AIM:To discuss the application value of retro-mode imaging by F-10 confocal scanning laser ophthalmoscope (cSLO) for detecting drusen in patients with age-related macular degeneration (AMD).METHODS:This was a retrospective case study.During the period of October 2015 to December 2016, 67 patients with unilateral AMD (67 affected eyes and 67 fellow eyes) were included in this study.All patients underwent color fundus photography, optical coherence tomography (OCT) and retro-mode imaging by F-10 cSLO.The features of drusen by color fundus photography, OCT and retro-mode imaging were comparatively observed in the affected eyes of patients with unilateral AMD.Positive numbers of drusen in the fellow eyes of patients with unilateral AMD detected by color fundus photography, OCT and retro-mode imaging were calculated and compared.RESULTS:Retro-mode imaging by F-10 cSLO gave easier to identify images of drusen than color fundus photography and OCT in the affected eyes of patients with unilateral AMD.In the fellow eyes of 67 patients with unilateral AMD, retro-mode imaging showed drusen in 56 cases(84%), color funds photography showed drusen in 36 cases(54%), OCT showed drusen in 48 cases(72%), the difference was statistically significant(χ2=14.31, P<0.05).The positive numbers of drusen detected by retro-mode imaging were significantly higher than color fundus photography, the difference was statistically significant(χ2=13.87, P′<0.0125).There was no statistically significant difference in the positive numbers of drusen detected by retro-mode imaging and OCT(χ2=2.75, P′>0.0125).CONCLUSION:Retro-mode imaging by F-10 cSLO provides a non-invasive technique and should be useful for detecting and monitoring drusen in AMD.

OBJECTIVETo investigate the effect of berbamine on human hepatoma cell line SMMC7721.

METHODSThe effects of 24 h and 48 h incubation with different concentrations (0 to approximately 64 microg/ml) of the berbamine on SMMC7721 cells were evaluated using 3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT) assay. Hoechst 33258 staining was conducted to distinguish the apoptotic cell, and the appearance of sub-G1 stage was determined by PI (propidium iodide) staining, the percentage of apoptotic cell was determined by flow cytometry following annexin V/PI staining. Flow cytometry was performed to analyze the cell cycle distribution and the mitochondrial membrane potential (psi(m)); the expression of activated caspase3 and caspase9 was analyzed by Western-blot.

RESULTSThe proliferation of SMMC7721 was decreased after treatment with berbamine in a dose- and time-dependent manner. Berbamine could induce apoptosis in SMMC7721 cells and could cause cell cycle arrest in G0/G1 phase, to induce loss of mitochondrial membrane potential (psi(m)) and activate caspase3 and caspase9. Berbamine-induced apoptosis could be blocked by the broad caspase inhibitor z-VAD-fmk.

CONCLUSIONBerbamine exerts antiproliferative effects on human hepatocellular carcinoma SMMC7721 cells. The anticancer activity of berbamine could be attributed partly to its inhibition of cell proliferation and induction of apoptosis in cancer cells through loss in mitochondrial transmembrane potential and caspase activation.

Alkaloids ; pharmacology ; Apoptosis ; drug effects ; Benzylisoquinolines ; pharmacology ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Enzyme Activation ; drug effects ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Medicine, Chinese Traditional ; Membrane Potentials ; drug effects ; Mitochondria, Liver ; drug effects ; metabolism ; ultrastructure

OBJECTIVETo Construction of P and NP genes eukaryotic expression vectors of Newcastle Disease Virus LaSota strain,study its reverse genetics and functional genome of NDV.

METHODSP, NP genes were amplified and cloned into pGEM-T easy vector and then subcloned into pcDNA3.1 (+) expression vector respectively, the recombinant plasmids were named pcDNA3.1 (+)-P and pcDNA3.1 (+)-NP, Recombinant plasmids were transfected into 293 and BHK-21 cells respectively and were detected using IE and Western blot analysis.

RESULTSExpression of P, NP genes were detected and confirmed by the IE and WB analysis.

CONCLUSIONThe recombinant eukaryotic plasmids pcDNA3. 1(+)-P, pcDNA3.1 (+)-NP were expressed in 293 and BHK-21 cells successfully. This research may be helpful for further study of reverse genetics and functional genome of NDV.

Animals ; Cell Line ; Cricetinae ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Humans ; Newcastle disease virus ; genetics ; metabolism ; Nucleoproteins ; genetics ; metabolism ; Phosphoproteins ; genetics ; metabolism ; Viral Proteins ; genetics ; metabolism

Tumor necrosis factor alpha (TNFalpha) is a pro-inflammatory cytokine, acting as a regulator of inflammation and immunity. TNFalpha plays a critical role in the pathogenesis of rheumatoid arthritis. Blocking of TNFa activity suppressed inflammatory tissue damage. In present study, the chimeric gene of soluble TNF receptor and IgG Fc fragment (sTNFR-IgG FC) was cloned into the mammalian cell expression vector pStar. When the plamid pStar/sTNFR-IgGFc-GFP was transfected into endothelial cells, a considerable expression of the sTNFR-IgG Fc fusion protein was detected. Moreover, the product in 100microL expression supernatant could completely antagonize the cytolytic effect of 1ng TNFa on L929 cells, even at 1/64 dilution. Then the plasmid was delivered into CIA-induced rheumatoid arthritis mice by tail vein injection. The expression of sTNFR-IgG Fc was detected in liver by RT-PCR. Animals in treatment group showed reduced symptoms of arthritis and more active. This treatment induced decrease of synovial incrassation and prevented the cartilage destruction of the mice RA model. These results show that tail vein injection is an effective way for gene therapy and sTNFR-IgGFc expression plasmid is potential for the treatment of rheumatoid arthritis.
Animals ; Arthritis, Rheumatoid ; chemically induced ; therapy ; Collagen Type II ; Endothelial Cells ; metabolism ; Escherichia coli ; genetics ; metabolism ; Etanercept ; Genetic Therapy ; Humans ; Immunoglobulin G ; biosynthesis ; genetics ; therapeutic use ; Male ; Mice ; Mice, Inbred DBA ; Receptors, Tumor Necrosis Factor ; biosynthesis ; genetics ; therapeutic use ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; therapeutic use ; Transfection ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo explore the role of BM2 protein in the life cycle of influenza B virus.

METHODSThe authors screened human kidney MATCHMAKER cDNA library for new binding partners of BM2 of influenza B virus by using the yeast two hybrid system with truncated BM2 (26-109 aa) as the bait.

RESULTSSix positive plasmids encoding N-acetylneuraminate pyruvate lyase, angiopoietin 3, zinc finger protein 251, ribosomal protein S20, protein arginine N-methyltransferase 1 variant 1 (PRMT) and transcription factor-like 1 (TCFL1) were obtained.

CONCLUSIONThe results suggest that BM2 may play an important role in the life cycle of influenza B virus.

Angiopoietin-like Proteins ; Angiopoietins ; genetics ; metabolism ; DNA-Binding Proteins ; genetics ; metabolism ; Gene Library ; Humans ; Influenza B virus ; genetics ; metabolism ; Kidney ; metabolism ; Oxo-Acid-Lyases ; genetics ; metabolism ; Plasmids ; genetics ; Protein Binding ; Protein-Arginine N-Methyltransferases ; genetics ; metabolism ; Repressor Proteins ; genetics ; metabolism ; Ribosomal Proteins ; genetics ; metabolism ; Transcription Factors ; genetics ; metabolism ; Two-Hybrid System Techniques ; Viral Proteins ; genetics ; metabolism ; Zinc Fingers ; genetics

OBJECTIVETo prepare monoclonal antibodies specific for M1 protein of H5N1 subtype human influenza virus, this work may provide new tool in rapid diagnosis and study of type A influenza virus.

METHODSBALB/c mice were immunized with purified recombinant H5N1 (A/Anhui/1/2005)/M1 protein expressed in E. coli. Spleen cells of the immunized mice were fused with sp2/0 cells to produce hybridoma cell lines. ELISA was performed to identify the monoclonal antibody against M1 protein of H5N1. Immunofluorescence assay (IFA) were applied to identify the specificity of these antibodies.

RESULTSThree hybridoma cell lines steadily secreting anti-H5N1/M1 McAb were obtained, and their cross reactivity was confirmed by cross-reaction test and IFA.

CONCLUSIONMonoclonal antibodies immunized with H5N1 subtype influenza virus M1 protein are cross-reactive, which can be used to detect different subtype of influenze virus type A.

Animals ; Antibodies, Monoclonal ; analysis ; immunology ; Antibodies, Viral ; analysis ; immunology ; Cell Line ; Cross Reactions ; Dogs ; Humans ; Influenza A Virus, H5N1 Subtype ; genetics ; immunology ; Influenza, Human ; immunology ; virology ; Mice ; Mice, Inbred BALB C ; Viral Matrix Proteins ; genetics ; immunology

OBJECTIVETo study the relationship between 500 kinds of commonly used Chinese herbal medicine and the classification of their efficacies in Chinese Materia Medica in relation to the common diseases listed in Internal Medicine.

METHODSDatabase retrieval frequency of the quantitative statistical method was adopted. First, the 8 980 kinds of Chinese herbal medicine recorded in Chinese Materia Medica were used as the original search objects, and 4 493 kinds which were cited in more than five articles were picked out and then rechecked for further title citations. Second, as judged based on the Criterion, the numbers of articles which included the medicines in the line of standards were examined. As a result, 500 species of Chinese herbal medicine were singled out based on their retrieval frequency and were then used for compilation of the classification statistics according to their efficacy and the common diseases in Internal Medicine.

RESULTSFrom the classification of Chinese medicines, herbs with wide efficiency and a meek nature had higher frequencies, but those which were not appropriate as decoctions had relatively lower frequencies. However, according to the average frequency, the Chinese herbal medicine for nourishing qi and tonifying blood, at 36,346 times and 34,544 times, respectively, were the most commonly used. Analyzed from the frequency of application of the Chinese medicine in the treatment of common diseases, most of the top 10 kinds of Chinese herbal medicine with the highest frequencies generally coincided with the 500 selected medicines. In addition, the Chinese medicines with clear pharmacological efficiency were easily isolated and purified to be made into injections, although other forms are more commonly used.

CONCLUSIONThe results of the research objectively reflected the current applications of Chinese herbal medicine, and could be used as references in teaching, research, clinical applications, and in compiling and increasing the drugs in textbooks and Pharmacopoeia.

Disease ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; utilization ; Research ; Treatment Outcome

Objective To investigate the epidemiological features of patients with nosoeomial invasive fungal infection. Methods Fungi in blood were identified by BaeT ALERT 3D, other clinical samples were cultured by Sabouraud' s dextrose agar (SDA) medium. Candidas were isolated and identified by CHRO Magar candida color medium. Fungus-cultured positive cases from Jan. 2004 to Nov. 2007 were analyzed on items as patients' age, underlying disease, sample, strain, and species distribution. All statistical analyses were carried out by SPSS 13.0. Results The overall incidence rate of invasive fungal infections was 4.12%. The average age of patients was 7-96 with most patients were male, with geriatric problems and different kinds of underlying diseases. Lower respiratory tract infection was the most frequent infection site, followed by urinary tract, gastrointestinal tract. The main pathogens of invasive fungal infections were Candidas (93.80%). Strains of Candida albicans were the most frequent organisms which accounted for 67.29% of all the isolates. Mould fungus infections accounted for only 6.20%. During the 4 years of observation, the detection rate of fungi, specimen sources and the distribution of species and compartment were different with significant differences (P<0.0083). Conduslon The epidemiological properties such as the source of specimen, the distribution of species and composition sections of invasive fungal infections were changing. Candida slaP. were still the main pathogens of invasive fungal infections but the sections of fungi changed. The incidence of Aspergillus infections had been increasing recently.

Objective To study the antiviral effect of Jin Qiao Tablets on influenza A H1N1 virus in vivo. Methods The mouse pneumonia model was established by nasal inhalation of 15 LD50 of influenza virus. After prophylactic or therapeutic medication for 5 d,mouse lung tissue was taken out and weighed. Viral load in lung tissue was measured by polymerse chain reaction(PCR),and the level of γ-interferon(γ-IFN)in rat serum and lung was detected by double antibody sandwich enzyme-linked immunosorbent assay (ELISA)for evaluating the effect of Jin Qiao Tablets on lung index, viral load and γ-IFN in rats. After prophylactic or therapeutic medication for 7 d,morbidity and mortality within 14 d of mice with pneumonia induced by nasal inhalation of 3 LD50 were observed to evaluate the action of Jin Qiao Tablets for protecting against death and prolonging life span. Results Jin Qiao Tablets markedly decreased the increased lung index,promoted the death-protection rate and life-prolongation rate, decreased viral load, raised the level of γ-IFN in mice (P < 0.05 or P < 0.01). Experimental results in vivo showed that Jin Qiao Tablets had better anti-influenza virus activity than Yinqiao Jiedu Tablets and Lianhua Qingwen Capsules, and the effect of Jin Qiao Tablets was equivalent to that of Tamiflu. The prophylactic effect of Jin Qiao Tablets was stronger than the therapeutic effect, but there was no significant difference between them. Conclusion Jin Qiao Tablets have obvious effect against influenza A H1N1 virus in vivo.

Based on the first isolated human H5N1 influenza virus strain A/Anhui/1/2005 in China, HA and HA1 genes were amplified and cloned into the eukaryotic expression vector pStar. The recombinant plasmids pStar-HA and pStar-HA1 were transfected into COS7 cells. Western blot and IFA showed that the two recombinant DNA plasmids were successfully expressed in eukaryotic cells. BALB/c mice were immunized with the plasmids DNA by intramuscular injection. Anti-HA specific antibody in peripheral blood of immunized mice was tested by ELISA. The results showed that the recombinant plasmids successfully induced the anti-HA humoral immune response, and there was no significant difference between HA and HA1 as immunogen. This work provides basis for future development of novel avian flu vaccine.
Animals ; Antibodies, Viral ; blood ; COS Cells ; Cercopithecus aethiops ; Female ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; immunology ; Immunoglobulin G ; blood ; Influenza A Virus, H5N1 Subtype ; immunology ; Influenza Vaccines ; immunology ; Mice ; Mice, Inbred BALB C ; Vaccines, DNA ; immunology