Objective To study the effect of cellulase on the extraction rate of medlar total saccharide, considering three factors of temperature, time and quantity of cellulose. The best extracting condition was determined according to the require of sphere symmetry design. Methods Add cellulase liquid into the medlar slurry and determine the content of medlar total saccharide by DNS method. Result The extraction rate of medlar total saccharide was increased 1.66% by cellulase at temperature of 60 ℃, time of 300 s and cellulase volume of 5 mL. Conclusion Cellulase can promote the extraction rate of medlar total saccharide.

Objective To evaluate the clinical value of endovascular therapy based on Angiosome concept in critical limb ischemia (CLI) patients.Methods A retrospective study was undertaken in 62 patients (62 limbs) diagnosed as critical limb ischemia (Rutherford class 5/6) and received percutaneous transluminal angioplasty from July 2011 to October 2013.Patients were divided into direct (35 patients) and indirect (27 patients) groups depending on whether feeding artery flow to the site of ulcer was successfully achieved or not based on the angiosome concept,and the between-group clinic outcomes including preoperative and postoperative ankle-brachial index (ABI),ulcer healing,and the rates of freedom from amputation were compared and analyzed.Results There were no significant difference between two groups for ABI pre-operatively (0.16 ±0.26 vs.0.15 ±0.28; P =0.885) and post-operatively (0.82 ± 0.26 vs.0.81 ±0.24; P =0.877).During follow-up,the healing rate of ischemia ulcer in patients without amputation at 1 year (91％ vs.74％ ; P =0.027) were significantly higher in the direct group than in the indirect group.There was no difference between two groups for healing time (162 ±49) d vs.(160 ±46) d; P =0.950).The rates of freedom from amputation at 1 year (84％ ±3％ vs.76％ ±4％ ; P =0.025) and 2 years (79％ ±4％ vs.72％ ±4％ ; P =0.031) were significantly higher in the direct group than in the indirect group.Conclusions Endovascular therapy based on Angiosome concept in CLI patient is an effective procedure with satisfactory clinic outcomes,contributing to healing of ischemia ulcer and increase amputation-free rate.

OBJECTIVE: To establish the droplet digital PCR (ddPCR) assay for the detection of NPM1 type A mutation in patients with acute myeloid leukemia (AML), and to evaluate its specificity, sensitivity and its value in clinical application. METHODS: NPM1 mutant and wildtype plasmids were used to verify the performance of ddPCR. Both ddPCR and Sanger sequencing were used to detect the bone marrow samples of 87 AML patients, which were confirmed by next generation sequencing (NGS). Moreover, NPM1 mutation burden was dynamically monitored in five patients by ddPCR. RESULTS: The limit of blank (LOB) of ddPCR established for NPM1 mutation detection was 1.1 copies/μl, and the limit of detection (LOD) was 2.43 copies/μl, which had good linearity. Among the 87 newly diagnosed AML patients, ddPCR identified seventeen cases positive for NPM1 mutation (19.5%)， which was consistent with Sanger sequencing. NGS confirmed 12 positive cases, including 8 of type A mutations, 2 of type D mutations, and 2 of rare type mutations. The results of dynamic monitoring of NPM1 mutation burden in 5 patients showed that the NPM1 mutation burden decreased obviously even close to 0, when patients achieve complete remission after chemotherapy. However, the mutation burden was increased again at the time of relapse. CONCLUSION In this study, we established a ddPCR method for detection of NPM1 mutation with good sensitivity and repeatability, which can be used for screening NPM1 mutation in newly diagnosed AML patients and for minimal residual disease monitoring after remission in positive AML patients to guide treatment.
Humans ; Leukemia, Myeloid, Acute/therapy* ; Mutation ; Nuclear Proteins/genetics* ; Nucleophosmin ; Polymerase Chain Reaction ; Prognosis

: AbstractObjective: To identify the expression and methylation patterns of lncRNA CASC15 in bone marrow (BM) samples of acute myeloid leukemia (AML) patients, and further explore its clinical significance. METHODS: Eighty-two de novo AML patients and 18 healthy donors were included in the study. Meanwhile, seven public datasets from Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) were included to confirm the expression and methylation data of CASC15. Receiver operating characteristic (ROC) curve analysis was applied to determine the discriminative capacity of CASC15 expression to identify AML. The patients were divided into CASC15high group and CASC15low group by X-tile method, and the prognostic value of CASC15 was identified by Kaplan-Meier method and univariate and multivariate Cox regression analysis. RESULTS: The expression level of CASC15 was significantly decreased in BM cells of AML patients compared with healthy donors (P<0.001). ROC curve analysis suggested that CASC15 expression might be a potential biomarker to discriminate AML from controls. The expression of CASC15 was high at the early stage of hematopoiesis, and reached a peak at the stage of multipotent progenitors differentiation, then decreased rapidly, and was at a range of low level fluctuations in the subsequent process. Among FAB subtypes, CASC15 expression in M0 was significantly higher than that in M1-M7. Clinically, CASC15low patients were more likely to have NPM1 mutations than CASC15high patients (P=0.048), while CASC15high patients had a significantly higher frequency of IDH1 and RUNX1 mutations (P=0.021 and 0.014, respectively). Moreover, CASC15low group had a shorter overall survival (OS) in patients with NPM1 mutations. Furthermore, multivariate analysis confirmed that CASC15 expression was a significant independent risk factor for OS in NPM1 mutated AML patients. In addition, CASC15 methylation level in BM samples of AML patients was significantly decreased compared with healthy donors. Patients with CASC15 high methylation had poor OS and disease-free survival. CONCLUSION The expression of CASC15 is decreased in AML, and low CASC15 expression may predict adverse prognosis in AML patients with NPM1 mutations. Moreover, CASC15 methylation level in AML is significantly decreased, and high CASC15 methylation may predict poor prognosis in AML.
Humans ; Leukemia, Myeloid, Acute/metabolism* ; Mutation ; Nuclear Proteins/genetics* ; Nucleophosmin/genetics* ; Prognosis ; RNA, Long Noncoding/genetics*

In this study, HPLC-ESI-MS and HPLC methods were established to explore the differences in the main chemical components and content of Mori Cortex with(mulberry root bark) and without(Mori Cortex) the phellem layer from both qualitative and quantitative aspects. The HPLC-ESI-MS method was used for quality analysis in positive and negative ion modes, and 33 compounds were identified in mulberry root bark, 22 compounds in Mori Cortex, and 26 compounds in phellem layer; mulberry root bark and Mori Cortex shared 22 components, and mulberry root bark has 11 unique compounds; Mori Cortex and its phellem layer shared 15 components, while Mori Cortex has 7 unique compounds. HPLC method was used to simultaneously determine 7 major constituents, including mulberroside A, chlorogenic acid, dihydromorin, oxyresveratrol, moracin O, kuwanon G, and kuwanon H, and the developed method showed good linearity(r>0.998 9) within the concentration range and the recoveries varied from 99.88% to 103.0%, and the RSD was 1.7%-2.9%. The HPLC results showed that the contents of the 7 compounds have great differences in 13 batches samples, compared with mulberry root bark, the contents of mulberroside A, chlorogenic acid, dihydromorin and moracin O of Mori Cortex were increased, while the contents of oxyresveratrol, kuwanon G and kuwanon H were decreased after peeling process. These results can provide a basis for the rationality and quality control of Mori Cortex required to remove the phellem layer.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; Mass Spectrometry ; Morus ; Plant Bark

In this study, the thermoresponsive micelles were synthesized with random copolymerization method and the photosensitizer indocyanine green (ICG) was loaded on micelles through the physical adsorption. The light energy was converted into heat energy to increase the temperature after irradiation with near-infrared light. When the phase transition temperature was reached, the micelle was disassembled and the targeted therapy was achieved. The nanoparticles were characterized with a transmission electron microscopy, Fourier transform infrared spectrometer, nuclear magnetic resonance spectrometer and other characterization were used to investigate. The critical micelle concentration (CMC), upper critical solution temperature, the photothermal properties of the carrier and the release of drug triggered by light were investigated after the doxorubicin (DOX) loaded. The carrier was evaluated for toxicity, cellular uptake, the effect of photothermal, the combination of photothermal and chemotherapy; the p(AAm-co-AN)-g-PEG (PAAP) was spherical in shape with a particle size of about 45 nm and a phase transition temperature was about 43℃. The critical micelle concentration was 24 μg·mL^-1. The particle size increased to 88 nm after loaded with ICG and DOX which the photothermal effect was obvious. The cumulative release of the drug under the irradiation of near-infrared light (808 nm, 2 W·cm^-2, 2 min·h^-1) was increased to 59.4% (pH 5.0) after 5 h. The results of the cell experiment indicated that ICG-PAAP was almost non-toxic and uptaken by the lysosomal pathway. The cell killing effect was stronger with combination of chemotherapy (DOX as 20 μg·mL^-1) with more than 70% of the cells killed. The results showed that the prepared micelle with low toxicity was thermoresponsive and could be used in combined therapy of tumor under the irradiation of near-infrared light.

Objective: To construct and identify an overexpression recombinant adenovirus vector carrying the mouse PTG gene(NM＿016854), and to lay a foundation for in-depth study of the function of PTG. Methods: The coding sequence of the mouse PTG gene was chemically synthesized, amplified by polymerase chain reaction(PCR), digested with restriction enzymes, and inserted into the GV314 vector(CMV-MCS-3 FLAG-SV40-EGFP) to obtain the recombinant shuttle plasmid pGV314-PTG. BamHⅠ/AgeⅠ double enzyme digestion was further carried out, and the product was transferred into linearized expression vector pDC315 to construct recombinant adenovirus Ad-PTG, which was transfected into HEK293 T cells and packaged into recombinant virus particles. After repeated amplification of several generations of HEK293 T cells, the recombinant adenovirus was purified and titer detected. Finally, PCR, Western blot and sequencing were used to verify the recombinant adenovirus. Results: After PCR, Western blot and sequencing, the results showed that the pGV314-CMV-MCS-3 FLAG-SV40-EGFP-PTG overexpression adenovirus vector(Ad-PTG) was successfully constructed, and the virus titer measured by end-point dilution method was 4×1010PFU/ml, Western blot and RT-qPCR showed that the protein and mRNA expression levels of PTG increased significantly. Conclusion The recombinant adenovirus vector carrying mouse PTG gene is successfully constructed, and the expression of PTG gene in hepatocytes is effectively up regulated.

For long-term application of the rotary pumps, it is necessary to solve the problems of bearing wear and thrombosis along the bearing. Currently, many investigators choose the magnetic bearing to realize zero-friction and no contact between the rotor and stator; the former avoids the mechanical wear and the latter eliminates the possibility of thrombus formation. We tried and found that it is difficult to apply a magnetic bearing to rotary pump without disturbing its simplicity, reliability and implantable; therefore, we have developed a much simpler and much more creative approach to achieving the same results. Instead of the sliding bearing, a rolling bearing has been devised for the pump; its friction is about 1/15 of the sliding bearing. Furthermore, a wear-proof material of ultra-high-molecular weight polythene has been adopted in making the rollers, their anti-wear property in 8 times better than that of metal. Thereby, the service life of the bearing has extended to several years. For preventing the thrombus formation along the bearing, the impeller reciprocation axially as the impeller changes its rotating speed periodically to produce a pulsatile flow. The reciprocation is a result of the effects of a magnetic force between the motor rotor and stator, and a hydraulic force between the blood flow and the impeller. Similar to piston pump, the oscillating impeller can make the blood in and out of the bearing, resulting in wash-out once a circle. This is obviously beneficial to preventing thrombosis along the bearing and in the pump. The endurance tests with saline of this novel pump demonstrated a durability of the device. It promises to be able to assist the circulation of the patients permanently and to be able to replace the heart transplantation in the future.
Heart-Assist Devices ; Magnetics ; instrumentation ; Prosthesis Design ; Thrombosis ; prevention & control

Magnetic bearing has no mechanical contact between the rotor and stator. And a rotary pump with magnetic bearing has therefore no mechanical wear and thrombosis due to bearing. The available magnetic bearings, however, are devised with electric magnets, need complicated control and remarkable energy consumption. Resultantly, it is difficult to apply an electric magnetic bearing to rotary pump without disturbing its simplicity, implantability and reliability. The authors have developed a levitated impeller pump merely with permanent magnets. The rotor is supported by permanent magnetic forces radially. On one side of the rotor, the impeller is fixed; and on the other side of the rotor, the driven magnets are mounted. Opposite to this driven magnets, a driving motor coil with iron corn magnets is fastened to the motor axis. Thereafter, the motor drives the rotor via a rotating magnetic field. By laboratory tests with saline, if the rotor stands still or rotates under 4,000 rpm, the rotor has one-point contact axially with the driving motor coil. The contacting point is located in the center of the rotor. As the rotating speed increases gradually to more than 4,000 rpm, the rotor will detache from the stator axially. Then the rotor will be fully levitated. Since the axial levitation is produced by hydraulic force and the driven magnets have a gyro-effect, the rotor rotates very steadly during levitation. As a left ventricular assist device, the pump works in a rotating speed range of 5,000-8,000 rpm, the levitation of the impeller hence is ensured by practical use of the pump.
Equipment Design ; Heart-Assist Devices ; Magnetics ; instrumentation

OBJECTIVESTo evaluate the clinical significance of the detection of IgG and IgM antibodies against Chlamydia trachomatis (CT) in semen of asymptomatic infertile patients.

METHODSOne hundred and sixteen asymptomatic infertile patients and eighteen fertile males were selected randomly. The routine parameter analysis of semen was fulfilled by computer aided semen analysis(CASA). Then the seminal plasma was separated and the IgG, IgM antibodies against CT in seminal plasma were determined with ELISA method.

RESULTSIgG and IgM antibodies against CT were present in 13.8% (16/116) and 3.4% (4/116) of the semen of infertile patients, while for the fertile males the percentages were 11.1% (2/18) and 0, respectively. There were no differences between the two groups(P > 0.05). In the infertile patients, 22 patients were azoospermia. And in the rest 94 infertile patients, the percentages of IgG and IgM antibodies in abnormal sperm density group were 21.4% (6/28) and 7.1% (2/28), which were higher than those in normal group, but there were no statistical differences(P > 0.05). Similarly, the IgG, IgM antibodies were not correlated with the sperm motility(P > 0.05). The positive percentage of CT in 116 patients was 25.9% (30/116).

CONCLUSIONSThe percentages of IgG and IgM antibodies against CT in semen of asymptomatic infertile patients are similar to that in fertile males, which do not correlate with the changes of semen parameters, and may not be used for indication of CT infection.

Adult ; Antibodies, Bacterial ; blood ; Chlamydia trachomatis ; immunology ; Humans ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Infertility, Male ; microbiology ; Male ; Sperm Count ; Sperm Motility