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

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

Microscopy, Electron, Transmission ; methods ; Negative Staining ; methods ; Viruses ; ultrastructure

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.

Objective To investigate the prevalence of clonorchiasis among residents of Huachuan Country,Heilongjiang Province and to provide a basis for development of control strategies.Methods From 2011 to 2012,cluster random sampling was performed to survey the incidence of clonorchiasis in Huachuan Country.Fecal specimens were collected and examined the clonorchis sinensis eggs by Kato-Katz method.A questionnaire survey was conducted to collect related information such as age,gender,occupation and eating habits.The infection characteristic was analyzed.Results Totally 884 patients with clonorchiasis were found among 2248 residents,and the infection rate was 39.32％.The infection rate in male[47.15％(611/1296)] was significantly higher than females [28.68％(273/952),x2 =34.55,P ＜ 0.01].The infection rate increased with age,which was higher in the 20-69 years old people,with the highest infection rate in the 50-59 years old groups[45.34％ (219/483)].Of the occupational distribution,farmers had the highest infection rate [47.24％ (420/889)],followed by cadres and staffs[38.38％(190/495)].Of residents with fresh fish eating history,the prevalence of clonorchiasis was 53.38％(150/281).Conclusions The prevalence of clonorchiasis is still high in Huachuan County.To reduce the prevalence of clonorchiasis,comprehensive prevention measures,health education and group chemotherapy should be carried out.

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 results of intraventricular tunnel procedure for double outlet right ventricle(DORV) with subaortic ventricular septal defect(VSD).Methods Nine children with DORV complicated with subaortic VSD underwent intraventricular tunnel procedure.Five of them had stenosis of pulmonary valve or subpulmonary stenosis,and one had double-chambered right ventricle. An intraventricular tunnel procedure was performed under cardiopulmonary bypass. Right ventriculotomy was made to repair the VSD with teflon patch. An internal tunnel was made between the left ventricle and the aorta.The right ventricle connected with the main pulmonary artery. Results All children survived and recovered finally. Echocardiography showed that the internal tunnel function was well.Conclusion With correct diagnosis and selection of procedure, intraventricular tunnel procedure is a satisfactory method for the treatment of DORV with subaortic VSD.

Objective To report the clinical characteristics and surgical treatment of prosthetic valve endocarditis (PVE).Methods A retrospective review of 20 consecutive patients,who underwent surgery for PVE between Jan 2003 and Dec 2012,was conducted.Excision of infected prosthetic valves and radical debridement of the infected tissues were completed under hypothermic cardiopulmonary bypass.Six patients were treated by the Bentall procedure,4 patients by the modified Cabrol procedure,4 patients by aortic valve replacement,3 patients by mitral valve replacement and 3 patients by double valve replacement.Results One patient died within 30 days after surgery due to severe sepsis complicated with multiple organ failure and other 19 patients discharged smoothly.A further 2 patients died 2 and 4 months after surgery due to recurrence of fungal infection.Fourteen patients were followed up for 20-124 (59.0 ± 31.8) months.No patient died and no relapse of endocarditis occurred during the period of follow-up.Conclusion Optimal timing of surgical intervention and radical debridement of all infected tissues are the keys to success.

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.