main death causes of extremely severe burn patients. The death risk nomogram predicting model established on the basis of use of ventilator, use of CRRT, and burn index have good predictive ability for death of extremely severe burn patients.

The oral cavity of each person is home to hundreds of bacterial species.While taxa for oral diseases have been studied using culture-based characterization as well as amplicon sequencing,metagenomic and genomic information remains scarce compared to the fecal microbiome.Here,using metagenomic shotgun data for 3346 oral metagenomic samples together with 808 published samples,we obtain 56,213 metagenome-assembled genomes(MAGs),and more than 64％of the 3589 species-level genome bins(SGBs)contain no publicly available genomes.The resulting genome collection is representative of samples around the world and contains many genomes from candi-date phyla radiation(CPR)that lack monoculture.Also,it enables the discovery of new taxa such as a genus Candidatus Bgiplasma within the family Acholeplasmataceae.Large-scale metagenomic data from massive samples also allow the assembly of strains from important oral taxa such as Por-phyromonas and Neisseria.The oral microbes encode genes that could potentially metabolize drugs.Apart from these findings,a strongly male-enriched Campylobacter species was identified.Oral sam-ples would be more user-friendly collected than fecal samples and have the potential for disease diagnosis.Thus,these data lay down a genomic framework for future inquiries of the human oral microbiome.