To improve the inulinase application in biotechnology, the characteristic of inulinase from Kluyveromyces marxianus YX01 was investigated. The inu gene of K. marxianus YX01 was transformed into Pichiapastoris GS115 host cells with molecular biology techniques. Then we achieved the heterologous expression of exo-inulinase whose molecular mass was about 86.0 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Furthermore, six His-tag was added to the inulinase and a two-step method was applied in the purification of inulinase, including concentration via dialysis by polyethylene glycol 20 000 and metal Ni-NTA Agarose affinity adsorption. The purification factor of purified protein was 3.6 and the recovery rate of enzyme activity was 33.1%. We characterized the purified inulinase. The optimum temperature was 60 degrees C and pH was 4.62. When inulin and sucrose were used as substrates, the K(m) and V(max) values were 80.53 g/L vs 4.49 g/(L x min) and 183.10 g/L vs 20.20 g/(L x min), respectively. In addition, metal ions including Mn2+, Ca2+, Cu2+, Zn2+ and Fe2+ exhibited different degrees of inhibition on the enzyme activity, and Cu2+, Zn2+ and Fe2+ exhibited the most significant inhibition. Our findings might lay a good foundation for industrial application of inulinase.
Glycoside Hydrolases ; chemistry ; genetics ; Industrial Microbiology ; Inulin ; Kluyveromyces ; enzymology ; genetics ; Pichia ; Sucrose ; Temperature

Objective To investigate the ultrasound diagnostic value for nipple discharge disease by comparison with fiberoptic ductoscopy. Methods Fiberoptic ductoscopy and ultrasonic inspection were performed on fifty-seven patients with spontaneous nipple discharge,and the results were analysed. Results In all patients, 55 cases (96. 49 % ) were abnormal in high-frequency ultrasound inspection:27 mammary duct expansion(47.36%),19 occupying lesion with mammary duct expansion or cyst (33.33%),6 cystic echo ( 10. 52% ) ,3 only solid echopattern (5.26%) ,and 2 nothing found (3.51% ). In ultrasonic inspection,color and pulse Doppler signal were found in 7 cases. On the other hand, 26 galactophoritis (45.61%), 12mammary duct ectasia (21.05 %), 17 intraductal papilloma (29.82%), 2 breast carcinoma (3.50%) were diagnosed by fiberoptic ductoscopy. In all 19 cases with occupying lesion,ultrasonography's sensitivity was 52. 63% ,specificity was 89. 47%, misdiagnosis rate was 10. 50%, rate of missed diagnosis was 47.36%.Conclusions To nipple discharge disease, high-frequency ultrasound has high sensitivity, easy to find abnormal indication,but is not accurate enough in the diagnosis of smaller occupying lesion.

The poor chemical stability and solubility, easy volatile and other shortcomings of TCM volatile oils can be improved after they are included by cyclodextrin. Therefore, cyclodextrin inclusion technology is widely used in the preparation process of volatile oil. This article reviewed the domain of cyclodextrin inclusion of volatile oils, the types of cyclodextrin, inclusion technologies, characterization for inclusion, components changes before and after inclusion, and dissolution characteristics of cyclodextrin inclusion.

Objective To investigate the factors affecting the stability of the volatile oil extracted fromYinqiaosan Decoction.Methods The main chemical compositions and the extraction repetitiveness of the compound volatile oil were determined by GC-MS, and the stability of multiple extracted volatile oil was studied. Absorbance of the compound volatile oil was used as the evaluation index, and the factors affecting the stability of the of the compound volatile oil were investigated, such as illumination, temperatures and pH values of volatile oil solution and metal ions.Results The results of the GC-MS chromatograph indicated that the main chemical compositions of the compound volatile oil extracted fromYinqiaosan Decoction twice were the same. The results of the stability of the volatile oil showed that the preservation temperature and illumination affected the stability of the volatile oil to a certain extent. The absorbance values of the compound volatile oil changed slowly when it was stored at a relatively low temperature (4℃) and shielded from light, and it was less stable when stored in normal temperature and under illumination. Meanwhile, the absorbance of the compound volatile oil changed quickly in acid or alkaline solutions and was in instability. The metal ions, such as Cu2+ and Fe3+, have chemical reactions with the compositions of the compound volatile oil and there was a big change in the UV-Vis spectrum of the compound volatile oils.Conclusion The compound volatile oil should be stored at a relatively low temperature (4℃) and shielded from light. At the same time, it should be stored avoiding acids, alkaline and the metal ions, such as Cu2+ and Fe3+, to guarantee its stability. This study provides a reference for the preservation conditions and the preparation conditions of the compound volatile oil extracted fromYinqiaosan Decoction.

Objective:To investigate the enantiomer separation for atropine by capillary electrophoresis .Methods:Capillary elec-trophoresis was used with an elastic quartz capillary column (60 cm ×75 mm, effective length of 40 cm).The concentration of phos-phate buffer was 30 mmol· L-1 .The high and time of injection was 10 cm and 5 s, respectively.The detection wavelength was 225 nm.The best separation conditions were selected including the type and concentration of chiral resolving agent , pH of the buffer solu-tion, operating voltage and organic solvent.Results:The optimum conditions of separation were as follows:the pH of buffer solution was 7.0, the concentration of S-β-CDP was 10 mg· ml-1 , and the operating voltage was 12 kV.Conclusion: The method is simple and fast, which can be used to se parate the optical isomers of atrpo ine.

Objective To observe the impact of the five-emotion restrictive therapy on coronary heart disease. Methods109 hospitalized patients with coronary heart disease were randomly divided into a experimental of 56 patients and a control group of 53 patients according to length of stay. The control group was treated with the conventional therapy; the experimental group was imposed five-emotion restrictive psychological intervention on the basis of conventional therapy. The investigation was done using self-rating anxiety scale(SAS) and self-rating depression scale(SDS) on 1 day after admission and ld before discharge respectively. SAS and SDS scores were surveyed. At the same time Holter monitoring and normal ECG test were compared. Results Compared with control group, the experimental group 1 d before discharge of the SAS and SDS scores were (41.9±7.4) and (38.6±8.4), and the difference was statistically significant (P＜0.01) . The total effective rate of Holter monitoring and normal ECG was 64.15％ and 92.85％ in the control and experimental group, and the difference was significant (P＜0.01) . ConclusionThe pathological seven emotions have a certain affection on the incidence of coronary heart disease, treatment and prognosis of the course. Five-emotion restrictive psychological intervention can adjust timely psychological state of patients, eliminate negative emotions, and significantly help the incidence, treatment and prognosis of the disease.

Objective To investigate the epidemic status of endemic fluorosis in Shandong Province,Jining City,and to provide a basis for prevention and control of the disease.Methods Based on Shandong Provincial Project Technical Solutions for Endemic Fluorosis,Rencheng,Jinxiang,Yutai,Jiaxiang and Liangshan Counties in Jining were selected as monitoring sites.According to the illness situation of mild,moderate or serious districts,one village was selected as a major survey site from each county(district).There were a total of 15 such villages selected.Survey content included drinking water fluorine level; dental fluorosis of children,adults' clinical skeletal fluorosis and urinary fluorine levels; water and urinary fluoride content were determined by the method of fluoride ion selective electrode; dental fluorosis of children was diagnosed by Deans method and clinical diagnosis was based on the Diagnostic Criteria of Endemic Skeletal Fluorosis (WS 192-2008).Results Sixty-one water samples from 15 villages of five counties (districts) were tested.Fluoride levels of 9 out of the 61 samples were exceeded the national standard (＞ 1.0 mg/L),and the rate was 14.75％; 1 sample ＞ 2.0 mg/L,and the maximum water fluoride was 2.25 mg/L.Seven hundred and seventeen people's real time urinary fluoride was detected in the 15 villages,including 420 children and 297 adults,and the geometric mean were 1.53 and 1.69 mg/L,respectively.Clinical examination of 755 children aged 8 to 12 showed that the detection rate of dental fluorosis was 26.89％ (203/755); defect rate was 9.12％(29/755) and dental fluorosis index weres 0.65.The detection rate of clinical skeletal fluorosis of 11 565 adults was 4.76％(550/11 565),including 303 moderate or serious cases.Conclusions The situation of excessive water fluorine in outside environment in Jining City has been controlled at a certain degree; groups urinary fluoride level is closed to the normal upper limit; the prevalence of dental fluorosis or skeletal fluorosis has been suppressed at a certain degree,therefore,the results of control should be further consolidated and expanded,in order to completely eliminate the fluoride hazard.

OBJECTIVE: The present study was aimed to assess the ability of tone recognition performance for postlingually deafed cochlear implanted adults using no-duration tone test materials. METHOD: Sixteen cochlear implanted users have used their devices at least six months. Ten monosyllabic syllables were selected, and combined with four lexical tones which were made up of 40 tone tokens. Then, these original words were recorded using one adult male and one adult female native Beijing Mandarin speaker. The speakers were asked to record these 40 monosyllabic words multiple times, and the 80 tokens in which the durations of four tones in each monosyllabic word were within 5 ms precision were chosen as the original tone tokens. The subjects were asked to perform a four alternative forced-choice study and select the tone they had heard. RESULT: The mean score of lexical tone perception tasks for 16 cochlear implanted users is (70.7 ± 22.0)%, and Tone 4 was the easiest to perceive, while Tone 2 was the hardest for cochlear implanted subjects to perceive. Tone 1 was more often misperceived as Tone 2 relative to other tones. Tone 2 was more often misperceived as Tone 1 and 3. Tone 3 was more often misperceived as Tone 2. The tone perception performance was positively correlated with duration of hearing aid use. CONCLUSION Duration cues may impact on the ability of tone recognition performance for cochlear implant subjects, especially for tone 3. Due to the limited ability for cochlear implant subjects on pitch perception, tone l and tone 2, and tone 2 and tone 3 are most often confused with each other without duration cues. The experience of hearing aid use may benefit for postlingual deafness cochlear implant recipients on tone perception performance.
Adult ; Asian Continental Ancestry Group ; Cochlear Implantation ; Cochlear Implants ; Female ; Hearing ; Hearing Aids ; Humans ; Language ; Male ; Pitch Perception ; Speech Perception ; Tongue

OBJECTIVE: This study was aimed at investigating the ability of lexical tone perception in listeners with sensorineural hearing impairment and auditory. neuropathy spectrum disorder. METHOD: Three groups of subjects were recruited in this study, including 11 subjects with normal hearing, 14 subjects with sensorineural hearing impairment, and 25 subjects with auditory neuropathy spectrum disorder. Ten monosyllabic syllables were selected, and combined with four lexical tones which were made up of 40 tone tokens as the original test materials. Then, these original words were recorded using one adult male and one adult female native Beijing Mandarin speaker. The speakers were asked to record these 40 monosyllabic words multiple times, and the 80 tokens in which the durations of four tones in each monosyllabic word were within 5ms precision were chosen as the test tone tokens. The subjects were asked to perform a four-alternative forced-choice study and select which tone they had heard. RESULT: The mean and standard deviation of the tone perception correct scores for normal-hearing subjects, subjects with sensorineural hearing impairment, and subjects with auditory neuropathy spectrum disorder were (97.3 ± 2.8)%, (88.0 ± 9.9)%, and (65.7 ± 17.1)%, respectively. Significant differences in tone perception scores were found to be between subjects with normal hearing and subjects with sensorineural hearing impairment (P < 0.01) and between subjects with auditory neuropathy spectrum disorder (P < 0.01). In addition, a significant difference was found to be between subjects with sensorineural hearing impairment and auditory neuropathy spectrum disorder (P < 0.05). A significantly negative correlation was observed between tone perception score and pure tone hearing thresholds for both subjects with sensorineural hearing loss (r = -0.756, P < 0.01) and subjects with auditory neuropathy spectrum disorder (r = -0.546, P < 0.01). CONCLUSION As the hearing loss became more severe, the ability to perceive lexical tone for both subjects with sensorineural hearing loss and auditory neuropathy spectrum disorder reduced. Subjects with auditory neuropathy spectrum disorder had more degraded ability to perceive lexical tone and a larger individual difference, in comparison with subjects with sensorineural hearing loss.
Adult ; Audiometry, Pure-Tone ; Case-Control Studies ; Female ; Hearing ; Hearing Loss, Central ; physiopathology ; Hearing Loss, Sensorineural ; physiopathology ; Humans ; Language ; Male ; Speech Perception

Objective This study was aimed at investigating the ability of temporal gap detection (TGD) in listeners with auditory neuropathy spectrum disorder (ANSD ) .Methods The temporal gap detection thresholds were measured for subjects using a TGD program developed by University of California ,Irvine .Three groups of subjects were recruited in this study ,including 12 subjects with normal hearing ,12 subjects with sensorineural hearing loss (SNHL) ,and 15 subjects with ANSD .The differences of TGD thresholds among the three groups of subjects were analyzed using a one - sample ANOVA method .Results No significant difference of the TGD thresh‐old was observed between two ears for all the subjects .The mean and standard deviation of the gap - detection threshold for both ears was about 3 .9 ± 1 .1 ms ,3 .6 ± 1 .1 ms ,and 13 .7 ± 9 .9 ms for normal - hearing subjects , subjects with SNHL ,and subjects with ANSD ,respectively .The TGD threshold in subjects with ANSD was sig‐nificantly higher than those of in normal hearing group (P< 0 .01) and SNHL group (P< 0 .01) ,while no signifi‐cant difference was found in TGD thresholds between normal - hearing subjects and subjects with SNHL .Conclusion For listeners with auditory neuropathy spectrum disorder ,their ability to resolve temporal information was de‐graded with a large individual variability .