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

Objective To investigate the clinical effect of lentinan and thymopentin injection combined with PTP chemotherapy in oral squamous cell carcinoma patients, and to observe the effect of lentinan on serum tumor specific growth factor ( TSGF) , IgG, IgA and IgM and quality of life in patients with oral squamous cell carcinoma .Methods The clinical data of 80 cases of oral squamous cell carcinoma ( postoperative lymph node metastasis) confirmed by operation and pathology were analyzed retrospectively, and the diagnosis and treatment were analyzed.The control group were treated with PTP regimen, 80 ~120 mg/m2 of cisplatin, 50 ~80 mg/m2 of teniposide on the first day, the second to fourth day, once daily +pingyangmycin 4~6 mg/m2 , once daily, 3 to 12 days, 3 weeks for a course of treatment, a total of 6 courses.In the control group based on the use of thymopentin injection 20 mg and lentinan 2 mg+5% glucose solution 250 mL intravenous infusion, chemotherapy 2 d before the start, infusion 14 d, 3 weeks for a course of treatment, a total of 6 courses, for the observation group;two groups were 40 cases.The levels of serum TSGF, IgA, IgG, IgM and CD3 +, CD4 +, CD8 +, NK cells and TSGF in the two groups before and after treatment were measured.The quality of life of the patients was investigated by hospital-made questionnaires.The clinical efficacy and side effects were analyzed.Results TSGF levels in the two groups were significantly lower than that before treatment, and the TSGF level in the observation group was (39.1 ±4.9)U/mL, which was significantly lower than that in the control group (52.3 ±5.8) U/mL(P<0.05).After treatment, the TSGF level in observation group was lower than that in control group [(39.1 ±4.9) U/mL vs. (52.3 ±5.8) U/mL](P<0.05).The CD3 +, CD4 +, NK, IgG levels in observation group were higher than those in control group and the CD8 +level in observation group was lower than that in control group (P<0.05).The scores of physical symptoms, sleep quality, mental state and social affection levels in observation group were higher than those in control group(P<0.05).The total efficacy in observation group was lower than that in control group (77.5% vs.45.0%, P<0.05).The adverse reaction rate was 20.0% in the observation group, which was significantly lower than that in the control group (P<0.05).Conclusion The use of lentinan and thymopentin injection combined with PTP chemotherapy in the treatment of oral squamous cell carcinoma has higher clinical efficacy and less adverse reactions.

Objective To investigate the effects of application of pseudoreplica technique in electron microscope observation of A?.Methods With the modification of classical pseudoreplica technique,ADDLs were spotted and concentrated on the gel before negative staining.Results The structures of 5nm to10nm spherical granules and 20 to 30nm coils were visualized by transmission electron microscopy(TEM).Conclusion Pseudoreplica is rapid and effective in ADDLs condensation,negative staining especially in further ultra miero structure observation and research.

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

Humans ; Immunotherapy ; methods ; Lung Neoplasms ; immunology ; therapy ; Vaccines ; therapeutic use

ObjectiveTo investigate the value of full-volume three-dimensional echocardiography (FV3DE) in evaluating left ventricular systolic function in normal pregnancy.MethodsFifty-five healthy pregnant women,including 18 cases in first trimester (first trimester group),18 cases in second trimester (second trimester group) and 19 cases in third trimester (third trimester group),and 20 healthy nonpregnant women(control group) were chosen.The hemodynamic indexes and left ventricular systolic function parameters were detected by FV3DE.ResultsThe imaging satisfactory rates of FV3DE of control group,first trimester group,second trimester group and third trimester group were 100.0％ (20/20),100.0％ (18/18),88.9％ (16/18) and 78.9％(15/19) respectively.Compared with control group,heart rates (HR) of first,second and third trimester groups were significandy quicker (P ＜ 0.05);total peripheral resistance (TPR)declined gradually (P ＜ 0.05);LVM,LVMI,LVEDV,LVSV and LVCO increased gradually (P ＜0.05) ; MAP,LVEF,PER of four groups had no significant differences (P＞ 0.05 ).LVCO in healthy pregnant women had negative correlation with TPR (P ＜0.01),and positive correlation with HR,LVM,LVMI,LVEDV,LVSV,LVEF and PER (P ＜0.01),PER had positive correlation with LVEF.ConclusionThe application of FV3DE has significant clinical value in early evaluation of left ventricular systolic function in normal pregnancy.

Objective To investigate the effects of permissive hypercapnia ventilation on cerebral oxygen metabolism and postoperative cognitive function in elderly patients.Methods One hundred and twenty ASA Ⅰ-Ⅲ patients,aged 65-80 yr,undergoing elective lower abdominal surgery under general anesthesia,were randomly divided into 2 groups (n=60 each): routine ventilation group (group R) and permissive hypercapnia ventilation group (group H).In group H,VT=6-8 ml/kg,RR=12-14 bpm,I: E=1: 2,and PaCO2 was maintained at 45-65 mm Hg and pH value ＞ 7.2,while in group R,VT=10-12 ml/kg,RR=14-16 bpm,I:E=1:2,and PaCO2 was maintained at 35-45 mm Hg.Blood samples were taken from the radial artery and jugular bulb for blood gas analyses at 0,5,15 and 30 min after tracheal intubation (T0.3).Cerebral A-V O2 content differences (Da-jvO2)and cerebral O2 extraction rate (CERO2) were calculated at the same time.Cognitive function was assessed by Mini-Mental State Examination (MMSE) at 1 day before operation,and 24 h,48 h,1 week and 2 weeks after operation.Results Compared with group R,PEr CO2 andPaCO2 were significantly increased,and pH value,Da-jvO2 and CERO2 were significantly decreased at T1.3,MMSE score was significantly increased after operation,and the incidence of post-operative cognitive dysfunction was significantly decreased after operation (P ＜ 0.05 or0.01).Compared with the baseline value at T0,Da-jvO2 and CERO2 were significantly decreased at T1-3 in both groups,PETCO2 and PaCO2 were significantly increased,and pH value,Da-jvO2 and CERO2 were significantly decreased at T1-3 in group H,and MMSE score was significantly decreased at 24 h,48 h,1 week and 2 weeks after operation in both groups (P ＜ 0.01).Conclusion Permissive hypercapnia ventilation can improve the cerebral oxygen metabolism during operation,and reduce post-operative cognitive dysfunction in the elderly patients.

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 analyze and compare the clinical characteristics of primary sclerosing cholangitis (PSC) with IgG4-related sclerosing cholangitis (IgG4-SC).Methods The clinical data of 32 PSC patients and 72 IgG4-SC patients who were hospitalized in Peking Union Medical College Hospital (PUMCH) from January 2004 to December 2014 were retrospectively analyzed.Results Of the 32 PSC patients,there were 16 male and 16 female.Of the 72 IgG4-SC patients,there were 61 male and 11 female,(ratio =5.5∶ 1).The average ages were 44.9 (11 ～ 77) and 59.8 (28 ～ 83) years,respectively (P ＜0.05).The most common symptoms of PSC and IgG4-SC were abdominal pain and jaundice,and the incidences of abdominal pain and jaundice were 50.0％ and 68.1％,78.1％ and 81.9％,respectively.The serum IgG4 level of the IgG4-SC patients was significantly higher than the PSC patients (P ＜ 0.05).The total protein in serum of the IgG4-SC patients was higher than the PSC patients (P ＜ 0.05).The rate of bile duct wall thickening as detected on endoscopic ultrasonography (EUS) was higher than by abdominal ultrasound and abdominal CT,which were 91.2％,11.5％ and 33.3％,respectively (P ＜0.05).12 PSC patients were followed up for over 2 years,including 2 patients who underwent liver transplantation after failure of conservative treatment,5 patients who died from hepatic failure and infection,and 3 with stable condition.43 IgG4-SC patients were followed up for over 2 years,including 16 patients with relapse.The recurrence rate was 37.2％ (16/43).The more the extrabiliary organs or bile duct segments were involved,the higher was the recurrence rate.Conclusions Both PSC and IgG4-SC are cholestatic diseases,and they have many similarities in clinical and imaging manifestations.However,they still have unique features.IgG4-SC is sensitive to glucocorticoids therapy and has good prognosis.Thus,it is important to differentiate PSC from IgG4-SC.

We wished to study the intracellular transport of adenoviruses. We constructed a novel recombinant adenovirus in which the structural protein IX was labeled with a mini-singlet oxygen generator (miniSOG). The miniSOG gene was synthesized by overlapping extension polymerase chain reaction (PCR), cloned to the pcDNA3 vector, and expressed in 293 cells. Activation of miniSOG generated sufficient numbers of singlet oxygen molecules to catalyze polymerization of diaminobenzidine into an osmiophilic reaction product resolvable by transmission electron microscopy (TEM). To construct miniSOG-labelled recombinant adenoviruses, the miniSOG gene was subcloned downstream of the IX gene in a pShuttle plasmid. Adenoviral plasmid pAd5-IXSOG was generated by homologous recombination of the modified shuttle plasmid (pShuttle-IXSOG) with the backbone plasmid (pAdeasy-1) in the BJ5183 strain of Eschericia coli. Adenovirus HAdV-5-IXSOG was rescued by transfection of 293 cells with the linearized pAd5-IXSOG. After propagation, virions were purified using the CsC1 ultracentrifugation method. Finally, HAdV-5-IXSOG in 2.0 mL with a particle titer of 6 x 1011 vp/mL was obtained. Morphology of HAdV-5-IXSOG was verified by TEM. Fusion of IX with the miniSOG gene was confirmed by PCR. In conclusion, miniSOG-labeled recombinant adenoviruses were constructed, which could be valuable tools for virus tracking by TEM.
Adenoviruses, Human ; chemistry ; genetics ; metabolism ; Arabidopsis Proteins ; chemistry ; genetics ; metabolism ; Flavoproteins ; chemistry ; genetics ; metabolism ; Humans ; Phototropins ; chemistry ; genetics ; metabolism ; Singlet Oxygen ; chemistry ; Staining and Labeling ; Transfection