BACKGROUND:To explore the methods for secondary limb revascularization after endovascular repair is one of the most urgent research topics.
OBJECTIVE:To investigate the biological mechanism about the transplantation of autologous peripheral blood stem cel s to lead and perfect the effect of ischemic tissue for secondary revascularization.
METHODS:Forty-two patients with critical low limb ischemia were selected and treated with endovascular repair as first revascularization and transplantation of autologous peripheral blood stem cel s as secondary revascularization. The secondary revascularization was carried out at 3 months after the first blood flow remodeling. The patients receiving secondary revascularization were fol owed for 4 years.
RESULTS AND CONCLUSION:After transplantation of autologous peripheral blood stem cel s, the patients complained that the rest pain of the lower leg was relieved obviously and intermittent claudication distance was significantly lengthened. Limb salvage rate was 100%. Skin temperature index was 1.6±0.3, transcutaneous oxygen pressure was (5.00±1.26) kPa, ankle-brachium index was 0.9±0.2, photoplethysmograpy index was 0.8±0.1, saturation of blood oxygen was (79.4±20.4)%, and digital subtraction angiography score was (1.3±0.2)
points. After transplantation, local blood flow indicators of the low limbs were positively correlated to local symptom indicators of the limbs (0.6

Objective To explore the expression of(hyopxia inducible factor-α,HIF-1α)and gax genes on the intimal hyperplasia for venous autografts in rat.Methods Sixteen Wistar rats were randomLy divided into 2 groups,a control group and an experimental group,8 rats in each group.A rat model of venous autografts was established by transplanting the right external jugulars vein into right common carotid artery in 16rats.The transplanted vein in the experimental group was immersed in solution with recombinant adeno-associated virus(rAAV)and HIF-1α by incubation for 45 minutes at 0℃-4℃ before anastomosis and coated using glue gel in which rAAV-gax added just after anastomosis,the control ones were immersed only in the physiological solution in which rAAV solution without HIF-1α gene added,nothing to be coated except for glue gel only.The vein grafts were taken at 14 days after operation,RT-PCR technique,Western blotting and electric microscopy were used to detect the expression of HIF-1α mRNA and protein,gax mRNA and protein,phenotypic switch of VSMC.Results The expression of HIF-1α mRNA,the experimental group(93.1±22.9)bP,the con trol group(42.7±15.9)bP,P<0.01;the exprossion of gax mRNA,the experimental group(106.9±38.7)bP,the contrd group(50.7±25.8)bP,P<0.01;the exprossion of HIF-1α protein,the experimental group(10.86±2.76)kD,the control group(4.52±1.90)kD,P<0.01;the expression of gax protein,the experimental group(13.65±3.35)kD,the control group(5.40±2.26)kD,P<0.01;VSMC proliferation,the experinental group(21.3±5.4)%,the control group(41.2±8.6)%,P<0.05.Conclusion The present study demonstrated that HIF-1α and gax genes transfection could inhibit proliferation of VSMC and their phenotypic conversion from contractile phenotype to synthetic phenotype in venous autografts.

Objective To investigate the efficiency of autologous transplantation of peripheral blood stem cell for treatment of patients with chronic lower limb ischemia. Methods Forty-six patients with chronic lower limb ischemia were treated by autologous peripheral blood stem cell transplantation. Results Fortytwo patients had pain relief on legs, and cold and cool feelings in lower limbs disappeared. Thirty-five patients had pain relief on feet, and 29 of them had disappearance of cold and cool feelings. Due to necrosis in middle and lower part of leg, 4 patients took extremity amputation after 4 weeks of the transplantation. In the 42 patients who kept their legs 3 months after transplantation, the distance of anginacruris extended from (87. 45 ±41.22) m to (348.52 ± 147.24) m, skin temperature increased from(28.52 ±0.51 ) ℃ to(33.56±0.62) ℃ ,and ankle-brachial index (ABI) increased from(0.48 ±0.06)to(0.75 ±0.07). There were statistical differences, and the scores of the distance of anginacruris, skin temperature, and ABI after transplantation were better than before. six months after transplantation autobiography of lower extremity was used, and neonatal lateral vessels of different degrees were found in 37 patients. Conclusion Autologous transplantation of peripheral blood stem cell is a simple, safe, and effective method, especially in treating patients with lower limb ischemia in which no arterial reconstruction is feasible.

Objective To discuss the safety and advantages of prone-straddle position applied in transvaginal hysteromyomectomy of the posterior wall of the uterus. Methods The clinical data of patients who were admitted into our hospital from March 2013 to Janaury 2015 and received transvaginal hysteromyomectomy were retrospectively analyzed. They were divided into group A ( prone-straddle position, 30 cases) and group B (traditional lithotomy position,24 cases). The exposure of operative field, convenience of operation, time of the placement, time of operation, patient satisfaction, as well as patients’ heart rate, blood pressure, and oxyhemoglobin saturation were observed and ana-lyzed. Results Compared with the lithotomy position, there were significant differences in the time of the placement and the time of opera-tion in the patients treated by prone-straddle position. The patients with prone-straddle position cooperated well, and there was no obvious discomfort. Their vital signs were stable during the operation. The operative field during prone-straddle position exposed better and it was more convenient which make the operation became easier for both the operators and nurses. Conclusion Prone-straddle position applied in transvaginal hysteromyomectomy of the posterior wall of the uterus is safe and practicable, and it is valuable for clinical application.

Objective: Extracting ephedrine and berberine from the water extract of Ephedra Herba and Rhizoma Coptidis. Methods: Modified PVA was used as separating membrane and separated alkaloids. Results: A colorless ephedrine aqueous solution was obtained when extraction time was less than 8 hours; while a three constitution solution was obtained when time of extraction is over 8 hours. So is Rhizoma Coptidis. The membrane is hardly fouled. Conclusion: Purity of alkaloids from Herba Ephedra and Rhizoma Coptidis by membrane separation is Superior to that by alcoholic sediment but amount of alkaloids is reverse.

Adult ; Animals ; Asthma ; epidemiology ; etiology ; Female ; Humans ; Immunoglobulin E ; blood ; Male ; Occupational Diseases ; epidemiology ; etiology ; Pyroglyphidae ; Skin Tests

In order to confirm the role that the 49th amino acid residue plays in enzymatic inactivity of Glutamine 49 phospholipase A2(Gln49-PLA2),site-directed mutagenesis of its 49th amino acid gene codon was conducted using PCR.Aspartic acid 49 phospholipase A2(Asp49-PLA2-Q49D-PLA2),the mutant of Gln49-PLA2 was expressed in E.coli with pET32a+ vector.The fusion protein,expressed as inclusion body,after being denatured,was on-column refolded and purified by immobilized metal affinity chromatography(IMAC),and then cleaved by Factor Xa.The mature Q49D-PLA2 mutant was obtained by Hitrap SP cation exchange and Superdex 75 gel filtration chromatography,with the recovery rate of 1.3%,and the specific activity of the mature Q49D-PLA2 mutant was 72 U/mg.It has been demonstrated that the 49th glutamine amino acid residue is the main reason in enzymatic inactivity of Gln49-PLA2 and the results are helpful for denatured protein refolding,especially in rich disulfide bonds conditions.

Objective To explore the value of the spectral CT iodine-based material decomposition technique in differential diagnosis of central pulmonary carcinoma from obstructive pneumonia and atelectasis.Methods 25 cases with central pulmonary carcinoma complicating with obstructive pneumonia and atelectasis underwent CT plain scan and spectral contrast scan including pulmonary-arterial-phase (PAP) and bronchial-arterial-phase(BAP).Conventional CT images and iodine base images were generated in each phase by GSI viewer.The 4 groups of images differences between the tumor and the obstructive pneumonia and atelectasis were analyzed.Results The margin of the tumor was ill-defined in plain scan.The cases that showed difference between the tumor and the obstructive pneumonia and atelectasis in 4 groups of images were respectively as follows,10 in conventional CT images of PAP,1 6 in conventional CT images of BAP,1 9 in iodine base images of PAP,and 23 in iodine base images of BAP.The difference of the 4 groups was significant (χ2 =16.54,P<0.05).There was the highest accuracy in iodine base images of BAP in defining the tumor margin (χ2 = 12.83,χ2 = 4.20,χ2 =1.34,P <0.05). The accuracy between conventional CT images of BAP and iodine base images of PAP had no significant differences (χ2 =0.86,P >0.05),but were better than that of the conventional CT images of PAP(χ2 =6.65,P <0.05).Conclusion Spectral CT iodine-based material decomposition technique is helpful to improve subjective diagnosis of central pulmonary carcinoma complicating with obstructive pneumonia and atelectasis,especially in iodine base images of BAP,it can provide accurate information for clinical stage and treatment.

BACKGROUND:Primary human lung epithelial cel s are difficult to be isolated and cultured in vitro, which is characterized as limited sources, low cel viability, slow proliferation capacity, and lacking of differentiation capability.
OBJECTIVE:To establish an air-liquid interface model of lung epithelium by in vitro proliferation of human bronchiolar epithelial cel s, which is used for research on function of lung epithelial cel s.
METHODS:Primary human bronchiolar epithelial cel s were isolated using Pronase and DNase I combined digestive methods, and then proliferated using medium containing ROCK kinase inhibitor. The proliferated cel s were used for establishment of the air-liquid interface epithelium model. Cel differentiation was identified using scanning electron microscope, phase contrast microscope and immunofluorescent staining.
RESULTS AND CONCLUSION:The primary human bronchiolar epithelial cel s could be expanded successful y using medium containing ROCK kinase inhibitor, and the basal cel marker Cytokeratin14 was preferential y expressed in the proliferated cel population, indicating that these basal cel s might be the main subpopulation of human lung epithelial stem cel s. Subsequently, the proliferated cel s under the air-liquid interface could differentiate into ciliated cel s and non-ciliated column cel s. The results suggest that the proliferation and differentiation of human bronchiolar epithelial cel s were maintained in the presence of ROCK kinase inhibitor, and the air-liquid interface could promote the differentiation of human bronchiolar epithelial cel s.

Objective To investigate the reproducibility and consistency in contouring parotid gland volume based on computed tomography (CT) and magnetic resonance (MR) images in patients with nasopharyngeal carcinoma during radiotherapy.Methods Twenty-seven patients with nasopharyngeal cancer in Shandong Cancer Hospital from December 2012 to May 2013 were randomly enrolled and underwent intensified CT and MR imaging before radiotherapy.The parotid gland were contoured with unified standard on both CT and T1-MR images by 11 radiotherapists.Specifically,one radiotherapist sketched the parotid gland on CT and MR images for ten times as intra-group comparison,the other ten were asked to sketch the parotid gland on CT and MR images only once as inter-group comparison.The intra-and inter-group's variations of parotid gland volumes were analysed.Results The average volumes of intra-group on CT and MR images were (33.8 ±9.4) cm3(L),(33.2±7.6) cm3(R) and (24.4 ±7.6) cm3(L),(22.5 ±7.4) cm3(R).As well,for inter-group the average volumes were (34.6 ± 12.1) cm3 (L),(34.3 ± 9.0) cm3 (R) and (24.6 ± 7.6) cm3 (L),(23.2 ± 8.1) cm3(R),respectively.The volume variable ratios on CT images were (6.8 ± 1.5)％ (L),(6.3 ± 1.5) ％ (R) for intra-group and (18.0 ± 4.8) ％ (L),(17.4 ± 4.6) ％ (R) for inter-group.Similarly,the intra-and inter-group ratios for contouring on MR images reached (2.3 ±0.4)％ (L),(2.1 ±0.7)％ (R) and (4.7 ±0.7)％ (L),(5.0±0.6)％ (R),respectively.Conclusion Parotid gland contouring based on MR images has a better reproducibility and consistency than that based on CT images.It is beneficial to get a more objective and true indicator to estimate the radiation injury of parotid gland.