1.Enzymatic cyclization of peptides using immobilized sortase A.
Shu-xiang ZHANG ; Min-zhi LIU ; Yan YANG ; Ke-di CHENG ; Jian-qiang KONG ; Wei WANG
Acta Pharmaceutica Sinica 2015;50(5):627-632
Peptide cyclization, a pivotal approach to modifying linear precursors of proteins and pepticles, has been used to enhance their biological activities and serum stabilities. Recently, sortase A (SrtA) from Staphyloccus aureus becomes a promising new technology for efficiently incorporating site specific modifications into proteins, conjugating the cell surface and cyclizing the linear peptides. In this study, we constructed two recombinant expression systems, one with chitin binding domain and the other with six-histidine tag and chitin binding domain on the N-terminal of SrtA, separately. The results of enzymatic kinetics indicate that the two recombinant tags do not impair the transpeptidase activity of SrtA compared with the standard reaction reported under the same reaction condition. The two synthesized peptides with N-ternimal three glycines and C-terminal penta-amino acid motif, LPETG, were cyclized using immobilized and recycled SrtA. The SrtA-based cyclization promises to represent a simple method for easy and efficient enzymatic synthesis of large cyclic peptides.
Aminoacyltransferases
;
metabolism
;
Bacterial Proteins
;
metabolism
;
Cyclization
;
Cysteine Endopeptidases
;
metabolism
;
Enzymes, Immobilized
;
metabolism
;
Kinetics
;
Peptides
;
metabolism
;
Peptides, Cyclic
;
biosynthesis
;
Staphylococcus aureus
;
enzymology
2.Application of medical image three dimensional visualization system in the evaluation of resectability of pancreatic and periampullary cancer
Chihua FANG ; Wen ZHU ; Yingfang FAN ; Nan XIANG ; Jian YANG ; Ning ZENG ; Deshuai KONG ; Huaizhi WANG
Chinese Journal of Digestive Surgery 2012;11(4):366-370
Pancreatic cancer is malignant with a poor prognosis,and its incidence is rising worldwide in recent years.Multiple slices spiral computed tomography and computed angiography are the first choice for the diagnosis of pancreatic cancer,while misdiagnosis of pancreatic cancer still exists.From August 2009 to October 2011,80 patients with pancreatic or periampullary cancer were diagnosed using the medical image three dimensional visualization system (MI-3DVS).The threedimensional models of the liver,pancreas,vascular system and tumors were reconstructed successfully based on the 64-slice spiral computed tomography data.According to the analysis of the three dimensional models,4 patients underwent standard pancreaticoduodenectomy,1 received palliative surgery.MI3DVS plays an important role in the diagnosis and assessement of resectability of pancreatic and periampullary cancer.
3.Value of serum cholinesterase in diagnosis/treatment and prognostic evaluation of liver diseases
Journal of Clinical Hepatology 2017;33(9):1806-1809
Serum cholinesterase (ChE) is synthesized by hepatic parenchymal cells and is released into blood immediately after synthesis,and therefore,serum ChE can be used as an objective and sensitive indicator of liver function.Serum ChE has been used as an independent factor for the evaluation of liver function for a long time and can reflect the synthetic function of the liver and help to determine the severity of liver diseases.Combined measurement of serum ChE and serum prealbumin,total cholesterol,total bile acid,and prothrombin time has an important value in the diagnosis and treatment of liver diseases.At the same time,serum ChE combined with Child-Turcotte-Pugh score or MELD score can accurately evaluate liver reserve function,which may help with the prognostic evaluation of patients with liver diseases.
4.Preparation and Identification of A Peptide Monoclonal Antibody Against Human Papillomavirus 1 8E6
Qianqian KONG ; Zhenhua TANG ; Fenfen XIANG ; Yueping ZHAN ; Jian XU ; Rong WU ; Xiangdong KANG
Journal of Modern Laboratory Medicine 2016;31(3):30-32,35
Objective To prepare a peptide monoclonal antibody (McAb)against human papillomavirus 18E6 separately,and identify its specificity and pathogenicity.Metheds The advantage epitope peptide was designed and synthesized by ABCpred and Bcepred,and then used to immunize BALB/c mice after coupling with bovine serum albumin (BSA).And the McAb was prepared by hybridoma technique.HPV18E6 gene was amplified from cervical swab specimen containing HPV18 and insert-ed into expression vector pET-28a.The constructed recombinant plasmid was transformed to E.coli BL21(DE3)for expres-sion under induction of isopropyl thio-β-D-galactoside.The expressed protein was used to identified the McAb had been pre-pared.Results The hybridoma cell lines could constantly produce MAbs against HPV18E6 peptides.Sequencing proved that recombinant plasmid pET-28a-HPV18E6 was constructed correctly.Western blotting showed that the anti-HPV18E6 pep-tides antibody could specifically recognize HPV18E6.Conclusion A monoclonal antibody against the advantage epitope pep-tide of human papillomavirus 18E6 prepared could specifically recognize HPV18E6 specifically.
5.Probiotics’ Preventive Effect on Pediatric Food Allergy:A Meta-analysis of Randomized Controlled Trials
Kong XIANG-YI ; Yang YI ; Guan JIAN ; Wang REN-ZHI
Chinese Medical Sciences Journal 2014;(3):144-147
Objective To investigate the preventive effect of probiotics on pediatric food allergy.
Methods From MEDLINE bibliographical database, we searched and reviewed all randomized controlled trials on the preventive effects of probiotics on pediatric food allergies up to September 2013 and excluded the studies that do not meet inclusion criteria and extracted the data. Meta-analysis for the results of homogenous studies was performed using RevMan 5.0 and the co-effect was pooled by using fixed-effects model of relative risk (RR) ratios.
Results Ten trials published between 2007 and 2012 including 2701 cases were included. Meta-analysis based on included data showed that the preventive effect of prenatal and postnatal probiotic supplementation on food allergies was not significant with the RR=0.88 (95%CI:0.76-1.03).
Conclusion Present evidences cannot show in unequivocal terms that prenatal and postnatal probiotic supplementation will prevent food allergic diseases.
6.Investigation of the hope level and its influencing factors in patients with chronic obstructive pulmonary disease
Lian-Shan TAO ; Jian-Xiang KONG ; Ding-Wei SU ; Cai-Hong KONG ; Gui-Mei WU
Chinese Journal of Modern Nursing 2011;17(31):3732-3735
Objective To investigate the hope level and its influencing factors in chronic obstructive pulmonary disease(COPD)patients.Methods Questionnaires were randomly applied to investigate the hope level and its influencing factors of 76 patients with chronic obstructive pulmonary disease with acute exacerbation (AECOPD).Results The hope level scores were in the range of 21-45 interval,and the mean score was (34.92 ±4.89).97.37% of AECOPD patients were in mid-high level.Univariate analyses showed that family relationship,monthly salary,activities of daily living(ADL),disease severity were influential factors(P<0.05).HHI of AECOPD with different respiratory training was different(F =3.76,P < 0.05),and the score of those never exercise was(34.29 ±5.12),(37.08 ± 1.00)in those exercise more often,and(41.50 ±3.54)of those exercise every day.HHI of patients with different oxygen-intake time was different(P < 0.01).Multivariate multiple linear regression analysis showed that monthly salary and ADL were influential factors,and the more salary,and the more hope-level.ADL,oxygenintake time month salary,and disease severity were related to hope-level of patients by multivariate analyses in oxygenintake patients(P < 0.01).Conclusions For nursing care of the COPD patients,attention should be paid on improving the hope level,interfering in the correlated influencing factor as soon as possible so as to continuously improve the quality of life.
7.Rapid determination of thiabendazole residue in barbary wolfberry fruit by ion exchange chromatography and fluorescence detection.
Jie ZHAO ; Qiang HE ; Xiang-hong KONG ; Jian-hua LI
China Journal of Chinese Materia Medica 2008;33(15):1853-1855
OBJECTIVETo developed a method for the determination of thiabendazole residues in barbary wolfberry fruit by ion exchange chromatography (IEC) with fluorescence detection.
METHODThe samples were extracted with 0.1% H3PO4. Chromatographic analysis was performed on a Supelcosil LC-SCX (4.6 mm x 250 mm, 5 microm) column eluted with 0.1 mol x L(-1) KH2PO4 (pH 3.0)-acetonitrile (70:30) and detection at lamdaex = 307 nm and lamdaem = 359 nm.
RESULTThiabendazole in measured samples was separated completely. The calibration curve was linear at the range of 0.0005-0.02 mg x L(-1) with good precision and accuracy.
CONCLUSIONThe proposed method was satisfactorily applied to the analysis of thiabendazole residues in barbary wolfberry fruit.
Chromatography, Ion Exchange ; methods ; Fruit ; chemistry ; Lycium ; chemistry ; Reproducibility of Results ; Spectrometry, Fluorescence ; Thiabendazole ; analysis ; chemistry
8.Discussing the safety application of Chinese materia medics in patients with cardiovascular diseases.
Jian ZHANG ; Min-Zhou ZHANG ; Xiang-Zhao KONG
Chinese Journal of Integrated Traditional and Western Medicine 2012;32(7):994-998
Along with the development of Chinese medicine and pharmacy (CMP), Chinese materia medica (CMM) has been used more and more widely at home and abroad. But we have to confront worsening problems such as lack of safety evidence, immature self -formulation technologies, lack of knowledge about their toxicities, and public misunderstanding, especially for patients with cardiovascular disease (CVD). Therefore, we cardiovascular physicians are requested improve knowledge for CMM preparations and their effects and side effects, supervise and identify the interactions between CMM and Westem medicine. Meanwhile, the researchers are also requested to assess the safety and efficacy of CMM through rigorous experimental designs, further improve the quality, safety, and efficacy of CMM, strictly formulate the specification of CMM products, guide the rational use of CMM by clinicians and the general public.
Cardiovascular Diseases
;
Drugs, Chinese Herbal
;
adverse effects
;
therapeutic use
;
Humans
;
Phytotherapy
;
adverse effects
;
Safety
9.Analysis of coronary arteriographic and fluorescence fundus angiographic characteristics in type 2 diabetic patients with coronary heart disease
Jian-Li GONG ; Wen-Hua LI ; Xiang-Yun KONG ; Xia TANG ; Xiu-Zhou ZHANG ; Yuan-Sheng TANG ; Xing-Lei ZHU
Chinese Journal of Endocrinology and Metabolism 2000;0(06):-
Objective To study the relationship between coronary arteriographic and fluorescence fundus angiographic characteristics in type 2 diabetic patients with coronary heart disease.Methods The study was carried out by the analysis of the data from coronary arteriography and fluorescence fundus angiography in 203 type 2 diabetic patients with coronary heart disease in different groups divided according to age or total cholesterol level. Logisitic regression analysis was applied to explore various risk factors to angiographic characteristics.Results With advancing age,there were more involvement of 3 coronary vessels or the left main branch along with stageⅢretinopathy,but less single vessel diseases in the coronary arteries and less stageⅠretinopathy.The difference in coronary angiographic and fluorescence fundus angiographic characteristics between groups with different total cholesterol levels was not significant.Logistic regression analysis suggested that coronary artery diaease was related to age,sex and blood glucose and triglyceride levels while diabetic retinopathy was related to blood glucose level and age.Conclusion There is great difference in coronary arteriography and fluorescence fundus angiography among different age groups.Aging may aggravate the lesions both in the coronary arteries and fundal vessels in type 2 diabetic patients with coronary heart diseease.
10.Autologous grafts of double-strut fibular cortical bone plate to treat the fractures and defects of distal femur: a case report and review of literature.
Xu CHEN ; Jian-Jun LI ; Zhan KONG ; Dong-Xiang YANG ; Xiang-Nan YUAN
Chinese Journal of Traumatology 2011;14(4):241-246
We reported a 23-year-old man who was involved in a high-speed motorcycle accident. He sustained a closed fracture at the right distal femur. The primary fracture happened on February 2008. He underwent open reduction and internal fixation with cloverleaf plate. And one hundred days after the surgery, the proximal screws were pulled-out, but the bone union was not achieved. Treatment consisted of exchanging the cloverleaf plate with a locking compression plate and using an auto-iliac bone graft to fill the nonunion gap. In July 2009, the patient had a sharp pain in the right lower limb. The X-ray revealed that the plate implanted last year was broken, causing a nonunion at the fracture site. Immediately the plate and screws were removed and an intramedullary nail was inserted reversely from the distal femur as well as a 7 cm long bone from the right fibula was extracted and longitudinally split into two pieces to construct cortical bone plates. Then we placed them laterally and medially to fracture site, drilled two holes respectively, and fastened them with suture. We carried on auto-iliac bone grafting with the nonunion bone grafts. The follow-up at 15 months after operation showed that the treatment was successful, X-ray confirmed that there was no rotation and no angular or short deformity. We briefly reviewed the literature regarding such an unusual presentation and discussed in details the possible etiology and the advantages of autologous double-strut fibular grafts to cope with such an intractable situation.
Bone Plates
;
Femur
;
Fibula
;
Fracture Fixation, Internal
;
Fractures, Bone
;
Humans