OBJECTIVE To establish SYBR Green Ⅰ fluorescent quantitative PCR assay for the detection of West Nile virus(WNV),which could be used for early laboratory diagnosis.METHODS A fragment of WNV gene was amplified by PCR,then cloned into pMD-18 T vector.The combinant plasmid was sequenced and analyzed by means of BLAST program,and used as the positive DNA in place of WNV.The SYBR GreenⅠfluorescent quantitative PCR assay was established based on positive plasmid.The sensitivity and specificity of the assay were performed.RESULTS The combinant plasmid was confirmed by sequencing and the fragment belonged to WNV.Ten copies of WNV RNA were detected by SYBR GreenⅠfluorescent quantitative RT-PCR assay.Results of the other members of Flaviviridae were negative,which indicated this assay was specific for WNV.CONCLUSIONS SYBR GreenⅠfluorescent quantitative PCR assay established in this study is highly sensitive and specific,and so it can be used for early diagnosis of WNV infection.

Objective To understand the risk factors for poor prognosis of severe adenovirus pneumonia (SAP)in children,and provide guidance for clinical diagnosis and prognosis.Methods Clinical data of 91 hospitalized children who diagnosed with SAP in Chongqing Children’s Hospital of Chongqing Medical University between January 2012 and January 2014 were analyzed retrospectively.Results Of 91 SAP children,23 (25.27%)had poor prognosis. Univariate analysis showed that risk factors for poor prognosis of SAP were age of onset,congenital heart disease and other serious underlying diseases,mechanical ventilation therapy,acute respiratory distress syndrome (ARDS), atelectasis and other serious radiological changes,and emergence of two and more extra-pulmonary complications (all P <0.05).Multivariate regression analysis showed that congenital heart disease and other serious underlying diseases,and emergence of two and more extra-pulmonary complications were independent risk factors for poor prognosis of SAP (all P <0.05).Conclusion Congenital heart disease and other serious underlying diseases,emer-gence of two and more extra-pulmonary complications are independent risk factors for poor prognosis of SAP,active intervention should be conducted during the process of clinical diagnosis and treatment,so as to improve the prognosis.

Objective To discuss the reasons and the best treatment of the encephalocele during severe head injury. Method Retrospective analysis 26 cases of the clinical data of patients with acute encephalo-cele during severe head injury used bilateral craniotomy method. Result Good recovery in 4 patients, mod-erate disability in 5 patients, severe disability in 2 patients, vegetative state in 3 patients, death in 12 pa-tients. Conclusion Delayed intracranial hematoma and acute diffuse brain swelling are the main reasons of craniotomy brain injury in acute encephalocele, and using bilateral craniotomy can significantly increase the survival rate and quality of life.

Transforming technology for semi-synthesized isocorydione from the natural product ofisocorydine was studied. The factors affecting on the reaction yield were investigated. UV spectrophotometry was used to indicate the semi-synthesized yield of isocorydione. The optimum reaction conditions were determined as following: reacting for 12 h in the solution of sodium dihydrogen phosphate at pH 10, the temperature was 25 degrees C and the ratio of isocorydine to Fremy's radical was 1 : 2. Under the optimum conditions, the yield could reach up to 50.0%. The molecular structure of isocorydione was elucidated by X-ray single-crystal diffraction analysis for the first time.

Objective To investigate the effects of sivelestat sodium on early inflammatory reaction in rats with smoke inhalation injury. Methods Forty SPF male SD rats were randomly divided into 5 groups:normal control group (A), injury group (B), smoke inhalation treated with 10 mg/kg sivelestat sodium group (C), smoke inhalation treated with 20 mg/kg sivelestat sodium group (D) and smoke inhalation treated with 30 mg/kg sivelestat sodium group (E), 8 rats for each group. After smoke inhalation injury model was established, the treatment groups were intraperitoneally injected sivelestat sodium 10 mg/kg, 20 mg/kg and 30 mg/kg separately. B group was treated with the same volume of physiological saline. After 24 hours,ELISA was used for detecting serum contents of neutrophil elastase (NE), myeloperoxidase (MPO), tumor necrosis factor-α(TNF-α) and interleukin-6 (IL-6) in five groups. Meanwhile the water content of lung tissue was measured, and the pathological changes were observed by HE staining. The thickness of alveolar septum was measured and compared between groups. Results Compared with control group, the serum levels of NE, MPO, IL-6, TNF-α, water content of the lung tissue and thickness of alveolar septum were significantly higher in other four groups (P<0.05). Compared with injury group, the serum levels of NE, MPO, IL-6, TNF-α, water content of the lung tissue and thickness of alveolar septum were significantly lower in treatment groups (P<0.05). Compared with 20 mg/kg treatment group and 30 mg/kg treatment group, the serum levels of NE, MPO, IL-6, TNF-α, water content of the lung tissue and thickness of alveolar septum were significantly lower in 10 mg/kg treatment group (P<0.05). Conclusion The result shows that sivelestat sodium can reduce the early inflammatory reaction of rats with smoke inhalation injury and attenuates the lung edema. In this experiment, the treatment effect of 10 mg/kg sivelestat sodium is better than other treatment doses.

Objective To explore if bacillus bifidus relieve CPFX-induced testosterone reduction in mouse testes.Methods Twenty-four male mices were divided into 4 groups, then administered saline for 6 days (Sal6 group), CPFX for 6 days (A6 group), CPFX for 6 days followed by bifidobacteria treatment for the next 6 days (A6+P6 group), CPFX for 6 days and then saline for the next 6 days (A6+Sal6 group).We detected serum levels of testosterone by RIA, as well as levels of steroidogenic enzymes mRNA [cholesterol side-chain cleavage enzyme (P450scc) and steroidogenic acute regulatory protein (StAR)] and NF-E2-related factor2 (Nrf2) mRNA in testes by real-time PCR, Nrf2, heme oxygenase-1 (HO-1), and 4-hydroxy-2-nonenal (4-HNE) by Western blot and4-HNE by Immunohistochemistry.Results The A6 group had significantly lower serum testosterone levels compared with the Sal6 group (P<0.001), the A6+P6 group had significantly higher compared with the A6 (P<0.001) and A6+Sal6 groups (P<0.01).The A6 group had significantly lower StAR mRNA compared with the Sal6 group (P<0.001), the A6+P6 group had significantly higher level compared with the A6 (P<0.01) and A6+Sal6 groups (P<0.01).The A6 group had significantly lower P450scc mRNA as compared with the Sal6 group (P<0.001), the A6+P6 group had significantly higher compared with the A6 (P<0.001) and A6+Sal6 groups (P<0.05).The A6 group had significantly lower Nrf2 compared with the Sal6 group (P<0.001), the A6+P6 group had significantly higher compared with the A6(P<0.01) and A6+Sal6 groups (P<0.05).The A6 group higher 4-HNE expression compared with the Sal6 group, the A6+P6 group had significantly lower compared with the A6 (P<0.01) and A6+Sal6 groups (P<0.05).Conclusions Bifidobacteria the reduction of CPFX-induced testosterone reduction, and these effects may potentially explained by Nrf2 inflammatory signaling pathway.

Objective To study comparatively the performance of rapid plasma reagin (RPR) test in syphilis detection among pregnant women and non-pregnant women to provide reference for detecting syphilis in pregnant women.Methods The women aged 20-40 years old were selected and divided into the pregnant group and the non-pregnant group.RPR and treponema pallidum particle assay(TPPA) were simultaneously adopted to conduct the syphilis detection.The positive cases were judged by the TPPA detection results combined with the contact history,clinical symptoms and treatment situation.The results were compared with those by RPR for determining the false negative and false positive in RPR.The false negative rate and false positive rate of RPR detection results were analyzed in the two groups.Results Among 117 pregnant women,15 cases were false negative in RPR and 9 cases were false positive in RPR;among 755 non-pregnant women,there were 44 cases of false negative RPR results and 8 cases of false positive RPR results.The false negative rates in the pregnant group and non-pregnant group were 25.0% and 8.8% respectively,the difference was statistically significant(χ2=14.739,P<0.05);the false positive rates in the pregnant group and non-pregnant group were 15.7% and 3.1% respectively,the difference was statistically significant (χ2=14.722,P<0.05).Conclusion There are many factors affecting RPR for detection syphilis,pregnant women are the specific group,so higher false positive rate and false negative rate exist than non-pregnant women,the detection results should be comprehensively judged by combining with clinical symptoms and disease history,if necessary,combining with other syphilis detection method for avoiding missed diagnosis and misdiagnosis.

Objective To compare a novel mirror-overlap method with the traditional manual measurement and the subjective assessment in assessing breast symmetry after oncoplastic surgery.Methods 59 patients with breast cancer who underwent conservation/reconstruction surgery were recruited.The post-operative assessment for breast symmetry was conducted using manual measurement,subjective assessment by the patient,their family member and a nurse,and the mirror-overlap method respectively.The latter method involved using the Photoshop to assess the area differences in breast outlines and shadows created by a flashlight when bilateral images were overlapped in the axis of midline.Results The distance differences between bilateral breasts measured using manual method were negatively correlated with symmetric coefficients in the mirror-overlap method (P＜0.05).There was a positive correlation between the average scores in the subjective assessment and symmetric coefficients of mirror-overlap method (P＜0.05).Conclusion The mirror-overlap method is an easy,feasible and cheap method which collects 3D breast information and provides an effective approach to assess breast symmetry after oncoplastic surgery.

AIM: In order to bridge the gap between pharmacogenomic research and its clinical application, we propose the concept of genetic electronic identity, named "GeneFace", and developed an electronic information system which integrated "drug-gene" interactions and recommendations for personalized medicine. METHODS: Based on the self-developed Precision Medicine knowledgebase, which concludes drug directions, guidelines or important literatures with high level of evidence, we developed GeneFace with Java-based open-resource application framework Spring Boot, further developed a mobile App with cross-platform framework Uni-APP. RESULTS: The App includes six modules: genetic testing appointment, genetic knowledge introduction, individualized medication advice, medication records, Geneface interpretation, and Precision Medicine knowledgebase. By detecting the genotype of more than 300 gene loci upon first use, users import the results to form a personal "drug-gene identity card". Then scan or enter the drug name in "GeneFace", the App would automatically give corresponding medication recommendations, including: risks for possible adverse drug reactions, risks for reducing the efficacy or even ineffectiveness, and possibility for dose adjustment, etc., which increase the safety of clinical drug use. People can obtain pharmacogenomics knowledge and basic drug information in the "GeneFace" app. CONCLUSION: Development as a digital therapeutic product, the expanded application of GeneFace can rapidly promote clinical applications of basic pharmacogenomics research and significantly improve drug use safety, which creating a new model for accelerating the clinical application of personalized medicine.

Objective:To characterize the histopathological subtypes and their clinicopathological parameters of gender and onset age by common, rare and sparse primary esophageal malignant tumors (PEMT).Methods:A total of 272 437 patients with PEMT were enrolled in this study, and all of the patients were received radical surgery. The clinicopathological information of the patients was obtained from the database established by the State Key Laboratory of Esophageal Cancer Prevention & Treatment from September 1973 to December 2020, which included the clinical treatment, pathological diagnosis and follow-up information of esophagus and gastric cardia cancers. All patients were diagnosed and classified by the criteria of esophageal tumor histopathological diagnosis and classification (2019) of the World Health Organization (WHO). The esophageal tumors, which were not included in the WHO classification, were analyzed separately according to the postoperative pathological diagnosis. The χ 2 test was performed by the SPSS 25.0 software on count data, and the test standard α=0.05. Results:A total of 32 histopathological types were identified in the enrolled PEMT patients, of which 10 subtypes were not included in the WHO classification. According to the frequency, PEMT were divided into common (esophageal squamous cell carcinoma, ESCC, accounting for 97.1%), rare (esophageal adenocarcinoma, EAC, accounting for 2.3%) and sparse (mainly esophageal small cell carcinoma, malignant melanoma, etc., accounting for 0.6%). All the common, rare, and sparse types occurred predominantly in male patients, and the gender difference of rare type was most significant (EAC, male∶ female, 2.67∶1), followed with common type (ESCC, male∶ female, 1.78∶1) and sparse type (male∶ female, 1.71∶1). The common type (ESCC) mainly occurred in the middle thoracic segment (65.2%), while the rare type (EAC) mainly occurred in the lower thoracic segment (56.8%). Among the sparse type, malignant melanoma and malignant fibrous histiocytoma were both predominantly located in the lower thoracic segment (51.7%, 66.7%), and the others were mainly in the middle thoracic segment.Conclusion:ESCC is the most common type among the 32 histopathological types of PEMT, followed by EAC as the rare type, and esophageal small cell carcinoma and malignant melanoma as the major sparse type, and all of which are mainly occur in male patients. The common type of ESCC mainly occur in the middle thoracic segment, while the rare type of EAC mainly in the lower thoracic segment. The mainly sparse type of malignant melanoma and malignant fibrous histiocytoma predominately occur in the lower thoracic segment, and the remaining sparse types mainly occur in the middle thoracic segment.