1. Construction of recombinant hING4 gene and its inhibitory effect on the growth of lung cancer NCI-H460 cells
Tumor 2008;28(11):942-945
Objective: To construct a recombinant human inhibitor of growth 4 (hING4) gene and observe its growth inhibition effect on lung adenocarcinoma NCI H460 cells. Methods: The GFP-labeled recombinant adenovirus vector hING4 (Ad-hING4) was constructed using pcDNA3. 0-mING4 plasmid as a template and site-specific mutagenesis technique. RT-PCR and fluorescence microscope were used to detect the expression of hING4 in NCI H460 cells. Bax and Bcl-2 protein expression were detected by Western blotting. Colony formation assay and flow cytometry were used to observe the effect of hING4 on the proliferation and apoptosis of H460 cells. Results: DNA sequence analysis and PCR indicated that Ad-hING4 were successfully constructed. Ad-hING4 was expressed in lung cancer H460 cells and had obvious cytopathic effect (CPE). Western blotting suggested that hING4 gene caused up-regulation of Bax expression and down-regulation of Bcl-2 expression. Colony formation assay and flow cytometry showed that hING4 inhibited the proliferation of H460 cells and induced apoptosis of lung cancer cells. Conclusion: Ad-hING4 gene was successfully constructed. It inhibited the growth of lung cancer H460 cells in vitro.
2.Genome amplification and sequence analysis of human bocavirus 2.
Huan-Di CUI ; Yu JIN ; Guang-Cheng XIE ; Wei-Xia CHENG ; Zhao-Jun DUAN
Chinese Journal of Virology 2014;30(3):257-262
To obtain the genome sequence of human bocavirus 2 (HBoV2), different regions of HBoV2 genome were amplified through PCR in fecal specimens which had been identified as single-positive for HBoV2 in 2010. A genome sequence of HBoV2 (HBoV2-NC, 5444 bp) was obtained after sequence assembly. The phylogenetic analysis showed that HBoV2-NC had the closest evolutionary relationship with HBoV2 Lanzhou strain. The predication of inverted terminal repeats of HBoV2-NC by DINAMelt showed that inverted terminal repeats were contained in HBoV2-NC 5' terminal, which had the typical stem-loop structure in other parvoviruses. Finally, some flanking sequences of HBoV2-NC were amplified by linker-PCR.
Base Sequence
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Gene Amplification
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Genome, Viral
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Human bocavirus
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chemistry
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classification
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genetics
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Humans
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Molecular Sequence Data
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Nucleic Acid Conformation
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Parvoviridae Infections
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virology
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Phylogeny
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RNA, Viral
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chemistry
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genetics
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Terminal Repeat Sequences
3.Progress on development and research of human bocavirus 1.
Huan-Di CUI ; Yu JIN ; Guang-Cheng XIE ; Zhao-Jun DUAN
Chinese Journal of Virology 2014;30(1):103-108
Human bocavirus 1 (HBoV1) is a novel virus that mainly causes respiratory tract infection, and it has the characteristic of genome of Parvovirus, containing three open reading frames that encode non-structural proteins NS1 and NP1 and structural proteins VP1 and VP2. Circular episome is present during the rolling circle replication of HBoV1, which provides the possibility of full genome amplification and infectious clone construction to save HBoV1. The recombination between HBoV1 and HBoV2-4 occurs frequently. With the three-dimensional culture, in vitro culture of HBoV1 provides a powerful tool for research on the pathogenesis of HBoV1. This review focuses on the molecular characteristics, association with diseases, in vitro culture, diagnosis and treatment of HBoV1.
Diarrhea
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virology
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Genomics
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Human bocavirus
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genetics
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isolation & purification
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physiology
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Humans
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Meningitis
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virology
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Respiratory Tract Diseases
;
virology
4.Current research on picornavirus 3C protease.
Hong WANG ; Guang-Cheng XIE ; Zhao-Jun DUAN
Chinese Journal of Virology 2014;30(5):579-586
The picornavirus family comprises many small viruses, several of which are important pathogens of humans and livestock. The 3C protease (3Cpro) of different species and genera of picornavirus contains the classic G-X-C-G motif and Cys-His-Asp/Glu catalytic triad. 3Cpro conducts maturation cleavage in the regions of VP2-VP3 and VP3-VP1 in P1, 2A-2B and 2B-2C in P2 and the whole P3. Picornavirus 3Cpro has been shown to have significant substrate preference in Q-G/S/A/V/H/R and E-S/G/R/M as well as species and genera specificity through analyses of the maturation cleavage of picornavirus polyproteins. Innate immune adaptors such as TRIF, MAVS, IRF3, IRF7 and NEMO have various potential cleavage sites in picornavirus 3Cpro (TRIF and NEMO show considerable diversity in their cleavage sites). Useful information will be provided for the development of broad-spectrum antiviral agents as well as evasion mechanisms of the innate immune system against picornavirus 3Cpro through continued research of picornavirus 3Cpro.
Cysteine Endopeptidases
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physiology
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Immunity, Innate
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Picornaviridae
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enzymology
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immunology
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Viral Proteins
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physiology
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Virus Replication
5.Research on pericellular matrix properties for chondrcytes.
Jun-liang HAN ; Wang-ping DUAN ; Guang-hua SHI ; Wei YUAN ; Xiao-chun WEI
China Journal of Orthopaedics and Traumatology 2015;28(6):576-579
Pericellular matrix (PCM) is a narrow tissue region surrounding chondrocytes, which "chondron" with its enclosed cells. A number of studies suggested that PCM is rich in proteoglycans, collagen and fibronectin, and plays an important role in regulating microenvironment of chondrocytes. Direct measures of PCM properties through micropipette aspiration technique showed that PCM was different from mechanical property of chondrocytes and nature extracellular matrix. However, the function of PCM is not clear, and need further study.
Animals
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Biomechanical Phenomena
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Chondrocytes
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chemistry
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cytology
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metabolism
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Extracellular Matrix
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chemistry
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metabolism
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Humans
6.Meroterpenoids and isoberkedienolactone from endophytic fungus Penicillium sp. associated with Dysosma versipellis.
Jun-Wei LI ; Rui-Gang DUAN ; Jian-Hua ZOU ; Ri-Dao CHEN ; Xiao-Guang CHEN ; Jun-Gui DAI
Acta Pharmaceutica Sinica 2014;49(6):913-920
Seven meroterpenoids and five small-molecular precursors were isolated from Penicillium sp., an endophytic fungus from Dysosma versipellis. The structures of new compounds, 11beta-acetoxyisoaustinone (1) and isoberkedienolactone (2) were elucidated based on analysis of the spectral data, and the absolute configuration of 2 was established by TDDFT ECD calculation with satisfactory match to its experimental ECD data. Meroterpenoids originated tetraketide and pentaketide precursors, resepectively, were found to be simultaneously produced in specific fungus of Penicillium species. These compounds showed weak cytotoxicity in vitro against HCT-116, HepG2, BGC-823, NCI-H1650, and A2780 cell lines with IC 50 > 10 micromol x L(-1).
Berberidaceae
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microbiology
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Cell Line
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Cell Line, Tumor
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Humans
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Lactones
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isolation & purification
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pharmacology
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Monoterpenes
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isolation & purification
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pharmacology
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Penicillium
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chemistry
8.Signal transduction of innate immunity to virus infection.
Guang-Cheng XIE ; Zhao-Jun DUAN
Chinese Journal of Virology 2012;28(3):303-310
The innate immune system is essential for the initial detection of invading viruses and subsequent activation of adaptive immunity. Three types pattern recognition receptors (PRRs) in innate immune cells play a pivotal role in the first line of host defense system. PRRs include Toll-like receptors (TLRs), RIG-I-like receptors(RLRs) and nucleotide-binding oligomerization domain (NOD)-like receptors (NLRs). PRRs recognize pathogen-associated molecular patterns(PAMPs) or danger-associated molecular patterns (DAMPs) to initiate and regulate innate and adaptive immune responses. Three types PRRs have their own features in ligand recognition and cellular location. Activated PRRs deliver signals to adaptor molecules (MyD88, TRIF, IRAK, IPS-1), which act as important messengers to activate downstream kinases (IKK complex, MAPKs, TBK1, RIP-1) and transcription factors (NF-kappaB, AP-1, IRF3), which produce effected molecules including cytokines, chemokines, inflammatory enzymes, and type I interferons. This review focuses on discussing PRRs signaling pathways and achievements in this field in order to provide beneficial strategies for human life and immune diseases prevention.
Animals
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Humans
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Immunity, Innate
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Receptors, Pattern Recognition
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genetics
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immunology
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metabolism
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Signal Transduction
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Virus Diseases
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immunology
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metabolism
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virology
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Virus Physiological Phenomena
9.Therapeutic evaluation of microvascular decompression in patients with trigeminal neuralgia associated with hypertension.
Yan-jun CHONG ; Jian CHEN ; Guang-ting ZHU ; De-yi DUAN ; Qi-long CHENG ; Qi-jie SHAO
Chinese Journal of Surgery 2005;43(5):331-333
OBJECTIVEThe aim of work was to evaluate the efficacy of microvascular decompression (MVD) in patients with trigeminal neuralgia (TN) associated with hypertension.
METHODSFive hundred and thirty-eight cases of TN associated with hypertension treated with MVD were retrospectively analyzed, all the cases were treated with MVD in the zone of the ventrolateral medulla oblongata and inspected postoperative blood pressure.
RESULTSAmong 538 cases, 341 cases (63.4%) had cure result, 71 cases (13.2%) had obvious effectiveness, 53 cases (9.8%) had effectiveness, and 73 cases had no effectiveness. Four hundred and eighty-three cases have been followed 1.0 approximately 15.8 years, mean 6.3 years. At the time of their follow-up, 313 cases had cure result, 63 cases had obvious effectiveness, 42 cases had effectiveness and 65 cases had no effectiveness.
CONCLUSIONSIt was pathogenesis of nedulla oblongata by abnormal vascular tab, long-term aching stimulation and emotional stress. It was an effective method to treat TN associated with hypertension with MVD in this zone.
Adult ; Aged ; Aged, 80 and over ; Decompression, Surgical ; methods ; Female ; Humans ; Hypertension ; complications ; Male ; Medulla Oblongata ; pathology ; Microsurgery ; Middle Aged ; Retrospective Studies ; Treatment Outcome ; Trigeminal Neuralgia ; complications ; surgery ; Vascular Surgical Procedures ; methods
10.Knee arthroscopic resection of articular capsule valvular treat popliteal cysts.
Rui HE ; Lin GUO ; Liu YANG ; Guang-xing CHEN ; Xiao-jun DUAN ; Chon-fu LUO
Chinese Journal of Surgery 2013;51(5):417-420
OBJECTIVETo retrospective study the results of arthroscopic resection of articular capsule valvular treated popliteal cysts and examined the incidence of complication.
METHODSFrom July 2007 to July 2010, 45 patients with symptomatic popliteal cysts such as knee pain and limitation of motion were treated with arthroscopic technique to resect the connecting valvular mechanism at posteromedial compartment of articular capsule. Through the posteromedial viewing portal and cyst resection working portal, resected the valvular mechanism by shaver and plasma cutter, associated intra-articular pathology was treated simultaneously. The functional outcome was evaluated by employed Rauschning and Lindgren score before the surgery and the last follow-up. By using of Wilcoxon rank sum test to statistics analyze the clinic outcome.
RESULTIn all cases, 40 patients were found the associated intra-articular pathology, the incidence was 88.9%.The intra-articular pathology was medial meniscus tear (27 cases, 60.0%), lateral meniscus tear (20 cases, 44.4%), cartilage lesion (28 cases, 62.2%), which composed with patellofemoral joint (12 cases, 26.7%),medial condyles of femur (10 cases, 22.2%),lateral condyles of femur (6 cases, 13.3%). The anterior cruciate ligment injury was found in 2 cases, the incidence was 4.4%. All the patients achieved symptoms relief postoperative, such as posterior knee discomfort and swelling after activities.The Rauschning and Lindgren score was improved 1-2 grade in average (Z = -6.092, P = 0.00). The recurrence rate was 2.2%. All the incision healed good. No major complications were encountered such as saphenous nerve, great saphenous vein, blood vessel in popliteal injury.
CONCLUSIONSArthroscopic resection of articular capsule valvular treatment of popliteal cyst and treatment of associated intra-articular pathology are effective and safe. This surgery technique has more minimally trauma and lower recurrence rate.The clinic results in short term are excellent.
Adolescent ; Adult ; Aged ; Arthroscopy ; methods ; Child ; Female ; Follow-Up Studies ; Humans ; Knee Joint ; surgery ; Male ; Middle Aged ; Popliteal Cyst ; complications ; surgery ; Retrospective Studies ; Treatment Outcome ; Young Adult