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.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.
5.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.
7.Advance in the management of the difficult-to-treat patients with chronic hepatitis C.
Chinese Journal of Hepatology 2009;17(7):484-486
Antiviral Agents
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administration & dosage
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therapeutic use
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Drug Therapy, Combination
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Genotype
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Hepatitis C, Chronic
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drug therapy
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Humans
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Interferon-alpha
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administration & dosage
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therapeutic use
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Polyethylene Glycols
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administration & dosage
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therapeutic use
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Protease Inhibitors
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therapeutic use
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Recombinant Proteins
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Ribavirin
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administration & dosage
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therapeutic use
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Treatment Outcome
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Viral Load
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Virus Replication
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drug effects
9.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.Factors Affecting Sterilization of HBV on Dental Hand Piece
Lei SHI ; Guangliang NIU ; Jinghua TIAN ; Hui ZHAO ; Ya LIU ; Yao XIE
Chinese Journal of Nosocomiology 2006;0(06):-
OBJECTIVE To investigate the factors influencing the effects of disinfection of HBV on dental hand piece. METHODS After polluted with HBV positive serum,the dental hand piece was disinfected with sterilizer at the condition of 134℃ for 5 minutes according different treatment procedures.The samples were then taken and detected HBsAg by the automatic enzyme immune analyzer using the third generation EIA agent.The efficacy of disinfection of HBV was presented by the reaction of HBsAg detection. RESULTS If the dental hand piece was sterilized directly not cleansed using clean water after pollution and dried in room temperature,there were 96.87% of the samples being HBsAg positive,and the polluted but dried hand piece,even cleansed there were 87.5% of positive result, even if prolonged the time of sterilization and the frequencies of vacuumization,the HBsAg positive rate of the sample was still up to 56.25%.But if the hand piece was irrigated and cleansed immediately after pollution and sterilized,all samples were HBsAg negative. CONCLUSIONS Whether or not cleansed immediately is the most important factor influencing the efficacy of hand piece sterilization after pollution.