BACKGROUND: Thrombus formation and neointimal hyperplasia limit its use for revascularization of small-caliber vessels (<6mm diameter) in the coronary or peripheral circulation.OBJECTIVE: To improve hemocompatibility, the luminal surface of a small diameter expanded polytetrafluoroethylene (Eptfe) vascular prosthesis was modified with alginate and recombinant hirudin.DESIGN: Observational experiment.SETTING: This study was performed in Nanomedicine and Biosensor Laboratory, Bio-X Center, Harbin Institute of Technology from Marcy 2006 to June 2007.MATERIALS: The GORE-TEX Epife vascular grafts (W. L Gore & Associates, Inc., Flagstaff, AZ) used in this study were 4mm in internal diameter. rHir was obtained from Calbiochem, Germany. Sodium alginate (also called alginic acid sodium salt; medium viscosity) was purchased from Sigma.METHODS: A p-diazonium diphenyl amine polymer (PA) was used as an interlayer between alginate and recombinant hirudin (rHir). The diazonium moieties were capable of covalently coupling with electron-rich aromatic systems such as histidine and tyrosine residues of hirudin. No need for chemical pretreatment took all advantage by preserving the bulk properties with almost no effect on stability and elasticity of the Eptfe vascular graft. rHir amount on Epife surface Was determined by the Micro BCA (Bicinchoninic acid) Protein Assay kit.MAIN OUTCOME MEASURES: ① Determination rHir amount on Eptfe surface; ② Static water contact angles; ③ Attenuated total reflectance fourier transform infrared spectroscopy (ATR-FTIR) was employed to confirm the change taking place in the Eptfe graft surface modification process; ④ Characterization of the surface morphology and platelet adhesion by SEM; ⑤ APTT and PT; ⑥ Percent hemolysis.RESULTS: ① The amount of rHir adsorbed onto the Eptfe vascular was deduced to be 16.35 μg/cm2. ② Surface analysis ATR-FTTR revealed the presence of new functional groups on the modified graft surfaces. ③ The water contact angle of the modified graft surface decreased. ④ The longer APTT and PT value lower than 5% hemolysis level and dramatically decrease of platelet adhesion assay showed that rHir modified graft had great improved blood compatibility.CONCLUS10N: Cross-linked Alg/rHir onto Eptfe can improve luminal surface and hemocompatibility.

OBJECTIVETo report and evaluate the clinical results of surgical treatment for long-segment iliofemoral arteriosclerosis obliterans, including external iliac-popliteal (EIP) and femoral-deep femoral (FDF) crossover bypass surgeries.

METHODSFrom July 1995 to December 2009, 85 patients (61 male, 24 female, aged from 64 to 91 years, mean age 75 years) with comprehensive unilateral iliac-superficial femoral arteriosclerosis obliterans were involved in this research. According to Fontaine classification, the 85 patients could be graded as 62 class IIb-III patients (72.9%), and 23 class IV patients (27.1%). In CT angiography or DSA examinations, the 85 patients were grouped into EIP (n = 49) and FDF (n = 36) surgical groups on the basis of visualizations in the affected deep femoral, supra-knee/infra-knee popliteal arteries. The healing time of ulcers and toe amputation wound, ankle-brachial index, and blood flow velocity in the affected limb together with accumulative patency rates in 1, 3 and 5 years and limb salvage rates in 3 and 5 years were analyzed.

RESULTSNone of the 85 patients died or had extremity amputated in perioperatively. Seventy-four patients (87.1%) had been followed up from 2 to 13 years (mean 5.7 years). Postoperative ankle-brachial index of FDF and EIP groups was 0.55 ± 0.11, and 0.94 ± 0.13 (t = -21.88, P = 0.000). Postoperative velocity of popliteal artery blood flow in FDF and EIP groups was (32 ± 9) cm/s, and (48 ± 4) cm/s (t = 16.76, P = 0.000); velocity of anterior or posterior tibial artery was (22 ± 7) cm/s, and (42 ± 4) cm/s (t = 10.50, P = 0.000). The primary and secondary patency rates of FDF and EIP groups were 87.8% and 88.3%, 80.7% and 81.2%, 68.4% and 57.4% at 1, 3 and 5 years, respectively. Limb salvage rates of FDF and EIP groups were 87.6% and 88.6%, 76.7% and 71.3%, at 3 and 5 years, respectively. There were no statistically significant differences in 1, 3, and 5 years' cumulative secondary patency rate and limb salvage rate between FDF and EIP groups.

CONCLUSIONSAs extra-anatomic bypass surgeries, FDF and EIP are both determined to be alternative procedures for unilateral common iliac-superficial femoral artery occlusive disease, especially suitable for high-risk patients. EIP group patients have better clinical outcomes than those in FDF group.

Aged ; Aged, 80 and over ; Arteriosclerosis ; surgery ; Arteriosclerosis Obliterans ; surgery ; Blood Vessel Prosthesis Implantation ; Female ; Femoral Artery ; surgery ; Humans ; Iliac Artery ; surgery ; Limb Salvage ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo investigate various activities of dissolved matter of glycyrrhizic acid of Radix Glycyrrhizae and the powder by supermicro-pulverization.

METHODThe contents of glycyrrhizic acid in different samples were tested.

RESULTThe dissolved matter of glycyrrhizic acid was greatly increased by supermicro-pulverization. The more time used for grinding, the smaller the size of the powder, and the easier the glycyrrhizic acid would be dissolved.

CONCLUSIONSupermicro-pulverization is helpful to the dissolved matter of glycyrrhizic acid of Radix Glycyrrhizae, and the size of powder exerts great influence on dissolved matter of glycyrrhizic acid.

Drug Combinations ; Drugs, Chinese Herbal ; chemistry ; Glycyrrhiza ; chemistry ; Glycyrrhizic Acid ; analysis ; Particle Size ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Powders ; Solubility

AIM To investigate the effects of the compatibility of Ephedrae Herba (Mahuang) and Glycyrrhizae Radix et Rhizoma (Gancao) on pharmacokinetic changes of the plasma levels of glycyrrhizic acid,glycyrrhetinic acid and liquiritin by UPLC-MS/MS.METHODS Twelve SD rats were randomly divided into two groups and orally administered with Glycyrrhizae Radix et Rhizoma and Ephedrae Herba-Glycyrrhizae Radix et Rhizoma (2 ∶ 1) Decoction.Blood samples were collected at different time points,and the main pharmacokinetic parameters were calculated by DAS3.2.2 software.RESULTS Compared with Glycyrrhizae Radix et Rhizoma group,the AUC0-t and Cmax were decreased,the MRT0-t and t1/2z were extended,the Vz/F was increased for glycyrrhizic acid,glycyrrhetinic acid and liquiritin;the CLz/F was increased for glycyrrhizic acid and liquiritin;the Tmax of glycyrrhizic acid was shortened and the Tmax of glycyrrhetinic acid was extended.CONCLUSION After combining Glycyrrhizae Radix et Rhizoma and Ephedrae Herba,the bioavailability of glycyrrhizic acid,glycyrrhetinic acid and liquiritin are reduced,the elimination of them delays,and the distribution volume increases.

Objective To investigate the ostengenie potential of adipose-derived stem cells(AD- SCs)when exposed to adenovirns containing hBMP-2 cDNA(Adv-hBMP-2)and offer a choice of cell source for gene therapy and tissue engineering.Methods Human adipose tissues were obtained from patients who received orthopaedic surgery or liposuction.ADSCs were obtained by digesting the adipose tissues.Firstly,flowcytometric analysis was performed for the confirmation of mesenchymal stem cell ori- gin and the surface markers including CD34,CD44,CD68,CD71,CD90,and CD105.The ADSCs were transfected by Adv-hBMP-2 and the effects were tested in vitro,lmmunoprecipitation and Western blotting and ELISA were performed for confirming BMP gone transduction and its stable expression.The transform of ADSCs was assessed by extracellular ALP staining,intracellular ALP spectrophotometry,von Kossa staining and RT-PCR.In the in vivo experiment ADSC-Adv-hBMP-2 cells were injected into the hind limb of nude mice and analyzed radiographically and histologically.Results ADSCs were successfully isolated from human adipose tissues.The isolated ADSCs expressed CD44,CD71,CD90 and CD105 and CD34 and CD68 were absent.The result confirmed the mesenchymal stem cell origin of the cells.West- ern blotting and ELISA confirmed successful and persistent hBMP-2 production by ADSC-Adv-hBMP-2 cells.Extracellular ALP staining,intracellular ALP spectrophotometry,yon Kossa staining and RT-PCR revealed that ADSCs treated with Adv-hBMP-2 had a tendency of transfering into osteoblast.X-ray and H&E sections from hind limb of nude mice injected with ADSC-Adv-hBMP-2 cells confirmed bone forma- tion at 2 weeks.Conclusions Liposuction aspirates contain abundant ADSCs that can be transduced with hBMP-2 gene,and the tranduced ADSCs differentiate into the osteoblast.ADSCs may be an ideal source of mesenchyme-lineage stem cells for gone therapy and tissue engineering.

OBJECTIVETo analyze anatomy data of popliteal veins (PV), with the purpose of selection of popliteal venous valves construction segment via venography, and to evaluate the surgical results.

METHODSFrom February 1998 to November 2010, after analyzing the popliteal vessel anatomy data of 39 limbs and related phlebography research of 862 cases, 102 patients (69 male and 33 female patients, aged from 48 to 71 years, mean 59 years) with severe deep venous insufficiency were selected for popliteal venous valve construction procedures. Doppler ultrasound, continuous dynamic venography, and intraoperative venous pressure measurements were used to assess the hemodynamic changes pre- and postoperatively. Venous clinical severity score (VCSS) were used to evaluate long-term results of deep venous valve construction procedures.

RESULTIn the 102 patients, 93.7% patients had one pair of valves in popliteal vein (PV), locating in the distal 1/3 segment of PV, with gastrocnemius veins (GV) joining with PV above PV valves. Postoperative blood flow volume of the PV was significantly higher than the preoperative volume (732.3 ml/min vs. 150.2 ml/min, t = 8.979, P < 0.001). The proximal pressure was significantly lower than the distal pressure ((12 ± 3) cm H(2)O vs. (15 ± 3) cm H(2)O, 1 cm H(2)O = 0.098 kPa, t = 8.049, P < 0.001). VCSS score was significantly lower after the surgery ((34 ± 15) cm H(2)O vs. (41 ± 14) cm H(2)O, t = 59.780, P < 0.001). Pre- and postoperative hemodynamic changes and VCSS scores were statistically significant (9.3 ± 1.9 vs. 1.8 ± 1.0, t = 59.780, P < 0.001). Mean follow-up were 8.9 years with an ulcer recovery rate of 96.3%, and a 3.7% ulcer recurrent rate.

CONCLUSIONSPopliteal vessel anatomy study and venography research provide critical information for the PV valve construction part selection, which stayed proximal to the communications of GV and PV. Restoration of gastrocnemius pump function and satisfactory long-term efficacy are received after valve construction.

Aged ; Female ; Hemodynamics ; Humans ; Lower Extremity ; blood supply ; Male ; Middle Aged ; Popliteal Vein ; anatomy & histology ; surgery ; Vascular Surgical Procedures ; methods ; Venous Insufficiency ; surgery ; Venous Valves ; surgery

Keratins are cytoskeleton proteins, which are involved in apoptotic resistance, growth and migration of cancer cells. The levels of some keratins are elevated in the serum of patients with cancers, or highly expressed in cancer tissues, and they are widely used in the diagnosis of tumor in clinic. The expressions of some keratins are negatively correlated with the prognosis of patients with cancers, and can be used as prognostic markers. Further clarifying the diagnostic and prognostic value of keratins combined with other markers detection can provide theoretical basis for the application of keratins in diagnosis and prognostic assessment of cancers.

Abstract: Objective　To develop a real-time fluorescent quantitative RT-PCR (qRT-PCR) method for qualitative and quantitative Chikungunya virus (CHIKV) analysis. Methods Based on the systematic analysis of the genomic sequences of Chikungunya and its related arboviruses, the specific nucleic acid sequences for Chikungunya virus were screened and identified, and then the primers and TaqMan probe were designed. Meanwhile, the human GAPDH gene was used as an internal reference. The reaction system for qRT-PCR was systematically optimized by L9(34) orthogonal design, and a rapid detection method for Chikungunya by qRT-PCR based on TaqMan probe methods was established. The sensitivity, specificity, reproducibility, and coverage of the established method were analyzed in detail. The standard curve was made, and the absolute quantitative method was established using the cloned nucleic acid fragments as positive samples. Results　A real-time fluorescent quantitative RT-PCR assay was developed for the qualitative and quantitative analysis of Chikungunya virus. The reaction system included Chikungunya virus and reference internal gene specific primers and probe, RT/Taq enzyme mixture, reaction buffer, and negative and positive reference. The established method obtained positive results with the ROSS strain of ECSA subtype, LR2006 strain of IOL branch, 181/25 strain of Asian type and Dongguan 2010 epidemic strains of Chikungunya virus, but there was no cross-reaction with other 18 arboviruses belonging to Flaviviruses, Alphaviruses and Bunyavirus. The minimum detection limit of the established method was 5.80 copies/mL, and a linear relationship was observed between the amount of input plasmid DNA and fluorescence signal value over a range of 5.80×102 copies/mL to 5.80×1010 copies/mL, and the correlation coefficient was 0.999 5. The qRT-PCR amplification efficiency was 91%, and the intra-assay variations and inter-assay variations were 0.01-0.07 and 0.03-0.11, respectively. Conclusions　The TaqMan qRT-PCR method developed in this study can qualitatively and quantitatively detect Chikungunya virus rapidly with specificity and sensitivity, providing a technical method for the prevention and control of this viral disease.

OBJECTIVETo investigate the clinical features of unexpected sudden death (SUD) clustered in families in Yunnan province.

METHODSThis retrospective study analyzed the clinical features of SUD occurred between July to September 2005 in 7 families in Yunnan province.

RESULTSAll 16 SUD patients shared common clinical features such as fatigue and repeated syncope and one group of SUD patients (n = 8 from 4 families) presented with the gastric intestinal tract manifestations including nausea, vomiting, abdominal pain and diarrhea with suspected dietary history and abnormal laboratory enzyme findings (GOT/GPT, CK/CKMB, LDH/LDH1 etc.). In SUD patients without gastric intestinal tract manifestations (n = 8 from 3 families), there were no clear symptoms before death and repeated ventricular tachycardia and ventricular fibrillation were recorded in one survivor. There was no clear evidence for the involvements of hereditary and infectious factors for observed SUD.

CONCLUSIONThe reason for the unexpected sudden death clustered in 7 families in Yunnan remains unclear. Repeated syncope and fatigue served as the common clinical features in the presence or absence of gastric intestinal tract manifestations in all SUD cases. Further studies are needed to clarify the pathology and detailed clinical manifestations of SUD occurred in this area.

Adolescent ; Adult ; Bias ; Cause of Death ; Child ; China ; epidemiology ; Death, Sudden ; epidemiology ; Family ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

OBJECTIVETo observe the in vivo inhibition effects of tumor necrosis factor-alpha (TNF-alpha) gene transduced tumor drainage node of lymphocytes (DNL) from tongue cancer on SCID mice transplanted tumor.

METHODS15 human tongue carcinoma models were established in SCID mice by subcutaneously injection of squamous cell carcinoma line Tca8113. TNF-alpha gene introduced DNL, combined with low dose Pinyancin (PYC), were locally injected into tumor site. The inhibition rate was determined by the weights at the 8th week after tumor dissection and fresh specimens were prepared and subject to histopathologic examination under transmission electron microscope, and in situ TUNEL was used to detect apoptosis.

RESULTSThe TNF/DNL and rIL-2 group, and the TNF/DNL and rIL-2 and PYC group both exerted a strong inhibition effect on the implanted tumor. Treated tumors of the TNF/DNL and rIL-2 and PYC group were significantly reduced in comparison with those of the TNF/DNL and rIL-2 group (P < 0.05). The apoptosis of tumor in the TNF/DNL and rIL -2 group was evidenced based on transmission electron microscope and TUNEL analysis, and the apoptosis index was higher than that of control group (P < 0.05).

CONCLUSIONLocal injection of DNL modified with TNF-alpha gene, combined with low dose PYC, exert a synergistic antitumor effect. Apoptosis may be an important mechanism of squamous cell carcinoma killed by TNF/DNL.

Animals ; Apoptosis ; Carcinoma, Squamous Cell ; Cell Line ; Drainage ; Humans ; Lymphocytes ; Mice ; Mice, SCID ; Neoplasm Transplantation ; Tongue Neoplasms ; Tumor Cells, Cultured ; Tumor Necrosis Factor-alpha