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.

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.

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

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

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

BACKGROUND:The principle of lyophilization is to sublimate the solvent of frozen materials in vacuum and retain the solute, thus making a pore structure. OBJECTIVE: To produce a chitosan tubular scaffold by lyophilization, and to test its physicochemical properties. METHODS: The chitosan tubular material was prepared by lyophilization method, folowed by gross observation and electron microscopic observation. The chitosan tubular material samples were placed into PBS solution and pure water for 50 days, respectively, and then immersed in trypsin liquid for 1 day folowed by embedded into the muscle and dorsal skin of neonatal Sprague-Dawley rats for 30 days. The degradation rate and porosity of the material were observed and calculated. The breaking strength and compressive strength of the material were determined both under drying and soaking conditions using tensile instrument and pressure meter, respectively. RESULTS AND CONCLUSION:The external form of the chitosan tubular material was normaly tubular. Under the electron microscope, it was composited by different size pores, and the pore size was 50-200μm. The degradation rates of the material were (5.33±0.12)% in PBS, (11.26±0.15) in water, 0.012% in the trypsin liquid and (35.2±3.7)in vivo. The porosity rate was (97.5±1.5)%. The breaking strength and compressive strength of the material was higher under the drying state than under the soaking state (P < 0.05). These findings indicate that the lyophilization method can produce the chitosan tubular material with good porosity rate and degradation rate as wel as good tensile ability and compressive capability.

Objective Non-traumatic and posteromedial subchondral osteonecrosis of trochlea tali (NTPSOTT) is a special type of necrosis of the talus , for which early diagnosis and treatment are particularly important .This study investigated the clinical ef-fectiveness of autogeneic cancellous bone transplantation in the treatment of NTPSOTT . Methods We retrospectively analyzed 21 cases of NTPSOTT treated by autogeneic cancellous bone transplantation and evaluated using the Clinical Rating System of the American Orthopedic Foot and Ankle Society ( AOFAS) , Visual Analogue Scale ( VAS) , and X-ray and CT examinations . Results The pa-tients were followed up for 12-26 months and all healed desirably , with the graft well integrated into the surrounding tissue , but no as-similation, collapse of the articular surface , or narrowing of the joint space .The last follow-up visit revealed significantly improved AOFAS score (90.55 ±6.73 vs 50.87 ±11.42, P=0.009) and VAS score (1.32 ±0.81 vs 6.43 ±1.66, P=0.027) as compared with the baseline . Conclusion Autogeneic cancellous bone transplantation is preferable for the treatment of NTPSOTT , which can effectively reduce the pain in the ankle , maintain the joint space , and protect the function of the ankle .

NSP4, as the diarrhea-related protein of rotavirus, is becoming an attractive candidate for vaccine development. To compare the immunogenicity of NSP4 from different genetic groups, we constructed eukaryotic expression plasmids comprising the NSP4 genes from four different genetic types using the pCI vector. The recombinant vectors were designated as pCI-97B6, pCI-97S36, pCI-97S34 and pCI-97SZ8, respectively. Following the conformation of the transient expression of the constructs in 293 cells, the plasmids were respectively subjected to the 5 round i. m. inoculation of BALB/c mice. The specific antibodies against NSP4 as well as the IgG1/IgG2a subclasses of immunoglobulin in mice sera were examined with indirect ELJSA after each immunization. The results showed that the immunization of plasmids expression NSP4s could elicit not only humoral but also cellular immunity, but the humoral immune response is dominant. There is a difference of immunogenecity among the NSP4 of different genetic type. Further studies were needed to focus on the relationship between the immunogenicity and protection effect.

RNA interference ( RNAi) is a process in which double-stranded RNA ( dsRNA) induces specific postranscriptional silencing of homologous transcripts. Systematic silencing of genes on a genome-wide scale using RNAi library targeting many thousands of genes provides a powerful research tool in functional genomics. RNAi libraries, which may be derived from plasmids cloning, virus packaging, PCR amplification, chemically synthesis or enzymatic digestion, have been successfully used to identify gene function, dissect signaling pathway and discover drug targets, etc. Promising progresses have been made in this field. The development, application and problems of RNAi library methodology and future prospects were reviewed.