Objective: Clone, express and purify human recombinant calreticulin (CRT). Methods: Human CRT cDNA was amplified from total RNA of human lung cancer cell line A549 cells by RT-PCR. Then, PCR product was subcloned into prokaryotic expression vector pET-15b. After sequencing, this recombinant plasmid was transformed into E.coli. Rossetta. Recombinant CRT was expressed in host cells by IPTG induction. Resulted protein was purified by Ni-NTA resin under denature condition and dialyzed to recover its native structure. SDS-PAGE and Western blot method were used to identify the expression and purification of reconbinant CRT. Results: Human CRT cDNA was cloned from total RNA of A549 cells. CRT prokaryotic expression vector pET-15b-crt was constructed. Reconbinant CRT was induced to express in E.coli and purified by Ni-NTA affinity chromatograph. Conclusion: A method for prokaryotic expression and purification of human recombinant CRT was successfully established. This method laid a foundation for the subsequent CRT research.

OBJECTIVE:To establish an HPLC method for the determination of the concentration of tetrahydropalmatine in Puyuan weikang capsule. METHODS: The HPLC separation was performed on Venusil XBP-C18 chromatographic column (150 mm?6.0 mm, 5 ?m) with mobile phase consisted of methanol-0.1% phosphoric acid solution(60∶40, with pH adjusted to 6.0 using triethylamine) with a flow rate of 1 mL?min-1. The detective wavelength was set at 280 nm and the column temperature was set at 25 ℃. RESULTS: The linear range of tetrahydropalmatine was 2.5～100 ?g?mL-1 (r=0.999 6) and the recovery rate was 94.33%(RSD=1.11%,n=6). CONCLUSION: The method developed in the study was proved to be simple, rapid and accurate, and applicable for the quality control of Puyuan weikang capsule.

Objective To analyze the clinical and MRI manifestations of neurosyphilis in order to improve the recognition and di‐agnosis of the disease .Methods The MRI and clinical data of 11 patients with neurosyphilis confirmed in clinic were collected .The patients were categorized into different phenotypic types according to the clinical manifestations ,and the MRI manifestations were al‐so analyzed .Results 5 patients with parenchymal types showed cerebral atrophy .3 patients with meningovascular types showed cer‐ebral infarction .1 patient with encephalitis showed brain signal changes and meningeal enhancement .1 patient without symptom showed signal changes in brain parenchyma and mild enhancement .1 patient of myelopathic type showed signal changes of thoracic spinal cord without enhancement .Conclusion There is an overlap of the MRI findings of neurosyphilis in different clinical pheno‐types ,exhibiting no specifity .Neurosyphilis can be diagnosed when the young patient is manifested as cerebral atrophy ,infarction , MR signal intensity abnormality of cerebral parenchyma and spinal cord ,as well as meningeal enhancement .

Objective To investigate the effects of a highly selective dopamine D4 receptor (ABT-724)on behaviors in Spontaneously hypertensive rats (SHR),a rat model of attention-deficit hyperactivity disorder (ADHD).Methods After intervention of methylphenidate(5mg/kg) and three different doses of ABT-724(0.04mg/kg,0.16mg/kg,0.64mg/kg),behaviors of SHR were verified by open-field test,Morris water maze and Làt maze.Results Number of square crossing after intervention of methylphenidate and ABT-724 in SHR( 70.67 ± 8.59,76.50 ± 10.75,79.17 ± 10.44,65.67 ± 20.62) was less than the saline control group( 130.33 ± 1 1.40) (P＜0.05).During Morris water maze,SHRs(52.50 ± 4.04,52.17 ± 2.99,61 ± 8.15,53.83 ± 9.87 ) after intervention of both had a better spatial memory ability than control group(38.83 ±7.17) (P＜0.05).In Làt maze,number of rearing after intervention of both in SHRs(57.17 ± 5.67,60.83 ± 8.28,55.17 ± 9.45,65.33 ± 9.50 ) was less than saline control group(78.00 ± 13.84) (P＜0.05).Conclusion ABT-724 could improve behaviors of spontaneous locomotor activity,cognitive ability,non-selective attention in SHR.

Objective To make a systematic review of pressure ulcers risk factors in critically ill patients. Methods We systematically reviewed all articles related to the pressure ulcers risk factors in critically ill patients. The Cochrane Library, PubMed, EMBASE, Web of Science Core Collection, CNKI, WANFANG and SinoMed were searched to August 2016. Results In total, 13 eligible articles were included. These studies included 18, 184 critically ill patients, six studies were classified as high quality, and seven were classified as moderate quality. Risk factors for the development of pressure ulcers include age, ICU stay, diabetes, mean arterial pressure<60-70 mmHg (1 mmHg=0.133 kPa), mechanical ventilation and mechanical ventilation, drugs, sedation and postural changes. Conclusions There is no single factor that can explain the occurrence of pressure ulcers. So it is in a variety of factors interaction, the occurrence of a significant increase in risk.

Objective To explore the effect of enhanced physical therapy with integrated traditional and western medicine on muscularweakness after selective posterior rhizotomy. Methods 44 children with muscular weakness after selective posterior rhizotomy were dividedinto the treatment group (n=28) and the control group (n=16). Exercise therapy, physical therapy, electroacupuncture and Chinese massagewere applied to the treatment group, while exercise therapy was applied to the control group only. The muscle strength of the leg of childrenwere compared right after the surgery and 2 weeks after the surgery. Results The muscle strength of the leg of children in treatment group recoveredbetter than the control group 2 weeks after the surgery (P<0.05), and it was almost at the same level with that before surgery (P<0.05). Conclusion The enhanced physical therapy with integrated traditional and western medicine method could rapidly recover the musclestrength of the leg of children after SPR.

Objective To explore the role of the autoantibodies against ?_1 and ?_1-adrenergic receptor(?_1-receptor)in the development of hypertension with renal failure.Methods The epitopes of the second extracellular loop of ?_1-receptor(197-222) and ?_1-receptor(192-218) were synthesized and used respectively to screen sera autoantibodies in patients with hypertension and renal failure(n=61),hypertension without renal failure(n=60) and healthy blood donors(n=40,control) by ELISA.Results The positive rates of the autoantibodies against ?_1-receptor(62.3%)and ?_1 receptor(50.8%) in patients with hypertension with renal failure were higher than those of patients with hypertension without renal failure(13.3% and10.0%)(P

This study was aimed to identify and distinguish Metacordyceps liangshanensis recorded by the Sichuan Province Traditional Chinese Medicine Standard from its adulterants and its relative species by combining ITS and COI barcode sequences in order to study the feasibility of this new method. After extracting DNA of 28 species of Cordyceps samples, DNA were amplified and sequenced. And then, ITS and COI sequences were received. Codon-Code Aligner V3.7.1 and Mega 5.0 were used to analyze the variable site and construct the N-J tree. The results showed that the minimum ITS inter-specific K-2P distance was relatively higher than the maximum intra-specific K-2P distance. The inter-specific sequence divergence between M. liangshanensis and its adulterants exhibited high while intra-specific sequence divergence exhibited low. And COI one was the same case. N-J tree of both ITS and COI indicated that same genus belonged together and each species belonged to relatively independent branch. It was concluded that based on the ITS and COI gene, the technology of DNA barcode can be an excellent identification of M. liangshanensis, its adulterants and its relative species. It provided technical support for the further research on species molecular identification and phylogenetics of Cordyceps .

OBJECTIVE: Different methods have been utilized to prevent enamel demineralization and other complications during orthodontic treatment. However, none of these methods can offer long-lasting and effective prevention of orthodontic complications or interventions after complications occur. Considering the photocatalytic effect of TiO2 on organic compounds, we hoped to synthesize a novel bracket with a TiO2 thin film to develop a photocatalytic antimicrobial effect. METHODS: The sol-gel dip coating method was used to prepare TiO2 thin films on ceramic bracket surfaces. Twenty groups of samples were composed according to the experimental parameters. Crystalline structure and surface morphology were characterized by X-ray diffraction and scanning electron microscopy, respectively; film thickness was examined with a surface ellipsometer. The photocatalytic properties under ultraviolet (UV) light irradiation were analyzed by evaluating the degradation ratio of methylene blue (MB) at a certain time. Antibacterial activities of selected thin films were also tested against Lactobacillus acidophilus and Candida albicans. RESULTS: Films with 5 coating layers annealed at 700℃ showed the greatest photocatalytic activity in terms of MB decomposition under UV light irradiation. TiO2 thin films with 5 coating layers annealed at 700℃ exhibited the greatest antimicrobial activity under UV-A light irradiation. CONCLUSIONS: These results provide promising guidance in prevention of demineralization by increasing antimicrobial activities of film coated brackets.
Candida albicans ; Ceramics* ; Crystallins ; Dental Enamel ; Esthetics ; Hope ; Lactobacillus acidophilus ; Methods ; Methylene Blue ; Microscopy, Electron, Scanning ; Ultraviolet Rays ; X-Ray Diffraction

Objective In our previous study, we had generated various zebrafish mutant lines with tissue?specific GFP expression by Tol2 transposon?mediated insertional mutagenesis. Among these mutants,the Tol2:20141221t line ex?presses GFP in nervous system, while the position within zebrafish genome where transposon inserted has not yet been iden?tified. The aim of this study was to identify and analyze this genetically modified mutant. Methods The transgenic inser?tion loci in the genome of Tol2:20141221t line was identified byTAIL?PCR and the spatial and temporal expression profile of the affected gene was examined by in situ hybridization. Homozygous mutant of Tol2:20141221t was generated for explo?ring related developmental defects. Results Tol2 transposon was inserted into the 8th intron region of sat1.a gene, and in?duced premature transcription termination. The maternal and zygotic mutants of Tol2:20141221t was generated, while with?out apparent developmental defects. Conclusions We have generated and identified the zebrafishsat1.a mutant mediated by Tol2 transposon. This gene insertion mutant exhibits no obvious developmental abnormalities, but may serve as a power?ful tool to study the development of nervous system.