1.Correlation between peak intensity of contrast enhanced ultrasound and injury severity after liver ischemia reperfusion
Hong LI ; Ying CHENG ; Lei YANG ; Yunfei ZHANG
Chinese Journal of Organ Transplantation 2017;38(4):234-238
Objective To discuss the correlation between peak intensity (PI) of contrast enhanced ultrasound and injury severity after liver ischemia reperfusion.Methods Pringle's maneuver was used to establish the liver 15-min ischemia model.Contrast enhanced ultrasound was used to observe the hepatic perfusion after reperfusion of 1 h,6 h and 24 h,and the PI was recorded.The correlation analysis on PI with alanine aminotransferase (ALT),aspartate aminotransferase (AST) and pathologic changes (Suzuki standard) was done.Results PI decreased after 1 h,6 h,and 24 h of liver reperfusion,most significantly at 6 h.ALT reached the peak after 1 h of reperfusion and then decreased after 6 h and 24 h.AST increased after 6 h of reperfusion and decreased after 24 h.After 1 h of reperfusion,mild injury appeared in pathology and the injury became severe after 6 h,then it got moderate-to-severe after 24 h of reperfusion.PI was negatively correlated to ALT,AST and pathologic injury degree.Conclusion The PI relates to the degree of hepatic injury after liver ischemia/reperfusion.The more severe the injury is,the lower the PI.
2.The efficiency of expressing human neprilysin by using lentiviral vector transduction in neural stem cells
Wen HUANG ; Xuean MO ; Chao QIN ; Jinou ZHENG ; Zhijian LIANG ; Daobin CHENG ; Yunfei WEI
Chinese Journal of Neurology 2013;(1):17-21
Objective To study the transduction efficiency of expressing human neprilysin by using a lentiviral vector (Lenti-NEP) in mouse embryonic neural stem cells (NSC) in vitro.Methods Primary NSC were harvested from C57BL/6J pregnant mouse at embryonic day 11.5 and transducted with LentiNEP.Immunofluorescent stainingand Western blot were performed to detect NEP protein expression in NSC.Degradation of amyloid beta 1-40 (Aβ1-40) by NEP protein transduced with Lenti-NEP in NSC was analyzed using ELISA and HPLC.Results Over 90% NSC were successfully transduced with Lenti-NEP via observation of fused protein green fluorescent protein under the microscopy.Expressions of NEP transduced with Lenti-NEP in NSC and of the markers of NSC (nestin) and neuron (MAP2).The enzyme activity of 2.5 μg (21.00 ± 2.51) and 1.0 μg (15.00 ± 0.54) NEP on degrading Aβ1-40 was shown to improve significantly compared to 0.5 μg NEP(8.00 ±0.81,t =40.4 and 12.7,respectively,both P <0.01).The activity of NEP was inhibited in the presence of 50 μmol/L phosphoramidon (0.5 pg:0.08 ±0.01 ;1.0 μg:0.04 ±0.01 ;2.5 μg:0.05 ±0.01,t =17.2,51.3 and 14.1,respectively,all P <0.01).The hydrolytic cleavage on degrading Aβ1-40 by NEP was 11.4%,28.4% and 93.7% with incubation for 1 h,4 h and 12 h,respectively.Conclusions Lentiviral vector successfully delivers NEP gene to NSC in vitro.Targeting on NEP and NSC may provide potential therapeutic tool for Alzheimer' s disease.
3.Treatment of reoperation on blood vessel prosthesis occlusion after arterial bypass graft in lower limbs
Yunfei JIANG ; Yongbin ZHU ; Yunfeng ZHU ; Yuanbing WU ; Hongwei GE ; Cheng CHEN
Chinese Journal of Postgraduates of Medicine 2011;34(20):18-20
Objective To review reoperation on blood vessel prosthesis occlusion after arterial bypass graft in lower limbs. Method The treatment effect of 21 patients with reoperation on blood vessel prothesis occlusion after arterial bypass graft in lower limbs was analyzed retrospectively. Results All the cases were followed up 6-36 (12 ±3) months. The limb salvage rate was 71.4%(15/21) ,the amputation rate was 28.6% (6/21). All 9 limbs that underwent revascularization from deep femoral artery reserved. Conclusions Endomembrane hyperplasy, occlusion of the inflow and outflow tracts are the major reasons for the occlusion of blood vessel prosthesis after arterial bypass graft in lower limbs. Appropriate procedures should be based on careful consideration of the occlusion reasons. Profundaplasty is an effective therapy for those who are treated by reoperation on blood vessel prosthesis occlusion in lower limbs.
4.Efficacy analysis after surgical treatment for gastric cancer with synchronous hepatic metastases
Cheng ZHANG ; Peng ZHANG ; Lei CHEN ; Yi LIU ; Shanglei NING ; Yunfei XU ; Zhaochen LIU ; Yuxin CHEN
Chinese Journal of Current Advances in General Surgery 2017;20(2):96-99
Objective:To clarify the indications and efficacy after simultaneous surgical treatment in gastric cancer patients with synchronous hepatic metastases (GCHM).Methods:A total of 21 GCHM patients who underwent D2 gastrectomy with R0 hepatic resection (11 cases) or non-R0 hepatic resection (10 cases) from March 2004 to April 2016 were analyzed retrospectively.The clinicopathological characteristics and survival were compared between the two groups.Results:Cumulative survival rate was improved in R0 hepatic resection group compared with non-R0 hepatic resection group:1-year (54.5% vs 48.0%),2-year(27.3% vs 0%),and 5-year(27.3% vs 0%,P=0.044).The median survival time in R0 hepatic resection group and non-R0 hepatic resection group were 16.2 and 5.9 months (P=0.008).Univariate analysis revealed that Bormann's classification (P=0.010) and state of regional lymph node (P=0.004) were significant predictive factors regarding cumulative survival rate.However,there was no significant prognostic factor (P=0.031) in multivariate analysis might partly owing to interaction among them and/or a small number of patients.Conclusion:Multidisciplinary treatments based on R0 hepatic resection combined with D2 gastrectomy could improve survival in selected GCHM patients.
5.Endovascular treatment of iliac vein compression syndrome:analysis of 33 cases
Yuanbing WU ; Yunfeng ZHU ; Hongwei GE ; Yongbin ZHU ; Yunfei JIANG ; Cheng CHEN ; Yiming HUANG
Chinese Journal of Postgraduates of Medicine 2014;37(32):47-49
Objective To evaluate the clinical effect of the endovascular treatment of iliac vein compression syndrome.Methods Thirty-three patients with iliac vein compression syndrome were treated with endovascular treatment.Of which,edema and varicose vein of the left lower extremity in 29 patients,complicated with acute deep vein thrombosis of left lower extremity in 3 patients,post deep venous thrombosis syndrome in 1 patient.Balloon dilatation and stent implantation were performed in all 33 patients.The diameter of balloon was 10-12 mm,diameter 12-14 mm Bard self expandable stent.Five patients with varicose vein and ulcer of left lower extremity were treated with two stage operation.Results The diagnosis was confirmed by left lower extremity deep veins angiography.There was no death patient,and no hematoma of hematoma locus.Follow-up for 3-30 months,the rate of follow-up was 100%(33/33).The edema of the lower extremity was markedly reduced or disappeared in 28 patients.Color Doppler ultrasound and left lower extremity angiography showed that the stent was unobstructed,no stent occlusion and new onset thrombosis cases.Conclusion Endovascular treatment is safe,effective with few complications,and is the first choice for the treatment of iliac vein compression syndrome.
6.Cloning and induced expression analysis of 4-hydroxy-3-methyl-but-2-enyl diphosphate reductase gene (smHDR) of Salvia miltiorrhiza.
Qiqing CHENG ; Yunfei HE ; Geng LI ; Chao JIANG ; Yuan YUAN ; Wei GAO ; Luqi HUANG
Acta Pharmaceutica Sinica 2013;48(2):236-42
This study reported the obtainment of the full-length cDNA of Salvia miltiorrhiza hairy roots (Abbr: SmHDR, GenBank number: JX233817), via extracting Salvia miltiorrhiza hairy roots total RNA, designing specific primers according to the transcriptome data and using the RACE strategy, and then analyzed it with bioinformatics approaches. On this basis, using the real-time PCR to detect SmHDR gene expression after Ag+ induction, and testing tanshinones contents of corresponding samples by UPLC. SmHDR has 1 647 nucleotides, and an open reading frame (ORF) encoding a protein of 463 amino acid residues. The deduced protein has isoelectric point (pI) of 5.72 and a calculated molecular weight about 51.88 kD. In the secondary structure, the percentage of alpha helix, beta turn and random coil were 35.64%, 20.30% and 44.06%, respectively. Sequence alignment and phylogenetic analysis demonstrated that SmHDR had relative close relationship to the HDR of Picrorhiza kurrooa, similar to HDR from other species of plants. Real time PCR results indicated that elicitor of Ag+ stimulated the increase of mRNA expression of SmHDR. At the same time, results of ultra performance liquid chromatography (UPLC), used to examine the accumulation of diterpenoid tanshinones in hairy roots, showed that the contents of diterpenoid tanshinones in hairy roots of Salvia miltiorrhiza were increased dramatically at 12 h after treated with Ag+, and then decreased significantly. This result showed a positive correlation between the levels of mRNA expression and tanshinones accumulation in Salvia miltiorrhiza stimulated by Ag+. The content of tanshinones was gradually raised, and it had an obvious increase at 120 h. The bioinformatics analysis and gene expression indicated that SmHDR might be involved in tanshinones biosynthesis, which laid the foundation for further study of secondary metabolic regulation mechanism of tanshinones.
7.Construction and characterization of host-vector balanced lethal system of attenuated salmonella choleraesuisΔcrpΔcyaΔasdC78-1 (pYA3493)
Ke SHANG ; Junfeng ZHANG ; Xiangchao CHENG ; Chunjie ZHANG ; Yinju LI ; Guihua CHEN ; Yunfei YAN ; Chongkai ZHAI ; Zhanqin ZHAO
Chinese Journal of Immunology 2015;(3):358-363
Objective:In order to develop an oral live vaccine vector of swine that can stably carry exogenous genes.Methods:Mutant ΔcrpΔcyaΔasdC78-1 was constructed by the method of suicide plasmid pREasd-mediated bacteria homologous recombination on the basis of attenuated Salmonella choleraesuisΔcrpΔcyaC78-1.Complementary plasmid pYA3493 with asd was electrotransformed into the mutant,and thenΔcrpΔcyaΔasdC78-1(pYA3493) host-vector balanced lethal system was constructed.Its biological characteristics were analyzed further.Results:The results of PCR and sequencing showed thatΔcrpΔcyaΔasdC78-1(pYA3493) was constructed suc-cessfully.Biological characteristics showed that the serotype of ΔcrpΔcyaΔasdC78-1(pYA3493) was identical to ΔcyaΔasdC78-1 and vaccine strain C500 and it can stably carry theΔasd gene in vitro;its growth speed was a little slower than ΔcrpΔcyaC78-1 strain,but both of their growth speeds were significantly slower than vaccine strain C500;the biochemical characteristics of ΔcrpΔcyaΔasdC78-1 ( pYA3493 ) were basically the same with ΔcrpΔcyaC78-1 strain.Oral virulence test in mice showed that the virulence ofΔcrpΔcyaΔasdC78-1 ( pYA3493 ) was similar with ΔcrpΔcyaC78-1, but its median lethal dose is 412 times of vaccine strain C500.Conclusion:These results demonstrated that attenuated Salmonella choleraesuisΔcrpΔcyaΔasdC78-1(pYA3493) strain had the potential to be used as an oral live vaccine vector for expressing foreign genes efficiently.
8.Study on the Mechanism of Celosia cristata N-butanol Extracts in the Improvement of Dysfunctional Uter-ine Bleeding of Rats
Yuanyuan GU ; Runqin ZHAO ; Lang SHI ; Yunfei ZHANG ; Cheng FENG ; Fang FANG ; Shuxiang ZHANG ; Lianzhi WANG ; Dazhong CHEN
China Pharmacy 2017;28(19):2617-2619
OBJECTIVE:To study the improvement effect of Celosia cristata n-butanol extracts on dysfunctional uterine bleed-ing of rats,and explore its mechanism. METHODS:60 pregnant SD rats were randomly divided into blank group,model group, Gongxuening capsule group (positive control,0.07 g/kg) and C. cristata n-butanol extracts high-dose,medium-dose,low-dose groups(4.32,2.16,1.08 g/kg),10 in each group. Except for the blank group,rats in other groups were intragastrically given mife-pristone and misoprostol on 7th of pregnancy for resulting incomplete abortion to induce models of dysfunctional uterine bleeding. Then rats in administration groups were intragastrically given relevant medicines,rats in blank group and model group were intra-gastrically given normal saline once every morning and evening,for 7 d. On 8th d of pregnancy,uterine bleeding amount,and thromboxane (TXA2),prostacyclin (PGI2) and tumor necrosis factor α(TNF-α) contents in serum were determined. RESULTS:Compared with blank group,uterine bleeding amount in model group was significantly increased(P<0.01),TXA2 content in se-rum was significantly reduced,PGI2 and TNF-α contents were significantly increased(P<0.01). Compared with model group,uter-ine bleeding amounts in administration groups were significantly reduced,TXA2 content in serum was significantly increased(P<0.01);PGI2 and TNF-α contents in serum in Gongxuening capsule group and C. cristata n-butanol extracts high-dose group and TNF-α content in serum in C. cristata n-butanol extracts medium-dose group were significantly reduced (P<0.01). CONCLU-SIONS:C. cristata n-butanol extracts show obvious improvement effect on incomplete drug abortion-induced dysfunctional uterine bleeding of rats,and the mechanism may be related to the regulation of TXA2/PGI2 dynamic balance and inhibition of TNF-α tran-sient secretion.
9.Construction and characterization of invasion protein B gene deleted mutant of Salmonella typhimurium SL1344 strain
Songbiao CHEN ; Jing LI ; Chuan YU ; Lei HE ; Chunjie ZHANG ; Xiangchao CHENG ; Yinju LI ; Yunfei YAN ; Xiuzhe JIN ; Guihua CHEN ; Zhanqin ZHAO
Chinese Journal of Immunology 2015;(2):215-220
Objective:In order to develop a safer vaccine strain exploit Salmonella typhimurium vaccine strain .A ΔsipB mutant of Salmonella typhimurium SL 1344 strain was constructed.Methods: Firstly, the recombinant suicide plasmid containing the missing 585 bp sipB ( PREΔsipB ) was built by homologous recombination , and screenned by two-step method.Results: PCR and sequencing results showed that SL 1344ΔsipB was successfully constructed.It was no significant changes compared with SL 1344.But compared with the parent strains SL 1344 , the mutant strain had obvious change in its virulence , oral challenge of bacteria in mice revealed that LD50 of the mutant strain was 1.70 ×108 CFU,the toxicity reduced about 1.4%.The protection rate induced by the sipB mutant was 50%,and the serum antibody peaked 14 d post-immunization.Conclusion:The SL1344ΔsipB mutant was constructed suc-cessfully,and genetic stability ,significantly reduced virulence.The study provides a new approach for further study of the relationship between the gene and pathogenicity of Salmonella typhimurium.It is likely that the ΔsipB mutant could be adapted to develope attenuated Salmonella vaccine.
10.Reconstruction method of language pathways in the preoperative planning of brain tumor surgery.
Jing YAN ; Junfeng LU ; Jingliang CHENG ; Jinsong WU ; Jie ZHANG ; Chaoyan WANG ; Yunfei NIE ; Beibei PANG ; Xianzhi LIU
Chinese Journal of Surgery 2015;53(5):362-367
OBJECTIVETo propose a clinically practical and simple fiber tracking method for language pathways, and to explore its feasibility in preoperative planning for brain tumors adjacent to the language cortex.
METHODSDiffusion tensor imaging was examined in 18 healthy subjects and 13 patients with brain tumors adjacent to the language cortex between December 2013 and June 2014. The associated fibers of language pathways were reconstructed using a commercial software (Syngo workstation). Firstly, the feasibility of fiber tracking method for language pathways in healthy subjects were studied, and then its application was assessed in patients with brain tumors. The anatomic relationship between tumors and the associated fibers was analyzed.
RESULTSBy selecting appropriate regions of interest, the associated fibers in the dorsal pathways (superior longitudinal fasciculus/arcuate fasciculus, including both direct and indirect pathways) and ventral pathways (uncinate fasciculus, middle longitudinal fasciculus, inferior longitudinal fasciculus and inferiorfronto-occipital fasciculus) were reconstructed in all 18 healthy subjects. In patients with brain tumors, the relationship between the tumors and adjacent associated fibers were divided into two types: adjacent associated fibers could be displaced or separated, and involved the superior longitudinal fasciculus/arcuate fasciculus (n=6), middle longitudinal fasciculus (n=4), uncinate fasciculus (n=3), inferior longitudinal fasciculus (n=3) and inferiorfronto-occipital fasciculus (n=2); alternatively, the adjacent associated fibers were infiltrated or destroyed, and involved the inferiorfronto-occipital fasciculus (n=10), uncinate fasciculus (n=8), middle longitudinal fasciculus (n=5), inferior longitudinal fasciculus (n=4) and superior longitudinal fasciculus/arcuate fasciculus (n=3).
CONCLUSIONSThe associated fibers of language pathways could be visualized rapidly and in real-time by fiber tracking technology based on diffusion tensor imaging. This is feasible for preoperative planning regarding brain tumors adjacent to the language cortex.
Brain Neoplasms ; surgery ; Cerebral Cortex ; surgery ; Diffusion Tensor Imaging ; Humans ; Language ; Neural Pathways ; Postoperative Complications ; prevention & control