Purpose] To detect respiratory virus antigen by direct immunofluorescence method and provide evidence for early diagnosis of children with viral infection of the respiratory tract disease.[Methods] Select 398 cases of respiratory tract infection in hospitalized children, pharynx in exfoliated cells in respiratory syncytial virus by direct immunofluorescence(RSV), adenovirus(ADV), influenza virus type A(IFVA),influenza virus type B(IFVB), parainfluenza virus type 1(PIV1), parainfluenza virus type 2(PIV2) and parainflue-Nza virus type 3(PIV3) of 7 common viral antigens were detected, and carried on statistics analysis.[Results] In 398 cases of children with respiratory tract infection, 91 cases were positive, the positive rate was 22.86%. The highest for respiratory syncytial virus in 37 cases(40.66%) followed by parainfluenza virus type 3 in 19 cases(20.88%), influenza virus type B in 14 cases(15.38%); The positive rates of acute tonsillitis, capillary bronchitis, pneumonia, acute bronchitis and upper respiratory were 46.67%(14/30),40.00%(20/50), 26.67%(40/150), 18.42%(7/38), 7.69%(10/130) respectively; With the growth of age, the respiratory tract virus infection rate gradually decreased(P<0.05), one year old the viral infection rate was 76.92%;it occurred in the winter season of respiratory tract infection in children .[Conclusion] The main virus in children with acute respiratory infection in the local area is RSV. Virus infection decreases with the increase of age, treatment should be careful in the use of antibiotics.

Objective To compare the compression properties of articular cartilage and polyinyl alcohol hydro-gel(PVA-H)as artificial carlage.Method Unconfined compression tests were conducted on articular card-lage and PVA-H,including stress-strain tests,creep tests and stress relaxation tests.The stress-strain rela-tionship of articular cartilage and PVA-H were measured.Result The compression modulus of articular card-lage was higher than that of PVA hydrogel.The average compression modulus of articular cartilage and PVA hydrogel was(3.6492±0.6199)Mpa and(1.5951±0.1469)Mpa,respectively.Condusions The experimental results revealed the differences existed between articular cartilage and PVA-H and this would be useful to fur-ther improve the mechanical properties of artificial cartilage.

Objective To determine the relationship between stressors and mental health among postgraduates in military medical universities. Methods This article analyzed population variable, mental health, the correlation of personality, coping styles and pressure based on Military Academy Graduate Student Stress Scale, Symptom Checklist-90, Simplified Coping Style Questionnaire and Eysenck Personality Questionnaire. Independent sample t-test and single factor analysis variance were used to compare the feel-ing of pressure differences among demographic variables groups and mental health differences between high and low pressure group. Pearson product-moment correlation and multiple step-wise regression analysis were performed to explore the relationship of personality, coping styles and feeling of pressure. Results ①There was significant difference on stressors among postgraduates in military medical universities (F=77.397, P=0.000), and LSD test showed that academic pressure> distribution pressure>economic pressure>marriage pressure>family pressure (P<0.05). ②Except phobic factors, SCL-90 total score and each dimension points of high stress group were significantly higher than that of low stress group (P<0.05).③Negative coping style and EPQ multiple dimensions significantly related to feeling of pressure values in post-graduate students. Neuroticism, lie and extroversion entered the regression equation (F=16.959,P=0.001) and together explained 19.0% of the total variance. Conclusion The pressure from teachers, academic, graduation distribution and application for jobs are the main source of pressure. And the students with high pressure sensitivity get poor mental health and tend to have neurotic personality traits and negative coping styles.

Objective:To elucidate the effect of hsa-microRNA-218(hsa-mir-218)on exogenous granulysin (GLS) expression in 293T cells.Methods:Total RNA was extracted from THP-1 cells induced by phorbol 12-myristate 13-acetatefor (PMA), and GLS gene was amplified by RT-PCR, and then cloned into pDsRed-Express-C1 to construct the GLS expression vector pDsRed-GLS.Then 293T cells were co-transfected with pDsRed-GLS and pGenesil-mir-218 (pGenesil-mir-control) and laser confocal microscopy was per-formed 36 h later to detect their co-expression .Total RNA and protein were extracted 48 h post transfection , and RT-PCR and Western blot were performed to detect the effect of hsa-mir-218 on exogenous GLS expression .Results:The GLS expression vector pDsRed-GLS was constructed successfully and laser confocal microscopy indicated that it was co -expressed with the interference vector .Compared with that of cells transfected with pGenesil-mir-control, Western blot showed a markedly decrease of GLS protein expression (50%) in the cells transfected with pGenesil-mir-218.However, GLS mRNA expression remained unchanged .Conclusion: hsa-mir-218 nega-tively regulates GLS expression at a post-transcriptional level , and this provides an experimental basis for future study of mechanism of GLS expression regulated by mir-218 .

OBJECTIVETo establish an UPLC-MS/MS method for simultaneous determination protocatechuic acid, isoorientin and scutellarin in rat tissues after a single intravenous administration of Compound Hongcao for injection, and investigate the distribution character of the three compounds.

METHODThree compounds were simultaneously determined by UPLC with Waters Acquity UPLC BEH C18 column. The mobile phase, consisting of acetonitrile and 0.1% aqueous formic acid, was programmed as a linear gradient. The flow rate was 0.35 mL x min(-1). The compounds were ionized in the electrospray ionization ion source of the mass spectrometer and detected in the multiple reaction model. The tissue samples were homogenated and the suspension were extracted with methanol for further use.

RESULTThe relationship between the concentration and the peak areas of the three compounds were all linear (r > 0. 99). The precisions, accuracy, extract recoveries and stability of the analytes meet the requirements. The method has been successfully applied to tissue distribution studies of the three compounds. The present study demonstrates that the higher tissues concentration of three components were obtained in kidney, lung and heart after a single intravenous administration of ones.

CONCLUSIONThe present study demonstrates that the three compounds have unequal distribution character. It was showed that the three compounds were mainly distributed in abundant blood-supply tissues such as kidney, lung and heart. It was also found that protocatechuic acid, isoorientin and scutellarin exceretion rapidly and have no long-term accumulation. The method was shown to be effective, convenient, and suitable for simultaneous study the distribution of the three compounds in rat.

Animals ; Apigenin ; analysis ; Chromatography, Liquid ; methods ; Drugs, Chinese Herbal ; pharmacokinetics ; Female ; Glucuronates ; analysis ; Hydroxybenzoates ; analysis ; Injections, Intravenous ; Luteolin ; analysis ; Male ; Rats ; Rats, Wistar ; Tandem Mass Spectrometry ; methods ; Tissue Distribution

OBJECTIVETo establish a UPLC-MS/MS analysical method for simultaneous determination of concentrations of isoorientin, scutellarin and cynaroside in rat plasma and to study their pharmacokinetic characteristics after intravenous injection of 3 doses of Fufang Hongcao in rats.

METHODAcidified plasma samples were precipitated for protein with methanol. Waters Acquity BEH C18 column was adopted for spectrum, with mobile phase as 0. 1% formic acid acetonitrile-0. 1% formic acid-water gradient elution. Detection was carried out by the multiple reaction monitoring (MRM) positive ion mode with ESI ionization source.

RESULTThree flavonoids show a good linear relationship, with the extraction recovery ranging between 78.56% and 101.91% and a high intra-and inter-day precisions and accuracy. The MRT of the three flavonoids were all lower than 22 min in rats.

CONCLUSIONThe above men tioned method is so specific, rapid, sensitive that it is suitable for pharmacokinetic studies of Fufang Hongcao injection in rats.

Animals ; Apigenin ; blood ; pharmacokinetics ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; pharmacokinetics ; Female ; Glucosides ; blood ; pharmacokinetics ; Glucuronates ; blood ; pharmacokinetics ; Luteolin ; blood ; pharmacokinetics ; Male ; Rats ; Tandem Mass Spectrometry ; methods ; Time Factors

OBJECTIVETo identify the compounds in herb of Polygonum orientale by ultra-performance liquid chromatography with PDA and electrospray ionization tandem mass spectrometry (UPLC-PDA-ESI/MS).

METHODThe chromatographic separation was carried out at 45 degrees C on a BEH C18 column (2.1 mm x 100 mm, 1.7 microm) eluted in the gradient program. The mobile phase consisted of acetonitrile and water containing 0.1% formic acid. The detecion of PDA was set between 190 nm and 400 nm. The mass spectra was obtain by Waters ACQUITY triple quadrupole tandem mass spectrometer in negative ion mode with ion spray voltage 3 kV. The temperature of the source and desolvation were set at 120 degrees C and 350 degrees C, separately. Nitrogen was used as the desolvation gas (750 L h(-1)) and cone gas (50 L h(-1)). For collision induced dissociation (CID), argon was used as the collision gas at a flow rate of 0.18 mL min(-1). The mass range recorde, m/z 100-1000.

RESULTOne phenolic acid, eleven flavonoids were well separated and identified by the method.

CONCLUSIONThe method is simple and rapid for the compound identification in the herb of P. orientale.

Chromatography, High Pressure Liquid ; methods ; Plant Extracts ; analysis ; Polygonum ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods

A simple and selective ultra performance liquid chromatography-electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS) assay was developed for the determination of the human plasma protein binding of four bioactive flavonoids (such as orientin and vitexin) in Polygonum orientale. Protein precipitation was used for sample preparation. Equilibrium dialysis technique was applied to determine the plasma protein binding under physiological conditions. The separation was achieved through a Waters C18 column with a mobile phase composed of 0.1%formic acid in acetonitrile and 0.1%aqueous formic acid using step gradient elution at a flow rate of 0.35 mL/min. A Waters ACQUITY?TQD system was operated under the multiple reaction monitoring (MRM) mode of positive electrospray ionization. All of the recovery, precision, accuracy and stability of the method met the requirements. Good correlations (r40.99) of the four compounds were found, which suggested that these compounds can be simultaneously determined with acceptable accuracy. Results showed that the plasma protein bindings of the four bioactive flavonoids were in the range of 74-89% over the six concentrations studied. The binding parameters containing protein binding affinity, protein binding dissociation constant, and protein binding site were studied. The maximum ability to bind with protein was also determined in the assay in order to understand the drug-protein binding of each compound better.

Objective:To analyze the epidemiological and etiological characteristics of a dengue fever outbreak in Hunan province in 2018.Methods:Real-time PCR assay was performed for the laboratory diagnosis of 8 suspected dengue fever cases. Etiological surveillance was performed in 186 suspected dengue fever cases and fever cases who had close contacts with dengue fever patients. C6/36 cells was used for the virus isolation from acute phase serum. By sequencing the full length of E genes of 15 dengue virus strains, phylogenetic analysis was performed based on the sequences obtained, including reference sequences from the NCBI GenBank database, the serotypes and gene subtypes of the virus were analyzed to trace the possible source of transmission. An emergency monitoring of vector density and a retrospective survey of sero-epidemiology in healthy population were conducted in the epidemic area.Results:In the serum samples of 8 suspected patients, 6 were dengue virus RNA positive, and 4 were NS1 antigen positive. In 186 suspected patients, 96 were dengue virus nucleic acid, NS1 antigen or antibody positive in etiological test. A total of 64 dengue virus strains were isolated. The phylogenetic analysis showed that all the dengue virus strains belonged to type 2, which might be from Guangdong or Zhejiang provinces. The Bretub index was up to 65, indicating an extremely high risk of transmission. The positive rate of the dengue virus IgG antibody was 0.53%(2/377) in retrospective survey of 377 healthy people.Conclusion:The field epidemiologic and the molecular genetics analyses showed the outbreak of dengue fever in Hunan in 2018 was caused by imported cases and dengue virus 2.

Objective: To explore the value and disadvantage of micro-mirror in the intracranial aneurysm surgery. Methods: Micro-mirror was used to assist microsurgical clipping to 36 intracranial aneurysms in 31 cases, of which 3 were carotid-ophthalmic artery aneurysms, 3 anterior choroidal artery aneurysm, 11 were posterior communicating artery aneurysms, 7 were middle cerebral artery aneurysms, 10 anterior communicating artery or anterior cerebral artery aneurysms, and the others were a posterior cerebral artery aneurysm and a posterior inferior cerebellar artery aneurysm. The micro-mirror was used before and after clipping to observe the anatomic features of necks hidden behind and medial to aneurysms, to visualize surrounding neurovascular structures, and to verify the optimal clipping position. Intraoperative indocyanine green fluorescein angiography confirmed the success of sufficient clipping. Results: All aneurysms were clipped successfully. The parent arteries were occluded temporarily in 26 cases, and 9 aneurysms ruptured during the operation. Postoperative follow-up lasted from 2 weeks to 1 year. After operations digital subtraction angiography (DSA) were repeated in 10 cases, computed tomography angiography(CTA) in 21 cases, and no insufficient clipping or occlusion of parent arteries were revealed. Conclusions Micro-mirror assisting microsurgical clipping to the intracranial aneurysm is conducive to accurate clipping, and is a convenient and practical operation.