2.Construction of a cDNA Library and Cloning of an Arabinosidase cDNA from Armillariella tabescens
Dongsheng YAO ; Hui HUANG ; Long ZHAO ; Chunfang XIE ; Daling LIU
China Biotechnology 2005;25(6):65-70
The expression cDNA library of A. tabescens was constructed by SMART technique, which useλTriplEx2 as a vector. The titer and the percentage of the constructed library were about 1.0 × 106pfu/mland 98.3% respectively, and the titer and the capacity of the amplified library were about 3.1 × 108pfu/mland 4.2 × 1010. The library was used to provide expressed sequence tags (ESTs). 147 Expressed SequenceTaqs (ESTs) were gained from 176 clones, which were selected randomly and sequenced at the 5'end. Thesequences were submitted to the EMBL database. Blasting the sequences in the GenBank, 43 of them werefound that they have significant similarity with data in GenBank. EST AJ620046 was has significantsimilarity with the arabinosidase of Bacteroides thetaiotaomicron. Using SMART-RACE a full-length cDNA ofAJ620046 was successfully obtained. In order to initially characterize the biochemical properties ofAJ620046, the ORF of AJ620046 named AF was cloned and expressed in Pichia Pastoris yeast.Recombinant pHIL-S1-AF constructed by inserting AF into pHIL-S1 was transformed into Pichia PastorisGS115. Preliminary experiments indicated that AJ620046 was expressed as a 32 kDa protein in recombinantyeast.
3.Anatomical locking compression plate augmented with anchor nail versus calvicular hook plate for Neer Ⅱb distal calvicular fractures
Sanfu LIN ; Xuedong YAO ; Zhangsheng DAI ; Hui YE ; Jinzhi YAO ; Junjie XIE
Chinese Journal of Orthopaedic Trauma 2017;19(1):41-46
Objective To compare the clinical efficacy of anatomic locking compression plate augmented with anchor nail versus calvicular hook plate in the treatmeut of Neer type Ⅱb distal calvicular fractures.Methods The data of 65 patients were retrospectively analyzed who had undergone surgery in our hospital from January 2012 to January 2015 for Neer type Ⅱb distal calvicular fractures.They were 39 men and 26 women,aged from 18 to 58 years(average,42.6 years).Open reduction plus anatonical locking compression plating augmented with anchor nailing was performed in 33 patients (group A) and open reduction plus calvicular hook plating was conducted in 32 (group B).The internal fixation was removed oue year after surgery in all.Constant Scale was used to evaluate shoulder function at 1,3 and 6 months postoperatively.X-ray examination on the shoulder was also conducted to observe fracture healing.Complications and percentage of the patients who resumed their job 3 months postoperatively were documented.Results The 2 groups were comparable because they showed no significant differences in general clinical information preoperatively (P > 0.05).All the 65 patients were followed up tor 12 to 18 months (average,15.2 months).The fracture clinical healing time in group A (23.9 ± 2.3 weeks) was significantly shorter than in group B (26.1 ± 3.0 weeks) (P < 0.05).The mean Constant-Murley scores at 1,3,6 months and the last follow-up postoperatively in group A (91.2 ±3.6,95.2 ±2.4,96.1 ±5.1 and 97.3 ± 1.6) were significantly higher thau those in group B (89.2 ± 6.1,91.1 ± 1.1,91.2 ± 6.2 and 92.1 ± 3.1) (P < 0.05).The rate of total complications in group A (6.1%,2/33) was significantly lower than in group B (25.0%,8/32) (P < 0.05).At postoperative 3 months,31 patients (93.9%) in group A resumed their job while 23 (71.9%) ones did in group B,showing a significant difference (P < 0.05).Conclusions For Neer lⅡb distal calvicular fractures,anatomical locking compression plate augmented with anchor nail is obviously superior to calvicular hook plate,because the former can avoid damage to the soft tissue surrounding the acromion,leading to satisfactory functional recovery of the affected shoulder.
4.Cloning, Expression and Property Analysis of Arabinosidase in Pichia pastoris
Dong-Sheng YAO ; Hui-Mei TAN ; Hui HUANG ; Da-Ling LIU ; Chun-Fang XIE ;
China Biotechnology 2006;0(04):-
SMART-RACE was performed after isolating the total RNA of Armillariella tabescens to amplify the full-length cDNA of arabinosidase (GenBank Accession No. AJ620046). Bioinformatics analysis was used to analyze the code frame of arabinosidase, to predict its structure and function. Recombinant plasmid pPIC9-AF was constructed and then electroporated into methylotrophic yeast Pichia pastoris GS115. The secreted 6 ? His fusion protein was purified to analyze its enzymology property. This arabinosidase had high activity at 30-35℃ under acid condition, and was stable within wide range of pH and temperature. It maintained about 80% activity at the range of pH4. 0-8.0 and 20-40℃,wider than many other cloned arabinosidase. So it was worthy to go step further to study this enzyme, and recombinant expression provided a chance of highly expressing arabinosidase.
5.Infliximab treatment of intravenous immunoglobulin-resistant Kawasaki disease in 2 children and literature review
Hui YAN ; Yonghong CHEN ; Qingyou ZHANG ; Wanzhen LI ; Yao XIE ; Qin HUI ; Junbao DU
Chinese Journal of Applied Clinical Pediatrics 2015;30(5):365-369
Objective To investigate the clinical effect and the prospect of Infliximab in treatment of intravenous immunoglobulin (IVIG)-resistant Kawasaki disease (KD) patients.Methods Clinical features,inflammatory markers and coronary changes were observed in 2 cases of IVIG-resistant KD patients hospitalized in Peking University First Hospital,who were treated effectively by Infliximab.Relevant researches on the mechanism and progress of the Infliximab treatment for IVIG-resistant KD in the last 10 years were reviewed at the same time.Results Two KD patients hospitalized in Peking University First Hospital had been treated with 2 g/kg IVIG for 2 times and followed by methylprednisolone treatment.However,fever and other clinical manifestations occurred again after 2 days and 6 days when temperature returned normal.They both defervesced and all the symptoms were improved after 1 dose of Infliximab (5 mg/kg) by laboratory examinations.Four published literatures of the basic research and 9 retrospective or prospective clinical researches of Infliximab treatment of KD showed that Infliximab alleviated the inflammatory level in the KD patients significantly.Complete remission was up to 72.73%-92.11%.Those KD patients defervesced within 12 h,with dramatic improvement of symptoms and signs.Arthralgia also disappeared in the patients with arthritis.Only 1 case was complicated with hepatitis in the acute phase and cholecystitis in recovery time.A phase 3 randomised,double-blinded,placebo-controlled trial had been done to assess the addition of Infliximab to the standard therapy.Conclusions Infliximab is a feasible choice for IVIG-resistant KD patients.Efficacy and safety of Infliximab for KD treatment have been proved in the literature.However,Infliximab for KD treatment has not been indicated in the drug instruction,so the informed consent from the guardians and Ethics Committee is needed.
6.Universality Assessment of matK Primer Pairs in Seed Plants
Linchun SHI ; Jinxin LIU ; Hui YAO ; Xiaolan XU ; Tianyi XIN ; Caixiang XIE ; Jingyuan SONG
World Science and Technology-Modernization of Traditional Chinese Medicine 2013;(3):381-386
matK is one of the core DNA barcode markers for plant DNA barcode identification and its universality using single makers has been in controversy. However, the universalities of different matK primer pairs in same seed plant group (order) and same matK primer pairs in different seed plant groups (order) are lack of systematic research. In this study, we collected 14563 full-length matK sequences of 11429 species of 3292 genera in 239 families belonging to 36 orders in seed plants. The universalities of 13 matK primer pairs and its 78 primer com-binations have been assessed using bioinformatics methods. The results indicated that xf/5r, 1F/8R, 390F/1326R and 3F_KIM/1R_KIM were the four most universality primer pairs. The four markers' universalities were 91.18%, 84.65%, 79.81% and 80.94% respectively in all 11429 seed plants. The most universality primer pairs in different orders were different. For each order, the primer pair with maximum universality was different. the xf/5r was the basal primer pair for primer combination and 1F/8R, 1F/1R, M3/M4 and 3F_KIM/1R_KIM could be the complementary primer pairs. This study could be a valuable resource for the primer selection of the research DNA barcoding identification in seed plants.
7.Controlled comparison of two supplemented aerobic and anaerobic blood culture bottles to detect simulated bacteremia specimens
Yao WANG ; Ying-Chun XU ; Xiu-Li XIE ; Hui WANG ; Min-Jun CHEN ;
Chinese Journal of Laboratory Medicine 2001;0(05):-
Objective To compare BACTEC Plus aerobic and anaerobic bottles with BacT/ALERT FA aerobic bottles and FN anaerobic bottles in the ability of detecting simulated bacteremia specimens.Methods The 202 pairs of specimens were composed of 5ml sterile blood and defined loads of microorganisms.112 pairs of specimens in them also contained defined doses of antibiotics to simulate the patients undertaking antibiotic therapy.Time-to-detect(TTD)and positive percentages were evaluated in four groups,including aerobic bottles detecting aerobic bacteria,anaerobic bottles detecting anaerobic bacteria and facultative anaerobic bacteria,and aerobic bottles contained antibiotics detecting aerobic bacteria.Results The positive percentages of two kinds of aerobic bottles were both 100%.For the specimens with bacterial concentration of 10~2 and 10~3 CFU/ml,average TTDs of BACTEC Plus aerobic bottles[(13.69?3.74)h,(11.54?2.87)h]were faster than those of BacT/ALERT FA bottles [(16.76?5.62)h,(14.47?4.30)h;t=-5.674,-7.294,P
10.Health management system in outpatient follow-up of kidney transplantation patients
Hong ZHANG ; Jinliang XIE ; Hui YAO ; Ling LIU ; Jianwen TAN ; Chunmi GENG
Journal of Central South University(Medical Sciences) 2014;(7):739-744
Objective: To develop a health management system for outpatient follow-up of kidney transplant patients. Methods: Access 2010 database sotfware was used to establish the health management system for kidney transplantation patients in Windows XP operating system. Database management and post-operation follow-up of the kidney transplantation patients were realized through 6 function modules including data input, data query, data printing, questionnaire survey, data export, and follow-up management. Results: The system worked stably and reliably, and the data input was easy and fast. The query, the counting and printing were convenient. Conclusion: Health management system for patients after kidney transplantation not only reduces the work pressure of the follow-up staff, but also improves the effciency of outpatient follow-up.