Objective:To explore any effect of upper limb swing training guided by rhythmic auditory stimulation (RAS) on the walking ability of stroke survivors.Methods:Eighty stroke survivors were randomly divided into an observation group and a control group. Both groups received conventional rehabilitation treatment, including neuromuscular facilitation, muscle strength training, balance training and gait training, but the observation group was additionally provided with RAS-guided upper limb swing training for 20min once a day, 5d per week for 6 weeks. Before and after the intervention, balance and lower limb function were quantified in both groups using Holden′s walking function classification, the Fugl-Meyer lower extremity motor function scale (FMA-LE), the Berg Balance Scale (BBS) and the 10m walk test (10MWT). Limits of stability were also quantified.Results:After the treatment, the average Holden, FMA-LE and BBS scores, as well as the average 10MWT time were significantly better in the observation group than in the control group. The average stability limits and their maintenance were also superior.Conclusion:RAS-guided upper limb swing training can improve the gait, walking ability, walking stability, walking speed and balance of stroke survivors.

Objective:To document any effect of clinical rehabilitation pathway management on intubation time, dysfunction and medical expenditure associated with tracheotomy after a stroke.Methods:A total of 154 stroke survivors undergoing tracheotomy were randomly divided into an observation group and a control group, each of 77. Both groups were given routine rehabilitation, while the observation group was additionally provided with clinical rehabilitation pathway management during the rehabilitation intervention. Kaplan-Meier analysis was performed before the experiment and after 2, 4 and 6 weeks of treatment. Clinical pulmonary infection scores (CPISs), scores on the Chelsea Physical Function Assessment Scale (CPAx) and hospitalization cost were compared between the two groups.Results:The median extubation time of the observation group (2d) was significantly shorter than that of the control group (10d). After 2, 4 and 6 weeks of treatment, the average CPIS scores of the observation group were in each case significantly lower than those before treatment and the control group′s averages at the same time points, even though after 4 and 6 weeks of treatment the control group′s average CPIS scores had improved significantly. After 2, 4 and 6 weeks of treatment, the average CPAx scores of the observation group were significantly higher than those before treatment and better than the control group′s averages, even though the control group too had improved significantly compared with before the treatment. Hospitalization days, total hospitalization cost, antibiotic cost and laboratory examination cost of the observation group were, on average, significantly lower than those of the control group.Conclusion:Rehabilitation path management can shorten the period of intubation, prevent pulmonary infections, relieve dysfunction, and reduce medical expenses for stroke survivors after a tracheotomy. It is worthy of clinical promotion.

Objective:To observe any effect of supplementing breathing training with electromyographic biofeedback in treating the swallowing function soon after a cerebral infarction.Methods:A total of 96 cerebral infarction survivors with dysphagia were randomly divided into a control group, a biofeedback group and a comprehensive treatment group, each of 32. All received conventional rehabilitation treatment for dysphagia, while the biofeedback group was additionally provided with EMG biofeedback training and the comprehensive treatment group received both breathing training and the biofeedback training. Before and after 6 weeks of treatment, all were subjected to a swallowing contrast examination to observe the opening of the upper esophageal sphincter, the forward displacement of the hyoid bone, and the upward displacement of the hyoid bone. The Rosenbak Penetration-Aspiration Scale and Functional Imported Food Scoring were also used to evaluate their swallowing function.Results:Before, there were no significant differences between the groups in terms of any of the measures. After the 6 weeks of treatment, all of the measurements had improved significantly in both groups, but the improvement in the biofeedback and comprehensive treatment groups on all of the measures was significantly greater than in the control group. The average values of the comprehensive treatment group were then superior to the biofeedback group′s averages.Conclusion:Supplementing breathing training with EMG biofeedback can significantly improve the swallowing function of stroke survivors and reduce their risk of aspiration.

Objective:To investigate the effect of nonalcoholic fatty liver (NAFLD) at different ages of onset with new-onset diabetes mellitus.Methods:The cohort study was conducted in Kailuan Group Company. Active and retired employees were used as study subjects. After excluding NAFLD diagnosed at baseline, previous history of diabetes mellitus, and long-term history of heavy drinking, 43 317 cases were finally included in the cohort. The study subjects were divided into five groups according to age (<30 years old as group 1, 30-39 years old as group 2, 40-49 years as group 3, 50-59 years as group 4, and ≥60 years as group 5). The prevalence and incidence density of new-onset diabetes mellitus were compared between each NAFLD and non-fatty liver population group. The effect of NAFLD at different ages of onset with new-onset diabetes mellitus was analyzed by multivariate Cox's regression model. Statistical analysis was performed using one-way ANOVA, χ2 test or multivariate Cox's regression model. Results:The prevalence and incidence density of diabetes mellitus was significantly higher in NAFLD than non-fatty liver population. The prevalence of diabetes mellitus in different age groups were 6.45%, 6.88%, 9.94%, 10.83%, and 11.43%, respectively. The incidence density of each age group was 9.21/1 000 person-years, 11.10/1 000 person-years, 16.17/1 000 person-years, 18.72/1 000 person-years, and 22.13/1 000 person-years, and the differences were statistically significant ( P<0.001). Multivariate Cox's regression model result showed that after adjusting for confounding factors such as gender, systolic blood pressure, and fasting blood glucose, the HRs (95% CI) for diabetes mellitus in each age group were 3.992 (1.897, 8.400), 2.321 (1.589, 3.392), 2.041 (1.667, 2.500), 2.007 (1.708, 2.360), and 1.908 (1.570, 2.319), and the differences were statistically significant ( P＜0.001). Conclusion:Newly developed NAFLD is an independent risk factor for new-onset diabetes mellitus. Early exposure to NAFLD increases the risk of developing diabetes mellitus compared with the same age group. Younger age of onset of NAFLD should be given attention and active treatment.

Agrobacterium tumefaciens-mediated transformation (ATMT), as a simple and versatile method, achieves successful transformation in the yeast-like cells (YLCs) of Tremella fuciformis with lower efficiency. Establishment of a more efficient transformation system of YLCs is important for functional genomics research and biotechnological application. In this study, an enzymolysis-assisted ATMT method was developed. The degradation degree of YLCs depends on the concentration and digestion time of Lywallzyme. Lower concentration (≤0.1%) of Lywallzyme was capable of formation of limited wounds on the surface of YLCs and has less influence on their growth. In addition, there is no significant difference of YLCs growth among groups treated with 0.1% Lywallzyme for different time. The binary vector pGEH under the control of T. fuciformis glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter was utilized to transform the enzymolytic wounded YLCs with different concentrations and digestion time. The results of PCR, Southern blot, quantitative real-time PCR (qRT-PCR) and fluorescence microscopy revealed that the T-DNA was integrated into the YLCs genome, suggesting an efficient enzymolysis-assisted ATMT method of YLCs was established. The highest transformation frequency reached 1200 transformants per 106 YLCs by 0.05% (w/v) Lywallzyme digestion for 15 min, and the transformants were genetically stable. Compared with the mechanical wounding methods, enzymolytic wounding is thought to be a tender, safer and more effective method.
Agrobacterium ; Blotting, Southern ; Digestion ; Genome ; Genomics ; Methods ; Microscopy, Fluorescence ; Oxidoreductases ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction ; Wounds and Injuries

In the post-genomic era, gene function analysis has attracted much attention. Transformation is often needed to investigate gene function. In this study, an easy, rapid, reliable, and cost-effective colony polymerase chain reaction (PCR) method for screening mushroom transformants was developed: picking up a suitable amount of transformant's tissue (1–10 μg) to 20 μl 0.25% Lywallzyme solution, and vortexing for 10 s followed by incubation at 34 °C for 15 min. Finally, 2 μl of the suspension was used as templates to perform PCR and single target bands were successfully amplified from respective transformants of Tremella fuciformis, Pleurotus ostreatus, and Pleurotus tuber-regium. This procedure could be widely employed for screening transformants in mushroom transformation experiments.

An improved method for extracting high quality and quantity RNA from a jelly mushroom and a dimorphic fungus—Tremella fuciformis which is especially rich in polysaccharides, is described. RNA was extracted from T. fuciformis mycelium M1332 and its parental monokaryotic yeast-like cells Y13 and Y32. The A260/280 and A260/230 ratios were both approximately 2, and the RNA integrity number was larger than 8.9. The yields of RNA were between 108 and 213 µg/g fresh wt. Downstream molecular applications including reverse transcriptional PCR and quantitative real-time PCR were also performed. This protocol is reliable and may be widely applicable for total RNA extraction from other jelly mushrooms or filamentous fungi rich in polysaccharides.
Agaricales* ; Fungi ; Humans ; Methods* ; Mycelium ; Parents ; Polymerase Chain Reaction ; Polysaccharides* ; Real-Time Polymerase Chain Reaction ; RNA*

Objective To analyze the application value of T-SPOT .TB and PPD test in the diagnosis of bone and joint tuberculo-sis .Methods A total of 94 patients with tuberculosis of the bone and joint receiving surgery or conservative treatment from January 2014 to August 2016 were collected in this study ,50 patients with osteoarthritistreated by surgical treatment of bone surgery were recruited as objects too .All the objects received T-SPOT .TB and PPD test ,and the diagnosis value were compared .Results The diagnostic sensitivity ,specificity ,positive predictive value ,negative predictive value and accuracy of PPD test were 46 .81% , 98 .00% ,97 .78% ,49 .49% ,64 .58% ,those of T-SPOT .TB were 93 .62% ,100% ,100% ,89 .29% ,95 .83% .The sensitivity ,nega-tive predictive value and the coincidence rate of PPD test were less than those of T-SPOT .TB and joint test ,the differences were statistically significant(P<0 .05) ,T-SPOT .TB ,combined with the diagnostic sensitivity ,specificity ,positive predictive value ,nega-tive predictive value ,with rate no significant difference(P>0 .05) .Six cases were misdiagnosed by T-SPOT .TB ,they were all in the early stage of spinal tuberculosis ,showed cavities ,but no spinal nerve defect .One cases were erroneous diagnosis ,while 50 cases were misdiagnosed by PPD test ,all the symptoms of misdiagnosed patients were relatively mild ,with rheumatoid arthritis and other autoimmune diseases .Conclusion T-SPOT .TB is an ideal method for diagnosis of bone and joint tuberculosis ,but present interfer-ence factors ,the operation of PPD testis simply ,but lack of sensitivity ,however it has certain value in screening .

Objective To explore the effect of intracavitary therapy on acute pulmonary embohsm.Methods Fifteen patients were selected as our subjects,who suffered acute pulmonary embolism and received percutaneous catheter thrombus crashing and catheter directed thrombolysis in Beijing Military.Region General Hospital from January 2009 to June 2011.Local injection of Urokinase was performed with a total amount of 500 000 U in catheter directed thrombolysis.After thrombolysis,low molecular Heparin was administered to patients for 7-10 days and oral administration of Warfarin was performed for 3-6 months.Clinical symptoms,improvement of physical signs,complications,changes of mean pulmonary arterial pressure(mPAP) and arterial partial pressure of oxygen(PO2),and the opening of pulmonary artery were recorded.Results The pulmonary arteries of the 12 patients were completely opened,and partially opened in 3 patients.The effective rate was 100％ (15/15).mPAP was reduced from (40.07 ±5.97) mmHg to (20.00 ±4.66) mmHg (t =-1.128,P ＜ 0.05),PO2 was increased from (50.26 ± 9.30) mmHg to (80.49 ± 9.04) mmHg (t =1.246,P ＜ 0.05).Patients were followed-up for 3-6 months and no recurrence case was seen.Conclusion The interventional therapy is effective,safe and practicable in the treatment of acute pulmonary embolism.

[ ABSTRACT] AIM: To study the autophagy of prostate cancer PC-3 cells induced by CD147 in vitro.ME-THODS:Themethod of amino acid starvation to induce autophagy was used.The expression of CD147 was detected by Western blotting.To study the functional effects of CD147 on autophagy in prostate cancer PC-3 cells, the down-regulation of CD147 expression was induced by the technique of RNAi.The conversion of autophagic marker protein LC3-I to LC3-II was determined by Western blotting.The cell death after starvation-induced autophagy was analyzed by trypan blue exclu-sion assay.RESULTS:The CD147 expression gradually increased in starvation-induced autophagy.The down-regulation of CD147 significantly increased the expression of autophagy-related protein LC3-II compared with control group.Mean-while, the cell death rates increased from (19.3 ±3.1)%and (22.3 ±3.5)%in control groups to (38.4 ±3.1)%in si-lencing the expression of CD147 in the PC-3 cells (P<0.05).CONCLUSION:CD147 inhibits starvation-induced auto-phgy and autophagy death in the prostate cancer PC-3 cells.