Objective To develop a diagnostic test based on indirect immunofluorescence assay(IFA) to detect special antibodies in the serum of SARS patients, thus to provide a reference material for confirmation of the clinical diagnosis of SARS. Methods SARS coronavirus GZ01 and BJ01 strains isolated in our laboratory were used to infect Vero E6 cells. When CPE reached 25%, cells were trypsinized and transferred to 10 well slides in a quantity of 40?l with a cell density of 2?10 7 /ml. After 4 hour incubation at 37℃,the slides were fixed with acetone, and IFA was used to detect antibodies in serum samples, which were obtained from 154 SARS patients and 14 non SARS patients with respiratory disease, as well as 100 healthy volunteers. Results IFA method for detecting antibodies of SARS coronavirus was developed. Sera from one hundred and forty two out of 154 clinically diagnosed patients were IFA positive, with a positive rate of 92 3%. Sera from 14 non SARS patients with respiratory disease and 100 healthy persons were all IFA negative. Conclusion The IFA method we developed was sensitive and specific in detecting SARS antibodies in serum, and was a reliable test for laboratory diagnosis of SARS coronavirus.

Air Pollutants, Occupational ; analysis ; Carbon Tetrachloride ; analysis ; Chromatography, Gas ; Environmental Monitoring ; methods ; Workplace

Objective To explore the behavior characteristics in children with benign epilepsy combined with centro-temporal spikes(BECTS).Methods Eighty-two children with BECTS aged 2.5-3.0(2.65?2.31)years old,51 male,31 female,who were free of mental retardation assessed with Gesell developmental schedules,untreated with antiepileptic drugs,and were investigated 15 days after the latest seizure.Eighty-two healthy children with sex and age matched to the cases,53 male,29 female,aged 2.5-3.0(2.6?0.4)years old.The behavior characteristics of infants in BECTS group and control group were assessed with CBCL,including 6 behavior factors which were sleep problem,social flinches,depression,physical aspect,attacking,act of sabotage and the infants-middle school student social ability scale.Results The total scores of behavior characteristics and the scores of depression,sleep problem,attacking and act of sabotage in BECTS group were all higher than those in control group,the differences were statistically significant.However,scores of social flinches,and physical aspect in BECTS group had no significant differences compared with those of control group.There were no significant difference of social adaptive component between the BECTS and control group.Conclusions Children with BECTS have behavior disorders to some extent,but their social adaptive capacity are the same as normal children.

Air Pollutants ; analysis ; Chromatography, High Pressure Liquid ; methods ; Paraquat ; analysis ; Workplace

A quail-origin subtype of the influenza virus was isolated from a human-infecting H7N9 subtype of the avian influenza virus found in a live poultry market and was given the name A/Quail/Hangzhou/1/ 2013 (H9N2). We analyzed the whole genome of this virus and its biologic characteristics. Sequence analyses suggested that the: HA and NS genes belonged to a CK/BJ/1/94-like lineage; NA, NP, PA and PB1 genes belonged to a SH/F/98-like lineage; M and PB2 genes belonged to a G1-like lineage. Analyses of key amino acids showed that the cleavage site in HA protein was PSRSSR ↓ GL, and that the HA protein had a human receptor-binding site with Leu226. Deletion of amino acids 69 - 73 was detected in the stalk of NA protein, the M2 protein had an Asn31 mutation, and the NS1 protein had two mutations at Ser42, Ala149. The intravenous pathogenicity of this virus was 0.36. A study in chickens suggested that all inoculated birds shed the virus from the trachea and cloaca on the third day post-infection (p. i. ) until 11 days. All chickens that had direct contact shed the virus on the second day p. i. until 8 days. Results of virus reisolation suggested that lung and tracheal tissues could shed the virus in 5 days, whereas the other organs could shed the virus in 3 days. These results suggest that this virus strain is H9N2 subtype LPAIV, whose lineage is prevalent in mainland China. This research provides evidence on how to monitor and prevent the H9N2 subtype of the avian influenza virus.
Animals ; Chick Embryo ; Chickens ; China ; Genotype ; Influenza A Virus, H9N2 Subtype ; classification ; genetics ; isolation & purification ; Influenza in Birds ; virology ; Phylogeny ; Quail ; virology

Objective To explore the clinical value of 3.0T nuclear magnetic resonance susceptibility weighted imaging in diagnosis of neonatal hypoxic ischemic encephalopathy,thus to provide guidance for the clinical.Methods From December 10,2015 to December 10,2017,100 cases of neonatal hypoxic ischemic encephalopathy (observation group) and 100 cases of normal newborns in Lishui People's Hospital during the same period accepted health examination (control group) were selected in the research.All the cases received 3.0T magnetic resonance susceptibility weighted imaging,the diagnostic value of 3.0T MR susceptibility weighted imaging was observed.Results The ADC value of the observation group[(0.00 1 13 ± 0.000 01)mm2/s] was significantly lower than that in the control group[(0.001 98 ±0.000 02)mm2/s] (P ＜0.05).The neonatal ADC value of the mild group[(0.001 21 ± 0.000 01)mm2/s] was significantly higher than that in the moderate group[(0.001 12 ± 0.000 02)mm2/s] and the severe group[(0.001 02 ± 0.000 03)mm2/s] (P ＜ 0.05).ADC value was positively correlated with neonatal hypoxic ischemic encephalopathy,namely,the lower the ADC value,the more serious the neonatal hypoxic ischemic encephalopathy.The fractional anisotropy value and relative anisotropy value of newborn babies in the observation group were significantly higher than those in the control group (all P ＜ 0.05).Conclusion For neonatal hypoxic ischemic encephalopathy patients,3.0T nuclear magnetic resonance susceptibility weighted imaging in the diagnosis is feasible,it can help the clinician to analyze the disease,and has positive significance to carry out the treatment.

Objective To investigate the effect of incomplete cryoablation on the biological behavior of prostatic cancer RM-1 cells and its mechanism.Methods RM-1 cells of prostatic cancer were placed in -20℃ icebox to be frozen for 5 min.After the recovery of the cell state,the RM-1 cells were frozen again for 10 min and 15 min successively.After culture for one day,the cellular morphology was microscopically examined.A total of 20 C57/BL mice were used to establish the tumor-bearing models,which were randomly and equally divided into the control group and the incomplete cryoablation group with 10 mice in each group.At scheduled time points the tumor lesion size was measured for all mice.The mice were sacrificed at 14 days,the lung tissues were collected and were stained with lE;the numbers of metastatic lesions in the lung were calculated.Transwell assay was used to test the cell migration and invasion,immuno-blotting method was adopted to determine the epithelial-mesenchymal transition-related (EMT-related) protein expression level,and the enzyme-linked immunosorbent assay (ELISA) was employed to check the secretion volume of transforming growth factor-beta (TGF-β).Results After incomplete cryoablation,RM-1 cells became disorderly arranged,their morphology was changed,and antenna structure might be formed.At 3 and 7 days after cryoablation,the tumor size in the incomplete cryoablation group was slightly smaller than that in the control group,but only the difference at 7 days after cryoablation was statistically significant between the two groups (P=0.019).At 10 and 14 days after cryoablation,the tumor volume of the two groups was almost equal.The pulmonary metastatic lesions in the incomplete cryoablation group were obviously much more than those in the control group (P＜0.001).Transwell assay indicated that the cell migration and invasion ability in the incomplete cryoablation group was stronger than that in the control group (P＜0.05).Immuno-blotting test revealed that,when compared with the control group,in the incomplete cryoablation group the expressions of N-cadherin,MMP-9 and Vimentin were up-regulated,while the expression of E-cadherin was downregulated.ELISA test showed that increased secretion of TGF-β was observed in the incomplete cryoablation group.Conclusion Incomplete cryoablation can enhance the migration and invasion ability of RM-1 cells,increase the number of pulmonary metastatic lesions in tumor-bearing mice,and affect the EMT-related protein expression level.

BACKGROUND: External ventricular drainage (EVD) is an important procedure for draining excessive cerebrospinal fluid (CSF) and monitoring intracranial pressure. Generally, EVD is performed in the operating room (OR) under aseptic conditions. However, in emergency circumstances, the operation may be performed in the intensive care unit (ICU) to save neuro-critical time and to avoid the unnecessary transfer of patients. In this study, we retrospectively analyzed the risk of EVD-induced CNS infections and their outcomes according to the operating place (ICU versus OR). In addition, we compared mortalities as well as hospital and ICU days between the CNS infection and non-CNS infection groups. METHODS: We reviewed medical records, laboratory data and radiographic images of patients who had received EVD operations between January, 2013 and March, 2015. RESULTS: A total of 75 patients (45 men and 30 women, mean age: 58.7 +/- 15.6 years) were enrolled in this study. An average of 1.4 catheters were used for each patient and the mean period of the indwelling catheter was 7.5 +/- 5.0 days. Twenty-six patients were included in the ICU group, and EVD-induced CNS infection had occurred in 3 (11.5%) patients. For the OR group, forty-nine patients were included and EVD-induced CNS infection had occurred in 7 (14.3%) patients. The EVD-induced CNS infection of the ICU group did not increase above that of the OR group. The ICU days and mortality rate were higher in the CNS infection group compared to the non-CNS infection group. The period of the indwelling EVD catheter and the number of inserted EVD catheters were both higher in the CNS infection group. CONCLUSIONS: If the aseptic protocols and barrier precautions are strictly kept, EVD in the ICU does not have a higher risk of CNS infections compared to the OR. In addition, EVD in the ICU can decrease the hospital and ICU days by saving neuro-critical time and avoiding the unnecessary transfer of patients. Therefore, when neurosurgeons decide upon the operating place for EVD, they should consider the benefits of ICU operation and be cautious of EVD-induced CNS infection.
Catheter-Related Infections ; Catheters ; Catheters, Indwelling ; Cerebrospinal Fluid ; Drainage* ; Emergencies ; Female ; Humans ; Intensive Care Units* ; Critical Care* ; Intracranial Pressure ; Male ; Medical Records ; Mortality ; Operating Rooms* ; Retrospective Studies ; Ventriculostomy

In traditional Chinese medicine(TCM),abnormal and diseased conditions have been defined as Zheng Hou, a unique disease definition system in the context of holism. For over 3000 years the main clinical treatment method for TCM therapeutics has been so called Fang-ji, a TCM medicinal formula usually composed of several herbs and medical materials. The compositions of Fang-ji are based on the clinical practice under the guidelines of "bian-zheng-lun-zhi" and the principles of "Jun-chen-zuo-shi". Each Zheng is treated with a correspondingly-individualized Fang-ji.The modern approach to the study of Fang-ji pharmacology,however,has been focusing on the isolation and identification of individual active components for cellular and molecular targets. Although this approach has led to the development of many new monomers purified from Fang-ji as new drugs widely used in clinical practice such as the an-timalarial artemsinin,which has earned a Nobel Prize,the pharmacological bases of these purified effective monomers or active components have lost the TCM characteristics and are far different from the phar-macological theory and clinical applications of Fang-ji,in terms of the principles of"bian-zheng-lun-zhi"and "Jun-chen-zuo-shi". Here we introduce the emerging pharmacophenophenics as a systematical paradigm for the pharmacological study of Fang-ji.Pharmacophenomics studies the orchestrated multi-target pharmacology of combination therapy.With well-defined molecular mechanisms of Zheng Hou at the level of multi-omics and a suite of new phenomics technologies and platforms, the pharmacophe-nomics may be used to characterize the drug-response phenome of Fang-ji and to identify the corre-sponding multiple therapeutic targets according to the TCM theory of Jun-chen-zuo-shi.Pharmacophe-nomic study of Fang-ji will also lay a theoretical foundation for the new science of precision medicine.

Objective:To investigate the expressions of miR-20a-5p and lysine (K) demethylase 6B (KDM6B) in osteosarcoma tissues and the effects of miR-20a-5p targeting KDM6B on the proliferation, migration and invasion of osteosarcoma cells and tumor growth.Methods:The clinicopathological and paracancerous tissues of 20 patients with osteosarcoma admitted to the First Affiliated Hospital of Chinese Medical University from January 2017 to March 2019 were collected. Quantitative real-time PCR (qRT-PCR) was used to detect the expression levels of miR-20a-5p and KDM6B mRNA in tissues. The osteosarcoma MG63 cells were divided into control group, mimic NC group, miR-20a-5p mimic group, and NC+ empty vector group, miR-20a-5p+ empty vector group, miR-20a-5p+ KDM6B group. The expression levels of miR-20a-5p and KDM6B mRNA of all groups were detected by qRT-PCR. Western blotting was used to detect the expression level of KDM6B. CCK-8 assay, cell scratch test and Transwell test were used to detect cell proliferation, migration and invasion ability. According to the random number table method, nude mice were divided into NC+ empty vector group, miR-20a-5p+ empty vector group and miR-20a-5p+ KDM6B group, with 5 mice in each group. Tumor growth ability was detected by tumor xenograft nude mouse models.Results:The relative expression level of miR-20a-5p mRNA in osteosarcoma tissues was 0.55±0.27, and that in paracancerous tissues was 1.22±0.28, with a statistically significant difference ( t=7.701, P<0.001). The relative expression level of KDM6B mRNA in osteosarcoma tissues was 1.66±0.19, and that in paracancerous tissues was 1.00±0.15, with a statistically significant difference ( t=12.219, P<0.001). After transfection of miR-20a-5p, KDM6B mRNA and protein expression levels decreased with the increase of miR-20a-5p expression level. After miR-20a-5p transfection for 48 h, the cell proliferation abilities of the blank control group, mimic NC group and miR-20a-5p mimic group were 0.83±0.04, 0.81±0.03 and 0.52±0.01 ( F=89.655, P<0.001), compared with the blank control group and mimic NC group, the cell proliferation ability was significantly inhibited in the miR-20a-5p mimic group (both P<0.001). The cell proliferation abilities of NC+ empty vector group, miR-20a-5p+ empty vector group and miR-20a-5p+ KDM6B group were 0.83±0.05, 0.52±0.01 and 0.67±0.05 ( F=43.919, P<0.001), compared with the NC+ empty vector group, the cell proliferation ability was significantly inhibited in the miR-20a-5p+ empty vector group ( P<0.001); compared with the miR-20a-5p+ empty vector group, the cell proliferation ability of miR-20a-5p+ KDM6B group increased significantly ( P<0.001). The scratch healing rates of the blank control group, mimic NC group and miR-20a-5p mimic group were (32.51±2.73)%, (30.26±3.22)% and (13.52±1.77)% ( F=46.314, P<0.001), compared with the control group and the mimic NC group, the scratch healing rate of the miR-20a-5p mimic group was significantly decreased (both P<0.001). The scratch healing rates of NC+ empty vector group, miR-20a-5p+ empty vector group and miR-20a-5p+ KDM6B group were (31.34±3.11)%, (12.15±1.64)% and (28.93±2.89)% ( F=47.511, P<0.001), compared with the NC+ empty vector group, the scratch healing rate of the miR-20a-5p+ empty vector group was significantly decreased ( P<0.001); compared with the miR-20a-5p+ empty vector group, the scratch healing rate of miR-20a-5p+ KDM6B group was significantly increased ( P=0.001). The numbers of transmembrane cells in the blank control group, mimic NC group and miR-20a-5p mimic group were 114±16, 108±11 and 42±6 ( F=36.282, P<0.001), compared with the control group and mimic NC group, the number of transmembrane cells of the miR-20a-5p mimic group was significantly decreased (both P<0.001). The numbers of transmembrane cells in the NC+ empty vector group, miR-20a-5p+ empty vector group and miR-20a-5p+ KDM6B group was 143±11, 39±4 and 139±12 ( F=112.120, P<0.001), compared with the NC+ empty vector group, the number of transmembrane cells of the miR-20a-5p+ empty vector group was significantly decreased ( P<0.001); compared with the miR-20a-5p+ empty vector group, the number of transmembrane cells of the miR-20a-5p+ KDM6B group was increased significantly ( P<0.001). The tumor volumes of mice for 21 d in the NC+ empty vector group, miR-20a-5p+ empty vector group and miR-20a-5p+ KDM6B group were (1 667.50±250.40) mm 3, (129.20±21.00) mm 3 and (775.41±77.51) mm 3 respectively, with a statistically significant difference ( F=77.651, P<0.001). The tumor weights of the 3 groups were (1.35±0.18) g, (0.12±0.01) g and (0.61±0.03) g respectively, with a statistically significant difference ( F=104.191, P<0.001). Conclusion:The expression of miR-20a-5p is significantly decreased in osteosarcoma tissues, and the expression of KDM6B is significantly increased in osteosarcoma tissues. Overexpression of miR-20a-5p may inhibit the proliferation, migration and invasion of osteosarcoma cells and tumor growth by targeting to reduce the expression of KDM6B.