In 2013, the World Health Organization reported the first case of human infection with a new influenza A (H7N9) virus in China. This has caused damage and panic within certain areas in China. Therefore, analysis of this virus with bioinformatics technology is very necessary. Neuraminidase (NA) is one of the most important antigens of the influenza virus and an important target for anti-flu drugs. In this study, the nucleotide and protein sequences of NA gene of A/H7N9 influenza viruses were retrieved from the NCBI database, and MEGA 5.0 software was employed to construct a phylogenetic tree based on the nucleotide coding sequence; BioEdit software was used to align the nucleotide and protein sequences of NA and calculate the homologies of nucleotides and amino acids and then to analyze the important mutation sites of NA gene. The results demonstrated that the spread of influenza virus H7N9 showed certain geographical and temporal relations. The H7N9 virus isolated from China in 2013 belonged to Euroasiatic serotype, and its NA stalk region hadobvious variation, which may be one of the reasons that this virus infects human. These analyses may be very helpful for understanding the evolutionary relationship and mutation trend of A/H7N9 influenza viruses.
Databases, Genetic ; Evolution, Molecular ; Humans ; Influenza A Virus, H7N9 Subtype ; enzymology ; genetics ; Mutation ; Neuraminidase ; chemistry ; genetics ; Phylogeny ; Sequence Analysis

Objective To observe the clinical effect of sling exercise therapy(S-E-T)combined with drug treatment for cervical vertigo in elderly patients.Methods Forty-nine elderly patients with cervical vertigo admitted to our hospital between January 2011 and July 2014 were randomly divided into an observation group(n=27)and a control group(n=22).The observation group was given 80 mg Ginaton(Extract of Ginkgo Biloba Leaves Tablets)produced by German Dr.Willmar Schwabe GmbH & Co.KG three times a day,combined with S-E-T,including cervical stability and stretching training for 40min,focusing on the neck global muscle and local stabilize muscle rehabilitation,once every other day.The control group was provided with the same drug treatment.During the 6-month intervention,both groups were given health education by the same therapist.Both groups were assessed using the neck disability index(NDI),visual analogue scale(VAS)and evaluation scale for cervical vertigo(ESCV) before and after the intervention,as well as at the last follow-up visit.Before the treatment and at the last follow-up visit,the cervical X-ray examination and trigger point check were also conducted for both groups.Results All the forty-nine patients were followed up for 4.83 to 6.70 months,with an average of(6.01 ± 0.49)months.Significant improvement was observed in the average ESCV score for both groups after the treatment.Compared with before the treatment,there was significant improvement in the average NDI and VAS right after the treatment and at the last follow-up visit in the observation group,but only at the last follow-up visit in the control group.From the cervical X-ray,no significant differences were found in the vertebral osteophyte formation,facet joints and uncovertebral joint degeneration between the 2 groups(P＞0.05),while significant differences were observed in the number of the neck trigger points(P＜0.05).Conclusion The sling exercise therapy combined with drug treatment can significantly improve cervical function,relieve pain and vertigo symptoms in elderly patients with cervical vertigo.The effect is better than drug treatment alone.

OBJECTIVETo investigate the effect of Huogu II Formula (II) with medicinal guide Radix Achyranthis Bidentatae (Ach) on bone marrow stem cells (BMSCs) homing to necrosis area after osteonecrosis of the femoral head (ONFH) frozen by liquid nitrogen in rabbit as well as to explore the mechanism of prevention and treatment for ONFH.

METHODSThe animal model of ONFH was established by liquid nitrogen frozen on the rabbit left hind leg. Forty-eight Japanese White rabbits were randomly assigned to sham-operated group, model group, Huogu II group, and Huogu II plus Ach group, with 12 rabbits in each. During the course of ONFH animal model establishment, all rabbits were subcutaneously injected with recombinant human granulocyte colony-stimulating factor [rhG-CSF, 30 μg/(kg·day) for continuous 7 days]. Meanwhile, normal saline and decoction of the two formulae were administrated by gavage, respectively. White blood cells (WBC) were counted in peripheral blood before and after injection of rhG-CSF. Materials were drawn on the 2nd and 4th weeks after model built; bone glutamine protein (BGP) and bone morphogenetic protein 2 (BMP2) levels in serum were tested. Histopathologic changes were observed by hematoxylin and eosin (HE) staining. BMP2 mRNA levels were detected with in situ hybridization (ISH) staining. 5-Bromo-2'-deoxyuridine (BrdU) and stromal cell derived factor 1 (SDF-1) were measured by immunohistochemical assay in femoral head of the left hind leg.

RESULTSCompared with the shamoperated group, the ratio of empty lacuna, serum BGP, and SDF-1 level in the model group increased significantly, and BMP2 in both serum and femoral head decreased significantly. However, in comparison with the model group, the empty lacuna ratio of Huogu II group and Huogu II plus Ach group decreased obviously in addition to the levels of serum BGP and BMP2, and the expressions of BMP2 mRNA, BrdU, and SDF-1 increased significantly. Above changes were particularly obvious in Huogu II plus Ach group. BGP and SDF-1 on the 2nd week and empty lacuna rate and serum BMP2 level on the 4th week in Huogu II group significantly exceeded their counterparts. On the 2nd week, only in Huogu II plus Ach group that the BrdU counting rose significantly. On the 4th week, empty lacuna rate and serum BMP2 level in Huogu II plus Ach group exceeded those in Huogu II group distinctively.

CONCLUSIONSTo a certain extent, the medicinal guide Ach improves the preventive and therapeutic effects of Huogu II Formula on experimental ONFH model. The possible mechanism of this is related to its promoting effect on directional homing of BMSCs to the necrosis area.

Achyranthes ; Animals ; Bone Marrow Cells ; cytology ; drug effects ; Bone Morphogenetic Protein 2 ; blood ; genetics ; Bromodeoxyuridine ; metabolism ; Cell Movement ; Chemokine CXCL12 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Enzyme-Linked Immunosorbent Assay ; Femur Head ; drug effects ; pathology ; Femur Head Necrosis ; blood ; genetics ; pathology ; therapy ; Gene Expression Regulation ; drug effects ; Granulocyte Colony-Stimulating Factor ; administration & dosage ; pharmacology ; Humans ; Leukocyte Count ; Male ; RNA, Messenger ; genetics ; metabolism ; Rabbits ; Radioimmunoassay ; Stem Cell Transplantation ; Stem Cells ; cytology ; drug effects

OBJECTIVETo observe effects of blood circulation promoting compounds combined with medicinal guides on content of bone glaprotein (BGP), bone morphogenetic protein-2 (BPM-2) and expression of BMP-2 mRNA in rabbits with femoral head necrosis, and explore its mechanism.

METHODSNinety-eight healthy Spragur-Dawley male rabbits were collected and weighted 2.2 to 2.8 kg. Eighty-four rabbits were built femoral head necrosis model by freezing left femoral head in liquid nitrogen, then randomly divided into 6 groups, 14 in each group. The 6 groups included model group,promoting blood circulation to remove meridian obstruction group,promoting blood circulation to remove meridian obstruction combined with achyranthes bidentata group,radix angelicae pubescentis, asarum group, and platycodon grandiflorum group,other 14 rabbits were sham operation group. While drug groups were administrated corresponding Chinese herb after molding,model group and shamp operation group were given saline. Recombinant human granulocyte-colony stimulating factor ( 30 microg x kg(-1) x d(-1))were injected into all rabbits for 7 days. Samples were taken on the second and fourth week,the content of BGP and BMP-2 were detected by enzyme-linked immunosorbent assay (ELSA) and radioimmunoassay (RIA), histopathological changes of left femoral head were observed by Hematoxylin and Eeosin staining (HE), and expression of BMP-2 mRNA were tested by fluorescence in situ hybridization.

RESULTSCompared with sham operation group, the rate of empty lacunae femoral head were obviously increased in model group, and the content of BGP were increased on the second week, and BMP-2 and BMP-2 mRNA were decreased on the fourth week. Compared with model group, the content of BGP, BMP-2 and BMP-2 mRNA were higher both of the second and fourth week in promoting blood circulation to remove meridian obstruction group. The rate of empty la- cunae femoral head were lower in achyranthes bidentata group, BGP, BMP-2 and BMP-2 mRNA were higher on the fourth week. The rate of empty lacunae femoral head were lower in platycodon grandiflorum group, and BGP were decreased on the second and fourth week, BMP-2 were lower on the second week ,while BMP-2 mRNA were decreased on the fourth week; the content of BMP-2 and BMP-2 mRNA were increased in radix angelicae pubescentis group on the second week; while there was no change in asarum group.

CONCLUSIONRadix angelicae pubescentis can increase the content of BGP, BMP-2 and expression of BMP-2 mRNA ,which is an effective mechanism of preventing femoral head necrosis.

Animals ; Blood Circulation ; drug effects ; Bone Morphogenetic Protein 2 ; analysis ; genetics ; Femur Head Necrosis ; drug therapy ; pathology ; physiopathology ; Male ; Meridians ; Osteocalcin ; blood ; Osteogenesis ; drug effects ; Rabbits

Distribution behavior of lipophilic drugs in the oil-in-water (O/W) microemulsions was studied. Fluorescence spectra analysis was performed to investigate the effect of the compositions of microemulsions on the fluorescence spectra of armillarisin and ofloxacin which were used as the model drugs. The fluorescence spectra of the model drugs in the microemulsions with various amount of the compositions were compared. The results showed that the armillarisin were both localized in the interfacial film of microemulsion systems with both phenylmethanol and PEG 400 as the co-surfactants, separately. Ofloxacin was localized in the interfacial film of microemulsion systems with Gradamol GTCC as the oil phase, but in the oil pool of microemulsion systems with oleic acid/olive oil (OA/OO) (1:1) as the oil phase. Besides, it was found that the drug would have the tendency to locate in the microenvironment where the composition with the largest solubility to model drug is located, and its actual localized position would be dependent on the amount of this composition. The results indicate that the localized region of lipophilic drug in the O/W microemulsion systems is related with the solubility of the model drug in various compositions.
Benzopyrans ; chemistry ; Benzyl Alcohols ; Emulsions ; Ofloxacin ; chemistry ; Oils ; Polyethylene Glycols ; Solubility ; Spectrometry, Fluorescence ; methods ; Surface-Active Agents ; Water

OBJECTIVETo observe the effect of Huogu II formula compatible with different channel ushering drugs on the homing of bone marrow stem cells of osteonecrosis of the femoral head (ONFH) induced by liquid nitrogen freezing in rabbits and discuss the mechanism for preventing and treating ONFH.

METHODThe ONFH model was established by liquid nitrogen freezing of 84 rabbits. They were randomly assigned to the model group and the Huogu II formula group and groups of Huogu II formula combining with Achyranthis Bidentatae Radix, Asari Radix et Rhizoma, Angelicae Pubescentis Radix, Platycodonis Radix. The remaining 14 rabbits were sham-operated. During the course of ONFH modeling, all of the rabbits were subcutaneously injected with recombinant human granulocyte colony-stimulating factor (rhG-CSF)(30 microg x kg(-1) x d(-1), for consecutively 7 days). Meanwhile, normal saline and decoction of the formulae were orally administrated respectively. WBC was counted in peripheral blood before and after the injection of rhG-CSF. HE stainings at the 2nd and the 4th weeks after the modeling were adopted to observe histopathological changes, vascular morphology was observed by ink perfusion, BrdU and SDF-1 were determined by immunohistochemical assay in femoral heads of the left hind leg.

RESULTCompared with the sham-operated group, the Huogu II formula group showed decrease in the ratio of empty lacuna and increase in vessel area, number of BrdU positive cells and SDF-1 level. In comparison with the model group, the Achyranthis Bidentatae Radix group displayed decreasing empty lacuna ratio and increasing vessel area at the 4th week and increasing SDF-1 at the 2nd week; the Platycodonis Radix group revealed a notably increasing empty lacuna ratio and a sharp decrease in the number of BrdU positive cells at 4th week; Asari Radix et Rhizoma and Angelicae Dubescentis Radix groups showed no remarkable change.

CONCLUSIONHuogu II formula can promote the directional homing of bone marrow stem cell to the necrosis area. Channel ushering drug achyranthes can further boost above effects of Huogu II formula.

Animals ; Bone Marrow Cells ; cytology ; Bromodeoxyuridine ; metabolism ; Chemokine CXCL12 ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Femur Head Necrosis ; blood ; drug therapy ; metabolism ; pathology ; Gene Expression Regulation ; drug effects ; Humans ; Leukocyte Count ; Male ; Rabbits ; Stem Cells ; cytology ; drug effects

Objective To explore the efficacy and safety of polyethylene glycol recombinant human granulocyte colony-stimulating factor (PEG-rhG-CSF) in preventing chemotherapy-induced neutropenia in locally advanced non small cell lung cancer (NSCLC) patients at nutritional risk. Methods A total of 337 locally advanced NSCLC patients at nutritional risk were selected. They were randomly divided into three groups: 112 cases in the non-prophylactic drug group (control group), 112 cases in the prophylactic use of rhG-CSF treatment group (rhG-CSF group), and 113 cases in the prophylactic use of PEG-rhG-CSF treatment group (PEG-rhG-CSF group). The incidence and duration of neutropenia after chemotherapy and the ratio of CD4⁺/CD8⁺T cells in peripheral blood were observed. Results The incidences of neutropenia in the control group, rhG-CSF group, and PEG-rhG-CSF group were 67.97%, 41.57%, and 38.98% (P < 0.05), respectively. The incidences of grade Ⅲ-Ⅳ neutropenia in the three groups were 22.39%, 14.25%, and 11.14% (P < 0.05); moreover, the incidence of febrile neutropenia in the three groups was 3.55%, 1.84%, and 1.21% (P < 0.05); in addition, the ratios of CD4⁺/CD8⁺T cells in peripheral blood were 1.27±0.44, 1.41±0.52, and 1.49±0.42 (P < 0.05). The duration of grade Ⅲ-Ⅳ neutropenia and the time required for the neutrophil value to reach 2.0×10⁹/L from the lowest value in the PEG-rhG-CSF group were lower than those in the control and rhG-CSF groups (P < 0.05). Conclusion The PEG-rhG-CSF preventive treatment used in the course of chemoradiotherapy in locally advanced NSCLC patients at nutritional risk can reduce the incidence of neutropenia and improve immunologic function. PEG-rhG-CSF preventive treatment is worthy of clinical recommendation.

Objective To observe the levels of serum interleukin-1β(1L-1β)and interleukin-1 receptor antagonist(IL-1Ra),the IL-1Ra/IL-1β ratio and the relationship between the levels of thern and the restenosis in patients with intra-and extra-cranial arteriostenosis after stent-assisted angioplasty.Methods Thirty-one patients with cerebral artery stenosis,admitted to our hospital from April 2003 to March 2006,were treated with stent-assistant angioplasty and followed up for 6-12 months.The relationship was analyzed between the restenosis of cerebral artery and both the levels of serum IL-1β,IL-1Ra and the IL-1 Ra/IL-1β ratio before and 1 h,1,3 and 5 d after stent-assisted angioplasty.Results The condition of 31 stents in 31 blood vessels of cerebral arteries was observed and followed up 6 to 12 months after the operation.Restenosis was noted in 6(19.3%)with 3 restenosis superior to 50% and 3 restenosis from 10%-30%.No obvious difference of the levels of serum IL-1β and IL-1Ra,the IL-1Ra/IL-1β ratio between restenosis group and non-restenosis group before the operation was found (P>0.05);IL-11β and IL-1Ra levels were positively correlated before the operation.The levels of serum IL-1β and IL-1Ra 1h,1,3 and 5 d after the operation were superior to those before the operation,respectively(P<0.05).The levels of serum IL-1β and IL-1Ra in the restenosis group showed no significant difference to those in the non-restenosis group 6 to 12 months alter the operation(P>0.05);however,the IL-1Ra/IL-1β ratio in the restenosis group was significantly lower than that in the non-restenosis group (P<0.05);positive correlation between the levels of IL-1β and IL-1Ra was observed.Conclusion The levels of serum IL-1β and IL-1Ra were associated with the inflammatory process after the operation and the immune maladjustment of IL-1β and IL-1Ra might be correlative to the restenosis,indicating that the IL-1Ra/IL-1β ratio might be an available index for monitoring the restenosis.

Objective To discuss mechanism and control measures of stent fracture and restenosis after percutaneous transluminal angioplasty and stenting (PTAS) for symptomatic ostial vertebral/subclavian artery stenosis. Methods A retrospective analysis was performed on 3 patients with stent fracture after receiving PTAS for symptomatic ostial vertebral/subclavian artery stenosis.Simple radiographic, ultrasonographic and clinical follow-up examinations were estimated. Related articles on coronary stent fracture were gone over, consulting in the types, cumulative incidence and occurrence time of adverse events, risk factors and preventive measures. Results Stent fractures of 3 patients with symptomatic ostial vertebral /subclavian artery stenosis were associated with in-stent restenosis and occlusion. Two of the 3 patients treated with the balloon angioplasty and after balloon dilatation, and the patients exhibited relief of symptoms. One patient was only managed for vascular disease risk factors, and no developing recurrent symptoms were noted during the follow-up period.Conclusions Stent fracture might appear in patients performed PTAS for symptomatic ostial vertebral /subclavian artery stenosis, and regular check is needed. Individual treatment was emphasized in case of serious symptoms appeared.

OBJECTIVETo study isolation, identification and differentiation characteristics of dermal multipotent stem cells from human of different age in vitro culture.

METHODSSkin samples( 1 cm x 1 cm) were harvested from fetus, infant, adult and elderly. The original clones were screened in stem cell medium. The diameter and number of clones were recorded. Analysis of each clone and determination of the expression of various related proteins were carried out.

RESULTSThe number of suspended clones from normal skins of fetus, infant, adult and the elderly were (20. 1 +/-2. 5) x 102 , (15. 8 +/-5. 7) x 102, (10. 8 +/-1.3) x 10(2), (6.2 +/- 1.4) x 10(2), respectively ( P <0.01), while the diameter of the clones from them were (83 +/-12) microm, (55 +/- 10) microm, (46 +/- 12) Lm, (42 +/-8) microm, respectively ( P <0.05). Cloned cells from fetus, infant, adult and elderly could differentiate into neuron cell , neuroglia cell, smooth muscle cell, and adipocyte. The clones from fetus were inclined to differentiate into neuron cells, but those from infant were inclined to differentiate into neuroglia cells, and those from adult and elderly were inclined to differentiate into adipocytes. After 1 month of culture, the clone forming rate of the cells from fetus, infant, adult and elderly were 41. 1% , 25.5% ,17.7% ,15.2% , respectively. The individual clone cells also showed ability of multidirectional differentiation. Nestin, fibronectin, c-Myc, STAT3 and hTERT protein were expressed in all clones.

CONCLUSIONMultipotent stem cells with multi-direction differentiation and proliferation can be efficiently isolated from dermis of human of different age in stem cell culture medium. The number, proliferation and differentiation of dermal multipotent stem cells can be affected by age.

Aborted Fetus ; cytology ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Cell Differentiation ; Cell Separation ; Cells, Cultured ; Child ; Child, Preschool ; Dermis ; cytology ; Female ; Humans ; Infant ; Male ; Middle Aged ; Multipotent Stem Cells ; cytology ; Pregnancy ; Pregnancy Trimester, Second