Application of the sorting technology for decommissioned radioactive waste in an urban radioactive waste repository / 中国辐射卫生

Objective To establish a radioactive waste sorting process. Methods According to the current laws, regulations, standards, waste minimization requirements, and domestic good practices, the non-radioactive waste and radioactive waste were distinguished, and the radioactive waste was further sorted according to the different subsequent disposal methods of radioactive waste. The precautions for each step of the sorting process were described in detail. Results This process has been used in sorting decommissioned radioactive waste in an urban waste repository and achieved good results. Approximately 83 tons of radioactive waste and 1173 waste radioactive sources were sorted. Conclusion Good radioactive waste sorting technology can not only distinguish between radioactive and non-radioactive waste, but also facilitate the subsequent disposal of radioactive waste, which minimizes radioactive waste and protects staff and the environment.

SARS-CoV-2 spike host cell surface exposure promoted by a COPI sorting inhibitor

Via an insufficient coat protein complex I (COPI) retrieval signal, the majority of SARS-CoV-2 spike (S) is resident in host early secretory organelles and a tiny amount is leaked out in cell surface. Only surface-exposed S can be recognized by B cell receptor (BCR) or anti-S therapeutic monoclonal antibodies (mAbs) that is the trigger step for B cell activation after S mRNA vaccination or infected cell clearance by S mAbs. Now, a drug strategy to promote S host surface exposure is absent. Here, we first combined structural and biochemical analysis to characterize S COPI sorting signals. A potent S COPI sorting inhibitor was then invented, evidently capable of promoting S surface exposure and facilitating infected cell clearance by S antibody-dependent cellular cytotoxicity (ADCC). Importantly, with the inhibitor as a probe, we revealed Omicron BA.1 S is less cell surface exposed than prototypes because of a constellation of S folding mutations, possibly corresponding to its ER chaperone association. Our findings not only suggest COPI is a druggable target against COVID-19, but also highlight SARS-CoV-2 evolution mechanism driven by S folding and trafficking mutations.

NDFIP1 limits cellular TAZ accumulation via exosomal sorting to inhibit NSCLC proliferation

NDFIP1 has been previously reported as a tumor suppressor in multiple solid tumors, but the function of NDFIP1 in NSCLC and the underlying mechanism are still unknown. Besides, the WW domain containing proteins can be recognized by NDFIP1, resulted in the loading of the target proteins into exosomes. However, whether WW domain-containing transcription regulator 1 (WWTR1, also known as TAZ) can be packaged into exosomes by NDFIP1 and if so, whether the release of this oncogenic protein via exosomes has an effect on tumor development has not been investigated to any extent. Here, we first found that NDFIP1 was low expressed in NSCLC samples and cell lines, which is associated with shorter OS. Then, we confirmed the interaction between TAZ and NDFIP1, and the existence of TAZ in exosomes, which requires NDFIP1. Critically, knockout of NDFIP1 led to TAZ accumulation with no change in its mRNA level and degradation rate. And the cellular TAZ level could be altered by exosome secretion. Furthermore, NDFIP1 inhibited proliferation in vitro and in vivo, and silencing TAZ eliminated the increase of proliferation caused by NDFIP1 knockout. Moreover, TAZ was negatively correlated with NDFIP1 in subcutaneous xenograft model and clinical samples, and the serum exosomal TAZ level was lower in NSCLC patients. In summary, our data uncover a new tumor suppressor, NDFIP1 in NSCLC, and a new exosome-related regulatory mechanism of TAZ.

Virus hijacking ESCRT system to promote self-replication: a review / 生物工程学报

Endosomal sorting complex required for transport (ESCRT) system drives various cellular processes, including endosome sorting, organelle biogenesis, vesicle transport, maintenance of plasma membrane integrity, membrane fission during cytokinesis, nuclear membrane reformation after mitosis, closure of autophagic vacuoles, and enveloped virus budding. Increasing evidence suggests that the ESCRT system can be hijacked by different family viruses for their proliferation. At different stages of the virus life cycle, viruses can interfere with or exploit ESCRT-mediated physiological processes in various ways to maximize their chance of infecting the host. In addition, many retroviral and RNA viral proteins possess "late domain" motifs, which can recruit host ESCRT subunit proteins to assist in virus endocytosis, transport, replicate, budding and efflux. Therefore, the "late domain" motifs of viruses and ESCRT subunit proteins could serve as promising drug targets in antiviral therapy. This review focuses on the composition and functions of the ESCRT system, the effects of ESCRT subunits and virus "late domain" motifs on viral replication, and the antiviral effects mediated by the ESCRT system, aiming to provide a reference for the development and utilization of antiviral drugs.

Enhancing the sensitivity of HIV DNA detection: a magnetic bead-based method / 中国输血杂志

【Objective】 To establish a magnetic bead enrichment strategy for the detection of human immunodeficiency virus deoxyribonucleic acid (HIV DNA) in peripheral blood, and to verify the improvement of the sensitivity of this method for the detection of HIV DNA in HIV infected patients after early antiretrovital treatment (ART). 【Methods】 Peripheral whole blood was collected at 4 timepoints in one ART HIV window period (WP) patient. Peripheral blood mononuclear cells (PBMCs) were isolated on a Ficoll gradient. CD4+ T lymphocytes were enriched from total PBMCs by negative sorting. HIV DNA concentration in magnetic beads enriched group and whole blood group was detected by HIV DNA detection kit. 【Results】 CD4+ T cells were isolated by magnetic beads and identified by FCM for purity at (96.4 ± 2.6)%. The viability was (95.9 ± 2.9)%, as demonstrated by trypan blue staining. The person on continued ART treatment in this study had significantly greater reduction in HIV viral load and undetectable HIV plasma RNA at follow up timepoint 4. No HIV DNA was detected in the whole blood group at all 4 timepoints. The quantitative results of HIV DNA in the CD4+ T lymphocyte group of the magnetic bead enrichment group were 73.4, 429.3, 137.1, 449.9 copies/106 CD4+ T cell′s respectively. 【Conclusion】 The magnetic bead enrichment method can be more sensitive in detecting the limit low copy HIV DNA in blood samples, and provide early confirmatory data for HIV WP infection and breakthrough infection after ART treatment.

A Comparative Assessment of Comprehensive Trail Making Test and Wisconsin Card Sorting Test Among Alcohol Dependence Patients

@#Introduction: Alcohol, when used frequently, accelerates the ageing process, causes brain damage, and results in a reduced volume of grey and white matter, leading to frontal lobe abnormalities. The neurotoxicity resulting from alcohol overuse affects the higher functions of the brain. This study aimed to evaluate the effect of alcohol dependence on the executive functioning of the brain. Methods: This study was carried out as a case-control study among 60 patients with alcohol dependence and 60 controls. Assessment of executive function was carried out using the Comprehensive trail-making test (CTMT) and the Wisconsin card sorting test (WCST). Comparison between the alcohol dependence group and normal healthy controls were calculated using the Mann-Whitney U test as data followed a non-parametric distribution. Results: The mean age of the participants among the cases and controls was 38.3±5.5 years and 37.8±5.4 years, respectively. The results showed a significant difference in both WCST and CTMT between cases and controls (p<0.05). Conclusion: This study concludes that there was an impaired performance in executive functions in alcohol- dependence patients in early abstinence compared to normal controls showing frontal lobe impairment in alcohol-dependence patients.

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.

Inertial label-free sorting and chemotaxis of polymorphonuclear neutrophil in sepsis patients based on microfluidic technology / 生物医学工程学杂志

Reduced chemotactic migration of polymorphonuclear neutrophil (PMN) in sepsis patients leads to decreased bacterial clearance and accelerates the progression of sepsis disease. Quantification of PMN chemotaxis in sepsis patients can help characterize the immune health of sepsis patients. Microfluidic microarrays have been widely used for cell chemotaxis analysis because of the advantages of low reagent consumption, near-physiological environment, and visualization of the migration process. Currently, the study of PMN chemotaxis using microfluidic chips is mainly limited by the cumbersome cell separation operation and low throughput of microfluidic chips. In this paper, we first designed an inertial cell sorting chip to achieve label-free separation of the two major cell types by using the basic principle that leukocytes (mainly granulocytes, lymphocytes and monocytes) and erythrocytes move to different positions of the spiral microchannel when they move in the spiral microchannel under different strength of inertial force and Dean's resistance. Subsequently, in this paper, we designed a multi-channel cell migration chip and constructed a microfluidic PMN inertial label-free sorting and chemotaxis analysis platform. The inertial cell sorting chip separates leukocyte populations and then injects them into the multi-channel cell migration chip, which can complete the chemotaxis test of PMN to chemotactic peptide (fMLP) within 15 min. The remaining cells, such as monocytes with slow motility and lymphocytes that require pre-activation with proliferative culture, do not undergo significant chemotactic migration. The test results of sepsis patients ( n=6) and healthy volunteers ( n=3) recruited in this study showed that the chemotaxis index (CI) and migration velocity ( v) of PMN from sepsis patients were significantly weaker than those from healthy volunteers. In conclusion, the microfluidic PMN inertial label-free sorting and chemotaxis analysis platform constructed in this paper can be used as a new tool for cell label-free sorting and migration studies.

Optimization of expression yield in a stable cell line expressing a novel mutated chimeric tissue plasminogen activator (mt-PA)

Abstract The development of stable cell lines producing recombinant proteins is very time-consuming and laborious. One of the practical approaches successfully performed is Fluorescence-Activated Cell Sorting (FACS). A mutated chimeric tissue plasminogen activator (mt-PA) was developed by removing the first three domains of t-PA, insertion of GHRP sequence and mutation toward resistance to plasminogen activator inhibitor-1 (PAI-1). In the current study, a new stable CHO-DG44 cell line producing mt-PA was developed by two sequential clonal selections: FACS and clonal-selection by limiting dilution. Furthermore, the expression was more evaluated using two different expression media. Finally, the high-producing clones were selected based on the dot blot and amidolytic activity test. The transfection efficiency of CHO-DG44 cells was 38% as measured by flow cytometry on green fluorescent protein (GFP). After performing FACS on stable cell pools, the expression yield was increased to fifty-fold. In terms of growth profile, CD-DG44 showed higher viability and cell density results than ProCHO5 medium. The expression of mt-PA was significantly higher in CD-DG44 than in ProCHO5, 765 and 280 IU/mL, respectively. Our data indicated that selection of an appropriate expression medium played a critical role in the development of potent producing stable cells by FACS.

Continuous purification and culture of rat type 1 and type 2 alveolar epithelial cells by magnetic cell sorting / 中华创伤杂志(英文版)

PURPOSE@#The incidence of acute lung injury (ALI) in severe trauma patients is 48% and the mortality rate following acute respiratory distress syndrome evolved from ALI is up to 68.5%. Alveolar epithelial type 1 cells (AEC1s) and type 2 cells (AEC2s) are the key cells in the repair of injured lungs as well as fetal lung development. Therefore, the purification and culture of AEC1s and AEC2s play an important role in the research of repair and regeneration of lung tissue.@*METHODS@#Sprague-Dawley rats (3-4 weeks, 120-150 g) were purchased for experiment. Dispase and DNase I were jointly used to digest lung tissue to obtain a single-cell suspension of whole lung cells, and then magnetic bead cell sorting was performed to isolate T1α positive cells as AEC1s from the single-cell suspension by using polyclonal rabbit anti-T1a (a specific AEC1s membrane protein) antibodies combined with anti-rabbit IgG microbeads. Afterwards, alveolar epithelial cell membrane marker protein EpCAM was designed as a key label to sort AEC2s from the remaining T1α-neg cells by another positive immunomagnetic selection using monoclonal mouse anti-EpCAM antibodies and anti-mouse IgG microbeads. Cell purity was identified by immunofluorescence staining and flow cytometry.@*RESULTS@#The purity of AEC1s and AEC2s was 88.3% ± 3.8% and 92.6% ± 2.7%, respectively. The cell growth was observed as follows: AEC1s stretched within the 12-16 h, but the cells proliferated slowly; while AEC2s began to stretch after 24 h and proliferated rapidly from the 2nd day and began to differentiate after 3 days.@*CONCLUSION@#AEC1s and AEC2s sorted by this method have high purity and good viability. Therefore, our method provides a new approach for the isolation and culture of AEC1s and AEC2s as well as a new strategy for the research of lung repair and regeneration.

Current status and progress in the standardized surgical management of specimens after radical gastric cancer surgery / 中华胃肠外科杂志

Standardized surgical management of postoperative specimens of gastric cancer is an important part of the standardized diagnosis and treatment of gastric cancer. It can reflect the accurate number and detailed distribution of lymph nodes in the specimen and lay the foundation for accurate and standardized pathological reports after surgery. Meanwhile, it can evaluate the scope of intraoperative lymph node dissection, the safety of cutting edge, and the standardization of surgery (principle of en-bloc dissection), which is an important means of surgical quality control. It also provides accurate research samples for further research and is an important way for young surgeons to train their clinical skills. The surgical management of postoperative specimens for gastric cancer needs to be standardized, including specimen processing personnel, processing flow, resection margin examination, lymph node sorting, measurement after specimen dissection, storage of biological specimens, documentation of recorded data, etc. The promotion of standardized surgical management of specimens after radical gastrectomy can promote the homogenization of gastric cancer surgical diagnosis and treatment in medical institutions and further promote the high-quality development of gastric cancer surgery in China.

Does decreased snx10 serve as a novel risk factor in atrial fibrillation of the valvular heart disease? - a case-control study

Abstract Introduction: Atrial fibrillation (AF) is the most common sustained arrhythmia. Sorting nexin 10 (SNX10) has been reported to be an important regulator in embryonic development and human diseases, however, little is known about its role in cardiac disease. The aim of this study was to investigate the clinical significance of SNX10 expression in AF. Methods: Nineteen valvular heart disease patients with AF and nine valvular heart disease patients with sinus rhythm (SR) were enrolled. Atrial tissue samples from patients undergoing open heart surgery were examined. Atrial tissues of normal hearts were obtained from two cases' autopsies. The SNX10 expression and its associations with the degree of fibrosis were analyzed by immunohistochemistry and Masson's trichrome staining. Results: SNX10 expression was detected in the cytoplasm of cardiac cells in human myocardial tissue. The SNX10 expression level was higher in the SR group than in the AF group (P=0.023). SNX10 expression was negatively associated with the degree of fibrosis (P=0.017, Spearman rho=-0.447), the New York Heart Association degree (P=0.003, Spearman rho=-0.545), left atrial diameter (P=0.038, Spearman rho=-0.393), right atrial diameter (P=0.043, Spearman rho=-0.386), and the brain natriuretic peptide (BNP) level 24 hours after surgery (P=0.030, Spearman rho=-0.426), but not the BNP level before surgery and 72 hours after surgery. No statistical significance was observed between SNX10 and the level of troponin T and C-reactive protein. Conclusion: Decreased SNX10 might serve as a potential risk factor in AF of the valvular heart disease.

In vitro culture and purification of Schwann cells: A systematic review / 中国组织工程研究

OBJECTIVE: Schwann cells can promote the regeneration of damaged peripheral nerves and serve as seed cells in the engineering repair of peripheral nerve tissue. The effective culture and purification of Schwann cells are the basis of clinical treatment for peripheral nerve injury. This paper summarized the literature of culture of Schwann cells in vitro in recent ten years and made a descriptive review on the research progress of Schwann cell culture and purification, in order to provide references for culture of Schwann cells in vitro. METHODS: Literature related to Schwann cell isolation, culture and purification was retrieved from PubMed, Web of Science, Medline databases, CNKI, Wanfang, VIP and other databases. The keywords included “Schwann cells; isolation; culture; purification”. All the papers obtained from the search were read, analyzed and judged, and finally 62 papers meeting the standards were included. RESULTS: The classical methods of Schwann cell culture include tissue block culture and enzyme digestion. Purification methods include mitosis resistance, immune selection, specific adhesion, pre degeneration, cold jet, differential adherent, laminin package, low serum, stimulating factor, fluorescence activated cell sorting or magnetic activated cell sorting, immunopanning, and extracorporeal shock wave treatment. CONCLUSION: Schwann cells can be cultured and purified in various ways in vitro, and each method has its advantages and disadvantages. How to obtain high-purity Schwann cells quickly and efficiently is still a challenge. Multiple methods can be combined for purification and affective factors can be controlled for Schwann cell proliferation as much as possible.

Current status of research on standardized management of specimens after radical gastrectomy for gastric cancer / 中华胃肠外科杂志

The quality control and standardization of procedures in radical gastrectomy for gastric cancer, especially the standardized processing of specimens after radical gastrectomy for gastric cancer, is very important. It is not only the basis of accurate pathological staging, but also the evidence of surgical quality and the original data of clinical research, which plays a pivotal role. The examination and classification of lymph nodes, specimens processing records, and data uploading and archiving after radical gastrectomy for gastric cancer are indispensable. It is necessary for surgeons to participate in the processing of surgical specimens. This article will combine the current research status and progress at home and abroad to review the standardized processing of specimens after radical gastrectomy for gastric cancer.

Advances of high-throughput screening system in reengineering of biological entities / 生物工程学报

Enzymes and cell factories are the core of industrial biotechnology. They play important roles in various fields such as medicine, chemical industry, food, agriculture, and energy. Usually, natural enzymes and cells need to be engineered to improve the catalytic efficiency, stability and enantioselectivity. Directed evolution makes it possible to rapidly improve the properties of enzymes and cell factories. Sensitive and reliable high-throughput screening approaches are the key for successful and efficient engineering of enzymes and cell factories. In this review, we first summarize the advantages and disadvantages of different screening methods and signal generation strategies as well as their application scope; we then describe the latest advances of ultra-high throughput screening technology applied in the directed evolution of enzymes and cell factories in the past three years. On this basis, we discuss the limiting factors that need to be further improved for high-throughput screening systems and forecast the future development trends of high-throughput screening methods, hoping that researchers in various fields including biotechnology and instrument development can cooperate closely to enhance the reliability and applicability of the high-throughput screening techniques.

Role and application of exosomes-mediated miRNAs in the treatment of glioma / 中国组织工程研究

BACKGROUND: Exosomes have become more and more popular in the field of tumor research in recent years. They can carry a large number of bioactive molecules directly to the receptor cells and participate in the. information exchange between cells. As the highest content and most kinds of non-coding RNA in exosomes, miRNAs play an important role in the proliferation, invasion, drug resistance and immunity of glioma. In addition, miRNAs have potential application value in the auxiliary diagnosis, treatment orientation and prognosis evaluation of glioma. OBJECTIVE: To describe the mechanism of exosome miRNAs sorting, the regulatory role in gliomas and its clinical application, to provide literature and theoretical basis for further exploring the relationship between exosome miRNAs and gliomas, and to exert positive significance in the diagnosis and treatment of gliomas. METHODS: The first author searched CNKI, WanFang database, PubMed database and EI database. The key words were “glioma, exosomes, miRNAs, sorting mechanism, biomarkers” in Chinese and English. Totally 118 literatures were retrieved. According to the inclusion and exclusion criteria, and literature supplement, totally 52 literatures were selected for summary and induction, mainly regarding the sorting mechanism of exosome miRNAs, the regulatory role in glioma and clinical application. RESULTS AND CONCLUSION: At present, the effective treatment of gliomas cannot meet the clinical needs even though a variety of high-intensity combined treatment schemes are adopted in this study. In this case, it is an important supplement for the clinical treatment of gliomas to propose the application of exosomes-mediated miRNAs in gene therapy of gliomas. Exosomes-mediated miRNAs cannot only play a regulatory role in the occurrence and development of glioma, invasion and metastasis, radiochemotherapy resistance, and immune regulation, but also play a potential role as a biomarker in the grading diagnosis, prognosis evaluation, treatment and other aspects of glioma. This will provide a solid theoretical basis for us to further explore the relationship between exosome miRNAs and glioma, and also has a positive clinical significance in the innovative diagnosis and treatment of glioma.

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.

Twin-arginine signal peptide of Bacillus licheniformis GlmU efficiently mediated secretory expression of protein glutaminase

Background: Protein glutaminase specifically deamidates glutamine residue in protein and therefore significantly improves protein solubility and colloidal stability of protein solution. In order to improve its preparation efficiency, we exploited the possibility for its secretory expression mediated by twin-arginine translocation (Tat) pathway in Bacillus licheniformis. Results: The B. licheniformis genome-wide twin-arginine signal peptides were analyzed. Of which, eleven candidates were cloned for construction of expression vectors to mediate the expression of Chryseobacterium proteolyticum protein glutaminase (PGA). The signal peptide of GlmU was confirmed that it significantly mediated PGA secretion into media with the maximum activity of 0.16 U/ml in Bacillus subtilis WB600. A mutant GlmU-R, being replaced the third residue aspartic acid of GlmU twin-arginine signal peptide with arginine by site-directed mutagenesis, mediated the improved secretion of PGA with about 40% increased (0.23 U/ml). In B. licheniformis CBBD302, GlmU-R mediated PGA expression in active form with the maximum yield of 6.8 U/ml in a 25-l bioreactor. Conclusions: PGA can be produced and secreted efficiently in active form via Tat pathway of B. licheniformis, an alternative expression system for the industrial-scale production of PGA.

Obtaining vacuolar protein sorting 34 gene in Sporothrix globosa by RACE method and analysis of its expression during the mycelium-to-yeast transition / 中华微生物学和免疫学杂志

Objective To obtain the full-length sequence of the vacuolar protein sorting 34 coding gene (vps34) of Sporothrix globosa (S. globosa) and to investigate the role of vps34 gene during the phase transition from mycelium to yeast in S. globosa. -ethods The 3′ end and 5′ end of the vps34 gene of S. globosa were amplified by rapid amplification of cDNA ends ( RACE ) . The obtained sequences were spliced and analyzed by bioinformatics software. Quantitative reverse transcription PCR ( qRT-PCR ) was used to analyze the expression of vps34 gene in mycelial and yeast phases. Results The vps34 gene of S. globosa was 3228 bp in length. The coding sequence was 3000 bp and encoded 999 amino acids with a mo-lecular mass of 111. 49×103 and an isoelectric point of 6. 38. It contained three domains including C2 PI3K class Ⅲ, PI3Ka Ⅲ and PI3Kc Ⅲ. The results of qRT-PCR showed that the expression of vps34 gene in yeast-phase S. globosa was higher than that in mycelial phase at 24 h (P<0. 05), and the greatest difference between them was observed at 48 h (P<0. 01). Conclusions Vps34p participates in the process of dimor-phic transformation of S. globosa. The obtainment of the full-length vps34 gene of S. globosa lays the founda-tions for further study on the function of Vps34p.

Application value of carbon nanoparticle labeled lymph node staining in radical resection of adenocarcinoma of esophagogastric junction with preoperative chemoradiotherapy / 中华消化外科杂志

Objective To explore the application value of carbon nanoparticle labeled lymph node staining in radical resection of adenocarcinoma of esophagogastric junction with preoperative chemoradiotherapy.Methods The retrospective cohort study was conducted.The clinicopathological data of 56 patients with adenocarcinoma of esophagogastric junction who underwent preoperative chemoradiotherapy in the Peking University Cancer Hospital from January 2014 to November 2017 were collected.There were 52 males and 4 females,aged from 22 to 76 years,with an average age of 62 years.Among 56 patients undergoing total gastrectomy and D2 lymphadenectomy,17 using carbon nanoparticle lymph node staining and 39 using traditional lymph node sorting were respectively allocated into observation group and control group.Observation indicators:(1) treatment situations;(2) detection of lymph nodes;(3) perioperative complications;(4) follow-up.Followup using outpatient examination and telephone interview was performed to detect tumor recurrence or metastasis up to May 2019.Measurement data with normal distribution were represented as Mean±SD,and comparison between groups was evaluated by the independent sample t test.Measurement data with skewed distribution were represented as M (range),and comparison between groups was evaluated by the Mann-Whitney U test.Count data were described as absolute numbers,and comparison between groups was analyzed using the chi-square test or Fisher exact propability.Comparison of ordinal data was analyzed using the nonparametric rank sum test.Results (1) Treatment situations:patients in both groups were successfully treated with concurrent chemoradiotherapy based on intensity modulated radiotherapy before operation.Radical gastrectomy with D2 lymphadenectomy was successfully performed after chemoradiotherapy,and Roux-en-Y esophagojejunostomy was used to reconstruct the digestive tract during operation.The operation time and volume of intraoperative blood loss were respectively (217± 58)minutes and (112±60)mL in the observation group,and (235±65) minutes and (119±77)mL in the control group,with no statistically significant difference between the two groups (t =1.017,0.341,P＞0.05).(2) Detection of lymph nodes:the average number of harvested lymph nodes,average number of radiation target lymph nodes,and average number of peritarget lymph nodes were respectively 32± 10,21±8,and 7±4 in the observation group,and 22±7,16±5,5±3 in the control group,with statistically significant differences between the two groups (t=4.138,2.881,2.401,P＜0.05).The median number of positive lymph nodes harvested,median number of positive radiation target lymph nodes,and median number of positive peritarget lymph nodes were respectively 0 (range,0-2),0 (range,0-2),and 0 (range,0-0) in the observation group,and 0 (range,0-7),0 (range,0-3),and 0 (range,0-1) in the control group,showing no statistically significant difference between the two groups (Z=1.305,1.101,0.660,P ＞ 0.05).(3) Perioperative complications:6 and 18 patients in the observation group and the control group had complications,respectively,with no statistically significant difference between the two groups (x2=0.570,P＞0.05).Patients with complications were improved after drug treatment and local treatment without second operation.No local or systemic adverse reactions caused by carbon nanoparticles was observed during and after operation in the observation group.(4) Follow-up:56 patients were followed up for 5-65 months,with a median follow-up time of 32 months.There were 14 and 6 patients in the observation group and the control group with tumor recurrence or metastasis,respectively,showing no significant difference between the two groups (x2 =0.002,P＞0.05).Conclusion Carbon nanoparticle labeled lymph node staining in radical resection of adenocarcinoma of esophagogastric junction with preoperative chemoradiotherapy can increase the number of harvested lymph nodes.