The recombinant plasmid pPIC-gpd-bgl-hyg was constructed, which contained GPD2 promotor and terminator from industrial yeasts Saccharomyces cerevisiae, ?-glucosidase gene (BGL1) from Sac-charomycopsis fibuligera and hyg from hygromycin as the selected marker. With the yeast’s high efficiency of homologous integrated, the BGL1 gene was successfully integrated into industrial yeasts S. cerevisiae. The recombined yeast could grow on the cultures with the cellobiose as a sole carbon source, and the ?-glucosidase activity achieved 0.764 U/mL after 48 hours’ cultivation. In the experiments of VHG ethanol fermentation, the cellobiose concentration in broth of recombined yeast was 80% lower than that of indus-trial yeast.

Objective To evaluate in vivo tracking of swine mesenchymal stem cells (MSCs) la-beled with super paramagnetic iron oxide (SPIO) in intraportal transplantation by a clinical 1.5T MR.Methods MSCs were isolated from swine and cultured as well as expanded, which were then incuba-ted with SPIO (Feridex I. V.). Prussian blue staining was performed for showing intracelluar irons.To establish a swine model of acute liver necrosis, 0.5 g/kg of D-galactosamine was administrated to 10 pigs. MSCs(labeled cells in six, unlabeled cells in four)were injected into liver via portal veins. MR imaging was performed with a clinical 1.5T MR immediately before and at 6 h, 3 d, 7 d, 14 d after transplantation, respectively. Results Prussian blue staining of SPIO labeled MSCs could be effec-tively labeled and the labeling efficiency was almost 100%. Signal intensity loss in liver by SPIO labe-ling on FFE sequence persisted until 14 days after transplantation. Histological analysis by Prussian blue staining showed homing of labeled MSCs in liver after 14 days, primarily distributing in hepatic sinusoids and liver parenchyma. Conclusion MSCs can be labeled with SPIO in vitro successfully.MRI can monitor magnetically labeled MSCs transplanted into liver.

OBJECTIVE: To suppress NgR gene expression in neural stem cells and observe differentiation of neural stem cells in vitro after interfered which provide nutritional support for the facial nerve repair in vivo. METHOD: PCR amplification, restriction endonuclease digestion, T4DNA ligase connections were used to connected NgR with rector pGCsi, and constructed recombinant vector (NgR shRNA). Lipofectamine 2000 were used to transfect the NSC. The expression of NgR was examined by Western Blot. The proportion of neural stem cells transformed into neurons after transfection was tested by Immunocytochemistry. Neural stem cells were planted in PLGA tubes after transfected, and were scanned by electron microscopy. RESULT: NgR shRNA plasmid was constructed and infected neural stem cells successfully. Western Blot showed that the expression of NgR decreased in neural stem cells after interference. Immunocytochemistry showed that the rate of the neural stem cells transformed into neurons after interfered was significantly higher (P < 0.01). CONCLUSION Neural stem cells were transformed into neurons after NgR shRNA plasmid infected neural stem cells, which promoted axonal regeneration more effectively and provided a efficient and stable gene platform for facial nerve repair.
Animals ; Cell Differentiation ; Cells, Cultured ; Facial Nerve ; surgery ; GPI-Linked Proteins ; genetics ; metabolism ; Myelin Proteins ; genetics ; metabolism ; Neural Stem Cells ; cytology ; metabolism ; Nogo Receptor 1 ; RNA Interference ; Rats ; Rats, Sprague-Dawley ; Receptors, Cell Surface ; genetics ; metabolism

Objective To investigate the efficacy of statin on carotid plaque stability in elderly patients assessed by K trans (the combination of E and F) of dynamic contrast material enhanced (DCE) magnetic resonance (DCE MR) imaging.Methods 37 elderly patients with carotid artery of low echo or mixed echo plaque in carotid artery patches were screened by B type ultrasound.According to the standard of 2011 ESC/EAS guidelines,all patients were divided into two groups:normal level of LDL-C group achieving LDL-C target and high level of LDL C group not achieving target of LDLC.Plaque area and size of lipid-rich necrotic core were measured by 3.0T enhancement magnetic resonance,and K trans was measured by DCE-MR dynamic contrast material enhanced (DCE) magnetic resonance (MR) imaging.and serum hs-C-reactive protein level was detected within 1 week.Results Plaque area,size of lipid-rich necrotic core,ratio of lipoid core over plaque area,K trans and serum level of hs-CRP were less in normal LDL-C level group than in high LDL-C level group [(2.06±0.45) mm2 vs.(3.63±0.62) mm2,(0.52±0.05) mm2 vs.(1.49±0.01) mm2,2 cases (11.8％) vs.14 cases (70.0％),(0.041±0.009) min1 vs.(0.079±0.011) min-1,(1.60±0.27) mmol/L vs.(2.80 ± 0.34),all P＜0.05].Linear regression analysis revealed that there was no significant correlations of K trans with the size of lipid necrotic core (r=0.19,P＞0.05) and hs-CRP (r=0.23,P＞0.05).Conclusions Dynamic contrast material enhanced (DCE) magnetic resonance (MR) imaging is a quantitative method assaying atheromatous plaque components; K trans may be a new indicator to measure the stability of plaques; Statin can stabilize plaques through inhibiting the proliferation of Vasa vasorum in plaque angiogenesis,anti-inflammatory and reducing the size of lipidrich necrotic core.

Objective To observe the clinical efficacy of thread embedding at the holographic point Kidney of the second metacarpal bone in treating tinnitus.Method Eighty-two patients with tinnitus were randomized into a treatment group and a control group, 41 cases in each group. The treatment group was intervened by thread embedding at the holographic point Kidney of the second metacarpal bone, while the control group was by intragluteal injection with Mecobalamin 0.5 mg. The Tinnitus Handicap Inventory (THI) was compared before intervention and after 1 treatment course.Result The THI scores were significantly changed in both groups after intervention (P<0.01). The change of THI score after intervention in the treatment group was significantly different from that in the control group (P<0.05).Conclusion Thread embedding at the holographic point of the second metacarpal bone in an effective approach in treating tinnitus.

Objective To observe the effect of acupuncture penetrated from Tiaokou (ST38) to Chenshan (BL57) combined with exer-cise on affected limbs of shoulder-hand syndrome (SHS) at stage I after stroke. Methods 40 inpatients with SHS at stage I after stroke from January, 2013 to December, 2014 were randomly divided into treatment group (n=20) and control group (n=20). The treatment group accept-ed acupuncture from Tiaokou to Chenshan on unaffected sides and exercise of affected upper limbs, and the control group accetpted routine acupuncture on Triple Shoulder Acupoints on affected sides. They were assessed with Pain Rating Index (PRI), Fugl-Meyer Assessment (FMA) and modified Barthel Index (MBI) before and 4 weeks after intervention. Results The incidence of improvement was 90% in the treatment group, and 80%in the control group (χ2=0.784, P=0.376). The scores of PRI decreased (t>5.37, P<0.001), and the score of FMA and MBI increased (t>4.66, P<0.001) significantly after intervention. The score of FMA and MBI improved more in the treatment group than in the control group (t>2.25, P<0.05). Conclusion Acupuncture from Tiaokou to Chenshan combined with exercise is more effective on SHS at stage I after stroke than conventional Triple Shoulder Acupoints.

[Abstract ] Objective To compare the accuracy between regular computed tomography colonography (CTC)and dual-energy CTC in lesion detection.Methods Twenty-eight patients with clinical suspicious space occupying lesions of the colon were selected.All patients were underwent dual-energy mode contrast-enhanced CT scan and the data were reconstructed with colonography and dual-energy iodine maps methods.The diameter,enhanced computed tomography (CT)value and iodine value were measured.The results of colonoscopy and pathology were taken as gold standard.The sensitivity, specificity,accuracy,positive predictive value and negative predictive value of regular CTC and dual-energy CTC were compared.Variance analysis was performed for measurement data comparison among groups and chi-square test was used for count data analysis.Results Among 28 patients,colorectal lesions were detected in 24 cases by regular CTC,of which four cases were false-positive and one case was false-negative confirmed by colonoscopy and pathology.Colorectal lesions were detected in 20 cases by dual-energy CTC,of which no false-positive and one case was false negative confirmed by colonoscopy and pathology.The contrast enhanced CT value of polyps,adenoma,adenocarcinoma and stool was (38.54± 6.82),(49.16±7.31 ),(52.61 ±5 .93 )and (34.00±1 .41 )Hu,respectively.The enhanced value of adenoma and adenocarcinoma was significantly higher than that of polyps and stool,the differences were statistically significant among groups (F = 10.760,P = 0.001 ).There was no significant difference between polyps and stool (t=1 .44,P =0.188).The sensitivity of regular CTC and dual-energy CTC in lesion detection was 95 .6% (95 %cofidence interval(CI ):77.9%-99.2%)and 95 .6% (95 %CI :77.9%-99.2%),respectively.The specificity was 42.8% (95 %CI :15 .4%-93.5 %)and 100.0% (95 %CI :47.9%-100.0%).Conclusion Compared with traditional CTC,dual-energy CTC would distinguish lesions from stool,help differentiate between benign and malignant tumors and further increase the accuracy of CTC diagnosis.

Objective This study was designed to observe the impact of cigarette cessation on anxiety and depression in patients with coronary heart disease (CHD).Methods A total of 690 cigarette smoking patients with CHD identified by coronary angiography (CAG) were included and analyzed in the study.The mental state were scored with Hamilton anxiety (HAMA) and depression (HAMD) scale both on admission and at 6-month follow-up.The patients were divided into two groups based on the cigarette cessation.The score of mental state between the two groups were compared.The patients were treated with percutaneous coronary intervention (PCI),coronary artery bypass grafting (CABG),or medicine therapy (MT).Results The clinic data and score of mental state were similar at the time of admission (HAMA:10.66±5.53 vs 11.09 ±5.61,P =0.311;HAMD:29.81 ± 10.13 vs 28.94 ± 10.22,P =0.266 4) between the two groups.After 6 months,the proportions of subjects in smoking cession group with anxiety (24.2％ vs 32.3％,P ＜0.05),depression (18.0％ vs 27.5％,P ＜0.05),and anxiety and depression (7.0％ vs 16.2％,P ＜0.001) decreased significantly compared with those in smoking group irrespective of the treatment strategy.Both the HAMA and HAMD scores were lower in smoking cessation group (HAMA:9.83±3.40;HAMD:24.91 ±7.90) than in smoking group (HAMA:10.98 ±4.87;HAMD:27.70 ± 11.16) (all P ＜ 0.001).Conclusions Smoking cessation is good for the relief of anxiety and depression in CHD patient.

OBJECTIVETo investigate the influence of seasons, blood types and semen parameters on the cryosurvival rate of frozen-thawed spermatozoa from sperm donors.

METHODSWe retrospectively analyzed the influence of seasons, blood types, abstinence period, semen volume, sperm morphology, and pre-freeze sperm motility and concentration on the cryosurvival rate of frozen-thawed sperm in 4 088 semen samples from Shaanxi Human Sperm Bank.

RESULTSThe cryosurvival rate of the post-thaw sperm was correlated negatively with the progressive motility of the pre-freeze sperm (r = -0.262, P < 0.01), but positively with pre-freeze sperm concentration (r = 0.247, P < 0.01), and it was significantly higher in the 6-day abstinence group ([70.2 +/- 5.4]%) than in the other abstinence groups (P < 0.01), so was it in the normal sperm morphology rate > 20% group ([71.4 +/- 5.1]%) than in the others (P < 0.01). The survival rate of the post-thaw sperm was not correlated with seasons, blood types, and semen volume (P > 0.05).

CONCLUSIONSperm motility, concentration and morphology and the abstinence period of sperm donors may be valuable predictors of the cryosurvival rate of post-thaw sperm, but no correlation was found between the survival rate and seasons, blood types and semen volume.

Objective: To investigate the applied feasibility of scaffold with modified PLA (Polymer of lactic acid) in tissue engineering. Methods:First, we adopted salting-in method to prepare porous foam scaffold. Then, we reconstructed tissue engineering skin by epidermal cells and fibroblasts combined with modified PLA. On the 14th day of cell culturing in vitro, we was a control. Results:The arfificial skin is composed of epidermis and dermis and similar to natural skin in appearance. The skin consists of fibroblasts and keratinocytes, which are in various proliferation and differentiation stages. Fibroblasts and keratinocytes distribute on the surface of polymer of lactic acid (PLA) and the number of fibroblast and keratinocyte increase. Conclusion:Dialdehyde starches (DAS) not only improve the function of PLA but also have good effects on cells. Moreover, it does not affect the growth and the metabolism of the cells. So it is feasible to use modified scaffold to construct tissue engineering skin.