1.Effect of overexpressing isocitrate lyase on succinate production in ldh(-1) Corynebacterium glutamicum.
Chao YANG ; Ning HAO ; Ming YAN ; Lu GAO ; Lin XU
Chinese Journal of Biotechnology 2013;29(11):1696-1700
Corynebacterium glutamicum SA001 is a mutant with lactate dehydrogenase (ldhA) deletion. In order to increase metabolic flux from isocitrate to succinate, and to improve the production of succinate under anaerobic conditions,we transducted the gene aceA coding isocitrate lyase (ICL) from Escherichia coli K12 into Corynebacterium glutamicum SA001 (SA001/pXMJ19-aceA). After 12 h aerobic induction by adding 0.8 mmol/L of IPTG, the recombinant strain was transferred to anaerobic fermentation for 16 h. Succinate reached 14.84 g/L, with a productivity of 0.83 g/(L x h). Compared to C. glutamicum SA001, the activity of ICL of the recombinant strain was increased 5.8-fold, and the succinate productivity was increased 48%. Overexpression of isocitrate lyase will increase the metabolic flux of glyoxylate bypass flowing to succinate.
Corynebacterium glutamicum
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genetics
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metabolism
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Escherichia coli
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enzymology
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genetics
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Gene Deletion
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Industrial Microbiology
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Isocitrate Lyase
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biosynthesis
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genetics
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L-Lactate Dehydrogenase
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genetics
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Succinic Acid
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metabolism
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Transduction, Genetic
2.Application of social media in medical field of China
Jun ZHU ; Lin MA ; Chao LU ; Tao SUO ; Yixin XU
Chinese Journal of Medical Library and Information Science 2014;(6):9-12
After the development of different medical social media in China was summarized , the advantages and trend of social media in medical field of China were analyzed , the challenges facing medical social media were pointed out and their countermeasures were proposed .
4.Application of Intra-Operative Cholangiography
Xing-yao, SHI ; Chao-hui, LIU ; Lin-xing, LU
Journal of Shanghai Jiaotong University(Medical Science) 2001;21(2):182-183
Objective To evaluate the application of intra-operative cholangiography in cholecystectomy. Methods A total of 442 patients with biliary calculi underwent cholecystotomy were reviewed. Results Of the 442 cases, 3 were false positive and 1 was false negative radiologically. The accuracy rate was 99.1%. Conclusion The results suggested that the application of intra-operative cholangiography could decrease the incidence of residual calculi and avoid unnessary exploration and trauma of the common bile duct.
5.Behavioral alterations and demyelization of the corpus callosum in the mouse model of MK-801 induced schizophrenia
Yun XIU ; Lei ZHANG ; Xuan QIU ; Lin CHEN ; Wei LU ; Chao PENG ; Guohua CHENG ; Fenglei CHAO ; Yong TANG
Chinese Journal of Nervous and Mental Diseases 2013;(11):641-645
Objective To explore the role of white matter injuries in the schizophrenia induced by the NMDA re-ceptor antagonist. Methods Adult male C57BL/6J mice (8 week old) were equally divided into four groups. One group was sub-chronically treated with saline solution, and the other three groups were intraperitoneally treated with MK-801 at dose of 0.025 mg/mL (M1), 0.050 mg/mL (M2) and 0.100 mg/mL (M3) in a volume 10 ml per kilogram body weight. All ani-mals were tested using Morris water maze at the 9th-15th day and using the Hole Board exploration as well as Rota Rod performance tests on the 16th day. The myelin basic protein (MBP) and the ultrastructure of the myelin sheaths in the cor-pus callosum were then examined using immunohistochemical methods, transmission electron microscope technique and stereological methods. Results The repeated sub-chronic MK-801 treatment did not induce impairment of spatial learning and memory in Morris water maze. The MK-801 treatment at 0.25 mg/kg and 1.00 mg/kg but not at 0.50 mg/kg resulted in less exploration to a new environment. The myelin staining with anti-MBP antibody was less intense in all three schizo-phrenic groups when compared to saline control group (P<0.01). Furthermore, MK-801 treatment caused pathological al-terations of the myelin sheaths including segmental demyelination of myelinated fibers and splitting of myelin sheath lamel- lae in schizophrenic groups. The ratio of the injured myelinated nerve fibers in the corpus callosum of MK-801 treated mice [M3 group, (22.42 ± 4.24)%] was significantly higher when compared to the control mice [(3.84 ± 1.35)%,P<0.01)]. Conclusions The present study demonstrated the white matter damages, mainly low MBP expression and segmental demye-lization in the corpus callosum in the mice sub-chronic treated with MK-801, indicating that the white matter changes might be involved in the schizophrenia induced by NMDA antagonist.
6.Clinical observation of 15° face-changing acetabular cup in total hip replacement for the treatment of developmental dysplasia of hip.
Wei-kun HOU ; Lin LIU ; Chao LU ; Kan PENG ; Zhi YANG ; Ke XU ; Peng XU
China Journal of Orthopaedics and Traumatology 2016;29(6):526-529
OBJECTIVETo explore early clinical effect of acetabular cup in total hip replacement for the treatment of Crowe II developmental dysplasia of hip.
METHODSEighteen patients (18 hips) with Crowe type II developmental dysplasia of hip were treated with total hip replacement from September 2001 to July 2013. Among them,including 13 males and 5 females aged from 42 to 60 years old with an average of 47.6 years old; the courses of diseases ranged from 9 to 22 years with an average of 13.5 years. All the patients had hip joint pain, limb shortening and limited hip function before operation. Harris score of hip joint were used to evaluate recovery of function at 1 day and 12 months after operation. Prosthetic coverage of acetabular cup at 1 week after operation was observed by using radiography.
RESULTSEighteen patients (18 hips) were followed up from 12 to 24 months with an average 17 months. All incisions were healed at stage I. No deep vein thrombosis, hip dislocation, periprosthetic joint infection and prosthesis loosening were occurred. No revision surgery during follow-up period. Prosthetic coverage of acetabular cup was more than 80% at 1 week after operation. Harris score were increased from 42.67 ± 5.06 before operation to 94.79 ± 3.27 at 12 months after operation (t = -45.269, P < 0.001).
CONCLUSIONFor type Crowe II developmental dysplasia of hip patients, 15° face-changing acetabular cups in THR could obtain higher actebular component coverage rate and satisfactory early clinical effects.
Acetabulum ; surgery ; Adult ; Arthroplasty, Replacement, Hip ; instrumentation ; Female ; Follow-Up Studies ; Hip Dislocation, Congenital ; surgery ; Hip Joint ; surgery ; Hip Prosthesis ; Humans ; Male ; Middle Aged
7.Investigation and analysis on the first case of human infected with avian influenza (H5N6) in Yunnan Province,China
Zhipeng MAO ; Yun LIN ; Yang ZHOU ; Xianxiang YOU ; Lu CHAO ; Jun WU
Chinese Journal of Zoonoses 2015;(10):978-981
We analyzed the epidemiological investigation results of the first case of human infected with avian influenza (H5N6) ,and summarized epidemiological characteristics of this case to provide reference to H 5N6 case screening and investiga‐tion in future works .We retrospectively described and analyzed the investigation progress ,specimen detection results of epi‐demic spot and intimate contact people ,and summarize epidemiological characteristics of this case .Result showed that the first case of human infected with H5N6 in Yunnan was a single case ,and did not showed human‐to‐human transmission .This case had contacts with wild bird and had birds’ market activity before diagnoses ;some environment specimen of birds market near patient’s home has detection results of H5N6 positive .This is a cases of local infection and did not show human‐to‐human transmission .From the investigation ,we could deduced that the transmission route is from birds to human or from birds to en‐vironment and then to human .
8.Change on composition of immune cells in spleen of miRNA-126 knockdown mice
Liangyu LEI ; Yan HU ; Mengmeng GUO ; Jia LU ; Wen ZHENG ; Hualin XU ; Chao CHEN ; Lin XU
Chinese Journal of Immunology 2016;32(4):460-464
Objective:To detect the change of composition of immune cells in the spleen of miR-126 knockdown(miR-126KD) mice and preliminarily explore its significance .Methods: The expression level of miR-126 in spleens of miR-126KD mice was deter-mined by Realtime PCR.And the total number of splenocytes was calculated.The pathologic morphology change of the spleen was observed by HE staining.And the changes on proportion of DCs ,macrophages cells ,γδ T cells and NK T cells,CD3+T cells and its subgroup ,as well as CD19+B cells in spleens of miR-126KD mice were analyzed by Flow cytometry and calculated respectively.The level of phosphorylated AKt and NF-κB was analyzed by Western blot assay.Results:Compared with those of WT mice ,the expression level of miR-126 decreased obviously ( P<0.05 ) and the total number of cells increased obviously in spleen of miR-126 KD mice ( P<0.05).Moreover,the pathologic morphology of miR-126KD mice was significantly changed.The proportion and number of NK T cells in the inherent cells were significantly increased ( P<0.05 ) , but the proportion of Mφcells were significantly decreased ( P<0.05 ) . Meanwhile,the proportion and number of CD3+T cells and CD4+T cells in the adaptive immune cells were significantly increased (P<0.05),while the total number of CD19+B cells were significantly decreased (P<0.05).Last,the level of phosphorylation Akt and NF-κB increased obviously in spleen of miRNA-126 knockdown mice.Conclusion: Change obviously on the composition of immune cells subsets in the spleen of miRNA-126 knockdown mice ,which it may be related to the altered level of phosphorylated AKt and NF-κB and provides a preliminary experimental basis for the further exploring the roles of miR -126 in immune response.
9.Dynamic expressions of interleukin-1 receptor associated kinase-4 and interleukin-1 receptor associated kinase-M in rats with endotoxin tolerance and their significance
Wenyuan LI ; Chao YE ; Zhuo LIN ; Qiao HONG ; Keyin WANG ; Mingqin LU ; Yongping CHEN
Chinese Journal of Infectious Diseases 2011;29(8):449-454
Objective To explore the dynamic expressions of interleukin-1 (IL-1)receptor associated kinase (IRAK)-M and IRAK-4 in rats with or without endotoxin tolerance (ETT)in acute liver failure (ALF).Methods Sixty-six male SD rats were divided into three groups:ALF group,ETT group and control group.The rats in ETT group received daily lipopolysaccharide (LPS)intraperitoneal injection for 5 days,while the rats in ALF group received daily injection with same volume of 0.75% NaCl solution.Both ETT group and ALF group received intraperitoneal injection with D-galactosamine (D-GalN)and LPS at 24 hours after the 5th injection of LPS or NaCl solution.At 2,6,12,24 and 48 h after D-GalN and LPS injection,the serum levels of tumor necrosis factor-α (TNF-α)and interleukin-6 (IL-6)were detected by enzyme-linked immunosorbent assay (ELISA).The liver pathologic changes were observed with HE staining by microscope.The mRNA expressions of IRAK-4,IRAK-M and NF-κB (p65)were detected by reverse transcriptase polymerase chain reaction (RT-PCR).The pairwise comparison between groups was done by lease significant difference (LSD)and Dunnet's t test.Results The liver pathologic changes in ETT group were much milder than those of ALF group.In ALF group,IRAK-4 mRNA/β-actin absorbance ratios at 2,6,12,24and 48 h after D-GalN and LPS challenge were 0.711 ±0.074,0.904±0.118,1.012 ±0.098,1.534±0.279 and 1.451±0.290,respectively,while the IRAK-M mRNA/β-actin absorbance ratios were 0.496±0.018,0.516±0.089,0.503±0.023,0.503±0.057 and 0.469±0.142,respectively.In ETT group,the IRAK-4 mRNA/β-actin absorbance ratios at 2,6,12,24 and 48 h after D-GalN and LPS challenge were 0.619±0.083,0.587±0.033,0.623±0.034,0.720±0.044 and 0.654±0.041,respectively,while the IRA'K-M mRNA/β-actin absorbance ratios were 0.929 ± 0.064,1.111±0.138,1.113±0.027,1.891±0.315 and 1.710±0.303,respectively.The IRAK-M mRNA expression level was significantly increased and IRAK-4 mRNA expression level was relatively decreased in ETT group compared to ALF group.The differences were all statistically significant at 2,6,12,24 and 48 h after D-GalN and LPS challenge (F= 17.305,54.921,121.031,67.607,55.279,respectively; F=19.506,43.777,110.823,302.681,202.822,respectively; F=172.003,59.519,987.055,68.463,96.601,respectively; all P<0.05).Conclusion LPS pretreatment may induce ETT in rat model by downregulating the expression of IRAK-4 and upregulating the expression of IRAK-M.
10.Development of microsatellites and genetic diversity analysis of Scutellaria baicalensis Georgi using genomic-SSR markers.
Lin-jie QI ; Ping LONG ; Chao JIANG ; Yuan YUAN ; Lu-qi HUANG
Acta Pharmaceutica Sinica 2015;50(4):500-505
A total of 12 775 SSRs were identified from Scutellaria baicalensis Georgi genomic database, accounting for 2.56% of the total genomic sequences. The result showed that S. baicalensis SSRs were based on 68.32% dinucleotide and 18.63% trinucleotide repeats; CT/GA and TTC/GAA were predominant in the dinucleotide motifs and the trinucleotide motifs respectively. Nine primers were selected to produce highly reproducible SSR bands and were used in studying the genetic diversity of S. baicalensis, 50 individuals from ten populations. 68 SSR polymorphic loci were detected, these loci were polymorphic and displayed 4 to 12 alleles per locus with a mean number of 7; the effect number of alleles was 3. Expected heterozygosities were 0.6 and were far more greater than the average in dicotyledonous plants. PIC (polymorphism information content) was 0.72, Shannon's information index was 1.32, these all proved that S. baicalensis had a high genetic diversity in general. Genetic differentiation among population Gst was 0.131, genetic variation among population accounted for 13.1% and genetic variation within population accounted for 86.9%. The cluster analysis showed that 10 populations S. Baicalensis were classified into 2 groups, but it was not associated with geographical distribution.
Alleles
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Cluster Analysis
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Genetic Variation
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Genomics
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Microsatellite Repeats
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Scutellaria baicalensis
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genetics
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Trinucleotide Repeats