ObjectiveTo identify the sites of thermal stability at Loop structure of xylanase Xyn ASP in Aspergillus saccharolyticus JOP 1030-1 and improve the thermal stability.MethodsThe amino acid sites related to thermal stability of xylanase were predicted,and beneficial mutation sites at Loop structure were screened by Fireprot online server.Singlepoint mutants Xyn(A79Y),Xyn(A79Y),Xyn(T81Q)and double-point mutants Xyn(T81Q)and double-point mutants Xyn(LQ)(A79Y/T81Q)were constructed by site-directed mutagenesis.The recombinant mutant plasmid was transformed to E.coli BL21(DE3)and induced by IPTG.The recombinant xylanase was purified by Ni-NTA protein purification kit,and determined for the optimum temperature,thermal stability,optimum p H and p H stability.ResultsBeneficial mutation sites A79Y and T81Q were screened at Loop structure,and the purified protein samples showed high purity.Compared with that of wild type Xyn ASP,the optimum temperature of mutants Xyn(LQ)(A79Y/T81Q)were constructed by site-directed mutagenesis.The recombinant mutant plasmid was transformed to E.coli BL21(DE3)and induced by IPTG.The recombinant xylanase was purified by Ni-NTA protein purification kit,and determined for the optimum temperature,thermal stability,optimum p H and p H stability.ResultsBeneficial mutation sites A79Y and T81Q were screened at Loop structure,and the purified protein samples showed high purity.Compared with that of wild type Xyn ASP,the optimum temperature of mutants Xyn(A79Y),Xyn(A79Y),Xyn(T81Q)and Xyn(T81Q)and Xyn(LQ)increased by 10℃，5℃and 5℃，respectively.After heat treatment at 40℃for 30 min,wild type Xyn ASP retained only 30.2%residual relative activity,while the mutant Xyn(LQ)increased by 10℃，5℃and 5℃，respectively.After heat treatment at 40℃for 30 min,wild type Xyn ASP retained only 30.2%residual relative activity,while the mutant Xyn(T81Q)retained 37.7%,Xyn(T81Q)retained 37.7%,Xyn(A79Y)and Xyn(A79Y)and Xyn(LQ )still retained more than 65%.After heat treatment at 60℃for 30 min,the enzyme activity of wild type was 7.1%,while that of Xyn(LQ )still retained more than 65%.After heat treatment at 60℃for 30 min,the enzyme activity of wild type was 7.1%,while that of Xyn(LQ)remained 26.4%.The thermal stability of mutants was improved compared with that of wild type Xyn ASP.The optimum p H of Xyn(LQ)remained 26.4%.The thermal stability of mutants was improved compared with that of wild type Xyn ASP.The optimum p H of Xyn(A79Y),Xyn(A79Y),Xyn(T81Q)and Xyn(T81Q)and Xyn(LQ)was 5.0,which was lower than that of wild type Xyn ASP(p H 6.0).The p H stability of Xyn(LQ)was 5.0,which was lower than that of wild type Xyn ASP(p H 6.0).The p H stability of Xyn(A79Y),Xyn(A79Y),Xyn(T81Q)and Xyn(T81Q)and Xyn(LQ)at pH 3.0～8.0 showed no significant change compared with the wild type.ConclusionSite-directed mutagenesis(A79Y and T81Q)was carried out at Loop structure,and xylanase mutants with obviously improved thermal stability were obtained,which laid a foundation of the later research on the structure,function and relationship of the enzyme and its industrial application.

PurposeTo investigate clinical significance of echocardiography combined with Flash CT in diagnosis of partial anomalous pulmonary venous connection (PAPVC). Materials and Methods Twenty-eight cases of PAPVC were recruited in the study. All the subjects underwent both preoperative echocardiography and coarse pitch Flash CT scanning. Imaging findings were compared with the results of surgery, including the type of PAPVC, size of atrial septal defect (ASD), and other combined malformations. Results Among 28 patients, 16 cases and 5 suspicious cases (4 cases of PAPVC and 1 case of pulmonary vein embolism) were diagnosed by echocardiography. Six cases were missed, and 1 case was misdiagnosed. CT misdiagnosed 1 case. The accurate diagnosis rate of echocardiography (71.4%, 20/28) was lower than that of Flash CT (96.4%, 27/28) (P<0.05). There were 44 combined malformations in 28 cases, 43 lesions were diagnosed by echocardiography, and 34 lesions were diagnosed by Flash CT. The accurate diagnosis rate of echocardiography (97.7%, 43/44) was higher than that of Flash CT (77.3%, 34/44) (P<0.05). The average DLP was (15.5±3.2) mGy·cm and average effective dose (ED) was (0.354±0.058) mSv.Conclusion Flash CT has a higher diagnosis accuracy of PAPVC but a lower diagnostic accuracy of combined malformations when compared with echocardiography. Moreover, echocardiography can make comprehensive evaluation in valve disease, pulmonary artery pressure and cardiac function. Flash CT combined with echocardiography may improve preoperative diagnosis.

A total of 89 elderly patients with heart failure and preserved ejection fraction were randomly divided into 2 groups:patients in control group received conventional medication,those in exercise group receive conventional medication plus aerobic exercise training,which was lasted for 6 weeks.The changes in VO2max,maximum heart rate (HRmax),VO2max/HRmax,exercise time,6-minute walk and quality of life (QOL) were evaluated after 16 weeks.The VO2max,HRmax,VO2max/HRmax,exercise time,6-minute walk and QOL had increased significantly in exercise group,compared with the control group.The results indicate that aerobic exercise can improve cardiopulmonary function of elderly patients with heart failure and preserved ejection fraction.

100 inpatients with heart failure were divided into two groups: 50 with preserved LVEF and 50 with reduced LVEF. Another 50 age-matched healthy subjects served as the control group. The severity and etiology of heart failure, tissue Doppler imaging indexes (Sm;Ea;Aa;E/Ea) and plasma BNP concentrations were compared among the three groups. There was no difference in terms of NYHA classification between groups of preserved LVEF and reduced LVEF. Hypertension and DM are more common in the group of preserved LVEF. The index of Sm decreased most significantly in the group of reduced LVEF, and was negatively correlated with the BNP concentration (r = -0. 35, P ＜ 0. 05 ). The ratio of E/Ea increased gradually among the three groups, and was positively correlated with the BNP concentration ( r = 0. 728, P ＜ 0. 05 ). The LV filling index E/Ea and the BNP concentration may be helpful in the diagnosis of heart failure patients with preserved LVEF.

Object The immunoregulation function was studied in experimental animals to observe the effect of single and/or combined use with Radix Aconiti (RA) and Radix Stephaniae Tetrandrae (RST). Methods The response of delayed type hypersensitivity (DTH) in mice, the phagocytosis of abdominal macrophages (M?) in mice, and the production of special antibodies in mice were observed. Results The two Chinese materia medica (CMM) in combination use could induce the increasing of phagocytosic function of M? hypoimmunitic model mice, the inhibiting on the responses of DTH in the hyperimmunitic model mice and the non immunosuppressive treated mice. On the production of special antibodies in mice, the enhance effect was found in RA used in single, and the suppressive effect was found when using the combination of the two CMM. Conclusion In accordance with the mechanism of the disorder between the anti inflammation effect and the induce inflammation effect to arthritis in the immune system, these data indicate that the bidirectional effect of combination of the two CMM used on the immune responses, may be the involved mechanism on treating arthritis.

Objective To investigate the effects of aerobic exercise on cardiorespiratory function in patients with obstructive sleep apnea/hypopnea syndrome (OSAHS).Methods A total of 84 patients with OSAHS were randomly divided into a control group and an aerobic exercise group (42 in each group).The control group was given only training in the activities of daily living (ADL).The acrobic exercise group was given ADL training combined with aerobic exercise training.The regime was maintained for 16 weeks.Each patient's VO2max,maximum heart rate (HRmax),exercise time to exhaustion,6-minute walking distance and quality of life (QOL) were evaluated before and after treatment.Results VO2max,HRmax,VO2max/HRmax,maximum exercise time,6-min walking distance and QOL all increased significantly more in the aerobic exercise group compared with the control group.Conclusion Aerobic exercise training can significantly improve cardiorespiratory function in patients with OSAHS.

The clinical features and lung CT findings of 174 heart failure patients,including 85 cases with normal left ventricular ejection fraction (LVEF) and 89 cases with reduced LVEF were reviewed.Patients with normal LVEF had a higher proportion of hypertension,diabetes and obesity than patients with normal LVEF; and cough and dyspnea were more common.The lung CT findings in patients with normal LVEF were frequently presented as interstitial lung edema,increased pulmonary texture,subpleural line,Kerley lines and diffuse ground-glass opacity.Patients with reduced LVEF had a higher proportion of coronary heart disease and clinical manifestations of acute left heart failure,and central alveolar edema presented in lung CT images.Results suggest that heart failure patients with normal LVEF usually have longer clinical history and occult symptoms,and have a different lung CT imaging features from those in heart failure patients with reduced LVEF.

Objective To investigate the activation of nuclear factor-B (NF-B)and hypoxia-inducible factor-1 (HIF-1)in patients with obstructive sleep apnea-hypopnea syndrome (OSAHS). Methods Subjects were classified into mild-to-moderate OSHAS group (n = 16), severe OSAHS group (n = 14) and the matched control group (n=30). Gene and protein expressions of NF-B and HIF-1 were measured by RT-PCR, Western blot in peripheral blood mononuclear cells (PBMC). Results NF-κB p65 mRNA and NF-κB p65 protein,HIF-1α mRNA and HIF-1α protein in PBMC were significantly higher in severe OSAHS group than those in mild–to-moderate group and control group (P<0.01). But there were no significant difference in NF-κB p65 mRNA and NF-κB p65 protein between mild-to-moderate group and control group (P=0.068, P=0.254 respectively). Only gene (P<0.05) not the protein (P=0.777) of HIF-1αwas higher in mild-to-moderate as compared with control group. Both NF-κB p65 mRNA and HIF-1αmRNA were positively correlated with AHI (r=0.493, P=0.006, r=0.508, P=0.004), while negatively correlated with nighttime lowest blood oxygen saturation (LSaO2)(r=-0.488, P=0.006, r=-0.46, P=0.011). There was a positive correlation between NF-κB p65 protein level and AHI (r=0.669, P<0.001). HIF-1αprotein level was positively correlated with AHI, ODI (r=0.628, P=0.001;r=0.480, P=0.018). There were positive correlations between NF-κBp65mRNA and HIF-1αmRNA (r=0.543, P=0.002), NF-κBp65 and HIF-1αprotein respectively (r=0.716, P<0.001). Conclusions As the gene and protein of NF-κB and HIF-1 were up-regulated in patients with OSAHS, and also positively correlated with the severity of sickness. We conclude that both NF-κB and HIF-1 were involed in the pathogenesis of OSAHS.

Objective To investigate the effect of narrative medicine mode in perioperative medical and nursing care for patients underwent radical nephrectomy. Methods A total of 124 patients received radical nephrectomy from February 2014 to October 2016 were selected and divided into experimental group and control group by block randomization grouping method with 62 cases each. Narrative medical treatment and care was applied in experimental group while control group received conventional treatment and care. The psychological status and social support of the patients were evaluated by Zung Self-rating Anxiety Scale(SAS), Self-rating Depression Scale (SDS) and social support rating scale(SSRS), as well as rehabilitation indexes including complications, first feeding time, hospitalization time, costs, etc. Results Anxiety and depression status were existed in both experimental and control groups before operations, and there were no significant difference in all pre-operative scales between two groups (P>0.05). The score of SAS, SDS, SSRS was (44.71 ± 4.67), (40.84 ± 4.05), (43.21 ± 3.60) points after operation and (54.98 ± 5.93), (46.52 ± 5.07), (31.87 ± 3.22) points before operation in experimental group. The score of SAS, SDS, SSRS was (53.24±5.97), (44.63±4.37), (32.61± 2.72) points after operation and (54.92 ± 5.83), (46.53 ± 4.89), (32.16 ± 2.48) points before operation in control group. Significant differences were found between before and after operation in experimental group (t=18.73, 8.85, 18.18, all P<0.01). However, no significant difference could be found between before and after operation in control group (t=1.590, 1.400, 1.910, P>0.05). After operation, the complications, first feeding time, hospitalization time and costs was 22.58%(14/62), (2.50±0.92) d, (11.16±3.72) d, (24.6±4.0) thousand yuan in experimental group, which was 48.39%(30/62), (3.98 ± 1.44) d, (15.48 ± 5.44) d, (40.2 ± 3.1) thousand yuan in control group. Significant differences were found between experimental group and control group (χ2=9.02, t=2.07, 8.06, 3.93, P<0.05 or 0.01). Conclusions Narrative medicine mode can provide medical care filled with respect, empathy, and humanistic concern, andimprove patients′psychological status, thus promoting rehabilitation.

OBJECTIVE: To investigate the characteristics of adductor spasmodic dysphonia phonatory break in mandarin Chinese and select the stimuli phrases. METHOD: Thirty-eight patients with adductor spasmodic dysphonia were involved in this study. Standard phrase " fù mŭ xīn" and a speech corpus in mandarin Chinese with 229 syllables covering all vowel and constant of mandarin Chinese were selected. Every patient read the phrases above twice in normal speed and comfortable voice. Two auditory perpetual speech pathologists marked phonatory break syllables respectively. The frequency of phonatory break syllables and their located phrases were calculated, rated and described. The phrases including the most phonatory break syllables were selected as stimuli phrases, the phonatory break frequency of which was also higher than that of standard phrase "fù mŭ xīn". RESULT: Phonatory break happened in the reading of all patients. The average number of phonatory break syllables was 14 (3-33). Phonatroy break occurred when saying 177 (77.3%) syllables in the speech corpus. The syllables "guŏ, rén, zāng, diàn, chē, gè, guăn, a, bā, ne, de" broke in 23.1%-41.0% patients. These syllables belonged to the phrases "pĭng guŏ, huŏ chē, shì de, nĭ shì gè hăo rén, wŏ mén shì yŏu zŏng shì bă qĭn shì nong dé hĕn zāng, wŏ mén nà biān yŏu wăng qiú yùn dong chăng, cān gŭan, jiŭ bā hé yī gè miàn bāo dìan, tā shì duō me kāng kăi a,wŏ yīng gāi zài xìn lĭ xiĕ yī xiē shén mē ne?". Thirty-seven patients (97.3%) had phonatory break in above mentioned words. Ratios of these words phonatory break also were more than "fù mŭ xīn". CONCLUSION Adductor spasmodic dysphonic patients exhibited different degrees of phonatory break in mandarine Chinese. The phrases" shì de, pĭng guŏ, huŏ chē, nĭ shì gè hăo rén, wŏ mén nà biān yŏu wăng qiú yùn dong chăng, cān gŭan, jiŭ bā hé yī gè miàn bāo dìan, tā shì duō me kāng kăi a" were recommended as stimuli phrases for adductor spasmodic dysphonia evaluation.
Dysphonia ; diagnosis ; physiopathology ; Female ; Humans ; Language ; Male ; Phonation ; Spasm ; diagnosis ; physiopathology ; Voice