Objective : To construct triggering receptor expressed on myeloid cells 2(TREM2) gene knockout(TREM2-/-) mice using CRISPR/Cas9 technology, to breed TREM2-/- mice and to analyze the genotype of TREM2-/- mice. Methods : CRISPR/Cas9 technology was used to selectively knock out exon 2-3 regions of TREM2 gene to construct a TREM2-/- mouse model, and the genetic background of all mice was C57BL/6J. Polymerase chain reaction(PCR) was used to identify the genotype of mice. Quantitative real-time PCR(qPCR) and Western blot were used to detect the expression level of TREM2 in major tissues of mice, and the authenticity and scientific nature of PCR identification results were verified from mRNA level and protein level. According to the sgRNA sequence, the possible off-target sites were predicted on the CCTop website, and the tail DNA of mice was extracted and amplified by PCR and then Sanger sequencing was performed to detect whether there was off-target effect in TREM2-/- mice. Results : TREM2-/- mice were successfully constructed by CRISPR/Cas9 technology, and the mice were genotyped. The results of agarose gel electrophoresis showed that the mouse genotype with only 415 bp band amplified was wild type(WT), the mouse genotype of the 449 bp band amplified only was TREM2-/-, and the mouse genotypes amplified with 415 bp and 449 bp double bands were heterozygous. qPCR results showed that compared with WT mice, the mRNA expression of TREM2 in heart and brain tissues of TREM2-/- mice was down-regulated(P-/- mice was reduced(P-/- mice. Conclusion TREM2-/- mice are successfully constructed and bred, a reliable genotype identification method is established, the genetic stability of the mouse model is verified, which will provide an important genetic animal model for the study of TREM2 gene function.

To explore how the joint-teaching mode of tomographic-radiologic anatomy and topographic anatomy in a way of case-based learning (CBL) promotes the education of medical students.

177Lu-prostate specific membrane antigen(PSMA)radio-ligand therapy has been approved abroad for advanced prostate cancer and has been in several clinical trials in China.Based on domestic clinical practice and experimental data and referred to international experience and viewpoints,the expert group forms a consensus on the clinical application of 177Lu-PSMA radio-ligand therapy in prostate cancer to guide clinical practice.

68Ga-DOTATATE/18F-FDG two-day low-dose total-body PET/CT imaging is increasingly employed to facilitate the diagnosis,prognosis,and heterogeneity assessment of neuroendocrine neoplasms.We present a consensus on operational guideline for a two-day combined imaging from experts in low-dose/ultra-low-dose total-body PET/CT from several domestic medical institutions.

Objective:To investigate correlation between neutrophil lymphocyte ratio(NLR),IL-6 and lymphocyte subsets and prognosis of viral pneumonia complicated with acute respiratory distress syndrome(ARDS).Methods:A total of 98 patients with viral pneumonia complicated with ARDS admitted to Xinxiang Central Hospital/the Fourth Clinical College from May 2017 to June 2019 were selected,NLR,IL-6 and lymphocyte subsets expressions in patients with different severity were analyzed.Correlation between NLR,IL-6 and lymphocyte subsets levels in prognosis of viral pneumonia complicated with ARDS was analyzed by Pearson correlation.Logistc regression was used to analyze prognostic risk factors of viral pneumonia complicated with ARDS.Results:With aggravation of viral pneumonia complicated with ARDS,NLR and IL-6 levels were significantly increased(P<0.05),while CD3+T,CD4+T and CD19+T cells levels were significantly decreased(P<0.05).Pearson correlation analysis results showed that NLR and IL-6 were positively correlated with prognosis of viral pneumonia complicated with ARDS(P<0.05),while CD3+T,CD4+T,CD19+T and CD8+T cells were negatively correlated with prognosis of ARDS with viral pneumonia(P<0.05).Multivariate Logistic analysis showed that NLR,IL-6,CD3+T and CD19+T cells were risk factors for viral pneumonia complicated with ARDS(P<0.05).Conclusion:Serum NLR,IL-6,CD3+T and CD19+T cells are closely related to severity and prognosis of patients with viral pneumonia complicated with ARDS.

Fibroblast activation protein inhibitors(FAPI)can specifically bind to fibroblast activation protein,which is highly expressed in tumor stroma,and 68Ga-FAPI do not get involved into the procedure of glucose metabolism in tumor cells.68Ga-FAPI PET/CT and PET/MR can be used for displaying malignant tumors at various sites and play a unique role for diagnosis and differential diagnosis.The research progresses of 68 Ga-FAPI-based PET/CT and PET/MR for diagnosing malignant tumors were reviewed in this article.

Objective:To explore the value of the imaging nomogram model based on preoperative CT features of patients with small intestinal stromal tumor (SIST) in predicting pathological risk classification.Methods:This was a cohort study. The patients who were diagnosed as primary SIST by postoperative pathology in Cancer Hospital, Chinese Academy of Medical Sciences from January 2014 to October 2023 were retrospectively included. According to the modified 2008 National Institutes of Health classification criteria, the patients were divided into a pathological intermediate/high-risk group (86 cases) and a very low/low-risk group (56 cases). The features of preoperative enhanced CT images of SIST were analyzed, including tumor boundary, necrosis, intra-tumoral hemorrhage, intra-tumoral calcification, growth pattern, enhancement pattern, enhancement degree, enlarged vessels feeding or draining the mass (EVFDM), and tumor location. Patients were followed up to determine the recurrence-free survival (RFS). Univariate and multivariate logistic regression were used to screen the independent predictors of SIST with pathological medium/high-risk group. The independent predictors were combined to construct an imaging prediction model, and a nomogram was drawn. The receiver operating characteristic curve was used to evaluate the predictive efficacy of the model. The Kaplan-Meier method was used to draw the survival curve, and the log-rank test was used to compare the differences in RFS.Results:Univariate logistic regression results showed that tumor shape, necrosis, intra-tumoral hemorrhage, EVFDM, and tumor location were potentially related to medium/high-risk SIST. Multivariate logistic regression results showed that tumor shape ( OR=3.92, 95% CI 1.58-9.71, P=0.003), necrosis ( OR=4.60, 95% CI 1.91-11.09, P<0.001), and EVFDM ( OR=6.25,95% CI 1.74-22.47, P=0.005) were independent predictors of pathological intermediate/high-risk SIST. The area under the curve of the imaging predictive model combining the three predictors to predict the intermediate/high-risk SIST was 0.835 (95% CI 0.769-0.901), the sensitivity was 0.810, the specificity was 0.839, and the accuracy was 0.789. Taking the cut-off value (0.810) as the boundary value, the patients were divided into the high-risk group (74 cases) and the low-risk group (68 cases) according to the prediction results. The median RFS of the predicted high-risk group was poorer than that of the predicted low-risk group, and the difference was statistically significant ( χ2=5.20, P=0.023). Conclusions:The imaging nomogram model based on preoperative CT image features shape, necrosis, and EVFDM can effectively predict the pathological intermediate/high-risk SIST before surgery and has important predictive value for postoperative recurrence.

Objective:To compare the predictive values of response evaluation criteria in lymphoma (RECIL)2017 and Lugano classification for the prognosis of patients with diffuse large B-cell lymphoma(DLBCL) at the end of treatment.Methods:A total of 107 patients (50 males, 57 females, age (49.3±17.4) years) with DLBCL who underwent PET/CT at the end of chemotherapy in the Fourth Hospital of Hebei Medical University between February 2014 and December 2021 were analyzed retrospectively. The RECIL2017 and Lugano classification were used to evaluate the response. Kaplan-Meier survival analysis was used to evaluate progression-free survival (PFS) and overall survival (OS). The Kappa test was used to evaluate the consistency of the two criteria after chemotherapy, and ROC curve (Delong test)was used to compare the predictive values of the two criteria for PFS and OS. Results:The median follow-up time was 47.5(33.4, 57.5) months. Kaplan-Meier analysis showed that the 5-year PFS rates (74.5%(35/47), 71.4%(15/21), 57.1%(12/21), 4/18; χ2=38.85, P<0.001) and OS rates (89.4%(42/47), 81.0%(17/21), 61.9%(13/21), 7/18; χ2=29.52, P<0.001) in complete metabolic response (CMR), partial metabolic response (PMR), no metabolic response (NMR) and progressive metabolic disease (PMD) groups evaluated by Lugano classification were statistically different, as well as those in complete response (CR), partial response (PR), minor response (MR), stable disease (SD) and progressive disease (PD) groups evaluated by the RECIL2017 (5-year PFS rates: 76.9%(40/52), 8/12, 6/11, 6/12, 30.0%(6/20), χ2=29.05, P<0.001; 5-year OS rates: 90.4%(47/52), 8/12, 6/11, 9/12, 45.0%(9/20), χ2=23.63, P<0.001). The RECIL2017 and Lugano classification had good consistency in the efficacy evaluation of DLBCL patients at the end of chemotherapy (70.1%(75/107); Kappa=0.57, P<0.001). The AUCs of Lugano classification for predicting PFS and OS were 0.730 (95% CI: 0.625-0.834, P<0.001) and 0.908 (95% CI: 0.845-0.970, P<0.001) respectively, and those of RECIL2017 were 0.717 (95% CI: 0.612-0.822, P<0.001) and 0.880 (95% CI: 0.812-0.949, P<0.001). The AUCs of the Lugano classification for PFS and OS were slightly higher than those of RECIL2017, without significant differences ( z values: 0.44, 1.09, both P>0.05) . Conclusion:Both RECIL2017 and Lugano classification can evaluate the prognosis of patients with DLBCL at the end of treatment, and Lugano classification is more accurate.

Production internship is an important teaching tache for undergraduate students to carry out engineering training by using professional skills, and it is a key starting point for fostering application-oriented talents in biotechnology. The Course Group of 'production internship of biotechnology majors' of Binzhou University is investigating application-oriented transformation for local regular colleges and universities, as well as fostering high-level application-oriented talents. By taking green fluorescent protein (GFP) polyclonal antibody as an example, the reform and practice on teaching content, teaching mode, assessment method, continuous improvement of curriculum were carried out. Moreover, the characteristics of the Yellow River Delta-Binzhou Biotechnology & Pharmaceutical Industrial Cluster were taken into account to intensify academic-enterprise cooperation. On one hand, this Course Group designed and rearranged the course contents, carried out essential training through online resources and platforms such as virtual simulation, and recorded, tracked and monitored the progress of production internship through practical testing and software platforms like 'Alumni State'. On the other hand, this Course Group established a practice-and application-oriented assessment method in the process of production internship and a dual evaluation model for continuous improvement. These reform and practices have promoted the training of application-oriented talents in biotechnology, and may serve as a reference for similar courses.
Humans ; Internship and Residency ; Curriculum ; Students ; Biotechnology

Objective To analyze the microflora structure and abundance of ileum and colon of SPF male SD rats by high-throughput sequencing technique. MethodsThe ileal and colonic lumen contents of 30 SPF male SD rats were collected, the total bacterial DNA in the contents was extracted and amplified by PCR. The V3-V4 region of bacterial 16S rRNA in the samples was sequenced using Illumina NovaSeq sequencing platform. Then the species structure and abundance of the intestinal flora were analyzed at the phylum and genus level based on validated data. The diversity and differences between ileal and colonic flora were analyzed using the QIIME software (amplicon) analysis tool. The Tax4Fun program was applied to predict the dominant gene enrichment pathways of ileal and colonic flora. ResultsAt the phylum level, the dominant microflora in the ileum of male SD rats were Firmicutes and Actinobacteria, accounting for more than 98%. The dominant colonic microbial community were mainly Firmicutes and Bacteroidetes, accounting for more than 95%. At the genus level, Lactobacillus and Alistipes were the predominant bacteria in the ileum. The colon was dominated by Lactobacillus and Romboutsia. In terms of flora diversity, the alpha diversity of colonic flora including richness index (Chao1 index) and diversity index (Shannon index) were significantly higher than those of ileum (P < 0.01), and the structural variability of the composition of its flora species was smaller than that of ileum; the ileal flora with significant structural variability were mainly of the phylum Firmicutes, Romboutsia, Peptostreptococcaceae, while the colonic flora had significant structural differences in Bacteroidales. In terms of flora function, the ileal flora dominant genes were significantly enriched in lipid-like metabolism, polyketide metabolism, membrane transport, biodegradation and other pathways, while the colonic flora dominant genes were significantly enriched in glycan biosynthesis metabolism, energy metabolism, biosynthesis of cofactor and vitamins and other products, and other pathways. ConclusionThere are significant differences in structure and abundance between the ileal and colonic flora of SPF male SD rats, and the abundance and diversity of colonic flora are higher than those of ileal flora.