Since 2000,most of H5N1 subtype influenza A virus had a unique mutation of NS gene with 15base pair deletion from 263 to 277. In order to investigate the bio-characteristics of this mutation,two different NS recombinants,RWSN-248 and RWSN-m248,were generated via plasmid rescue from A/WSN/33(H1N1) and A/SD/04(H5N1). RWSN-248 had a higher viral titer than RWSN-m248 in MDCK and COS-1 cells that have an IFN response,but they had the similar growth ability in Vero cells that lack an IFN response. Both of two recombinants grew well in embryonated chicken eggs and had the similar viral titer and MDT. The results above revealed that the deletion from 263 to 277 sites of NS gene did not influence viral virulence to but decreased viral anti-IFN ability of H5N1.

It has been showed that there were obvious obstacle effects of Panax notoginseng replanting. Crop rotation was the main effective technique to overcome the obstacle. To find a reasonable crop rotation system for P. notoginseng, aqueous extracts from root, stem and leaf of P. notoginseng were analyzed for allelopathic effect on three maize varieties (which are often grown in regions where P. notoginseng grown). The main results were as follows: (1) Allelopathic effect of P. notoginseng stem and leaf extracts on the three other tested plants was stronger than that of root extracts; (2) Corn was more vulnerable to the effects of allelochemicals at seedling stage than at germination stage, and the corn root was more sensitive than aerial part to allelochemicals; (3) Lusan No. 3 and Yunrui No. 1 showed resistance to P. notoginseng allelopathy, with respective comprehensive sensitivity indexes (M3) of - 0.089 3 and -0.159 2, while Bainuo No. 1 is sensitive at M3 = -0.261 0. It then can be concluded that Lusan No. 3 and Yunrui No. 1 may be an alternative rotation plants for overcoming P. notoginseng continuous cropping obstacle.
Allelopathy ; Panax notoginseng ; chemistry ; growth & development ; Pheromones ; pharmacology ; Plant Extracts ; pharmacology ; Plant Leaves ; chemistry ; Plant Roots ; chemistry ; Plant Stems ; chemistry ; Zea mays ; drug effects ; growth & development

OBJECTIVETo probe into clinical therapeutic effect of acupuncture on diabetic nephropathies and the mechanism.

METHODSUsing multi-central, randomized and blind methods, 130 cases of diabetic nephropathy were divided into an observation group and a control group, 65 cases in each group. They were treated by routine diabetic therapy, and in the observation group, acupuncture at Quchi (LI 11), Zhigou (TE 6), Hegu (LI 4), Xuehai (SP 10), Zusanli (ST 36), Yinlingquan (SP 9), Fenglong (ST 40), Diji (SP 8), Sanyinjiao (SP 6), Taichong (LR 3), Tianshu (ST 25), Gaohuang (BL 43), Shenshu (BL 23), Zhongwan (CV 12), Zhongji (CV 3) were added with needling method of harmonizing spleen-stomach. While in the control group, acupuncture at Shenshu (BL 23), Taixi (KI 3), Sanyingjiao (SP 6), Yanglingquan (GB 34), Xuanzhong (GB 39), Guanyuan (CV 4), Shousanli (LI 10), Waiguan (TE 5), Yangxi (LI 5), Liangqiu (ST 34), Shangjuxu (ST 37), Neiting (ST 44), Huaroumen (ST 24), Dachangshu (BL 25). The treatment was given twice a day in the two groups. Clinical therapeutic effects were assessed according to clinical symptoms and signs, blood sugar, blood lipids, urinary albumin excretion rate, urinary monocyte chemotactic protein-1 (MCP-1), glomerular filtration rate (GFR), renal blood flow, etc..

RESULTSThe needling method of harmonizing spleen-stomach not only could improve symptoms and signs of the patients, and also had benign regulative action on metabolism of blood sugar and lipids, and GFR, renal blood flow and urinary albumin level, with significant or very significant differences as compared with the control group ( P < 0.05 or P < 0.01).

CONCLUSIONThe needling method of harmonizing spleen-stomach is an effective method for diabetic nephropathies, which can improve progressive renal lesion induced by abnormal metabolism of blood sugar and lipids, improve renal blood flow and GFR, decrease urinary albumin secretion, inhibit over expression of MCP-1, protect glomerulus and renal tubules, so as to improve renal function and delay renal lesion.

Acupuncture Therapy ; methods ; Aged ; Blood Glucose ; analysis ; Blood Urea Nitrogen ; Diabetic Nephropathies ; therapy ; Female ; Glomerular Filtration Rate ; Humans ; Kidney ; physiopathology ; Lipids ; blood ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Spleen ; physiopathology ; Stomach ; physiopathology

Objective To observe the effect of Tetramethylpyrazine combined with cisplatin on Lewis lung adenocarcinoma by the change of hypoxia-inducible factor 1 alpha ( HIF -1α) , basic fibroblast growth factorb ( b-FGF) ,vasohibin-1 ( VASH-1 ) in mice tumor tissues and explore the mechanism of anti -tumor.Methods Forty -eight mice were randomly divided into 4 groups, control group, cisplatin group, Tetramethylpyrazine group and combined group, 12 rats in each group, cisplatin at a dose of 2 mg· kg -1 , Tetramethylpyrazine at a dose of 100 mg· kg-1 , combination group( the doses as above).All mice were sac-rificed after treatment for two weeks and these tumors were obtained.The weighed, and the tumor inhibitory was calculated.The expressions of HIF-1α, b -FGF and VASH -1 were determined by immunohisto-chemistry after two weeks administration.Results Compared with the control group, the tumor inhibition rate in experimental groups was signi-ficantly increased ( P<0.05 ) , and was highest in combination group.The expression of HIF -1α, b -FGF and VASH -1 in the cisplatin group, Tetramethylpyrazine group and combined group were decreased obviously compared with control group( P<0.05).And the levels of the expression were the lowest in the combined group and the highest in control group.HIF -1αis positively correlated with b -FGF ( P <0.05 ) , b -FGF is positively correlated with VASH-1 ( P <0.05 ) , HIF-1αis uncorrelated with VASH-1.Conclusion Tetramethylpyrazine combined with cisplatin decreased the expression of HIF-1α, thereby reducing the expression of b-FGF and VASH-1 , inhibiting the tumor growth synergistically.

Objective To investigate the effect of Tetramethylpyrazin ( TMP) combined with cisplatin( DDP) on expression of a disintegrin and metalloproteinase with thrombospondin typeⅠmotif ( ADAMTS1 ) and vascular endothelial growth factor ( VEGF ) of Lewis lung cancer rats.Methods The Lewis lung carcinoma model of c57 bl/6 were produced by intraperitoneal injection of Lewis lung cancer cells in right axillary, per 0.2 mL.Then the 56 tumor-bearing mice were randomized into 4 groups: model group ( 0.9%NaCl, 0.2 mL ) , TMP group ( 100 mg· kg-1, 0.2 mL),DDP group(2 mg· kg -1, 0.2 mL),TMP+DDP group( doses as above, n=14).Different administration were served from day 5 transplantation and all mice were sacrificed after 19 days.The weight and volume of transplanted tumors were recorded.And the expressions of VEGF and ADAMTS1 were detected by immunhistochem-isty.Results Compared with model group, the growth of transplanted tumors in TMP group, DDP group, TMP+DDP group were significantly inhibited.The weight of tumor body in the three groups were markedly lower than that of model group.The inhibitory effect of TMP combined with DDP on tumor growth was enhanced com-pared with TMP or DDP alone treatment.The expression levels of VEGF were significantly lower but ADAMTS1 was sig-nificantly higher in TMP group,DDP group,TMP+DDP group than that in model group ( P<0.05 ).Pearson correla-ted analysis showed that the expression VEGE and ADAMTS1 negatively correlated(P<0.05).Conclusion TMP has synergetic inhibitory effect with DDP on transplanted tumor growth of Lewis lung carcinoma in mice.The action mecha-nism may be due to down-regulate the VEGF expression and up-regulate the ADAMTS1 expression,inhibiting tumor angiogenesis.

To investigate the angiotensin I-converting enzyme (ACE) inhibitory activity of beta-chain hemoglobin fragments, 17 fragments were synthesized by microwave-assisted solid-phase synthesis method. Wang resin or Trt(2-Cl) resin, Fmoc and HBTU-HOBt were used as solid carrier, N-terminal amino acid protecting groups and coupling reagents, respectively. The ACE inhibitory, alpha-glucosidase inhibitory, antibacterial and antitumor activities of the synthesized fragments were assayed. In vitro, Val-Val-Tyr-Pro-Trp-Thr showed high ACE inhibitory activity (IC50 = 7.42 micromol x L(-1)). The results indicate that there are two active sites in Val-Val-Tyr-Pro-Trp-Thr-Gln-Arg-Phe, one consists of Val-Val-, and the other -Gln-Arg-Phe. Peptides showed high ACE inhibitory activity when the N-terminal was hydrophobic amino acid such as Val and C-terminal tripeptide contained Phe, Trp or Arg. Some of the fragments showed low a-glucosidase inhibitory activity. No antibacterial activity or antitumor activity was detected in vitro. The results indicate that these peptides have a potential antihypertensive effect and possible application in the treatment of hypertension.
Amino Acid Sequence ; Angiotensin-Converting Enzyme Inhibitors ; pharmacology ; Anti-Bacterial Agents ; pharmacology ; Antihypertensive Agents ; pharmacology ; Antineoplastic Agents ; pharmacology ; Cell Line, Tumor ; Glycoside Hydrolase Inhibitors ; Humans ; Peptide Fragments ; chemical synthesis ; chemistry ; pharmacology ; Peptidyl-Dipeptidase A ; drug effects ; Solid-Phase Synthesis Techniques ; methods ; alpha-Glucosidases ; drug effects ; beta-Globins ; chemical synthesis ; chemistry ; pharmacology

OBJECTIVETFo compare the efficacy and complications rate of intramedullary (IM) nailing or K-wire versus plating fixation for clavicular fractures.

METHODSPubmed, Embase, Cochrane Library databases, CNKI, VIP and Wangfang databases were searched to find all randomized or quasi-randomized controlled trials of clavicle fractures using plating versus IM nailing or K-wire. The methodologic quality of the studies was assessed. After independent study selection by 2 authors ,data were collected and extracted independently. Outcomes of postoperative shoulder functional measurement, the efficacy and information of the operation and complications rate were meta-analyzed using RevMan 5 software.

RESULTSNine hundreds and seventy-six patients in 10 randomized controlled trials (RCTs) and 3 quasi-RCTs were involved in the meta-analysis,of which 5 studies compared the K-wire and the plating fixations and 8 studies compared the IM nailing and the plating fixations. The overall odds ratio(OR) (with 95% CI) of the operation efficacy for K-wire versus the plating was 3.79 (1.93, 7.46). The overall weighted mean difference (with 95% CI) of Constant Shoulder score for plating versus IM fixation was -1.39 (-3.43, 0.65) in 6 studies. The overall OR of the plating versus IM nailing was 9.34(2.70, 32.32) for the overall major complications in 5 studies and 5.04 (1.52,16.77) for the revision rate in 5 studies.

CONCLUSIONThe current limited evidences suggested that the IM fixation could reduce the incidences of the overall major complications and the revision surgery, while the post-operative efficacy of the plating was superior to the K-wire. More high quality RCTs are still needed in the future.

Bone Nails ; Bone Wires ; Clavicle ; injuries ; surgery ; Fracture Fixation, Intramedullary ; instrumentation ; methods ; Fractures, Bone ; surgery ; Humans ; Randomized Controlled Trials as Topic

S1 nuclease (from Aspergillus oryzae) is a specific enzyme to degrade single stranded DNA or RNA molecules. It has been reported to be able to convert superhelical circular DNA molecules into open circle or linear forms under certain conditions, but this function has not been well explored. In order to use the action of S1 nuclease to linearize circular DNA and develop a novel way of cloning microcircular DNAs, the pUC19 was used to investigate the relationship between the linearization efficiency of S1 nuclease and the amount of enzyme used. By this way the optimal conditions for linearization of circular DNAs by S1 nuclease would be determined. 0.3u to 17u S1 nuclease per 100ng pUC19 DNA was added into a 25 microL system, respectively, to perform the reaction. The effectiveness of enzyme digestion was realized by electrophoresis in a 1.2% agarose gel. The results showed that along with the increase in enzyme amount from 0.3u to 17u a gradual decrease in the superhelical form, a gradual increase in the linear form and then in the circular form was obvious. The conversion from superhelical form to linear and circular form was directly related to the enzyme amount used. A higher proportion of linear DNA molecules was achieved by using 5 to 17u S1 nuclease per 100ng DNA. Besides, electrophoretic mobility of the S1 nuclease-linearized pUC19 was the same as that of the linear form produced by restriction enzyme digestion. According to the result of phiX174 digested by S1 nuclease it has been proposed that the enzyme cleaves first randomly on one site of one strand, thus converting the superhelical molecules into open circle form, and then on the same site of the complementary strand to produce the linear form. Therefore, the S1 nuclease-linearized DNA molecules are intact in the sense of their length and can be used for cloning. The plasmid-like DNA pC3 from cucumber mitochondria is a double stranded circular DNA molecule with about 550bp and the smallest known plasmid-like DNA in eukaryotic mitochondria. Many attempts have been made to linearize the molecule by using restriction enzymes but failed. Therefore, S1 nuclease was used to linearize pC3 based on the results obtained with pUC19. The linearized pC3 DNA molecules formed a very sharp band in a 2.5% agarose gel after electrophoresis. They were then recovered from the gel, added an "A" tail and ligated with T-vector. After transformation into E. coli JM109 cells, the positive clones were, screened by the blue-white selection. The insert was then cut using restriction enzymes EcoRI and Pst I. The result of electrophoresis shows that the electrophoretic mobility of the insert is just the same as that predicted. A 32 P-labled probe was synthesized using pC3 as the template and Southern blot analysis was carried out. The result shows that the inserted DNA is hybridized to the probe, which indicates that the cloned DNA fragment is from pC3. The sequence information of the insert shows that the plasmid-like DNA pC3 was 537bp in length. The nucleotide sequence was deposited in the GenBank (the accession number is AF522195).
Blotting, Southern ; Cloning, Molecular ; methods ; DNA, Circular ; genetics ; metabolism ; Fungal Proteins ; genetics ; metabolism ; Molecular Sequence Data ; Single-Strand Specific DNA and RNA Endonucleases ; genetics ; metabolism

OBJECTIVETo investigate the influence of fluorosis on nicotinic acetylcholine receptors (nAChRs) in protein and gene levels in SH-SY5Y cells and the mechanism of the receptor modification.

METHODSSH-SY5Y cells, a human neuroblastoma cell line, were incubated with different concentrations of fluoride or with antioxidant for 48 hours. The functions of cells were measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) method, and protein oxidation detected by carbonyl content; the alpha3 and alpha7 nAChR subunits in protein level were measured by Western blotting and in mRNA level by RT-polymerase chain reaction (RT-PCR).

RESULTSIn high-dose group as compared to the control, the decreased MTT (49%), increased protein oxidation (72%), and lower expression of alpha3 (51%) and alpha7 (47%) nAChR subunit proteins were obviously observed in SH-SY5Y cells. There were no changes in expression of nAChR subunit mRNAs between the cells treated with fluoride and those un-treated in controls. Prior treatment with antioxidant resulted in preventing the decrease of nAChR protein in cells exposed to the high doses of fluoride.

CONCLUSIONFluorosis should result in damage of cells and the declined expression of nAChRs in protein levels, but no influences on gene expression of the receptors in human neuroblastoma neurons. The decreased nAChR proteins might be involved in the mechanism of oxidative stress induced by fluorosis.

Cell Line, Tumor ; Fluoride Poisoning ; metabolism ; Fluorides ; toxicity ; Humans ; Neuroblastoma ; metabolism ; pathology ; Protein Processing, Post-Translational ; drug effects ; Proteins ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, Nicotinic ; biosynthesis ; genetics

Objective To develop a new algorithm to reconstruct the distribution of acoustic sources of magnetoacoustic tomography with magnetic induction(MAT-MI)in the acoustic inhomogeneous media,which is developed on the basis of generalized finite element method (GFEM) and modified time inversion algorithm. Methods The acoustic and acoustic coupling theory and the basic equations of acoustics were used to study the forward and inverse problems of the acoustic inhomogeneous concentric sphere magneticacoustic coupling model. The solution of acoustic non-uniform media wave equation based on GFEM was proposed.The method solved the problem of acoustically inhomogeneous media sound source reconstruction and conductivity reconstruction.At the same time,the distribution of velocity was reconstructed by rotating the pairs of transducers and the time reversal algorithm. Results The proposed algorithm could accurately reconstruct the acoustic source distribution in acoustic inhomogeneous media,and could obtain the distribution of sound velocity during the reconstruction of sound source and recover the image well. Conclusion The proposed algorithm had its feasibility and effectiveness verified,and gains advantages in MAT-MI reconstruction of acoustic inhomogeneous media.