1.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
2.Effects of different doses of PcTx1 on global cerebral ischemia-reperfusion injury in rats
Xianming ZENG ; Yaomei CUI ; Huixian CHENG ; Yunhe ZHU ; Wei GAO ; Qiuting ZENG ; Manlin DUAN ; Jianguo XU
Chinese Journal of Anesthesiology 2012;32(6):708-712
ObjectiveTo explore the effects of different doses of PcTx1,a specific blocker of acid-sensing ion channel 1a,on global cerebral ischemia/repedfusion (I/R) injury in rats,MethodsSixty adult male Sprague Dawley rats (weighing 250-300 g) were randomly divided into 6 grups ( n =10 each):sham operation group (group S),I/R group,different doses of PcTx1 ( 10 ng/ml,group P1 ; 25 ng/ml,group P2 ; 50 ng/ml,group P3 ;and 500 ng/ml,group P4 ) groups.Global cerebral ischemia was induced by the modified procedure of Pulsinelli 4-vessel occlusion.In groups P1,P2,P3 and P4,different doses of PcTx1 ( 10,25,50 and 500 ng/ml),6 μl each,were respectively injected into the lateral cerebral ventricle at the initiation of reperfusion,while equal volume of double distilled water was injected instead in group I/R.Six rats in each group were sacrificed at 24 h of reperfusion,and the brains were immediately removed,Thereafter,the contents of malondialdehyde (MDA),reduced glutathione (GSH) and ritric oxide (NO),the activities of constitutive NO synthase (eNOS) snd inducible NO synthase (iNOS) were detected in hippocampus.Four rats in each group were sacrificed at 72 h of reperfusion,and hematoxylin and eosin staining was used to observe the pathomorphological changes of the hippocampal neurons.ResultsCompared with group S,the other groups showed decreases in the contents of GSH,while increases in the contents of MDA and NO and the activities of cNOS and iNOS ( P < 0.05 or 0.01 ).The contents of GSH increased,while the contents of MDA and NO and the activities of cNOS and iNOS decreased in groups P2,P3 and P4 compared with group I/R ( P < 0.05 or 0.01).Compared with group P1,the contents of GSH increased,the contents of MDA and the activities of cNOS decreased in groups P2,P3 and P4,and the contents of NO and the activities of iNOS decreased in groups P3 and P4 ( P < 0.05 or 0.01 ).Compared with group P2,the activities of iNOS decreased in groups P3 and P4(P < 0.05 or 0.01).The damage to neurons in hippocampal CAI was severe in groups I/R and P1,but it was attenuated in groups P3 and P4.ConclusionPcTx1 25,50 and 500 ng/ml (6 μl)injected into lateral cerebral ventricle can attenuate global cerebral I/R injury in rats,and the dose 50 ng/ml (6 μl) is more suitable.
3.Sodium houttuyfonate inhibits virulence related motility of Pseudomonas aeruginosa.
Da-qiang WU ; Wei-feng HUANG ; Qiang-jun DUAN ; Hui-juan CHENG ; Chang-zhong WANG
China Journal of Chinese Materia Medica 2015;40(8):1585-1588
Sodium houttuyfonate (SH) is a derivative of effective component of a Chinese material medica, Houttuynia cordata, which is applied in anti-infection of microorganism. But, the antimicrobial mechanisms of SH still remain unclear. Here, we firstly discovered that SH effectively inhibits the three types of virulence related motility of.Pseudomonas aeruginosa, i.e., swimming, twitching and swarming. The plate assay results showed that the inhibitory action of SH against swimming and twitching in 24 h and swarming in 48 h is dose-dependent; and bacteria nearly lost all of the motile activities under the concentration of 1 x minimum inhibitory concentration (MIC) (512 mg x L(-1) same as azithromycin positive group (1 x MIC, 16 mg x L(-1)). Furthermore, we found that the expression of structural gene flgB and pilG is down-regulated by SH, which implies that inhibitory mechanism of SH against motility of P. aeruginosa may be due to the inhibition of flagella and pili bioformation of P. aeruginosa by SR Therefore, our presented results firstly demonstrate that SH effectively inhibits the motility activities of P. aeruginosa, and suggest that SH could be a promising antipseudomonas agents in clinic.
Alkanes
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pharmacology
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Anti-Bacterial Agents
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pharmacology
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Bacterial Proteins
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genetics
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metabolism
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Biofilms
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drug effects
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Drugs, Chinese Herbal
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pharmacology
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Fimbriae, Bacterial
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drug effects
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genetics
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metabolism
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Houttuynia
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chemistry
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Pseudomonas aeruginosa
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cytology
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drug effects
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genetics
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pathogenicity
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Sulfites
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pharmacology
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Virulence
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drug effects
4.The distribution and related risk factors of femoro-popliteal artery chronic total occlusion lesions
Gangzhu LIANG ; Fuxian ZHANG ; Hailiang WEI ; Yongli DUAN ; Hui ZHAO ; Mingyi ZHANG ; Long CHENG
Journal of Interventional Radiology 2015;24(12):1052-1055
Objective To investigate the distribution of femoro-popliteal artery chronic totally occlusions (CTO) and to analyze the influence of different risk factors on the distribution of lesions. Methods A total of 47 patients with CTA and DSA proved femoro-popliteal artery CTO, who were admitted to the Affiliated Beijing Shijitan Hospital of Capital Medical University during the period from January 2013 and May 2015, were enrolled in this study. In order to make a clear description about the distribution characteristics of the lesions, the superficial femoral artery was averagely divided into three segments (F1-F3), and the popliteal artery was also divided into three segments (P1-P3) according to the bony landmarks. Based on the CTA images (including 3D reconstruction images) and DSA images, the distribution of CTO lesions was determined. The factors that might influence the distribution of CTO lesions were collected, and the results were analyzed using multivariable logistic regression analysis. Results A total of 59 femoro-popliteal artery CTO lesions detected in 47 patients were included in this study. The average length of lesion segment was (12.91 ± 10.13) cm. According to the 6-section division method of femoro-popliteal artery, the lesions involved F1 (n=23), F2(n=34), F3(n=48), P1(n=18), P2(n=6) as well as P3(n=5) segments. Multivariable logistic regression analysis reveals that F1 segment was most prone to be affected by CTO lesions in patients with hypertension (RR=3.21) and in patients who had cigarette smoking habit (RR=1.76). In male patients, P1segment was more easil y involved by CTO lesions (RR=1.98). In patients with diabetes mellitus, the RR value was gradually increased from the proximal to the distal end of the blood vessel, indicating that the lesion was more likely to involve distal vessels. Conclusion The distributions of femoro-popliteal artery CTO lesions have certain characteristics, which are closely associated with femoro-popliteal artery hemodynamics and atherosclerosis risk factors.
5.Adeno-associated vector mediated intracellular biological activity of human Kallistatin.
Xun-Wei DUAN ; Si-Yi CHEN ; Feng WANG ; Zhi-Yun CHENG ; Ming-Qing TANG ; Rui-An XU
Acta Pharmaceutica Sinica 2014;49(7):993-999
Human tissue kallikrein-binding protein (Kallistatin, KAL), a secretory protein that participates in the regulation of multiple signaling pathways by binding to the extracellular receptor, however, at present has not been reported about the intracellular activity, and whether it has the similar biological activity with extracellular activity. Here we constructed no signal peptide KAL (NSK) into the adeno-associated virus vector to explore the intracellular activity of KAL. Both the endothelial cell and lung cancer cells could express KAL, but not secreted after rAAV2-NSK transfection. The proliferation and migration of human umbilical vein endothelial cells (HUVECs) were inhibited, but the apoptosis rate was not affected. The proliferation rates, mobility and tubule formation of all the three tested lung cancer cells, such as NCI-H446, NCI-H460 and A549, were inhibited to different extents. This cellular study not only confirmed the intracellular activity, but also suggested it may serve as a kind of "balance factor" in multi-targeted controlling, which may provide a new train of thoughts to explain the regulatory contradiction in PI3K-Akt signaling pathways by KAL.
Apoptosis
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Cell Proliferation
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Dependovirus
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Genetic Vectors
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Human Umbilical Vein Endothelial Cells
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metabolism
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Humans
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Lung Neoplasms
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metabolism
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Serpins
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metabolism
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Signal Transduction
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Transfection
6.Effect of total flavonoids of acanthopanax senticosus on erythrocyte membrane fluidity in mice
Xiao-Xu WEN ; Li-Jing CHENG ; Wei LIU ; Duan-Hao FENG ; Ling SUN ;
Chinese Journal of Primary Medicine and Pharmacy 2006;0(08):-
Objective To observe contents and effect of total flavonoids of acanthopanax senticosus extract on erythrocyte membrane fluidity.Methods The contents of total flavonoids of acanthopanax sentieosus were deter- mined by spectrophotometry,and erythrocyte membrane fluidity and microviscosity were measured by fluorescence polarization technique.Results The total flavonoids contents of acanthopanax senticosus extract and injection were alike.Both of them increased erythrocyte membrane fluidity and lowered microviscosity,and the effect of acan- thopanax senticosus injection was more significant.Conclusion Total flavonoids of acanthopanax senticosus can in- crease erythrocyte membrane fluidity,so that blood viscosity is lowered and hemorheology factors are improved.
7.Effect of the anterior aspect of sacral nerve root tunnel on iliosacral screw placement on the standard lateral image of sacrum.
Hong-Min CAI ; Chuan-De CHENG ; Xue-Jian WU ; Wu-Chao WANG ; Jin-Cheng TANG ; Wei-Fang DUAN ; Chuan ZHANG ; Hong-Wei LI ; Wu-Yin LI
China Journal of Orthopaedics and Traumatology 2014;27(4):326-330
OBJECTIVETo introduce the location and course of S1, S2 sacral nerve root tunnel and to clarify the significance of the anterior aspect of sacral nerve root tunnel on placement of iliosacral screw on the standard lateral sacral view.
METHODSFirstly the data of 2.0 mm slice pelvic axial CT images were imported into Mimics 10.0, and the sacrum, innominate bones, and sacral nerve root tunnels were reconstructed into 3D views respectively, which were rotated to the standard lateral sacral views, pelvic outlet and inlet views. Then the location and course of the S1, S2 sacral nerve root tunnel on each view were observed.
RESULTSThe sacral nerve root tunnel started from the cranial end and anterior aspect of the vertebral canal of the same segment and ended up to the anterior sacral foramen with a direction from cranial-posterior-medial to caudal-anterior-lateral. The tunnel had a lower density than the iliac cortex and greater sciatic notch on the pelvic X-rays,especially on the standard sacral lateral view, on which it showed up as a disrupted are line and required more careful recognition.
CONCLUSIONIt can prevent the iliosacral screw from penetrating the sacral nerve root tunnel and vertebral canal when recognizing the anterior aspect of sacral nerve root tunnel and choosing it as the caudal-posterior boundary of the "safe zone" on the standard lateral sacral view.
Adult ; Aged ; Bone Screws ; Female ; Fracture Fixation, Internal ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Pelvic Bones ; diagnostic imaging ; injuries ; innervation ; surgery ; Radiography ; Sacrococcygeal Region ; diagnostic imaging ; innervation ; surgery ; Sacrum ; diagnostic imaging ; injuries ; innervation ; surgery ; Spinal Nerve Roots ; diagnostic imaging ; surgery ; Young Adult
8.Reversal of multidrug resistance in drug-resistant cell line EAC/ADR by cepharanthine hydrochloride and its mechanism.
Yu-cheng SONG ; Wei XIA ; Jin-hua JIANG ; Qing-duan WANG
Acta Pharmaceutica Sinica 2005;40(3):204-207
AIMTo investigate the correlation between reversal effect of cepharanthine hydrochloride (CH) on multidrug resistance (MDR) in drug-resistant cell line EAC/ADR and the nuclear transcription factor-KB (NF-KB).
METHODSCytotoxicity was determined by the tetrazolium (MTT) assay in vitro. An EAC/ADR cell homograft model was established to investigate the effect of CH on reversing MDR in vivo. The constitutive activity and activation of NF-KB by drugs were measured by Dot-Enzyme-linked Immune Sorbent Assay (Dot-ELISA).
RESULTSCH was shown to potentiate the cytotoxicity of ADR, a 13- fold reversal effect of resistance was achieved in vitro. In mice bearing EAC/ADR cell homografts, CH was found to prolong the survival time of animals bearing tumor. Increase in life span over control was 75. 37%. In addition, the constitutive activity of NF-KB and activation of NF-KB by chemotherapy were lowered by CH.
CONCLUSIONThe findings suggest that CH is able to reverse drug resistance and its mechanism may be related to suppressing the constitutive activity and activation of NF-KB by drugs.
Alkaloids ; isolation & purification ; pharmacology ; Animals ; Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Benzylisoquinolines ; Carcinoma, Ehrlich Tumor ; drug therapy ; metabolism ; pathology ; Cell Line, Tumor ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Female ; Male ; Mice ; NF-kappa B ; metabolism ; Neoplasm Transplantation ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Random Allocation ; Stephania ; chemistry
9.Influence of the included angle between anterior aspects of S2 and S1 vertebral bodies on pelvic inlet imaging in mid-line sagittal plane.
Hong-ming CAI ; Shu-tu GAO ; Chuan-de CHENG ; Xue-jian WU ; Wu-chao WANG ; Jin-cheng TANG ; Shou-ya CHANG ; Wei-feng DUAN ; Chuan ZHANG
China Journal of Orthopaedics and Traumatology 2014;27(8):645-649
OBJECTIVETo analyze the influence of included angle between the anterior aspects of S2 and S vertebral bodies on pelvic inlet imaging in the pelvic midline sagittal plane.
METHODSTotally 58 axial pelvic CT scans were chosen as study objects including 43 males and 15 females,with an average age of 40.7 years old (ranged,18 to 68 years old). The angles between the anterior aspects of S2 and S1, vertebral bodies and the horizontal plane on midline sagittal CT reconstruction were measured to simulate the optimal S2 and S1 inlet angles. The included angle between the anterior aspects of S2 and S1 vertebral bodies was calculated by subtrocting the S1,inlet angle from the S2 inlet angle defined as a base number. Then, the impact of the calculated included angles on the pelvic inlet imaging was analyzed. Results:The S2 inlet angles averaged (30.5±6.5) degrees; the S inlet angles averaged (25.7±5.9) degrees. The difference between them was significant (t=3.35, P=0.001). Ten patients had zero angle between the anterior aspects of S2 and S1 vertebral bodies; 14 patients had negative angle, averaged-(8.9±8.1) degrees; 34 patients had positive angle,averaged (11.8+6.4) degrees.
CONCLUSIONThe difference of included angle between the anterior aspects of S2 and S1 vertebral bodies leads to the difference between S1 inlet view and S2 inlet view in most cases, complicating the pelvic inlet imaging,and affecting the reliability of the application of pelvic inlet view. Utilizing the angles measured on the preoperative midlihe sagittal CT reconstruction to obatin the patient-customized S1 and S2 inlet views could accurately guide the S1 and S2 iliosacral screw insertion.
Adolescent ; Adult ; Aged ; Animals ; Bone Screws ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Image Processing, Computer-Assisted ; Male ; Middle Aged ; Pelvis ; anatomy & histology ; injuries ; Spine ; anatomy & histology ; Tomography, X-Ray Computed ; Young Adult
10.Classification of upper sacral segment based on continuous axial pelvic computed tomography scan.
Hong-min CAI ; Shu-tu GAO ; Chuan-de CHENG ; Xue-jian WU ; Wu-chao WANG ; Jin-cheng TANG ; Wei-feng DUAN ; Chuan ZHANG
China Journal of Orthopaedics and Traumatology 2014;27(10):866-869
OBJECTIVESTo introduce a classification system of upper sacral segment and its significance based on the continuous pelvic axial computed tomography scan.
METHODSThe whole pelvis 2.0 mm thick axial scan images of 127 cases were observed, the sacroiliac screw channel of S1 were measured, according to the size of the transverse screw channel the upper sacral segment were classified. Such as transverse screw channel existed and in at least 4 layer scan images its width was > 7.3 mm, it was defined as sacral segment of the normal type. Such as transverse screw channel existed and its maximum width was 7.3 mm or less on scanning level, it was defined as a transitional. Such as transverse channel did not exist, or its width on all scanning level was 0 mm or less, it was defined as dysplastic. Various cases,percentage, and the average of the transverse screw channel were calculated.
RESULTSThere were 58 normal (45.7%),42 transitional (33.1%), and 27 dysplastic (21.2%) upper sacral segments with an averaged width of the tansverse screw channel of 13.9 mm, 5.2 mm, and 0.9 mm, respectively. Each specimen could be defined as one of the three types of upper sacral segment without exceptions.
CONCLUSIONIt is possible to insert a transverse iliosacral screw into a normal upper sacral segment when indicated because of the capacious transverse screw channel. The transverse iliosacral screw placement into the transitional and dysplastic upper sacral segments was contraindicated because of the limited or none transverse screw channel. The transitional upper sacral segment was superior to the dysplastic segment due to its starting point location restriction on the true lateral sacral view.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bone Density ; Bone Screws ; Female ; Fracture Fixation, Internal ; Humans ; Male ; Middle Aged ; Pelvic Bones ; diagnostic imaging ; surgery ; Sacrum ; diagnostic imaging ; surgery ; Tomography, X-Ray Computed ; Young Adult