Method of transfection of STOP gene lentiviral vector into oligodendrocyte precursor cells of BTBR mouse model of autism / 解剖学报

Objective To investigate the effect of microtubule binding protein STOP on myelin formation of oligodendrocyte in BTBR mice spectrum disorder in vitro, a highly purified primary culture method of oligodendrocyte precursor cells from cerebral cortex of BTBR mice was established. Establishment of a highly efficient transfection method for overexpression of STOP gene in oligodendrocyte precursor cells of BTBR mice cerebral cortex using lentiviral vector. Methods BTBR mice were used as experimental objects, 6-10 suckling mice were taken each time, repeat 3 times independently. The single cell suspension was prepared by trypsin digestion, and the primary oligodendrocyte precursor cells were obtained by immunomagnetic bead cell sorting method . After 5 days of culture, the cell purity was identified by oligodendrocyte precursor cell marker staining. The primary cultured oligodendrocyte precursor cells were transfected with STOP gene vector constructed in the early stage of the project group. 72-96 hours after transfection, the fluorescence staining of oligodendrocyte precursor cells was observed under fluorescence microscope, and the transfection rate and cell survival rate were calculated. Results The oligodendrocyte precursor cells of BTBR mice extracted by immunomagnetic beads sorting method basically adhered to the wall completely after 48 hours, and the cells had strong ability of proliferation. On the fifth da)' of culture, the purity of the cells was more than 95% identified by immunofluorescence. A lentivirus transfection method for primary oligodendrocyte precursor cells of BTBR mice with high transfection efficiency was established. The fluorescence expression of the cells was obvious after being photographed by high connotation microscope, the lentivirus transfection rate of oligodendrocyte precursor cells was increased to 60%-70%. Conclusion The primaiy oligodendrocyte precursor cells of BTBR mouse cerebral cortex with high purity were successfull)' isolated and cultured. A method for lentivirus infection of primaiy oligodendrocyte precursor cells in the cerebral cortex of BTBR mice is successfully established.

Comparative of Forensic DNA Identification Using Cell Lysis Method and Magnetic Beads Method / 法医学杂志

OBJECTIVES@#To compare the effects of cell lysis method and magnetic beads method in forensic DNA identification and to explore these two methods in forensic DNA identification.@*METHODS@#The genome DNA of THP-1 cells in different quantities was extracted by the cell lysis method and magnetic beads method, and the DNA content was quantified by real-time quantitative PCR. The cell lysis method and magnetic beads method were used to type the STR of human blood with different dilution ratios.@*RESULTS@#When the numbers of THP-1 cell were 100, 400 and 800, the DNA content extracted by cell lysis method were (1.219±0.334), (5.081±0.335), (9.332±0.318) ng, respectively; and the DNA content extracted by magnetic beads method were (1.020±0.281), (3.634±0.482), (7.896±0.759) ng, respectively. When the numbers of THP-1 cells were 400 and 800, the DNA content extracted by the cell lysis method was higher than that by the magnetic beads method. The sensitivity of cell lysis method and magnetic beads method was similar in STR typing of human blood at different dilution ratios. Complete STR typing could be obtained at 100, 300 and 500-fold dilutions of blood samples, but could not be detected at 700-fold dilution. STR typing of undiluted human blood could not be detected by cell lysis method.@*CONCLUSIONS@#The cell lysis method is easy to operate and can retain template DNA to the maximum extend. It is expected to be suitable for trace blood evidence tests.

Esferas de hidrogel relacionadas con oclusión intestinal / Bowel obstruction related to hydrogel beads

Resumen Introducción: La obstrucción intestinal por ingesta accidental de cuerpos extraños se presenta muy rara vez en la edad pediátrica debido a que del 80 al 90% de los objetos pueden pasar libremente por el tracto gastrointestinal. Caso clínico: Se aborda el caso de una paciente de sexo femenino de 1 año 2 meses quien presentó obstrucción intestinal debido a la ingesta de esferas de hidrogel (poliacrilato de sodio). Dichas esferas, que son utilizadas como juguetes didácticos o sensoriales, aumentan de 200 a 400 veces su tamaño inicial mediante la absorción de agua. Se realizó radiografía abdominal en proyección decúbito supino, donde llamó la atención el hallazgo de defecto de llenado redondeado en asa de flanco derecho, ya que las esferas de hidrogel son radiolúcidas. El diagnóstico se estableció mediante ultrasonido abdominal, donde se reportó líquido libre peritoneal con datos de suboclusión por cuerpos extraños a nivel intestinal. Se indicó tratamiento conservador, encontrando persistencia de aumento de perímetro abdominal. Se realizó enterotomía y se encontraron las esferas impactadas a 30 centímetros de la válvula ileocecal. Conclusiones: Las esferas de hidrogel son peligrosas para la población pediátrica. La evolución de la paciente fue favorable debido al conocimiento del objeto extraño ingerido. Sobresale la conducta expectante que se tuvo que desempeñar debido a que se desconocía el crecimiento de las esferas y en qué momento no podrían continuar su paso por el tracto gastrointestinal.

Abstract Background: Bowel obstruction due to accidental ingestion of foreign objects occurs rarely in children because 80 to 90% of the objects can pass freely through the gastrointestinal tract. Case report: We report a case of a 14-month-old infant who presented bowel obstruction caused by the ingestion of hydrogel beads (sodium polyacrylate). Hydrogel beads are used as sensory and didactic toys that can increase their initial size 200 to 400 times by liquid absorption. An abdominal X-ray was perfomed in anteroposterior supine projection, where a round filling defect at the loop of the right flank was detected; this came to our attention because hydrogel beads are usually radiolucent. The diagnosis was established by abdominal ultrasound where free intraperitoneal fluid was reported with data of small bowel pseudo-obstruction by foreign objects. Conservative treatment was prescribed, finding persistence of increased abdominal perimeter, so an enterotomy was performed for their removal; finding impacted hydrogel beads 30 centimeters from the ileocecal valve. Conclusions: Hydrogel beads are dangerous for the pediatric population. The evolution of the patient was favorable thanks to the knowledge of the foreign objects ingested. The expectant behavior that had to be executed, stands out because we had no knowledge as to the maximum size of the hydrogel in the gastrointestinal tract.

A preparation strategy for protein-oriented immobilized silica magnetic beads with Spy chemistry for ligand fishing / 药物分析学报

Due to the complexity of bioactive ingredients in biological samples,the screening of target proteins is a complex process.Herein,a feasible strategy for directing protein immobilization on silica magnetic beads for ligand fishing based on SpyTag/SpyCatcher(ST/SC)-mediated anchoring is presented.Carboxyl functional groups on the surface of silica-coated magnetic beads(SMBs)were coupled with SC using the 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride/N-hydroxysulfosuccinimide method,named SC-SMBs.The green fluorescent protein(GFP),as the capturing protein model,was ST-labeled and anchored at a specific orientation onto the surface of SC-SMBs directly from relevant cell lysates via ST/SC self-ligation.The characteristics of the SC-SMBs were studied via electron microscopy,energy dispersive spectroscopy,and Fourier transform infrared spectroscopy.The spontaneity and site-specificity of this unique reaction were confirmed via electrophoresis and fluorescence analyses.Although the alkaline stability of ST-GFP-ligated SC-SMBs was not ideal,the formed isopeptide bond was unbreakable under acidic conditions(0.05 M glycine-HCl buffer,pH 1-6)for 2 h,under 20％ethanol solution within 7 days,and at most temperatures.We,therefore,present a simple and universal strategy for the preparation of diverse protein-functionalized SMBs for ligand fishing,prompting its usage on drug screening and target finding.

Establishment and Optimization of a Method for Rapid Extraction of Plant Virus RNA Based on Functional Magnetic Beads / 中国生物化学与分子生物学报

Plant virus diseases are one of the major diseases restricting erop production.Timely identification of their pathogen and development rules is the prerequisite for effective control of their large- scale spread.However, long cycle, tedious steps and strict detection environment were the disadvantages existing in the detection technology of plant virus disease.In this study, Tobacco Mosaic virus (TMV) was used as a model to he extract UNA based on CMBs-ACPtmv , which was design based on the principle of complementary base pairing.Meanwhile, the experimental conditions were optimized and analyzed, including the preparation conditions of functionalized magnetic beads, the reaction conditions during extraction, and the sensitivity, stability and other properties of the method.The results showed the ability to capture RNA of CMBs-ACPtmv were best when prepared with 4 fxmol capture probe (ACPTMV ) and 0.08 mg carboxyl magnetic beads (CMBs) ; After 3 min of extraction, CMBs-ACPtmv has the best RNA extraction effect, but when the extraction temperature of CMBs-ACPtmv was changed, its extraction capacity showed no significant change; In the comprehensive performance evaluation, the sensitivity of CMBs-ACPjjjv can reach 2.5 ng/fxL, and the detection stability is good.Compared with conventional RNA extraction technology, CMBs-ACPimv has outstanding advantages in detection time and sample consumption.The functional magnetic beads extraction method established in this study is fast, safe and simple.It can achieve rapid extraction of plant virus RNA with simple equipment, which has a broad application prospect.

Localization of small pulmonary nodules with magnetic beads: An animal experiment / 中国胸心血管外科临床杂志

@#Objective    To investigate the feasibility of using magnetic beads to locate small pulmonary nodules. Methods    Twelve rabbits were randomly divided into two groups, 6 in each group. One group underwent thoracotomy after anesthesia and the other group underwent percutaneous puncture under the guidance of X-ray. One and two cylindrical tracer magnets (magnetic beads) with a diameter of 1 mm and a height of 3 mm were injected adjacent to the imaginary pulmonary nodules in left lung in each group. The magnetic beads beside the imaginary nodules were attracted by a pursuit magnet with a diameter of 9 mm and a height of 19 mm. The effectiveness of localization by magnetic beads were determined by attraction between tracer and pursuit magnets. Results    All processes were uneven in 12 rabbits. There was micro hemorrhage and no hematoma in the lung tissue at the injection site of the magnetic beads. When tracked with the pursuit magnets, there was one bead divorce in cases that one bead was injected, but no migration or divorce of the magnetic beads in cases that two magnetic beads were simultaneously injected to localize the small pulmonary nodules. Conclusion    The feasibility of using magnetic beads to locate small pulmonary nodules has been  preliminarily verified.

Design of Detection Module in Coagulometers Based on Dual-magnetic Circuit Beads Method / 中国医疗器械杂志

Coagulometer, known as blood coagulation analyzer, is a product that can provide accurate test results for medical diagnosis and treatment analysis by detecting a series of items closely related to thrombosis and hemostasis in coagulation reaction. On the basis of previous traditional methods, and with our deep understanding about the principles of hemagglutination detection, we propose a hemagglutination detection method by using the dual-magnetic circuit beads method. Then, the corresponding hemagglutination detection module is designed. The coagulation time of plasma can be measured by detecting the movement of the magnetic beads when the magnetic field intensity is appropriate. The activated partial thromboplastin time(APTT) of plasma is tested when the most suitable magnetic field intensity is found. The results preliminarily show that this blood coagulation test method is valid and the corresponding test module has a potential value in business.

Determination of Immunoglobulin Content in Humoral Immunity Evaluation of Biomedical Material Products by Cytometric Beads Array / 中国医疗器械杂志

In this study, cytometric beads array(CBA) was used to determine the immunoglobulin content in humoral immunity evaluation of biomedical materials. The bovine-derived acellular dermal matrix was selected as a test sample and implanted into Balb/C mice subcutaneously to 4 weeks according to the high, medium and low dose groups. Four weeks later, IgG1, IgG2a, IgG2b, IgG3, IgA, and IgM were measured by CBA. The data of the test group and the control group were analyzed statistically. The results showed that compared with the negative control group, there was no significant difference in the IgG3, IgA content in the positive control group, while the IgG1, IgG2a, IgG2b, and IgM contents were significantly higher than the negative control group; no significant differences were seen in the sample groups. The results show that the method is suitable for analysis of immunoglobulin content in humoral immunity evaluation of biomedical materials.

Stability and Sensory Evaluation of an Innovative Antiaging Formulation Based on Marine Ingredients

Abstract The high consumption of antiaging cosmetics represents an outstanding opportunity for the development of new processes and attractive products in the cosmetic industry. Stability studies and sensory analyses are critical steps in the development process and production chain. Here we present a potential antiaging cosmetic product with innovative sensory characteristics. Caviar extract antioxidant properties were firstly evaluated by the DPPH method since it is an important mechanism against skin aging. Ca-alginate beads containing 2% of caviar extract and 0.2% of black pigment were prepared to obtain spheres similar to caviar. The beads were incorporated in a gel phase (hydroxyethylcellulose 2.5%) containing 3% of dimethylaminoethanol. Stability was evaluated in different storage conditions (sunlight exposure, 5 ± 2 °C, 37 ± 2 °C and r.t.) through the parameters: appearance, color and odor, pH (6-7), density (0.98-1.14 g.mL-1), centrifugation and average size. After approval by the Committee for Ethics in Research (n° 3.503.061), 30 volunteers tested the new formulation and answered a questionnaire. At 2%, caviar extract was able to scavenge 10.9% ± 0.58 of DPPH radical. Formulations showed good stability after 90 days, even considering the average size (7.47 ± 0.41 - 8.4 ± 0.65 mm2). 90% of the sensory test participants reported that they would buy the new product. Therefore, the new product developed demonstrates a promising potential as an attractive cosmetic product.

Survival of commercial probiotic strains and their effect on dark chocolate synbiotic snack with raspberry content during the storage and after simulated digestion

BACKGROUND: A key challenge for manufacturers of pro-health food containing active probiotic microorganisms is to develop a product with attractive sensory features along with maintenance of declared number of microorganisms during storage and transfer by alimentary tract. RESULTS: The highest concentration of polyphenols was observed in snacks without an additive of probiotics as well as those with an additive of L. rhamnosus and B. animalis bacteria and concentration of these compounds increased by 9.5% during six months of storage. None of the products distinguished itself in the sensorial assessment although each was assessed positively. The number of microorganisms was stable and comparatively high during six months of storage at a room temperature and in cooling conditions (108 cfu/g). In the digestion model, an influence of aggressive digestion conditions was examined in the alimentary tract on the number of microorganisms, which allowed to arrange strains from the most resistant (S. boulardii) to the most sensitive (B. breve). It must be noted that currently on the market there is no available snack containing probiotic yeast as well as there is no literature data on works on such formulation of food. CONCLUSIONS: In the newly developed snack made of chocolate, in which sugar has been replaced with maltitol, a raw material was added in the form of raspberry, prebiotic in the form of inulin and a strain of probiotic bacteria, including the unprecedented so far S. boulardii, which stands a high chance to occupy a good place on the market of functional food.

A comparison between drug-eluting bead-transarterial chemoembolization and conventional transarterial chemoembolization in patients with hepatocellular carcinoma: A meta-analysis of six randomized controlled trials

Objective: Transarterial chemoembolization (TACE) is the standard treatment for unresectable intermediate hepatocellular carcinoma. Drug-eluting beads (DEB)-TACE is a promising approach expected to improve the efficiency and safety of conventional (c) TACE. However, controversy remains whether DEB-TACE performs better than cTACE. This meta-analysis aimed to compare cTACE and DEB-TACE in terms of overall survival (OS), adverse events, and response rate. Literature search was performed in PubMed, Cochrane, Embase, and Web of Science. Complete response (CR), partial response (PR), disease control (DC), stable disease (SD), OS, and major complications were compared between these two modalities. The pooled relative risk and 95% confidence interval were calculated for assessment. Six randomized controlled trials were included for further analysis after a comprehensive search. No significant difference was found in overall response at 3, 6, 9, and 12 months, CR, PR, DC (SD), OS and complications between cTACE and DEB-TACE. Conclusion: DEB-TACE had similar therapeutic effects to those of cTACE. Furthermore, major complications in both therapies were similar. The superiority of DEB-TACE over cTACE remains unclear, and further research with high-quality evidence is needed

Isolation and Biological Characterization of Lung Cancer Stem Cells / 中国医学科学院学报

To explore the methods of screening and biological characteristics of lung cancer stem cells. We selected the ABCG2 and ABCG2 cells from SPC-A-1/adriamycin(ADM)cell line induced by ADM and analyzed the tumorigenicity of ABCG2 and ABCG2 cells by flow cytometry and transplantation in nude mice. The average fluorescence intensity of SPC-A-1 cells was(1.001±0.014)×10 ,which was significantly lower than that of SPC-A-1/ADM cells [(10.257±0.023) ×10 ](=17.320,=0.001);the difference was also statistically significant between the ABCG2/BCRP-FITC treatment group and the SPC-A-1 control group(=5.269,=0.021) and the SPC-A-1 control group(=6.869, =0.012) and between the SPC-A-1/ADM cell control group and the SPC-A-1/ADM cell homotype control group(=8.112,=0.015).The positive rate of SPC-A-1/ADM cells treated with ABCG2/BCRP-FITC was 9.8%,39.84 times higher than that of SPC-A-1 cells;it showed significant difference between the ABCG2/BCRP-FITC group and the SPC-A-1/ADM group(=9.120,=0.005) and the SPC-A-1/ADM group(=8.257,= 0.006).The positive rate of group B cells was 684 times that of group A cells,and the difference was statistically significant(=11.235,=0.001),and the fluorescence intensity of group B cells was strong.The average tumorigenic volume of the mice inoculated with SPC-A-1 cells,group A cells,and group B cells was(6.96±1.82),(6.70±2.55),and(9.17±2.41) mm ,respectively.Among them,group B was the highest,but there was no significant difference among these three groups(=2.362,=0.086).The tumorigenic rate of group B cells was 75.00%,which was significantly higher than that of SPC-A-1 cells and group A cells(=19.780,=0.002). ABCG2 cells from human lung adenocarcinoma SPC-A-1/ADM cell line can be isolated by ABCG2 antibody combined with immunomagnetic beads sorting method,and the tumor formation rate in nude mice can be observed to explore the identification and biological characterization of lung cancer stem cells.

Effect of arsenic trioxide-loaded CalliSpheres beads in the treatment of rabbits with VX2 liver tumor / 临床肝胆病杂志

ObjectiveTo investigate the effect of arsenic trioxide-loaded CalliSpheres beads (CBATO) in transarterial chemoembolization (TACE) in the treatment of rabbits with VX2 liver tumor. MethodsA total of 120 tumor-bearing rabbits were divided into control group, CalliSpheres beads (CB) group (blank beads for TACE), CBATO group, and conventional TACE (cTACE) group (arsenic trioxide lipiodol for TACE) using a random number table, with 30 rabbits in each group. Five rabbits in each group were sacrificed at 12 hours and on days 3, 7, and 14 after TACE, and immunohistochemistry was used to measure the proliferation index and apoptosis percentage of tumor cells in the residual tumor area. The tumor necrotic volume was measure on day 7 after TACE, and the growth rate and necrosis rate of tumor cells were calculated. Ten rabbits were randomly selected from each group for the observation of survival time. An analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups; the Kaplan-Meier survival analysis was used to evaluate survival time, and the log-rank test was used for comparison. ResultsOn day 7 after TACE, the CBATO group had a significantly lower growth rate and a significantly higher necrosis rate of tumor cells than the cTACE group, the CB group, and the control group (all P＜0.05). At each time point after TACE, there were significant differences in the proliferation index and apoptosis percentage of tumor cells between the CBATO group and the other three groups (all P＜0.05). The median survival time was 26 days in the CBATO group, 18.5 days in the CB group, 22 days in the cTACE group, and 15.5 days in the control group, and the CBATO group had a significantly longer survival time than the other three groups (χ2=3.95, 8.99, and 13.47, P=0.049, P=0.003, and P＜0.01). ConclusionCBATO has a better effect than cTACE and CB in the treatment of rabbits with VX2 liver tumor and can significantly improve tumor necrosis rate, promote the apoptosis of tumor cells, and prolong the survival time of experimental animals.

Advances in chromatography-based methods for screening active compounds from natural products / 药学学报

Natural products have been a major source of leading compounds in drug discovery. How to effectively screen active compounds from complex matrix remains an interesting topic. In this review, we comprehensively summarized advanced liquid chromatography based approaches in natural products screening, including pre-column, on-column and post-column screening methods. Their advantages, disadvantages and prospect are also discussed.

Short-term effect evaluation on auricular magnetic bead application for poor eyesight of primary school students / 上海预防医学

Objective To ascertain the short-term effects of traditional Chinese medicine(TCM) auricular magnetic beads application for treatment of poor vision in primary school students. Methods In 2017, a randomized sampling method was adopted in this study.A total of 458 students with a naked eye visual acuity ≤4.9 were screened from a primary school in the jurisdiction.Of them, 230 students were informed by the parent′s informed consent to receive the TCM ear acupoint application, and the remaining 228 students served as the control group. Results The average eyes visual acuity of intervention group was 4.842, increasing to 4.848 or 4.859 after 6 or 12 weeks′ treatment by auricular magnetic bead application.After statistically analysis, there is a significant difference in short-term effect of auricular magnetic bead application on eyesight improvement for primary school students. Conclusion The TCM auricular magnetic bead application proves to have a short-term effect on the poor eyesight of the students, and the operation is simple and safe.It is conducive to improving students′ participation in poor eyesight control and achieving family and school participation in TCM prevention and treatment of myopia.

Canna indica-chitin-alginate composite beads for the treatment of textile dye effluent containing Rose Bengal dye

Aim: To study the adsorption of Rose Bengal dye used in textile industries by Canna indica-chitin-alginate beads. Methodology: C. indica-chitin-alginate composite beads were prepared using root tubers of Canna indica, chitin from shrimp shells and sodium alginate. Batch adsorption of Rose Bengal dye was carried out with optimized parameters like pH, contact time, adsorbent dosage and dye concentrations. Characterization studies like SEM, FTIR and TGA and reusability of composite beads were also studied. Results: The maximum adsorption of 97.9% was obtained at pH 6, 80 min contact time with the optimized ratio of 1:1:0.05 C. indica-chitin-alginate towards 100 mg l-1 dye concentration. The SEM analysis showed a porous surface morphology whereas FTIR results exhibited the functional groups of Rose Bengal dye and composite beads, proving successful adsorption. Thermogravimetric analysis revealed that the composite beads could withstand a maximum temperature of 300oC. Interpretation: It is inferred from the study that a biosorbent prepared from a commonly available plant and shells of shrimp, which is considered as a waste, can be effectively used in the adsorption of harmful textile dyes from the effluent in an eco-friendly and cost-effective way.

A new method for rapid detection of dengue antigen based on microfluidic immune magnetic beads / 热带医学杂志

@# Abstract：Objective To establish a sensitive，rapid and convenient method for the detection of dengue antigen and assist clinical diagnosis of dengue. Methods In this paper，we developed a rapid detection method for dengue antigen based on microfluidic immune magnetic beads. Solidwork software was used to design microfluidic chip，which was prepared by mechanical processing and chemical sealing. Immunomagnetic beads of dengue antibody were prepared by chemical coupling reaction. Using HRP ⁃TMB ⁃H2O2 as color system，dengue NS1 antigen was detected on microfluidic chip carrier by double antibody sandwich method. Finally，57 clinical samples were tested by the novel method and traditional ELISA kit，and the accuracy of the method was analyzed，and the advantages and disadvantages of the two methods were compared. Results 20 minutes was needed to detect dengue NS1 antigen by using the novel ELISA method，and the reaction system only needed 10 μg beads and 10 μL samples. In the verification experiment，the method could distinguish the negative from the positive obviously. The positive sample had color rendering，while the negative and blank samples had no color rendering. In terms of detection performance，the coincidence rate between the new ELISA method and the traditional ELISA method reached 100%. Conclusion The novel ELISA detection platform had the advantages of simple，rapid，reagent and sample saving，high sensitivity，good stability and high accuracy，and could be used for the detection of dengue antigen.

Efficacy analysis of drug-eluting beads chemoembolization in the treatment of liver cancer patients who are waiting for fiver transplantation / 中华肝胆外科杂志

Objective To evaluate the value of DEB-TACE before liver transplantation for hepatocellular carcinoma patients.Methods From Jan.2016 to Jan.2018,23 patients received DEB-TACE before liver transplantation for hepatocellular carcinoma were induced.Complications evaluation was followed up after interventional therapy.4 weeks after the intervention,the imaging examination was performed to examine the tumor response rate depond on mRECIST,the pathological conditions and tumor free survival were studied in the patients who received liver transplantation.Results The achievement ration of operation was 100％ in 23 patients.23 patients received 24 times successfully,1 patient received DEB-TACE twice,and the remaining 22 patients received DEB-TACE once.No serious complications occurred.Eighteen patients (78.3％,18/23) had postembolic syndrome after interventional therapy,mainly fever and pain.Four weeks after DEB-TACE,the complete response rate was 47.8％ (11/23),partial response rate was 30.4％ (7/23),disease stability rate was 21.7％ (5/23).All the 23 patients were included in the waiting list for transplantation.Among them,15 cases received liver transplantation.Pathological results showed that the total necrosis rate was 53.3％ (8/15),and the tumour necrosis rate in 4 of them was less than 50％.The average tumour necrosis rate of the neoplasm was 75.0％.The 15 patients who received liver transplantation were alive with no tumor recurrence.Conclusion DEB-TACE is a safe and effective treatment for patients suffered from hepatocellular carcinoma in waiting for liver transplantation.However,due to the short time of DEBs in China,further research is needed.

Separation and forensic identification of sperm from cell mixtures using anti-hLCN6 monoclonal antibody coupled magnetic beads / 生物工程学报

Human lipocalin 6 (hLCN6) is an epididymis-specific secretory protein. It binds to sperm and plays important role in sperm maturation. To explore the feasibility for isolating spermatozoa from mixed cells using anti-hLCN6 monoclonal antibody-conjugated immunomagnetic beads (anti-hLCN6 IMBs) and establish a new method for the separation of sperms from mixed stains, 2 sets of 30 cases of cell mixture suspensions and stains containing different proportions of sperm and epithelial cells were prepared. Biotin-labeled anti-hLCN6 monoclonal antibody (mAb) was incubated with the cell mixtures, and the spermatozoa were then isolated with avidin-coated IMBs. Sperm DNA was extracted and analyzed by PCR-STR typing. Differential lysis was also conducted to compare the effect of the two different isolation methods. The dissociation constant (Kd) of anti-hLCN6 mAb was 3.47×10⁻⁹ mol/L measured by ELISA. Western blotting and immunofluorescence assays showed that hLCN6 was detectable on sperm cells and mainly located on the post-acrosomal region of the sperm head, but not in epithelial cells. Anti-hLCN6 IMBs could capture and separate the sperm cells successfully. Microscopic observation showed that the IMBs could bind to the head of sperm specifically. The success rate of STR typing (more than 13 STR loci, RFU>200) was 90% when the number of sperm cells was 10³/mL and 100% when the sperm cells number was equal to or more than 10⁴/mL. When the number of sperm cells was 10³/mL, 10⁴/mL and 10⁵/mL in mixed stain samples, the success rate of STR typing were 40%, 90% and 100%, respectively. Taken together, the anti-hLCN6 immunomagnetic beads (IMB) method described here could be effective for the isolation of sperm from mixed cells, and the success rate was higher than that of the traditional differential lysis strategy. IMB sorting is a simple and efficient method for the separation of sperms from sperm and epithelial cell mixture, and can be utilized as a supplementary method for forensic mixture samples analysis in sexual assault cases.

CELL SURFACE PROTEINS OF CRYPTOCOCCUS NEOFORMANS MEDIATE ADHERENCE AND INTERNALISATION INTO MAMMALIAN CELLS

@#Cryptococcus neoformans is an encapsulated fungal pathogen that causes severe disease primarily in immunocompromised patients. Adherence and internalisation of microbial pathogens into host cells often begin with engagement of microbes to the surface receptors of host. However, the mechanisms involved remain poorly understood. In this study, we investigated the association of cell surface determinants of C. neoformans with mammalian cells. Our results showed that treatment with trypsin, but not paraformaldehyde or heat killing, could reduce host-cryptococci interaction, suggesting the involvement of cell surface proteins (CSPs) of C. neoformans in the interaction. We extended our investigations to determine the roles of CSPs during cryptococci-host cells interaction by extracting and conjugating CSPs of C. neoformans to latex beads. Conjugation of CSPs with both encapsulated and acapsular C. neoformans increased the association of latex beads with mammalian alveolar epithelial cells, alveolar macrophages and monocyte-derived macrophages. Further examination on the actin organisation of the host cells implied the involvement of actin-dependent phagocytosis in the internalisation of C. neoformans in CSP-conjugated latex beads. We hypothesised that CSPs present on the cell wall of C. neoformans mediate the adherence and actin-dependent phagocytosis of cryptococci by mammalian cells. Our results warrant further studies on the exact role of CSPs in the pathogenesis of cryptococcosis