HLA-B Antigens ; genetics ; HLA-B27 Antigen ; genetics ; Humans ; Spondylitis, Ankylosing ; genetics

This work reports the properties of dextran-spermine polycation (DSP) as a gene vector and its gene transfection efficiency in vitro. Oxidized dextran was reacted by reductive amination with spermine to obtain DSP, the resulted polycation was then incubated with plasmid pEGFP to form polyplexes by electrostatic interactions. DSP formed stable polyplexes when the weight ratio (DSP/DNA) varied from 4 : 1 to 20 : 1. The particle size and zeta potential of polyplexes were in the range of 162.6 - 187.9 nm and increased from + 8.45 mV to + 39.6 mV, respectively. DSP could effectively protect condensed DNA from DNase I degradation, and it showed strong buffering capacity in a certain pH range. The highest yields of transfection efficiency were found to be as high as Lipofectamine 2000 when the polyplexes were transfected to SMMC-7721 and BHK-21 cells at the weight ratio of 8 : 1. This research indicates that dextran-spermine polycation is a highly active gene vector in vitro.
Animals ; Cell Survival ; Cricetinae ; DNA ; administration & dosage ; Dextrans ; administration & dosage ; Genetic Vectors ; Spermine ; administration & dosage ; Transfection ; methods

Adenocarcinoma ; metabolism ; pathology ; Adenocarcinoma, Mucinous ; metabolism ; pathology ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Signet Ring Cell ; metabolism ; pathology ; Erythropoietin ; metabolism ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Immunohistochemistry ; Lymphatic Metastasis ; Microvessels ; chemistry ; pathology ; Middle Aged ; Stomach ; chemistry ; pathology ; Stomach Neoplasms ; metabolism ; pathology ; Tissue Array Analysis ; methods

OBJECTIVETo study the conditions on separation and regeneration of protoplast from Phellinus igniarius.

METHODThe effects of enzymolysis conditions of P. igniarius mycelia on yield of protoplast and culturing conditons on regeneration ratio of protoplast were investigated.

RESULTWhen the 8 days-old mycelia was hydrolysed by 1.5% of lywallzyme adding to driselase of 0. 5% and at 30 degrees C for 3 h and enzymolysis was stablized by sucrose as a stablisher of osmotic pressure, higher yield of P. igniarius protoplast was obtained. If 10 days-old mycelia was used as raw material of enzymolysis and manntol was selected as stablisher of osmotic pressure of enzymolysis, higher regeneration ratio of P. igniarius protoplast also would be obtained in following regeneration step at same time keeping higher yield. For the regeneration processing, it was beneficial for the regeneration of P. igniarius protoplast that PDA plusing mulberry ramulus was used as the culture medium of regeneration and manntol was selected as the osmotic pressure establisher of regeneration culture medium.

CONCLUSIONThe method and conditions to keep both higher yield and regeneration ratio of P. igniarius protoplast were obtained.

Culture Media ; pharmacology ; Fungal Proteins ; pharmacology ; Glucan Endo-1,3-beta-D-Glucosidase ; pharmacology ; Glycoside Hydrolases ; pharmacology ; Mannitol ; pharmacology ; Multienzyme Complexes ; pharmacology ; Osmotic Pressure ; Peptide Hydrolases ; pharmacology ; Polyporaceae ; drug effects ; physiology ; Protoplasts ; drug effects ; physiology ; Regeneration ; drug effects ; Sucrose ; pharmacology ; Temperature

Objective:To study the effect of nerve growth factor(NGF)on bone fracture healing of inflicted T_(10)spinal cord injury(SCI)complicated with tibia fracture in rats.Methods:Totally 120 rats were randomly divided into tihia fracture group (F group,n=40),T_(10)SCI+tibia fracture group(FS group,n=40),and T_(10)SCI+tibia fracture+NGF group(FSN group,n=40).Four weeks later,the fracture sites in the 3 groups were subjected to CT scanning;the maximum transverse diameter of the fracture ends and the gray scales of non-osseous area were measured;the changes of biomechanics property of the fracture ends were determined by three-point bending test;the bone morphometry,bone density,and histomorphology of callus were determined;the expression of OCN was detected by immunohistochemical method;the osteoblast ultrastructure was observed by TEM and the expression ofⅠ,Ⅱtype collagen were examined by Western blotting.Results:The maximum transverse diameter of F group was less than those of FS group(P

Objective To examine expression of PTEN gene in ovarian cancer cisplatin-sensitive cell line OV2008 cells and cisplatin-resistant cell line C13K cells,and evaluate the effect of wild-type PTEN gene on reversing cisplatin-resistance of C13K cells and underlying mechanisms.Methods The expression of PTEN mRNA and protein in OV2008 and C13K cells were detected by semi-quantitative RT-PCR and western blot.Recombinant eukaryotic expression plasmid containing human wild-type PTEN gene was transfected into C13K cells by lipofectamine 2000.The expression of PTEN mRNA was monitored by RT- PCR and the expression of PTEN,protein kinase B(AKT),phospho-AKT(p-AKT)protein were analyzed by western blot in PTEN transfected and untransfected C13K cells.Proliferation and chemosensitivity of cells to cisplatin were measured by methyl thiazolyl tetrazolium(MTT),and cell apoptosis was detected by flow cytometry after treatment with cisplatin.Results(1)The expression of PTEN mRNA and protein(1.02 ?0.05,1.02?0.07)in OV2008 cells were significantly higher than those in C13K cells,which were 0.45 ?0.03 and 0.55?0.03 respectively(P

We investigated brucellosis incidence trends in Kashgar,Xinjiang in 2005-2016 for further prevention and control,and analyzed the brucellosis epidemiological characteristics there,by using the descriptive epidemiology method.A total of 767 cases were reported,with an average annual incidence of 1.68/100 000,and incidence of annual report showed a relatively stable trend (Z:29.49,P＜0.001).The maximum number of reported cases was mainly concentrated from May to July.Incidence ratio of the male and female was 1.81:1.Cases were identified in each age group,the minimum age was five months,and the maximum age was 85 years old,with the median of 39.The highest proportion of reported cases was peasant.The top five average annual incidence counties (cities) were the Markit County,Yopurga County,Tashikuergan Tajik Autonomous County,Bachu County and Kashgar City.The brucellosis incidence increased year by year,especially during 2012-2016.We need further analysis for the data from increasing brucellosis outbreak and further strengthen the prevention and control of that in Kashgar area.

OBJECTIVETo construct a chimeric SEA-hPLAP-1 cDNA with gene splicing by overlap extension.

METHODSThe SEA gene and a DNA fragment encoding the signal for GPI-anchor attachment of hPLAP -1 were amplified by PCR. The two amplified gene sequence was annealed to form a chimeric GPI- anchored SEA molecule with gene splicing by overlap extension. The resulting chimera was cloned in pGEM-T vector and verified by sequencing analysis.

RESULTA chimeric SEA-hPLAP-1 cDNA was successfully constructed with gene splicing by overlap extension.

CONCLUSIONGene splicing by overlap extension is a successful specific PCR technique for gene recombination.

Alkaline Phosphatase ; Base Sequence ; Enterotoxins ; genetics ; GPI-Linked Proteins ; Isoenzymes ; genetics ; Molecular Sequence Data ; Polymerase Chain Reaction ; RNA Splicing ; Recombinant Fusion Proteins ; genetics

OBJECTIVETo evaluate the efficacy of antifibrinolytic agents in coronary artery bypass grafting (CABG) patients receiving antiplatelet.

METHODSWe searched PubMed, EMbase, Highwire, CENTREN and its affiliated clinical trial registration data center, CBMdisc and CNKI databases from 2000 to 2010. Randomized controlled trials investigating the efficacy of anti-fibrinolytic agents (aprotinin and tranexamic acid) in CABG patients were identified. Study selection and meta-analysis were conducted according to the Cochrane Handbook for systematic reviews. Date were extracted from these trials by 2 reviewers independently and analyzed by RevMan 5.0 software.

RESULTEleven RCT trials (n = 725) were included and data confirmed the efficacy of antifibrinolytic therapy in terms of reducing bleeding within 24 hours after operation (MD = -306.5 ml, 95%CI: -351.52 to -261.52, P < 0.01), number of patients who need blood transfusion (OR = 0.37, 95%CI: 0.26 to 0.51, P < 0.01), amount of blood transfusion (MD = -0.59 U, 95%CI: -0.69 to -0.50, P < 0.01), surgical re-exploration(OR = 0.27, 95%CI: 0.09 to 0.78, P = 0.02), and thrombotic events (OR = 0.49, 95%CI: 0.25 to 0.97, P = 0.04) in CABG patients receiving antiplatelet, while compared with blank treatment.

CONCLUSIONThis analysis showed that antifibrinolytic agents are effective for reducing bleeding within 24 hours after operation, amount of blood transfusion, surgical re-exploration and do not increase the incidence of thrombotic events in CABG patients receiving antiplatelet before operation.

Antifibrinolytic Agents ; therapeutic use ; Coronary Artery Bypass ; Humans ; Perioperative Period ; Platelet Aggregation Inhibitors ; therapeutic use ; Randomized Controlled Trials as Topic ; Treatment Outcome

Objective To evaluate the cerebral autoregulation in patients with obstructive sleep apnea-hypopnea syndrome (OASHS) using transcranial Doppler (TCD)-CO2 test and head-upright tilt test (HUTT) from the aspects of nocturnal hypoxemia/hypercapnia and sleep structure. Methods Seventy-six patients with OSAHS visiting our hospital from February 2007 to May 2009 were chosen in our study and divided into severe OSAHS group (n=26), moderate OSAHS group (n=29) and mild OSAHS group (n=21) according to the apnea-hypopnea index (AHI), and the lowest oxygen saturation (LSaO2); 32 healthy controls, having snore history, were adopted too. Polysomnography monitor was used for night-7-h sleep monitoring and blood pressure monitoring; sleep-related indicators and blood pressure at different times were analyzed. Cerebrovascular reactivity was calculated in terms of the breath-holding index (BHI) and vascular motor reactivity (VMR) by TCD-CO2 test; Changes of cerebral blood flow velocity (CBFV), blood pressure (Bp), and the time from squatting-to-tilt position for the mean arterial pressure (TMAP) and the CBFV (TCBFV) returning to ＞90% of baseline levels were detected by HUTT to assess the cerebral pressure-autoregulation. Results The AHI, microarousal index (MI) and the percentages of S1 in the non-rapid eye movement sleep period in the severe, moderate and mild OSAHS groups were all significantly higher than those in the control group (P＜0.05); the LSaO2 and the percentages of S3+4 in the non-rapid eye movement sleep period in all the OSAHS groups were significantly lower than those in the control group (P＜0.05); no significant difference in blood pressure before apnea was noted between the OSAHS groups and the control group (P＞0.05), however, the systolic blood pressure while apnea in all the OSAHS groups was significantly higher than that in the control group (P＜0.05). As compared with the controls and mild OSAHS group (1.89±0.36, 1.75±0.41), severe and moderate OSAHS groups (0.71 ±0.17, 1.12±0.23, respectively) showed significantly decreased BHI (P＜0.05); As compared with the controls (0.68±0.11), and the mild, moderate and severe OSAHS groups (0.20±0.04, 0.34±0.07 and 0.55±0.17, respectively) showed significantly decreased VMR (P＜0.05); TMAP in the moderate and severe OSAHS groups was significantly longer than that in the controls and mild OSAHS group (P＜0.05); TCBFV in the mild, moderate and severe OSAHS groups was significantly longer than that in the controls (P＜0.05). Significant difference on the levels of Bp and CBFV during tilt was noted between the moderate and severe OSAHS groups (P＜0.05); Pearson analysis showed a linkage between Bp and CBFV changes (r=0.384, P=0.005). Conclusion Cerebrovascular autoregulation is impaired in patients with OSAHS, especially in the moderate and severe groups, which may increase the risk of stroke. The major risk factors for cerebrovascular autoregulation in patients with OSAHS are night hypoxemia, hypercapnia, blood pressure fluctuation and severe sleep disorders.