Ebola virus (EBOV) and Marburg virus (MARV) belong to the family Filoviridae. Filoviruses cause severe filovirus hemorrhagic fever (FHF) in humans, with high case fatality rates, and represent potential agents for bioterrorism and biological weapons. It is necessary to keep surveillance of filoviruses, even though there is no report of their isolation and patients in China so far. To characterize MARV morphology, the Lake Victoria marburgvirus--Leiden was stained negatively and observed under a transmission electron microscope which is one of important detection methods for filoviruses in emergencies and bioterrorism. MARV showed pleomorphism, with filamentous, rod-shaped, cobra-like, spherical, and branch-shaped particles of uniform diameter but different lengths. Pleomorphism of negatively stained MARV is summarized in this article, so as to provide useful information for possible electron microscopic identification of filoviruses in China.
Animals ; Humans ; Marburg Virus Disease ; virology ; Marburgvirus ; growth & development ; ultrastructure ; Microscopy, Electron, Transmission ; Virion ; growth & development ; ultrastructure

Aged ; Aged, 80 and over ; Corynebacterium ; isolation & purification ; Cross Infection ; epidemiology ; microbiology ; Female ; Humans ; Male ; Random Amplified Polymorphic DNA Technique

OBJECTIVETo establish a localization ultrathin section method through which target cytopathic cells could be sectioned in situ.

METHODSLab-Tek Chamber slide system (177402) was selected as resin embedding mould. Cells infected with Human adenovirus type 5 (Ad5) or A/HN/SWL3/ 2009 (H1N1) influenza virus were embedded in situ as models. Target cytopathic cells were exposed by trimming, sectioned and observed under transmission electron microscope (TEM).

RESULTSTarget cells could be sectioned in situ and virus particles could be found easily on sections.

CONCLUSIONA localization ultrathin sectioning method was established and this technique could be applied in virus detection in cytopathic cells to improve TEM detection efficiency.

Adenovirus Infections, Human ; pathology ; virology ; Adenoviruses, Human ; physiology ; ultrastructure ; Cell Line ; Humans ; Influenza A Virus, H1N1 Subtype ; physiology ; ultrastructure ; Influenza, Human ; pathology ; virology ; Microscopy, Electron, Transmission ; Microtomy ; methods

The systematic position of Fritillaria hupehensis has been in dispute. Phylogentic analyses were conducted on sequences of ITS, rpl16, matK sequences for species of F. hupehensis and allies. Lilium davidii was designed as outgroup. The analyses were performed using MP and ML methods. Conclusions could be achieved as follow. The topologies of MP and ML are consistent. The samples of F. hepehensis from different places form a supported clade with a strong bootstrap. And then form a strongly supported clade with F. anhuiensis, F. monantha. The results suggests that although F. hupehensis has a closet relation with the two ones, it exists some difference.
DNA, Plant ; chemistry ; genetics ; DNA, Ribosomal Spacer ; genetics ; Endoribonucleases ; genetics ; Fritillaria ; classification ; genetics ; Molecular Sequence Data ; Nucleotidyltransferases ; genetics ; Phylogeny ; Plant Leaves ; genetics ; Ribosomal Proteins ; genetics ; Sequence Analysis, DNA ; Species Specificity

Objective To study the recombination type of B/C genotype in hepatitis B virus.Methods The PCR was applied to amplifed the whole genes of HBV through the serums of four chronic HBV carriers who come from Jinghong distict,Yunnan province.The whole HBV genomes were ligated with pMD18-T vector and trasformed to E.coli JM109.After the positive colones were picked up,the HBV genotypes and recombinated sites were discoved through sequenced the acquired positive colones.Results All the acquired sixteen HBV sequences from the four HBV carriers were genotype B which were combinated with genotype C in some region.There are two ways of the combinations.For the first one,a 496 bp fragment from genotype C taked place the genotype B at the place of nt1825 to nt2320 of precore C/C region.For the second way,a 695 bp fragment of genotype B taked place at the both sites of nt822 to nt1020 of P gene region and nt1825 to nt2320 of precore C/C region.Conclusion A new recombination type of B/C genotype in hepatitis B virus was reported for the first time.The new Bj subgenotype was combinated with genotype C not only at the region of precore C/C but also at the place of P gene region.

Objective Functional mitral regurgitation (FMR) refers to the systolic regurgitation of mitral valve secondary to compromised cardiac function or geometry abnormity with non-organic change of leaflets and ancillary parts of the valve.Severe aortic insufficiency (AI) with left ventricular dilation and dysfunction is clinically a complex heart disease and its postoperative complications and mortality are higher than usual valvular surgery.And such patients are often accompanied by FMR.It is generally acknowledged that FMR may improve after aortic valve replacement(AVR).This study follow up AI patients with left ventricular dilation and dysfunction and preoperative 2 + ＜ FMR≤3 + to evaluate the outcome of FMR after AVR.Preoperative clinical data is assessed by regression analysis.Methods From January 2000 to April 2011 our hospital treated 74 cases of patients with severe aortic regurgitation combined with left ventricular dilation (LVEDds ≥70 mm) and dysfunction (left ventricle ejection fraction,LVEF≤0.35) accompanied by 2 + ＜ FMR≤3 +.Postoperative follow-up was performed.Results The perioperative mortality rate was 8.1％.Average follow-up time was (14.9 ± 7.7) months and follow-up rate of 83.6％.5 patients died during follow-up.6 months after surgery,the average of FMR was (2.64 ± 1.17) (+),compared with preoperative data.LVEDd,LAD,CPMA,all P ＞ 0.05 compared with the preoperative data.LVEF,PAH,both P ＜ 0.05 compared with preoperative data.3.Multiple regression analysis:FMR pre/FMR post ratio is not correlated with age,gender,weight,LVEDd ≥ 75 mm,LVEF≤0.30,hypertension,ventricular arrhythmia and FMR postoperative improvement.However,PAH ≥ 50 mm Hg(1 mm Hg =0.133 kPa),LAD ≥ 50 mm,PAH≥50 mm Hg,CPMA ≥ 15mm,preoperative atrial fibrillation was negatively correlated with FMR postoperative improvement.Conclusion Severe AI with left ventricular dilation and dysfunction is a critical clinical heart disease and its postoperative complications and mortality are high.PAH ≥ 50 mm Hg,LAD ≥ 50 mm,PAH ≥ 50 nun Hg,CPMA ≥ 15mm,preoperative atrial fibrillation was negatively correlated with FMR postoperative improvement.Since patients with 2 + ＜ FMR ≤3 + usually do not improve or even worsen after AVR,those who have these above conditions preoperatively,should be treated on FMR during AVR.

Objective Functional mitral regurgitation (FMR) refers to the systolic regurgitation of mitral valve secondary to compromised cardiac function or geometry abnormity with non-organic change of leaflets and ancillary parts of the valve.Severe aortic insufficiency (AI) with left ventricular dilation and dysfunction is clinically a complex heart disease and its postoperative complications and mortality are higher than usual valvular surgery.And such patients are often accompanied by FMR.It is generally acknowledged that FMR may improve after aortic valve replacement (AVR).This study follow up AI patients with left ventricular dilation and dysfunction and preoperative 2 + ＜ FMR ≤3 + to evaluate the outcome of FMR after AVR.Preoperative clinical data is assessed by regression analysis.Methods From January 2000 to April 2011,74 cases of patients were treated,who with severe aortic regurgitation combined with left ventricular dilation (left ventricle,LVEDD ≥ 70 mm) and dysfunction (left ventricle ejection fraction,LVEF ≤ 0.35) accompanied by 2 + ＜ FMR ≤3 +.Postoperative follow-up was performed.Calculation FMR preoperative/FMR postoperative ratio,the age,sex,weight,high blood pressure,ventricular arrhythmia,atrium fibrillation,LVEDD,LVEF,left atrium diameter(LAD),pulmonary artery pressure (PAH),mitral leaflet coaptation point and the mitral annular(CPMA).All factors for logistic multiple faotors regression analysis.Results The perioperative mortality rate was 8.1％.Average follow-up time was (14.9 ± 7.7) months and follow-up rate of 83.6％.5 patients died during follow-up.6 months after surgery,the average of FMR was 2.64 ± 1.17 (+),P ＞0.05 compared with preoperative data.LVEDD,LAD,CPMA,P ＞0.05 compared with the preoperative data.LVEF,PAH,both P ＜0.05 compared with preoperative data.3.Multiple regression analysis:FMR preoperative/FMR postoperative ratio is not correlated with age,gender,weight,LVEDD ≥75 mm,LVEF≤0.30,hypertension,ventricular arrhythmia and FMR postoperative improvement.However,PAH ≥50 mm Hg,LAD ≥50 mm,PAH ≥50 mm Hg,CPMA ≥ 15 mm,preoperative atrial fibrillation was negatively correlated with FMR postoperative improvement.Conclusion Severe AI with left ventricular dilation and dysfunction is a critical clinical heart disease and its postoperative complications and mortality were high.PAH ≥50 mm Hg,LAD ≥50 mm,PAH ≥50 mm Hg,CPMA ≥15 mm,preoperative atrial fibrillation was negatively correlated with FMR postoperative improvement.Since patients with 2 + ＜ FMR≤3 + usually do not improve or even worsen after AVR,those who have these above conditions preoperatively,should be treated on FMR during AVR.

To investigate the morphogenetic process of human adenovirus type 41 (HAdV-41), 293TE cells were infected with purified wild-type HAdV-41, and ultrathin sections of infected cells were prepared and observed under a transmission electron microscope. Results showed that HAdV-41 entered host cells mainly through three ways: non-clathrin-coated pit, clathrin-coated pit, and direct penetration of plasma membrane. In addition, cell microvilli might help HAdV-41 enter cells. After entering into cells, HAdV-41 virus particles could be found in vacuoles or lysosomes or be in a free state in cytoplasm. Only free virus particles could be found near nuclear pores (NP), suggesting that the virus needed to escape from lysosomes for effective infection and viral nucleoprotein entered the nucleus through NP. Progeny viruses were as-sembled in the nucleus. Three types of inclusion bodies, which were termed as fibrillous inclusion body, condense inclusion body, and stripped condense inclusion body, were involved in HAdV-41 morphogenesis. In the late phase of viral replication, the membrane integrity of the infected cells was lost and viral particles were released extracellularly. This study reveals the partial process of HAdV-41 morphogenesis and provides more biological information on HAdV-41.
Adenovirus Infections, Human ; virology ; Adenoviruses, Human ; genetics ; growth & development ; physiology ; ultrastructure ; Cell Membrane ; virology ; Cell Nucleus ; virology ; Humans ; Virus Release ; Virus Replication

Objective To develop a safe,practical,disposable slide culture medium for fungi culture.Methods Based on the principle of traditional slide culture medium with small steel ring,a plate of polyvinyl chloride (PVC) with high transparency was chosen as the bottom material of the new medium which held the size of the conventional slide (25.2 mm × 75.9 mm × 1.05 mm) with frosted design at both ends.The diameter of inoculation hole was 2.4 mm on the rounded culture dish which was designed with diameter of 19.4 mm and side height of 3.75 mm.The users could inject potato dextrose agar (PDA) or other medium into the culture dish as needed and then sealed it with a plug.The upper cover was prepared with toughened glass.The improved disposable slide culture medium should be kept in humidifying box with sponge strips in the water tanks of both sides and sealed with cap.The growth status,microscopic morphology and staining result of filamentous fungi in the totally enclosed slide culture medium were observed and the preservative status of the prepared medium was also monitored simultaneously.Results The effects of the improved slide culture medium were satisfactory in clinical practical application.The growing status of fungi could be observed visually and the pigment was clear.The original growth form of fungi could be monitored under microscope and dye material could be perfused directly to stain with good results.The appearance and the volume of packaged slide medium were unchanged after preservation at 4 ℃ for 3 months.Conclusion An improved slide culture medium was successfully developed,which should be easy to operate,high visible,satisfactory for sealing effects and reliable for culture performance with high biological safety.The growth status of fungi could be observed under microscope at any time,and the medium could also be monitored under oil immersion lens directly and stained with cotton blue.The improved medium could be used in morphological examination for fungi in different levels of medical laboratories since its favorable results in clinical application.

Objective To study the cell vitality and ultra-structure of in vitro cultured fetus chondrocytes exposed to different doses of fluoride.Methods Primary chondrocytes were obtained from articular cartilage of the 24-27 weeks,aborted and dead fetuses.The third generation of primary cultured chondmcytes were exposed to concentrations of 0,10-2,5 × 10-3,10-3,10-4,10-5,10-6,10-7 and 10-8 mol/L fluoride for 24,48 and 72 h.Cell vitality was detected with Cell Counting Kit-8 (CCK-8) and ultra-structure of chondrocytes was observed by transmission electron microscope.Results The cell vitalities of chondrocytes exposed to doses of fluoride (10-2,5 ×10-3,10-3,10-4,10-5,10-6,10-7 and 10-8 moL/L) for 24,48 and 72 h were(15.04 ± 0.55)％,(62.53 ± 1.03)％,(100.34 ± 5.19)％,(111.40 ± 3.69)％,(121.47 + 6.09)％,(129.95 ± 4.96)％,(121.81 ± 4.97)％,(111.00 ± 1.63)％;(10.35 ± 0.64)％,(35.23 ± 2.41)％,(110.30 ± 2.07)％,(113.66 ± 6.98)％,(120.36 ± 6.23)％,(133.40 ± 5.80)％,(126.06 ± 5.40)％,(115.62 ± 7.33)％; (6.19 ± 0.16)％,(18.44 ± 0.21)％,(120.83 ± 4.93)％,(123.77 ± 4.82)％,(129.09 ± 5.21)％,(140.44 + 4.18)％,(131.99 ± 7.00)％,(124.10 ± 3.68)％,respectively.The cell vitalities of 10-2,5 × 10-3 mol/L fluoride groups were significantly lower than that of the control group (all P ＜ 0.05).The cell vitality of 10-2 mol/L group was significantly lower than that of the 5 × 10-3 mol/L group (P ＜ 0.05).Doses of fluoride (10-2,5 × 10-3 mol/L) could inhibit the cell vitality and promote the apoptosis of chondrocytes in vitro with increasing doses and prolonged time.The cell vitalities of 10-3,10-4,10-5,10-6,10-7,10-8 mol/L of fluoride groups were significantly higher than that of the control group (except the 24 h 10-3 mol/L,P ＜ 0.05).Between 10-4 and 10-3 mol/L groups(the vitalities of 48 h and 72 h were higher,but not significantly); 10-5 and 10-4 mol/L groups (the vitality of 72 h was higher,but not significantly); 10-6 and 10-5 mol/L groups,the cell vitalities were significantly higher than that of the control group(all P ＜ 0.05).Between 10-7 and 10-6 mol/L groups,10-8 and 10-7 mol/L groups (the vitality of 72 h was lower,but not significantly),the cell vitalities were significantly lower than that of the control group(all P ＜ 0.05).Doses of fluoride(10-3-10-8 mol/L) could promote the cell vitality of chondrocytes in vitro with prolonged time.The optimal concentration for the promotion was 10-6 mol/L.The cells of the control group were characterized as regular morphology,the abnormal surface microvillis,abundant cytoplasm and mitochondrial,abundant and slightly expanded rough endoplasmic reticulums and low electron-dense materials.The cells of 10-6 mol/L fluoride group had the following changes,increased and swell mitochondrial,hypertrophy and expanded rough endoplasmic reticulums.The cells of 5 × 10-3 mol/L fluoride group had the following changes,decreased microvillis,invaginated cell membrane,pyknosis and apoptotic body.Conclusion Doses of fluoride (10-3-10-8 mol/L) can promote the proliferation of human chondrocytes cultured in vitro.Doses of fluoride (10-2,5 × 10-3 mol/L) can promote the apoptosis of human chondrocytes cultured in vitro.