[Objective] To explore the feasibility of using whole blood as a source of NK cells for allogeneic CAR NK cell therapy and activated NK cell reinfusion therapy, and initially construct a technical system for the separation and purification of NK cells from whole blood. [Methods] All peripheral blood mononuclear cells (PBMCs) were enriched from 400 mL of whole blood by manual separation and machine separation, respectively. The erythrocyte loss rate, PBMCs number, NK cell purity of the two methods were compared. NK cells were sorted from PBMCs by three separation and enrichment methods as immunomagnetic bead negative selection method, platelet lysate culture expansion and PERCOLL density gradient separation method, and the purity and yield of NK cells, the activity of NK cells and the tumor-killing ability of the three separation and enrichment methods were compared. [Results] The proportion of NK cells in the lymphocyte population was higher in the manual separation method than in the machine separation method[(13.16±5.16)% vs (8.56±3.92)%, P<0.05]; the number PBMCs was lower in the manual separation method than in the machine separation method[(4.09±1.80)×10⁸vs (6.49±2.16)×10⁸, P<0.05], and there was no difference in the red blood cell loss between the two methods (P>0.05). The purity of NK cells isolated and enriched from PBMCs by manual separation method using immunomagnetic was (96.77±2.31)%; the yield was (56.27±10.47)%; the inhibition of tumor proliferation was (38.67±14.05)%; and the tumor killing rate was (19.90±8.05)%. The purity of NK cells isolated and enriched from PBMCs by manual separation method using platelet lysis culture expansion method was the highest at day 7, which was (54.84±15.80)%; the cell expansion multiple could reach 16.92±6.28 at day 7; the in vitro tumor killing rate of NK cells was (15.83±5.5)%; the tumor inhibition rate was (44.33±13.5)%; and there was no difference in the toxicity and activity of NK cells between the two methods (P>0.05). The purity of NK cells isolated and enriched by PERCOLL density gradient separation method was (15.83±5.82)%, and the yield was (14±6.25)%, which was significantly lower than the other two methods. [Conclusion] PBMCs isolated from whole blood by manual separation and NK cells enriched by negative selection with immunomagnetic beads have the potential to provide NK cell materials for CAR-NK cell therapy, and NK cells enriched by platelet lysate-conditioned medium have the potential to provide NK cells for large-scale NK cell activation reinfusion therapy.

OBJECTIVE To evaluate the efficacy， safety and cost-effectiveness of dupilumab in the treatment of severe asthma with rapid health technology assessment （HTA）， and to provide evidence-based evidence for clinical treatment. METHODS Retrieved from PubMed，Cochrane Library，CNKI，Wanfang， VIP database， and other related websites of HTA. HTA reports， systematic review/meta-analysis， and economic studies of dupilumab in the treatment of severe asthma were collected. Researchers independently identified literature， extracted data， and assessed the quality of included studies. Qualitative description was performed. RESULTS A total of 15 pieces of literature were included， involving 9 systematic reviews/meta-analyses and 6 economic studies. In terms of effectiveness， dupilumab was significantly better than placebo. Compared with other biological drugs， in the patients with severe asthma aged 12 years and above， for those with eosinophil （EOS） ≥300 cells/μL， dupilumab ranked first in improving forced expiratory volume in one second （FEV1）， outperforming tezepelumab， benralizumab， and mepolizumab. It ranked second in reducing acute exacerbations， surpassed only by tezepelumab， while its effect on improving asthma control questionnaire score was relatively lower， being better only than benralizumab. For those with 150 cells/μL ≤EOS＜300 cells/μL， dupilumab was superior to mepolizumab in reducing asthma exacerbation， while the effect on FEV1 was weaker than benralizumab and mepolizumab. In terms of safety， there was no significant difference in the incidence of adverse events and serious adverse events between dupilumab and placebo or other biological drugs， while the incidence of injection site reactions of dupilumab was significantly higher than placebo. In terms of cost-effectiveness， the research results of different countries were not consistent， and there was a lack of research data from China. CONCLUSIONS Dupilumab is an effective and safe choice in the treatment of severe asthma， and its cost-effectiveness requires further research based on China’s medical environment to be determined.

Recent studies have indicated that the expression of ubiquitin-specific protease 51 (USP51), a novel deubiquitinating enzyme (DUB) that mediates protein degradation as part of the ubiquitin‒proteasome system (UPS), is associated with tumor progression and therapeutic resistance in multiple malignancies. However, the underlying mechanisms and signaling networks involved in USP51-mediated regulation of malignant phenotypes remain largely unknown. The present study provides evidence of USP51's functions as the prominent DUB in chemoresistant triple-negative breast cancer (TNBC) cells. At the molecular level, ectopic expression of USP51 stabilized the 78 kDa Glucose-Regulated Protein (GRP78) protein through deubiquitination, thereby increasing its expression and localization on the cell surface. Furthermore, the upregulation of cell surface GRP78 increased the activity of ATP binding cassette subfamily B member 1 (ABCB1), the main efflux pump of doxorubicin (DOX), ultimately decreasing its accumulation in TNBC cells and promoting the development of drug resistance both in vitro and in vivo. Clinically, we found significant correlations among USP51, GRP78, and ABCB1 expression in TNBC patients with chemoresistance. Elevated USP51, GRP78, and ABCB1 levels were also strongly associated with a poor patient prognosis. Importantly, we revealed an alternative intervention for specific pharmacological targeting of USP51 for TNBC cell chemosensitization. In conclusion, these findings collectively indicate that the USP51/GRP78/ABCB1 network is a key contributor to the malignant progression and chemotherapeutic resistance of TNBC cells, underscoring the pivotal role of USP51 as a novel therapeutic target for cancer management.

Objective:To evaluate the therapeutic effect of glycyrrhetinic acid on cough variant asthma (CVA) mice based on molecular docking technique; To explore the possibility of its treatment for cough variant asthma.Methods:The software of Autodock Vina was used for molecular docking. The mice were divided into control group, model group, prednisone acetate group, glycyrrhetinic acid high-, medium-, and low-dosage groups according to the random number table method, with 8 mice in each group. Except for the blank control group, all other groups were induced by egg protein to establish cough variant asthma models. Glycyrrhetinic acid high-, medium-, and low-dosage groups were orally administered glycyrrhetinic acid suspension at 20, 10, and 5 mg/kg, while the prednisone acetate group was orally administered prednisone acetate at 5 mg/kg. The blank control group and model group were orally administered equal volumes of physiological saline, once per day for 14 consecutive days. The animal asthma behavior was observed after drug administration. The secretion of bronchial mucus in lung tissue were observed by AB-PAS staining and the index of spleen were recorded. The protein expressions of Gata3, IL-4 and IL-13 in the spleen tissue were determined by Western blot.Results:Molecular docking results showed that glycyrrhetinic acid had good binding ability to Th2-related factors Gata3, IL-4 and IL-13. Results of animal experiment showed that compared with the model group, the mucus secretion decreased in glycyrrhetinic acid groups, the index of the spleen of mice obviously decreased, protein expression levels of IL-4 and IL-13 in the spleen tissue of mice in glycyrrhetinic acid high-, medium-, and low-dosage groups decreased ( P<0.05), and Gata3 in glycyrrhetinic acid medium- and low-dosage groups decreased ( P<0.05). Conclusion:Glycyrrhetinic acid can correct the shift of Th2 in the immune system of cough variant asthma mice and has a certain therapeutic effect.

Practicing the great educational ideology of establishing morality and cultivating people in classroom teaching is a strategic measure for medical colleges and universities to improve the quality of ideological and political education and personnel training.At the juncture of its transition from the exploration stage to the normalization stage,it is necessary to understand the current situation,summarize the experience,as well as discover and solve the problems faced by sustainable development through investigation and research.The empirical research mainly focuses on two related aspects.Firstly,for the stage of lesson preparation,examining and studying the design description of teaching and educating people in randomly selected teaching plans to understand the current situation of teachers'learning and understanding of the concept of establishing morality and cultivating people.Secondly,for the implementation of classroom teaching,utilizing the opportunity of peer listening and supervising class evaluation to conduct on-site inspections and record relevant data,so as to further understand the teacher's concept of establishing morality and cultivating people and their current situation of integrating into classroom.Data statistical analysis and on-site investigation indicated that the exploration of practicing the concept of establishing morality and cultivating people in classroom teaching was generally successful,but there were also some shortcomings.To turn to the normalization stage,it is urgent to solve three major problems,namely,the need to deeply learn and understand the ideology of establishing morality and cultivating people,the need to carefully design teaching ideas of establishing morality and cultivating people in teaching plans,and the need to careful optimization of implementing measures for establishing morality and cultivating people in classroom teaching.

Objective:To evaluate alteration of autograft function by pathological types after total parathyroidectomy (tPTX) plus autotransplantation (AT) in patients of hyperthyroidism.Methods:A total of 51 patients with end-stage chronic renal failure who underwent total parathyroidectomy with autologous forearm transplantation from Mar 2017 to Feb 2021 were divided into chief cell type (CC) and oxyphil cell type (OC) according to dominating graft cell type. iPTH, calcium, phosphorus and ALP levels were compared between the two groups from the perioperative period to 6 months and the 3D ultrasonography was performed at 6 months to cocalculate the size of the autograft.Results:Between the two groups, there were no statistically significant differences in iPTH, blood calcium, blood phosphorus, or ALP levels pre-,and 30 minutes, one month post surgery (all P>0.05). On three months, there was no significant difference in ALP levels [CC group: (99±23) U/L, OC group: (89±35) U/L, t=0.776, P=0.442]. At 6 months the PTH level [CC group: (290±77) pg/ml, OC group: (246±59) pg/ml, t=2.034, P=0.047], and blood calcium [CC group: (2.62±0.65) mmol/L, OC group: (2.21±0.20) mmol/L, t=2.531, P=0.015] blood phosphorus [CC group: (1.38±0.28) mmol/L, OC group: (1.68±0.34) mmol/L; t=-3.269, P=0.002], were all in favor of CC group. By 3D ultrasnography at 6 months the size of autograft was larger in CC group than in OC group [V cc=(2.17±0.37) cm 3,V oc=(1.85±0.29) cm 3, t=3.172, P<0.05]. Recurrences at 1 year after surgery were not significantly different between the two groups ( ncc=3, noc=1, t=0.277, P>0.05). Conclusion:The biological activity of CC-type grafts is higher than that of OC suggesting a longer functioning period as a parathyroid autograft.

Hepatocellular carcinoma(HCC)is a highly lethal cancer with significant incidence and mortality rates.The study of the biological features of liver cancer cells is critical for the development of novel treatment strategies for HCC.While traditional cell culture techniques fail to provide information on the growth and diffusion of cells in a threedim-ensional space(3D),3D bioprinting technology provides a new method to study the functional characteristics of HCC cells.With the development of 3D cell culture technology,researches are largely focused on exploring the function and behavior of cells in a three-dimensional environment,particularly in complex tumor models like liver tumors that comprise intricate cellular tissues and blood vessels.This article discussed about the cellular functions that need to be studied in the 3D bioprinting environment of HCC and other tumor cells by reviewing the research progress of 3D bioprinting in HCC and various other cancer cell lines,aiming to help researchers replicate the in vivo growth environment of tumor cells more accurately,expand application scenarios,and further explore cancer treatment methods.In addition,3D bioprinting technology is considered a promising tool for drug development,providing a more clinically relevant model for the pharmacological study of anti-cancer drugs,better revealing the drug sensitivity and resistance of tumors,reducing the need for animal experiments,and providing more possibilities for precision medicine.

Objective:To evaluate CT and or MRI imaging in the diagnosis of lateral lymph node metastasis in patients of middle and low rectal cancer.Methods:In this study, 112 lateral lymph nodes were harvested in 79 patients with middle and low rectal cancer. The relationship between the preoperative imaging features of the lateral lymph nodes and the postoperative pathology was evaluated.Result:Thirty-eight cases (48%) were pathologically confirmed to have lateral lymph node metastasis. The diameter of metastasis-positive lateral lymph nodes was significantly larger than that of metastasis-negative lateral lymph nodes ( P<0.01). Multivariate analysis of clinical features and imaging features found that, tumors poorly differentiated, mucinous adenocarcinoma, signet ring cell carcinoma ( P=0.006), and the largest short diameter of the lateral lymph node ≥7 mm ( P=0.024), uneven density or signal ( P=0.022) were independent risk factors for lateral lymph node metastasis. Conclusion:Poor tumor differentiation, lateral lymph node maximum short diameter ≥7 mm, density or signal unevenness are independent risk factors for lateral lymph node metastasis in middle and low rectal carcinoma.

Purpose: Platelet-related indices, including mean platelet volume (MPV) and plateletocrit (PCT), have been reported as new prognostic factors of overall survival (OS) in many cancers, but not yet in biliary tract cancer (BTC). We intended to assess these indices in predicting OS in BTC patients with the aim to build a new prognostic model for patients with BTC after surgical resection. Materials and Methods: Survival analysis and time receiver operating characteristic analysis were applied to screen the platelet indices. Univariate and multivariate Cox analyses were used to identify independent prognostic factors and develop a new prognostic model. Harrell’s C-statistics, calibration curves, and decisive curve analysis were used to assess the model. Results: MPV and platelet distribution width (PDW)/PCT showed the best prognostic accuracy among the platelet indices. In multivariable analysis, factors predictive of poor OS were presence of nodal involvement, Non-radical surgery, poor tumor differentiation, carbohydrate antigen 19-9 > 100 U/mL, MPV > 8.1 fl, and PDW/PCT > 190. The new model was found to be superior to the TNM staging system and our new staging system showed higher discriminative power. Conclusion MPV and PDW/PCT have high prognostic value in BTC patients, and the novel staging system based on these two indices showed good discrimination and accuracy compared with the American Joint Committee on Cancer 7th TNM staging system.

Purpose: Platelet-related indices, including mean platelet volume (MPV) and plateletocrit (PCT), have been reported as new prognostic factors of overall survival (OS) in many cancers, but not yet in biliary tract cancer (BTC). We intended to assess these indices in predicting OS in BTC patients with the aim to build a new prognostic model for patients with BTC after surgical resection. Materials and Methods: Survival analysis and time receiver operating characteristic analysis were applied to screen the platelet indices. Univariate and multivariate Cox analyses were used to identify independent prognostic factors and develop a new prognostic model. Harrell’s C-statistics, calibration curves, and decisive curve analysis were used to assess the model. Results: MPV and platelet distribution width (PDW)/PCT showed the best prognostic accuracy among the platelet indices. In multivariable analysis, factors predictive of poor OS were presence of nodal involvement, Non-radical surgery, poor tumor differentiation, carbohydrate antigen 19-9 > 100 U/mL, MPV > 8.1 fl, and PDW/PCT > 190. The new model was found to be superior to the TNM staging system and our new staging system showed higher discriminative power. Conclusion MPV and PDW/PCT have high prognostic value in BTC patients, and the novel staging system based on these two indices showed good discrimination and accuracy compared with the American Joint Committee on Cancer 7th TNM staging system.