Objective ToexploretheclinicalmanifestationsandMRIfeaturesofnervoussystemdiseasecomplicationsinpediatric EV71relatedhand-foot-mouthdisease(HFMD).Methods Theclinicaldataof17EV71relatedHFMDpatientswithnervoussystemdisease wereanalyzedretrospectively.Results 4patientswerediagnosedwith meningitisandthebrain MRIshowedasymmetricunilateral ventricularenlargement.9patientswerediagnosedwithbrainstemencephalitis,amongwhich8patientsshowedthecraniocerebral MRIfeaturesthatthelesionswerelocatedonthedorsalsideofthepontomedullaryjunction,andtheT2imageshowedahighsignal withasymmetrycharacteristic.1caseofbrain MRIdisplayedthatthelesionwaslocatedinthemidbrain,andT2imagewasahigh signalwithsymmetricalfeatures.4patientswerediagnosed withacuteflaccidparalysis.1casehadbrainstem encephalitis,andthe brain MRIshowedthatthelesion waslocatedonthedorsalsideofthepontomedullaryjunction,andthe MRIofthespinalcord showedthelesionintheC1-C7segment.1caseofspinalcordMRIshowedthatthelesionwaslocatedatT2-L1segment,1caseof spinalMRIshowedthelesionlocatedinT8-T12segment;1caseofspinalcord MRIshowedthatthelesionwaslocatedatT9-L1 segment,andsagittalT2 wasahighsignal,andthelesionconcentratedintheanteriorhornofthespinalcordand (or)theanterior rootofthespinalnerve.Byfollow-up,patientswithmeningitishadagoodprognosisandwithoutneurologicalsequelae.Thelesionsof brainstemencephalitiswerebasicallyabsorbedanddisappeared,theprognosiswasgood.Fortheacuteparalysis,theaffectedlimbs recoveredtovaryingdegrees,andthelesionswereabsorbedpartially.Conclusion HFMDpatientsclinicalmanifestationsand MRI characteristicsofconcurrentnervoussystemdiseasewerediversity.Whenthenervoussystemsymptomshappened,weshouldactively performtheMRIexaminationtoidentifytheinflammatoryinvasionarea.Onceinvolvingthebrainstemandspinalcord,highalert, earlyrecognitionandtimelyinterventionarekeytoreducethemorbidityandmortalityofacuteHFMD,toenableHFMDtobeoptimizedand comprehensivelytreated.

Objective To apply the MRI T2 signal intensity of hippocampal subfield in patients with amnestic mild cognitive impairment (aMCI) as early imageology. Methods From October, 2014 to August, 2015, 20 aMCI patients accepted cognitive training (training group), 20 aMCI patients accepted speech communication (speech group), and 20 age-and gender-matched healthy old people (control group) were scanned with MRI using FSE-T2 sequence. The margin of hippocampal subfields were outlined manually for each side to measure the T2 sig-nal intensity. The correlation between hippocampal T2 signal intensity and the scores of Mini-Mental State Examinatlon (MMSE) was ana-lyzed in the training group. Results Before treatment, T2 signal intensity in the bilateral hippocampal head was significantly high in the aM-CI patients compared with that in the control group (P<0.05). After treatment, T2 signal intensity in left hippocampal head decreased in the training group compared with that in the speech group (P<0.05), similar to the control group (P>0.05). There was negative correlation be-tween left hippocampal head's T2 signal intensity and the scores of MMSE in the training group before and after treatment (r=-0.61, r=-0.54, P<0.05). Conclusion The T2 signal intensity in left hippocampal head may respond to the cognitive function in patients with aMCI in the early stage, that could be used for diagnosis and evaluation in clinic.

Objective To investigate the value of the hippocampal subfield's MRI T2 signal intensity in evaluating the effect of the hydrochloric donepezil for mild cognitive impairment(MCI).Methods 20 MCI patients with hydrochloric donepezil (treatment group) and 20 patients with placebo (control group) were scanned by MRI using FSE-T2 sequence.The margin of hippocampal subfields was outlined manually for each side to measure the MRI T2 signal intensity.The relationship between hippocampal MRI T2 signal intensity and MMSE scores was analyzed in the treatment group.Results Before the treatment, there was no significant difference of the MRI T2 signal intensity between groups.After the treatment, the MRI T2 signal intensity in the bilateral head of the treatment group was significantly decreased compared with the control group(P<0.05).There was an inverse relationship between the MRI T2 signal intensity of the bilateral hippocampal head and MMSE scores in the treatment group(P<0.05).Conclusion The MRI T2 signal intensity in the bilateral hippocampal head could be regarded as a valuable marker in making clinical diagnosis and evaluating the effect of the treatment for MCI in its early stage.

Objective To investigate the value of MR T2* in evaluation of liver iron overload caused by long term blood transfusion.Methods Thirty one patients with long term blood transfusion were collected.Both serum ferritin(SF)and CRP levels were measured and the adjusted serum ferritin(ASF)concentration was calculated.MRI of the maximum cross-sectional liver was performed using a 1.5T scanner (Siemens Avanto).The T2* value of the liver was measured,and the correlation analysis was carried out.Twenty one normal volunteers served as controls.The transfusion group was divided into iron overload group and non overload group,and differences between groups were analyzed.Results The correlation coefficient in 31 patients between liver T2* value and SF,ASF and blood transfusion volume was-0.695,-0.728 and -0.657 respectively,and the difference was statistically significant(P<0.05).The correlation coefficient between SF and blood transfusion volume was 0.518,and the difference was statistically significant(P<0.05).There was difference in T 2* value between three groups.Conclusion MR T2* has some practical value in the evaluation of liver iron overload in patients with long term blood transfusion.

Objective To investigate the effects of exercise on knee cartilage tissue structure using quantitative MR T2 mapping. Methods Sagittal T2 maps of the knee joints of 26 healthy volunteers were obtained by using 3.0T MR before,immediately after, and 1 5 min after running.The original images were classified into three terms of knee cartilage T2 map after postreconstruction.The T2 values of regions of interest (ROC)(T2 pre ,T2 post ,T2 delay )in the superficial,middle and deep cartilage of femoral and tibial joint were measured.Statistical differences of cartilage T2 values of three terms after running were analyzed.Results For the tibial joint cartilage,the T2 pre ,T2 post ,T2 delay were (49.71 ± 1.95)ms,(44.30 ± 2.56)ms,(49.41 ± 1.62)ms in the superficial layer,respectively.The three terms T2 were (42.43 ± 2.23)ms,(39.01 ± 2.37)ms,(41.90±2.28)ms in the middle layer,respectively.The differences were statistically significant(F=55.673,16.759 respectively.P<0.001).While the three terms T2 were (19.39±2.13)ms,(19.20±2.22)ms, (19.49±2.05)ms in the deep layers cartilage,respectively.The differences were not statistically significant(F =0.122,P =0.886).And the differences between T2 pre and T2 post ,T2 post and T2 delay were statistically significant (all P <0.001)in superficial and middle alyers,but there were no significant difference between the T2 pre and T2 delay (P =0.610,0.403,respectively).For the femoral joint cartilage,the T2 pre ,T2 post ,T2 delay were (50.22 ± 1.47)ms,(45.60 ± 2.82)ms,(49.84 ± 1.84)ms in superficial layers,respectively.The three terms T2 were (42.67±2.23)ms,(39.36 ± 1.98)ms,(42.40 ± 2.57)ms in the middle layer,respectively.The differences were statistically significant (F=37.976,16.987 respectively,P<0.001).While the three terms T2 were (20.30±2.73)ms,(20.60±2.44)ms,(20.51± 2.24)ms in the deep layer,the differences were not statistically significant (F =0.098,P =0.907).And the differences between T2 pre and T2 post ,T2 post and T2 delay were statistically significant (all P <0.001)in superficial and middle layers,but there were no significant difference between the T2 pre and T2 delay (P=0.520,0.679,respectively). Spatial distribution of T2 values of articular cartilage from deep to superficial layers showed a ascending trend.T2 maps showed the spatial distribution trend of T2 value change.Conclusion T2 mapping can monitor quantitatively the changes of articular cartilage molecular structure after running.The change of articular cartilage T2 value after exercise is uneven and the change of articular cartilage structure after exercise is reversible.

Objective To analyze the MRI findings of Wallerian degeneration in pyramidal tract after cere-bral injury for improving the diagnosis accuracy.Methods The MRI findings of Wallerian degeneration in pyramidal tract in 25 cases were analyzed in association with the primary cerebral injury.Results All the primary diseases were above the tentorium of cerebellum in 25 cases,MRI in 2 cases demonstrated a continuous thin band -like long T1 or isometric T1 and long T2 abnormal changes in pyramidal tract,hyperintensify on FLAIR and DWI.2 cases showed hypointensify on T1 WI,hyperintensify on T2 WI and FLAIR,isointensity on DWI.21 cases showed hypointensify on T1 WI,hyperintensify on T2 WI and FLAIR,isointensity or hypointensify on DWI with brainstem atrophy.Conclusion MRI is valuable in the diagnosis of Wallerian degeneration in pyramidal tract after cerebral injury.

Objective To investigate the feasibility of hydrogen proton magnetic resonance spectroscopy (1 H-MRS)for detection of Wallerian degeneration of the pyramidal tract after cerebral infarction.Methods Multiple metabolic indices including NAA,Cho, Cr of the bilateral cerebral peduncle were detected by using 1 H-MRS in 1 5 patients with unilateral middle cerebral artery infarction and 1 5 age-matched healthy volunteers,NAA/Cr,Cho/Cr were also calculated.Comparing the difference between cerebral infarction group and the control group ,the ROC curve was analyzed.Results The values of NAA/Cr in the ipsilateral cerebral peduncle of cerebral infarction patients were significantly lower than that of the contralateral,and there were significant differences between the two groups(P <0.05).The values of NAA/Cr in the ipsilateral cerebral peduncle of cerebral infarction patients decreased (P <0.05),as compared with that of the control group.The area under the ROC curve was 0.947,the truncation point was 1.63.The NAA/Cr val-ues of the contralateral cerebral peduncle of cerebral infarction patients had no significant differences compared with the control group (P >0.05),the Cho/Cr values of the ipsilateral and the contralateral cerebral peduncle had no significant difference compared with the control group (P >0.05).Conclusion 1 H-MRS can detect Wallerian degeneration of the pyramidal tract after cerebral infarction.

Objective To improve the recognition and reduce the misdiagnosis of bipartite patella by analyzing and summarizing the imaging features.Methods X-ray,CT and MR images of 1 7 cases of bipartite patella were collected,imaging features were ana-lyzed and summarized.Results X-ray revealed the smooth interface of the main and accessory patella fragments with sclerotic mar-gins,the cleft was slightly wide but not sharp.CT showed the bone sclerosis of interface was mostly smooth,rough and vesicular interface could also be found in a few patients,low density soft tissue could be found in separation zone.MRI could detect the intact but thinned cartilage covering the main and accessory patella fragments,fibrous like tissues could be found in separation zone with MRI.Conclusion Imaging findings of bipartite patella are characteristic,these features are helpful in the diagnosis and differential di-agnosis of bipartite patella.

To establish a prokaryotic expression and purification protocol for nuclease P1 (NP1), we first obtained a synthetic NP1 by splicing 22 oligonucleotides with overlapping PCR. We constructed and transformed a secretory expression vector pMAL-p4X-NP1 into Escherichia coli host strains T7 Express and Origami B (DE3) separately. Then, the recombinant NP1 was purified by amylose affinity chromatography, and its activity, thermo-stability and metal-ion dependence were investigated systematically. The results indicated that the expressed fusion proteins MBP-NP1 (Maltose binding protein-NP1) existed mainly in soluble form both in host strains T7 Express and Origami B (DE3), but the specific activity of recombinant protein from Origami B(DE3) strain was higher than T7 Express strain (75.48 U/mg : 51.50 U/mg). When the MBP-tag was cleaved by protease Factor Xa, the specific activity both increased up to 258.1 U/mg and 139.2 U/mg. The thermal inactivation experiments demonstrated that the recombinant NP1 was quite stable, and it retained more than 90% of original activity after incubated for 30 min at 80 degrees C. Zn2+ (2.0 mmol/L) could increase enzyme activity (to 119.1%), on the contrary, the enzyme activity was reduced by 2.0 mmol/L Cu2+ (to 63.12%). This research realized the functional expression of NP1 in the prokaryotic system for the first time, and provided an alternative pathway for NP1 preparation.
Cloning, Molecular ; Enzyme Stability ; Escherichia coli ; genetics ; metabolism ; Fungal Proteins ; biosynthesis ; genetics ; metabolism ; Genes, Synthetic ; Genetic Vectors ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; metabolism ; Single-Strand Specific DNA and RNA Endonucleases ; biosynthesis ; genetics ; metabolism

Wheat grain peroxidase 1 (WP1) belonged to class III plant peroxidase with cofactor heme, which not only has antifungal activity, but also influences the processing quality of flour. In order to enhance functional expression of WP1 in prokaryotic system by increasing endogenous heme synthesis, we constructed a recombinant plasmid pACYC-A-L containing hemA and hemL of Esherichia coli. Then, we co-transformed it into host strain T7 Express with secretive expression vector (pMAL-p4x-WP1) or non-secretive expression vector (pET21a-MBP-WP1), respectively. The MBP-WP1 fusion protein was further purified by amylose affinity chromatography and its peroxidase activity was assayed using 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonate) (ABTS) as substrate. At 12 h after induction at 28 degree, the extracellular 5-aminolevulinic acid (5-ALA) production of T7 Express/pACYC-A-L was up to 146.73 mg/L, simultaneously the extracellular porphrins also increased dramatically. The peroxidase activity of functional MBP-WP1 obtained from T7 Express/ (pACYC-A-L + pMAL-p4x-WP1) was 14.6-folds of that purified from T7 Express/ pET21a-MBP-WP1. This study not only successfully enhanced functional expression of wheat peroxidase 1 in Esherichia coli, but also provided beneficial references for other important proteins with cofactor heme.
Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Heme ; biosynthesis ; genetics ; Peroxidases ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transformation, Genetic