Objective:To study the influence of 5′-untranslated region modification of pPIC9 on expression of LL-37 in Pichia pastoris.Methods:The sequence GGATCCAA was deleted from 5′-UTR of pPIC9 and the modified product was trans- formed into E.coli DH5?to construct a modified eukaryotic vector pPIC9-EDIT.After PCR and sequencing,pPIC9-EDIT was ligated with LL-37 sequence coded by the biased codon of yeast,the product was then transformed into E.coli DH5?to con- struct the recombinant expression vector pPIC9-EDIT-LL-37,the latter was transformed into P.pastoris GS115 by spheroplas- ting and the insert was confirmed by PCR.The bacteriolytic activity to E.coli.DH5?was analyzed to screen the highest ex- pressing strain and to determine the best inducing time and concentration of methanol.The fermentation product was analyzed by Tricine-SDS-PAGE and Western blotting.The antibacterial activities of expression products of pPIC9-LL-37 and pPIC9-ED- IT-LL-37 were compared,and the changes of LL-37 protein expression were determined before and after modification.Results: pPIC9-EDIT and pPIC9-EDIT-LL-37 were successfully constructed.Expression of LL-37 gene was confirmed by PCR in P.pastoris after pPIC9-EDIT-LL-37 transformation.The highest expressing strain was identified;the best inducing time was 72 h and the best concentration of methanol was 0.5%.Tricine-SDS-PAGE and Western blotting analysis showed that the ex- pression product was LL-37.The expression level of LL-37 protein increased by 35 times after modification.Conclusion:Modifi- cation of pPIC9 5′-UTR can obviously improve expression of LL-37 protein in P.pastoris;it is worth to be used in the research of other heterogenous protein.

Objective: : T o study the microanatomy of the facial nerve in the cerebellopontine angle (CPA) a nd the internal auditory meatus (IAM) for the preservation of the facial nerve i n the acoustic neurinomas surgery. Methods: Forty sides of CPA f acial nerve of 20 adult cadaver heads were examined. Results: In the pontomedullary sulcus the facial nerve (1.98? 0.10) mm anterior to the vestibulocochlear nerve,(8.76?1.42) mm lateral to the abducent nerve at the points where the nerves join the brain stem at the lateral end of the sulcu s, (8.15?2.18) mm above the junction of the Ⅸ nerve with the medulla. There were 62 nutritious artery supplying the facial nerve in the CPA, mainly from the anterior inferior cerebellaris artery (AICA) and its branches; and 17 in the IA M were mostly from the internal auditory arteria (IAA). Conclusion: To be familiar with the microanatomy of the facial nerve in CPA and IAM is h elpful for localizing the facial nerve and improve its preservation in acoustic neuroma surgery.

Objective To assess the feasibility of intestitial MR lymphangiography (MRL) with subcutaneous injection of a commercially available,non-ionic,extracellular paramagnetic contrast agent,to visualize lymphatic vessels in patients with primary lymphedema.Methods Forty lower extremities in 31 patients with clinically advanced stages of primary lymphedema were examined with magnetic resonance lymphangiography.A 1 ml mixed liquor of gadobenate dimeglumine and mepivacainhydrochloride were injected subcutaneously into the dorsal aspect of both feet For MRL,a 3D fast spoiled gradient-recalled echo T1-weighted images with a fat saturation technique (T1 high resolution isotropic volume excitation,THRIVE) were performed after subcutaneous application of the contrast material To outline lymphatic vessels,source images were used to reconstruct images of MIP.The SNR and CNR of enhanced lymphatic vessels and veins were measured and calculated respectively.The significance of the differences of the data comparisons was assessed using an unpaired student t teat.Results Of the 40 lower extremities,the beaded appearance of dilated lymphatic vessels was detected in 36 lower legs (90.0%) and 17 upper legs (42.5%).The numbers of the dilated lymphatic vessels displayed in all segments of lower extremities added up to 365 and its mean diameter was (3.4 ± 0.1) mm on MRL MIP image.The average SNR and CNR were 257 ±130,207 ± 113 in the dilated lymphatic vessels and 218 ± 129,152 ± 113 in the vein respectively,which was statistically significant (SNR t = -2.649,CNR t = -3.404,P <0.01).Contrast enhancement was observed in 30/40 inguinal lymph node groups (75.0%).In 26 lower extremities (65.0%) collateral vessels with dermal back-flow areas between lymphatic vessels were seen.Conclusions As a novel tool to image the pathologically modified lymphatic vessels in patients with clinically advanced stage of primary lymphedema,magnetic resonance lymphangiography is a safe,useful technique.

Objective To evaluate invitro antibacterial activity of Gymnema sylvestre extractant.Methods The antibacterial activity of Gymnema sylvestre extractant against staphyloccus aureus,bacillus coli,bacillus proteus,bacillus aeruginosus was detected by agar dilution method.Results The minimum inhibitory concentration were 0.65,1.25,1.25,2.5 mg/mL respectively.Conclusion Gymnema sylvestre extractant had strong antibacterial activity against various bacteria.

Objective To explore the relationship between structure and function in the diseases of posterior visual pathways as well as the anatomic mechanism of the abnormal visual responses.Methods Eleven cases of diseases of posterior visual pathways(3 gliomas,4 meningiomas,3 metastasis,1 stroke) involving either cortical or subcortical visual pathways were investigated by combining fMRI and DTI.fMRI was performed by using flashing checkerboard at 8 Hz.For imaging processing,fMRI analysis was performed with SPM99,and DTI and tractography with DTVⅡ.Fractional anisotropy(FA) of optic radiations and activated volume(VOXELs) of primary visual cortices(V1 and V2) were measured and analyzed at the affected side and the contralateral side.Relative FA(rFA) and relative activated volume(rVOXELs) were also calculated.3D tractography of optic radiations was performed successfully in 11 patients.Results In the patients with brain tumors,FA values in the affected side of optic radiations were significantly different with the contralateral side(P

The clinical laboratory equipment on hospital ship was introduced, whose status and common faults were analyzed during Philippines humanitarian medical rescue mission and 3 times of Mission of Harmony. The experience was summarized for the maintenance of the clinical laboratory equipment. The problems and countermeasures were proposed for the clinical laboratory equipment.

ObjectiveTo evaluate the ability of high resolution magnetic resonance (MR) imaging at 3.0 tesla to depict the characterization of human carotid arterial vessel wall and detect atherosclerotic lesions ex vivo in comparison to histopathologic results. MethodsEighteen carotid arteries obtained from 9 elderly donors underwent fat-suppressed T1 - and T2-weighted MR imaging at 3.0 tesla MR system with a mouse coil. Corresponding histological sections were obtained for the comparison. Correlation between MR images and histopathologic slices was obtained by Pearson or Spearman correlation coefficient.Cohen K was computed to quantify the agreement between MRI and histopathologic findings.Results Lumen area,intima and media area measured on fat-suppressed T2-weighted images showed stronger correlation with the corresponding histopathologic slices [ MRI vs.histopathology:( 27. 53 ± 6. 77 ) mm2 vs.( 25.83 ±6. 69 ) mm2, r2 ＝ 0. 91,P2 ＝ 0. 024, ( 12.31 ± 3. 31 ) mm2 vs.( 12. 28 ± 3.71 ) mm2, r2 ＝ 0. 70, P2 ＝ 0. 020,Median 12. 29 mm2(Min 1.12 mm2, Max 33.18 mm2) vs.Median 11.62 mm2(Min 0.89 mm2, Max 32. 84 mm2 ), r2 ＝ 0. 74, P2 ＝ 0. 016, respectively]. The Cohen K score between the MR imaging and American Heart Association classifications was 0. 74, which corresponds to a good agreement. Conclusions 3.0 T high-resolution multi-sequence MRI can clearly show the structure of ex vivo carotid artery wall and allow quantitative assessment.Fat-suppressed T2Wimaging has a greater advantagein presenting atherosclerotic lesions.

ObjectiveTo access the value of combined application of high-resolution interstitial MR lymphangiography (MRL) and heavily T2WI for the visualization of lymphatic vessels in patients with primary lymphedema.Methods Forty lower extremities in 31 patients with primary lymphedema were examined by heavily T2 WI and interstitial MRL with a 3.0 T MR unit (Philips Medical Systems,Best,the Netherlands).Two experienced radiologists analyzed the images and tried to determine the differences in number of lymphatic vessels,and their maximum diameters,SNR and CNR,and accumulated lymph fluid in the tissue.Statistical analyses were conducted by using Wilcoxon test and t test.ResultsDilated lymphatic vessels in 73 leg segments of 40 lower extremities were visualized on heavily T2WI (median 5,1 to 24),which were more than those on MRL ( median 3,1 to 16) (Z =-2.92,P ＜ 0.01 ).The maximum diameter of lymphatic vessels was (4.3 ± 1.5 ) mm on heavily T2 WI,whereas it was ( 3.4 ± 1.0) mm on MRL ( t =6.90,P ＜ 0.0 1 ).The average SNR and CNR in the dilated lymphatic vessels were 257 ± 130,207 ± 113 on MRL and 169 ± 91,135 ± 82 on heavily T2 WI,which was statistically significant ( SNR t =- 5.95,CNR t =-5.10; P ＜ 0.01 ).The visualization of regions of accumulated lymph fluid on heavily T2WI (median 1 ) had a higher score than that on MRL ( median 0 ) ( Z =- 5.64,P ＜ 0.01 ).Conclusions The heavily T2WI has greater sensitivity and the MRL image has better SNR and CNR.Combining these two MR techniques can provide adequate information for clinicians in the therapeutic planning of patients with advanced stages of lymphedema.

Objective To construct eukaryotic expression vector of human antimicrobial peptide LL-37 pPIC9-LL-37, and transform the plasmid pPIC9-LL-37 into P.pastoris GS115 to obtain the recombinant P.pastoris strains.Methods The full-length of antimicsobial peptide LL-37 gene was artificially synthesized by overlap extension method and was fused to pPIC9 and then the fused plasmid was transformed into E.coli DH5?.After analysis by PCR and sequencing,the plasmid pPIC9-LL-37 was transformed into P.pastoris.The colonies exhibiting the phenotype of His+Mut+ or His+Mut-were screened by means of MM and MD plates and the insertion was confirmed by PCR.Results The results of PCR and sequencing confirmed that the LL-37 gene was correctly inserted into pPIC9. The colonies of 10 His+Mut+ and 9 His+Mut-were obtained by means of MM and MD plates screening and were confirmed by PCR.Conclsion The recombinant P.pastoris strains containing LL-37 were successfully obtained.

Objective:To explore and compare the clinical efficacy of lyophilized recombinant human brain natriuretic peptide (Lrh-BNP) and dobutamine (Dob) in the treatment of patients with acute heart failure (AHF) and impacts on the plasma galectin (Gal)-3,Cystatin C (CysC) and endothelin (ET-)-1 levels.Methods:114 cases of patients with AHF in our hospital from February 2015 to February 2017 were selected as the research objectives and randomly divided into two groups.Dob group was treated by Dob,while Lrh-BNP group was treated by Lrh-BNP.The cardiac function parameters,plasma Gal-3,CysC,ET-1 levels before and after treatment,clinical comprehensive efficacy and incidence of adverse reactions were compared between two groups.Results:The FS,LVEF levels of both groups at 72 hours after treatment were significantly higher than those before treatment (P＜0.01),but the LVEDD,plasma Gal-3,CysC,ET-1 levels were obviously decreased (P＜0.01),the index mentioned above of Lrh-BNP group improved more significantly than those of the Dob group(P＜0.01).The overall effective rate of Lrh-BNP group was 89.5 ％,which was significantly higher than that of the Dob group (73.7％,P＜0.05).No significant difference was found in the incidence of adverse reaction between two groups(P＞0.05).Conclusion:Lyophilized recombinant human brain natriuretic peptide was more effective in the treatment of AHF than Dobutamine with equal safety,which might be related to the decrease of plasma Gal-3,CysC,ET-1 levels.