β-glucosidase has important applications in food, pharmaceutics, biomass conversion and other fields, exploring β-glucosidase with strong adaptability and excellent properties thus has received extensive interest. In this study, a novel glucosidase from the GH1 family derived from Cuniculiplasma divulgatum was cloned, expressed, and characterized, aiming to find a better β-glucosidase. The amino acid sequences of GH1 family glucosidase derived from C. divulgatum were obtained from the NCBI database, and a recombinant plasmid pET-30a(+)-CdBglA was constructed. The recombinant protein was induced to express in Escherichia coli BL21(DE3). The enzymatic properties of the purified CdBglA were studied. The molecular weight of the recombinant CdBglA was 56.0 kDa. The optimum pH and temperature were 5.5 and 55 ℃, respectively. The enzyme showed good pH stability, 92.33% of the initial activity could be retained when treated under pH 5.5-11.0 for 1 h. When pNPG was used as a substrate, the kinetic parameters K_m, V_max and K_cat/K_m were 0.81 mmol, 291.99 μmol/(mg·min), and 387.50 s^-1 mmol^-1, respectively. 90.33% of the initial enzyme activity could be retained when CdBglA was placed with various heavy metal ions at a final concentration of 5 mmol/L. The enzyme activity was increased by 28.67% under 15% ethanol solution, remained unchanged under 20% ethanol, and 43.68% of the enzyme activity could still be retained under 30% ethanol. The enzyme has an obvious activation effect at 0-1.5 mol/L NaCl and can tolerate 0.8 mol/L glucose. In conclusion, CdBglA is an acidic and mesophilic enzyme with broad pH stability and strong tolerance to most metal ions, organic solvents, NaCl and glucose. These characteristics may facilitate future theoretical research and industrial production.
beta-Glucosidase ; Sodium Chloride ; Temperature ; Glucose ; Ethanol/chemistry* ; Ions ; Hydrogen-Ion Concentration ; Enzyme Stability ; Substrate Specificity

Objective:To discuss the development and management of full-time research assistants in China′s large hospitals.Methods:Through the methods of policy sorting, literature research, summarization, and classification, this study focused on the policy background, development opportunities, and the needs of the times for the position establishment of research assistants in China′s large hospitals, comprehensively classified the practical exploration and existing problems of employing and managing full-time research assistants in typical large hospitals in the country, and discussed the relevant management strategies learned from international experience.Results:In addition to hospitals directly affiliated with colleges and universities, dozens of large local hospitals in more than ten provinces have carried out beneficial practices of establishing the employment, management, and assessment systems for research assistants positions and achieved some progress. However, there are still common problems such as limited cognition of ″temporary positions″, insufficient coverage of the positions, low salary and welfare, and lack of training and development mechanisms as well as promotion and career development paths.Conclusions:The development and management of research assistants in large hospitals have gradually shifted their purpose from alleviating the employment issue of the graduates to reserving talented people in medical teaching, researching and managing, forming a high-quality, professional, and full-time medical scientific team, and continuously innovating. This article puts forward some suggestions about establishing the management system of research assistants, strengthening the standardized training for talented people, and constructing a diversified incentive mechanism combining performance appraisal and target management for research assistants.

Objective:To analyze the current situation of human genetic resources management in a tertiary hospital, and propose improvement suggestions for the management of human genetic resources in tertiary hospitals.Methods:According to the review of laws and regulations of human genetic resources management in China, analyze the current situation, challenges and management suggestions of human genetic resources management in this particular tertiary hospital, and further management suggestions are proposed.Results:Main problems were identified in the tertiary hospital, including the knowledge of researchers needs to be improved, and due to lack of supervision and management on process of human genetic resources projects, the management of investigator initiated clinical trials needs to be further strengthened.Conclusions:The tertiary hospital needs to strengthen the researcher training on the management of human genetic resources, establish the whole-process supervision of human genetic resources project and pay more attention to the management of investigator initiated clinical trials.

Objective:To evaluate the therapeutic effect of 177Lu-prostate specific membrane antigen (PSMA)-I&T on prostate cancer. Methods:The culture medium of 1.85, 18.50, 185.00, 555.00 and 925.00 MBq/L 177Lu-PSMA-I&T was added into LNCaP cells (200 μl/well, 5 experimental groups and 1 control group, 3 replicates in each group) for 24 h, and the cell viability in each group was detected. The culture medium of 3.7 MBq 177Lu-PSMA-I&T was added into LNCaP cells (1 experimental group, 1 control group, 3 replicates in each group) for 48 h to detect the changes of cell cycle. LNCaP cells (3 experimental groups and 1 control group, 3 replicates in each group) were added into the culture medium of 3.7, 19.5 and 37.0 MBq 177Lu-PSMA-I&T for 48 h to detect cell apoptosis. Tumor-bearing mice models were established (BALB/c-nu/nu nude mice, n=32). The changes of tumor volume and body mass of tumor-bearing mice were observed within 20 d after treatment. On the 7th day after treatment, tumor tissues of tumor-bearing mice were stained with HE staining and fluorescently stained with Ki-67 protein, and apoptosis was detected by TdT-mediated dUTP nick end labeling (TUNEL) assay. On the 20th day after treatment, pathological analysis was performed on the main viscera of the tumor-bearing mice. One-way analysis of variance, the least significant difference t test and paired t test were used to analyze the data. Results:Compared with the control group ((100.00±12.35)%), the cell survival rates were significantly decreased after 177Lu-PSMA-I&T intervention in 185.00, 555.00, 925.00 MBq/L groups ((57.56±6.35)%, (38.65±3.39)%, (27.95±4.48)%; F=78.91, t values: 8.312-14.106, all P<0.01). Cell survival rates were significantly reduced in 185.00 MBq/L group at different time points (24, 48 and 72 h; F=78.28, t values: 6.628-14.384, all P<0.01). The proportion of LNCaP cells in G2/M phase was increased from (12.36±0.28)% to (19.92±0.48)% ( t=17.180, P<0.01). The apoptosis rates of cells were significantly increased in 18.5 and 37.0 MBq groups ( F=71.86, t values: -6.138, -13.050, both P<0.05). The difference of relative tumor volume (RTV%) was statistically significant among 3.7, 14.8 and 29.6 MBq groups and control group (136.7±7.4, 59.2±23.8, 47.3±13.8 vs 240.3±3.7; F=78.20, t values: 7.549-13.345, all P<0.01). But there was no significant difference in body mass of tumor-bearing mice among groups. Compared with the control group, the positive rates of Ki-67 staining cells ((37.23±3.04)% vs (14.89±3.80)%, (5.60±1.83)%, (3.46±0.71)%) and TUNEL-fluorescein isothiocyanate (TUNEL-FITC) staining ((0.74±0.18)% vs (1.61±0.30)%, (3.19±0.44)%, (3.54±0.47)%) in tumor tissues of 3.7, 14.8 and 29.6 MBq groups were statistically significant ( F=103.91, t values: 10.429-15.762; F=38.66, t values: from -9.312 to -2.881, all P<0.01). Conclusions:177Lu-PSMA-I&T has a good therapeutic effect on prostate cancer, with no obvious therapeutic side effects. Therefore, 177Lu-PSMA-I&T is expected to be an ideal drug for treating prostate cancer.

To investigate whether the laboratory specimens preserved in Beijing Hospital Biobank during a specific period had been contaminated by SARS-Cov-2 through a cross-sectional study, and to establish a retrospective biobank safety screening system. Laboratory specimens were collected from the Department of Respiratory and Critical Care Medicine and the Fever Clinic of Beijing Hospital from November 1, 2019 to January 22, 2020, nucleic acid and serological antibody testing were performed for SARS-CoV-2 in these specimens (including 79 serum, 20 urine, 42 feces and 21 bronchoalveolar lavage fluid specimens). The safety of the stored samples during this period was defined by negative and positive results. Both the nucleic acid test and serological antibody test showed negative for SARS-CoV-2, indicating that these specimens were safely stored in the biobank. High-risk specimens collected in our hospital during the early stage of the COVID-19 outbreak are free of SARS-CoV-2, and a safety screening strategy for the clinical biobank is established to ensure the biosafety of these samples.
Biological Specimen Banks ; COVID-19 ; Cross-Sectional Studies ; Hospitals ; Humans ; Retrospective Studies ; SARS-CoV-2

Objective:To build a scientifically applicable scientific research evaluation system based on the Science and Technology Evaluation Metrics (STEM) of Chinese hospitals.Methods:Twenty-four experts were invited to form an expert advisory group, scientific research evaluation indicators were collected according to literature review, panel discussion and expert consultation. Use Delphi method to carry out two rounds of expert consultation, calculate the authority level and positive factor of experts, and use AHP to calculate the weight of each indicator.Results:The experts invited for consultation had a high positive coefficient and a high degree of expert authority, and finally, a scientific research evaluation index system including 3 first-level indicators, 10 second-level indicators and 24 third-level indicators was established.Conclusions:The research based on STEM has developed an evaluation system well combined quantity and quality from three dimensions, including scientific research input, scientific research output and scientific research influence. Taking into full consideration of the regulatory requirement of clinical trials and the research administration of research achievements transformation, this research established an evaluation system and management model applicable to the Tertiary class A general hospitals, provided reference value for the development of the hospital's scientific research.

Objective: To explore the main layout and countermeasures of Scientific research projects during the public health emergency of COVID-19. Methods: Literature investigation method is applied to collect information of scientific research and emergency research projects of COVID-19 funded since January 20th by different entities including national, provincial and municipal administrative departments, public health agencies, research institutes, universities and industries. Results: Along with priorities identified for the emergency response and key research agendas, the national science and technology authorities at all levels and scientific research institutions have deployed a series of new science and technology projects, as well as a number of supporting policies and measures. Conclusions In the campaign of science and technology to deal with COVID-19, the leadership of Chinese government and their coordination with local agencies facilitated quick response in both R&D investment and supporting policies. Periodical achievement is recognized so far, and with the continuous development and in-depth research, the related scientific research results will be gathered into the core force of epidemic prevention. Moreover, it will improve China's capacity to deal with health emergencies and the level of medical health innovation, and better protect the health of the people.

Objective To investigate the fetal management of prenatally diagnosed fetal mediastinal masses and the initial experience of neonatal thoracoscopic minimally invasive treatment. Methods We per-formed a retrospective study from November 2015 to November 2016 of all newborns affected by mediastinal masses and treated by thoracoscopic surgery. This group of cases were found with mediastinal masses by pre-natal ultrasound. The earliest detection of abnormal time was 16 to 31 weeks of pregnancy,with an average of 25 weeks. In the fetal period,the patients were treated with multidisciplinary consultation and individual man-agement. Prenatal examinations helped us except for chromosomal abnormalities and other organ abnormali-ties. After birth,the patients underwent CT and MRI examination. The diameter of the tumor was 1. 7 to 5. 7 cm,with an average of 3. 2 cm. The operative age was 4 to 29 days,with an average of 12. 4 days. This group of newborns were performed thoracoscopic mass resection and confirmed by intraoperative pathological exam-ination. Results After individualized precise prenatal management,all children were born successfully and confirmed that prenatal diagnosis was accurate. All mediastinal masses were completely excised in the neo-natal period. Five mediastinal masses were completely excised. One posterior mediastinum immature teratoma was converted to open thoracotomy. The mean operative duration was 112 min(100 to 150 min). There was no operative complication with a minimal amount of blood loss. With a smooth recovery,the hospital stay was 11-17 days. Pathological results included:1 esophageal duplication,2 bronchogenic cysts,1 lymphangioma, 1 cystic teratoma of anterior mediastinum,1 immature teratoma of posterior mediastinum. During a mean fol-low-up period of 8-14 months,neither complication nor recurrence occurred. Conclusion These are the pre-conditions for early treatment of neonatal patients with mediastinal masses,including definite prenatal diagno-sis,multidisciplinary consultation system and individualized and accurate fetal management. Throcoscopic ex-cision of mediastinal masses is both feasible and safe in neonates. Proper preoperative case selection may pre-vent a conversion into thoracotomy due to huge solid mass.

[Abstract] Objective: To establish a chimeric antigen receptor（CAR）modified T cells specifically targeting CD19 molecule (CD19CAR-T cells) and to testify their in vitro killing effect on target cells. Methods: CD19-CAR fragments yielded by PCR were constructed into pCDH-GFP lentiviral vectors by molecular cloning technology. The packaged lentiviral particles were transducted into CD3+ T cells of donors. Transduction efficiency was measured by flow cytometry and PCR. The in vitro cytotoxicity of obtained CD19CAR-T cells against CD19+ Ramos cells was tested by 7-AAD staining. Results: The amplification folds of CD3+ T cells increased to (78.8± 23.2) folds after in vitro culture for 10 days, and about (58.3±5.4)% cells expressing GFP.About (57.4±9.3)% CD19+Ramos cells were specifically killed by the CD19-CAR-T cells in vitro at the E∶T ratio of 5∶1. Conclusion: This study successfully established an effective method for constructing and amplifying CD19-CAR-T cells in vitro, which showed profound efficiency and specific cytotoxity against CD19+ Ramos cells.And this report might provide an experimental evidence for clinical treatment of CD19+ B cell neoplasmas.

Objective To investigate the effects and mechanism of interleukin 33 (IL-33) on renal tubular injury in mice with lupus nephritis.Methods Twelve-week-old female MRL/lpr mice were randomly divided into model group,IL-33 group and solvent control group with 10 rats in each group.Ten female MRL/MP mice of the same age were used as normal control group.The mice in IL-33 group were intraperitoneally injected with 100 μL of phosphate buffer saline (PBS),containing 2 μg of recombinant mouse IL-33,once a day for 14 days.The mice in control group and the model group were intraperitoneally injected with the same dose of PBS.All the mice were sacrificed at 14 weeks of age.Serum creatinine (Cr) and urea nitrogen (BUN) concentrations were determined by serum separation.The urine in 24 hours was collected testing urinary protein creatinine ratio and urinary protein quantification.The contents of E-cadherin,α-SMA,and JAK/STAT pathway signaling proteins,including JAK2,p-JAK2,STAT1,and p-STAT1,were detected by Western blot.Results The BUN,urinary protein creatinine ratio and urine protein level of the IL-33 group were significantly higher than those of the model group (all P＜0.05).The expression of renal tubular epithelial cells o-SMA in the IL-33 group was higher than that in the model group,and the difference was statistically significant (P＜0.05).Compared with the model group,the expression of E-cadherin in the tubular epithelial cells of IL-33 group decreased and the expression of p-JAK2 and p-STAT1 protein increased,and the differences were all statistically significant (all P＜0.05).Compared with the model group,the levels of JAK2 and STAT1 in IL-33 group change little,and the differences were not statistically significant (all P＞0.05).Conclusions IL-33 can cause tubulointerstitial lesions in lupus mice,and its mechanism may be related to the activation of JAK/STAT pathway.