3.High level expression of PNGase F in Escherichia coli and its bioactivities.
Yi-Shan SU ; Sheng-Jun WANG ; Peng WANG ; Qing-Sheng QI
Chinese Journal of Biotechnology 2005;21(6):911-915
In order to obtain active recombinant PNGase F in Escherichia coli, a prokaryotic expression vector pET28a/PNGase F was constructed. Amplification of PNGase F was obtained using PCR technique employing suitable primers designed according to the PNGase F gene sequence from Flavobacterium nmeningosepticum. The expression of PNGase F gene in LB medium or M9 medium led to the formation of inclusion body and soluble protein, respectively. The refolding of the denatured inclusion body was successful by gradual dilution. Further purification of the refolded protein and soluble crude extract from M9 medium with Ni2+ -NTA argarose resulted a 90% purified PNGase F. The purified protein catalyzed the complete and intact cleavage of N-linked oligosaccharides from various glycoproteins. The efficiency of this cleavage was affected by the substrate status in the reaction system. In summary, we have developed an enzyme production system where PNGase F was over-expressed in recombinant E. coli. This system can provide more than 15 mg/L purified active PNGase F. This purified active PNGase F can be used as tools in analyzing the oligosaccharide structure.
Bacterial Proteins
;
genetics
;
metabolism
;
Escherichia coli
;
genetics
;
metabolism
;
Flavobacterium
;
enzymology
;
genetics
;
Glycosylation
;
Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase
;
genetics
;
metabolism
;
Recombinant Fusion Proteins
;
genetics
;
metabolism
4.Analysis of influencing factors of multi-site work-related musculoskeletal disorders in surgeons
HUANG Kai sheng HUANG Dang sheng WU Yun PENG Zhi heng SHI Yi ming WANG Zhong xu YANG Min
China Occupational Medicine 2022;49(05):493-
Objective - -
To analyze the prevalence and influencing factors of multi site work related musculoskeletal disorders
( ) Methods
WMSDs in surgeons. A total of 102 surgeons from four hospitals were selected as study subjects by convenient
sampling method. The Chinese version of Musculoskeletal Disorders Questionnaire was used to investigate the prevalence of
, Results
WMSDs in the past one year the related individuals and occupational factors. The total prevalence of WMSDs among
( ), ( ) ( )
surgeons was 54.9%. The top three sites were neck 48.0% lower back 35.3% and shoulder 32.4% . The prevalence of
( vs ,P )
WMSDs in multiple sites was higher than that in a single site 43.1% 11.8% <0.01 . Multivariate logistic regression
, ,
analysis showed that surgeons who smoked were tired at work and had a bent back had a higher risk of developing WMSDs
[ ( - ), ( - ), ( - ), P ]
odds ratios and 95% confidence intervals were 3.66 1.41 9.46 8.33 2.15 32.20 and 18.74 2.14 166.77 all <0.01
Conclusion -
after excluding the influence of confounding factors. The prevalence rate of multi site WMSDs among surgeons is
,
high and the influencing factors include bad living habits and occupational factors such as working load and working posture.
5.Application on small incision extracapsular cataract extraction in large - scale vision recovery action in Shaanxi Province
Juan, ZHANG ; Wen-Jun, ZHAO ; Yong-Kang, ZHANG ; Yi, GUO ; Jing, LI ; Yang-Sheng, PENG
International Eye Science 2014;(9):1659-1662
To investigate the characteristics of scale cataract operations and the effects and experiences of small incision extracapsular cataract extraction with intraocular lens ( lOL) implantation in large - scale vision recovery action.
●METHODS: Four thousand eight hundred ninety - two cases ( 4892 eyes ) of cataract were treated by small incision non-phacoemulcification cataract extraction from March 2010 to November 2011 in our hospital ( Fuming No. 1 surgery car of Shaanxi Province ) which were retrospectively analyzed. Visual acuity, intraoperative and postoperative complications, the recovery of postoperative inflammation were observed.
●RESULTS: Visual acuity reached 0. 3 or more in 4521 eyes (92. 42%) at 1d after the operation, at 3d after the operation in 4571 eyes (93. 44%), there were 4887 eyes with lOL implantation, implantation rate was 99. 90%. All the cases had lesser intraoperative and postoperative complications, and the postoperative inflammation recovered quickly.
●CONCLUSlON: Small incision extracapsular cataract extraction with lOL implantation is simple, effective, economical, safe and adapting for large - scale vision recovery action.
6.Anatomical comparison of morphology and structure of collagen fiber of the hilar bile duct between human and rat
Wengang LI ; Sheng CHEN ; Lei QU ; Jinjian XIANG ; Bin LI ; Yi JIANG ; Zhihai PENG ; Zhiqiang HUANG
Chinese Journal of Digestive Surgery 2008;7(4):303-306
Objective To explore the differences in morphology and structure of collagen fiber of the hilar bile duct between human and rat. Methods The morphology and structure of vertical and horizontal cross-section of human and rat collagen fiber of the normal and dilated hilar bile duct, and their changes under stress were observed after Masson trichrome staining. Results The morphology and structure of collagen fiber of hilar bile duet in human was similar to that in rat. The collagen fiber mainly distributed in the middle and outer layer of the hilar bile duct. The wave-like collagen fiber bundles were arranged in parallel, consistent with the longitudinal axis direction of the bile duct, and connected by the small branches. Conclusions The morphology and structure of collagen fiber of the hilar bile duct in human is similar to that in rat. The anatomical structure of the collagen fiber is adapted to its function.
7.Mechanism of genuineness of Glycyrrhiza uralensis based on SNP of β-Amyrin synthase gene.
Yi-mei ZANG ; Yan-peng LI ; Jing QIAO ; Hong-hao CHEN ; Chun-sheng LIU
Acta Pharmaceutica Sinica 2015;50(7):906-909
β-Amyrin synthase (β-AS) genes of Glycyrrhiza uralensis from 6 different regions were analyzed by PCR-SSCP and sequenced, then the correlationship between β-AS SNP and regions of Glycyrrhiza uralensis were determined. According to the 1 coding single nucleotide polymorphism on the first exon of β-AS gene at 94 bp site, Glycyrrhiza uralensis could be divided into 3 genotypes. In these genotypes, the percentage of 94A type in genuine regions was much higher, and it had significant differences with the percentage in non-genuine regions (P < 0.001). The results of the experiment proved that different β-AS genotypes at 94 bp site from different regions may be one of the important reasons to result in the genuineness of Glycyrrhiza uralensis.
Exons
;
Genotype
;
Glycyrrhiza uralensis
;
classification
;
enzymology
;
genetics
;
Intramolecular Transferases
;
genetics
;
Plant Proteins
;
genetics
;
Polymorphism, Single Nucleotide
;
Polymorphism, Single-Stranded Conformational
8.Characteristics of Electrocardiography in Pressure Overload-induced Cardiac Hypertrophy Rats
Yao-Sheng WANG ; Li-Peng HE ; Yi-Hua ZHOU ; Lu-Min XIAO ; Xiao-Shu CHENG ;
Chinese Journal of Hypertension 2006;0(10):-
Objective To analyze the electrocardiography(ECG)data of pressure overload-induced cardiac hy pertrophy rats.Methods Pressure overload cardiac hypertrophy was induced by abdominal aorta constriction. Echocardiogram and heart weight measurement demonstrated the occurrence of cardiac hypertrophy.Standardized ECG parameters of limb and chest were measured and statistically analyzed.Results Two weeks after hypertrophy models were established,echocardiogram showed greater LVPWTd,IVSTd,LVDd.ECG showed that left axis deviation and higher R waves in V_A,V_B,V_C(P
9.Expression of CD147,cyclophilin A and cyclophilin B in psoriatic lesions
Xiang CHEN ; Juan LI ; Juan SU ; Yi-Xi WU ; Jing CHANG ; Sheng PENG ; Ye-Hong KUANG
Chinese Journal of Dermatology 2003;0(10):-
Objective To study the expression and pathologic significance of CD147,cyclophilin A (CyPA)and cyclophilin B(CyPB)in psoriatic lesions.Methods Immunohistochemical method was ap- plied to detect the expression of CD147,CyPA and CyPB in skin specimens of 15 patients with psoriasis pustulosa(PP),20 patients with progressive psoriasis vulgaris(PPV),and 20 patients with inactive psori- asis vulgaris(IPV).Immunoreactivity intensity distribution index(IRIDI)was calculated to assess the expres- sion intensity of CD147,CyPA and CyPB.Results CD147,CyPA and CyPB were detected in all speci- mens.The IRIDI scores of CD147 and CyPA were significantly higher in keratinocytes of psoriatic lesions than in those of the control specimens(all P0.05).The IRIDI score of CyPB in T lymphocytes of PP lesions was significantly elevated than that in PPV lesions,which was in turn higher than that in IPV lesions(all P0.05).Conclusion CD147,CyPA and CyPB may play a role in the occurrence and development of psoriasis.
10.Hepatic Resection Combined with Radiofrequency Ablation versus Hepatic Resection Alone for Multifocal Hepatocellular Carcinomas: A Meta-analysis
XU LIANG-LIANG ; ZHANG MING ; YI PENG-SHENG ; ZHENG XIAO-BO ; FENG LEI ; LAN CHUAN ; TANG JIAN-WEI ; REN SHENG-SHENG ; XU MING-QING
Journal of Huazhong University of Science and Technology (Medical Sciences) 2017;37(6):974-980
This meta-analysis aimed to comprehensively assess the efficacy and safety of hepatic resection combined with radiofrequency ablation versus hepatic resection (HR) alone for the treatment of multifocal hepatoeellular carcinomas (HCC).A literature search was conducted from the database including MEDLINE,Embase,Cochrane Central Register of Controlled Trials (CENTRAL) and China Biology Medicine (CBM) disc.The primary outcomes included the 1-,3-,5-year overall survival (OS) and disease-free survival (DFS) rate.The secondary outcomes contained the intraoperative parameters and postoperative adverse events (AEs).These parameters were all analyzed by RevMan 5.3 software.After carefully screening relevant studies,four retrospective studies of high quality involving 466 patients (197 in the combined group and 269 in the HR group) were included in this study.The pooled results showed that the 1-,3-,5-year OS rate in the combined group were comparable with those in the HR group (OR=0.77,0.96,0.88;P=0.33,0.88,0.70,respectively).Similarly,there was no significant difference in 1-,3-,5-year DFS rate between the combined group and the HR alone group (OR=0.57,0.83,0.72;P=0.17,0.37,0.32,respectively).And the intraoperative parameters and postoperative AEs were also comparable between the above two cohorts.However,two included studies reported that tumor often recurred in the ablation site in the combined group.The present meta-analysis indicated that the HR combined with RFA could reach a long-term survival outcome similar to curative HR for multifocal HCC patients.And this therapy may be a promising alternative for these patients with marginal liver function or complicated tumor distribution.Furthermore,high quality randomized controlled trials (RCTs) are imperative to verify this conclusion.