Glucose oxidase (GOD) is an oxygen-consuming dehydrogenase that can catalyze the production of gluconic acid hydrogen peroxide from glucose, and its specific mechanism of action makes it promising for applications, while the low catalytic activity and poor thermostability have become the main factors limiting the industrial application of this enzyme. In this study, we used the glucose oxidase AtGOD reported with the best thermostability as the source sequence for phylogenetic analysis to obtain the GOD with excellent performance. Six genes were screened and successfully synthesized for functional validation. Among them, the glucose oxidase AhGODB derived from Aspergillus heteromorphus was expressed in Pichia pastoris and showed better thermostability and catalytic activity, with an optimal temperature of 40 ℃, a specific activity of 112.2 U/mg, and a relative activity of 47% after 5 min of treatment at 70 ℃. To improve its activity and thermal stability, we constructed several mutants by directed evolution combined with rational design. Compared with the original enzyme, the mutant T72R/A153P showcased the optimum temperature increasing from 40 to 50 ℃, the specific activity increasing from 112.2 U/mg to 166.1 U/mg, and the relative activity after treatment at 70 ℃ for 30 min increasing from 0% to 33%. In conclusion, the glucose oxidase mutants obtained in this study have improved catalytic activity and thermostability, and have potential for application.
Glucose Oxidase/chemistry* ; Enzyme Stability ; Aspergillus/genetics* ; Pichia/metabolism* ; Temperature ; Catalysis ; Fungal Proteins/metabolism* ; Hot Temperature

Lipids have been found to modulate tumor biology, including proliferation, survival, and metastasis. With the new understanding of tumor immune escape that has developed in recent years, the influence of lipids on the cancer-immunity cycle has also been gradually discovered. First, regarding antigen presentation, cholesterol prevents tumor antigens from being identified by antigen presenting cells. Fatty acids reduce the expression of major histocompatibility complex class I and costimulatory factors in dendritic cells, impairing antigen presentation to T cells. Prostaglandin E2 (PGE2) reduce the accumulation of tumor-infiltrating dendritic cells. Regarding T-cell priming and activation, cholesterol destroys the structure of the T-cell receptor and reduces immunodetection. In contrast, cholesterol also promotes T-cell receptor clustering and relative signal transduction. PGE2 represses T-cell proliferation. Finally, regarding T-cell killing of cancer cells, PGE2 and cholesterol weaken granule-dependent cytotoxicity. Moreover, fatty acids, cholesterol, and PGE2 can improve the activity of immunosuppressive cells, increase the expression of immune checkpoints and promote the secretion of immunosuppressive cytokines. Given the regulatory role of lipids in the cancer-immunity cycle, drugs that modulate fatty acids, cholesterol and PGE2 have been envisioned as effective way in restoring antitumor immunity and synergizing with immunotherapy. These strategies have been studied in both preclinical and clinical studies.

Mevalonate metabolism plays an important role in regulating tumor growth and progression; however, its role in immune evasion and immune checkpoint modulation remains unclear. Here, we found that non-small cell lung cancer (NSCLC) patients with higher plasma mevalonate response better to anti-PD-(L)1 therapy, as indicated by prolonged progression-free survival and overall survival. Plasma mevalonate levels were positively correlated with programmed death ligand-1 (PD-L1) expression in tumor tissues. In NSCLC cell lines and patient-derived cells, supplementation of mevalonate significantly up-regulated the expression of PD-L1, whereas deprivation of mevalonate reduced PD-L1 expression. Mevalonate increased CD274 mRNA level but did not affect CD274 transcription. Further, we confirmed that mevalonate improved CD274 mRNA stability. Mevalonate promoted the affinity of the AU-rich element-binding protein HuR to the 3'-UTR regions of CD274 mRNA and thereby stabilized CD274 mRNA. By in vivo study, we further confirmed that mevalonate addition enhanced the anti-tumor effect of anti-PD-L1, increased the infiltration of CD8⁺ T cells, and improved cytotoxic function of T cells. Collectively, our findings discovered plasma mevalonate levels positively correlated with the therapeutic efficacy of anti-PD-(L)1 antibody, and provided the evidence that mevalonate supplementation could be an immunosensitizer in NSCLC.

[This corrects the article DOI: 10.1016/j.apsb.2023.04.002.].

Objective:To analyze the correlation between the body composition and cardiorespiratory fitness (CRF) decline in physical examination population of different genders.Methods:Clinical data of the cardiopulmonary exercise testing (CPET) and body composition analysis of 439 people who received physical examination in the Medical Examination Center of Peking University Third Hospital from May 2021 to September 2021 were retrospectively analyzed. The general data, physical examination, biochemical parameters, body composition and CPET results were collected. The subjects were divided into normal group and decline group according to the percentage of peak oxygen uptake (VO 2peak) levels ≥ 85% or<85%. Multivariate logistic regression was applied to investigate the influencing factors of CRF decline in subjects of different genders. Results:Among men, total cholesterol and triglyceride in the decline group were significantly higher than those in the normal group [(5.097±0.890) vs (4.865±0.856) mmol/L, (1.778±1.200) vs (1.485±0.709) mmol/L], and the blood homocysteine (Hcy) and skeletal muscle index were significantly lower than those in the normal group [13.00 (11.30, 15.90) vs 13.80 (12.05, 17.10) μmol/L, (7.89±0.65) vs (8.08±0.64) kg/m 2] (all P<0.05). Among women, skeletal muscle index in the decline group was significantly lower than that in the normal group [(6.21±0.52) vs (6.53±0.56)kg/m 2], and percent body fat was significantly higher than that in the normal group [(32.83±4.92)% vs (31.21±4.55)%] (all P<0.05). The elevation of triglyceride level ( OR=1.487, 95% CI: 1.042-2.121) and visceral fat area ( OR=1.032, 95% CI: 1.014-1.051) were positively correlated with the decline of CRF in man, the decrease of skeletal muscle index ( OR=0.215, 95% CI: 0.106-0.435) and the increase of percent body fat ( OR=1.149, 95% CI: 1.060-1.245) were positively correlated with the decrease of CRF in women (all P<0.05). Conclusions:There is a correlation between body composition and CRF decline in physical examination population of different genders. Men should control visceral fat more effectively, and women should pay attention to increase muscle mass while reducing body fat, in order to improve CRF.

Objective:To evaluate the use of serosa muscular layers circumferential incision combined with mucosal layer cutting and closure by laparoscopic or robotic surgery for gastrointestinal mesenchymal tumors at difficult sites of the stomach.Methods:From Jul 2019 to Apr 2021, 18 gastric mesenchymal tumor patients undergoing serosa muscular layers circumferential incision combined with mucosal layer cutting and closure by laparoscopic or robotic surgery at the Department of Surgery, the Fourth Hospital of Hebei Medical University were retrospectively analyzed.Results:All 18 patients had successful surgery, including 7 cases of robotic surgery, 11 cases of laparoscopic surgery, and there was no conversion to open surgery. Tumors were at the gastric in cardia, 8 cases at the gastric body and lesser curvature in 4 cases, and at the gastric antrum in 6 cases, respectively. Eleven cases were of endogenous and 7 cases were of dumbbell type. The average operation time was (99±29) min, the intraoperative blood loss was (10±5) ml, the first time taking food per mouth was (2.0±1.0) d, and the postoperative hospital stay was (4.9 ± 1.2) d. Pathology showed gastrointestinal stromal tumor in 11 cases, leiomyoma in 5 cases and schwannoma in 2 cases. All were with negative margins. The average tumor diameter was (4.7±1.4) cm. The median follow-up time was 16.5 months, and there was no sign of tumor recurrence or metastasis.Conclusion:The serosa muscular layers circumferential incision combined with mucosal layer cutting and closure technique in laparoscopic or robotic surgery is a safe and feasible procedure for treating gastrointestinal mesenchymal tumor at difficult sites of the stomach.

Objective:To investigate the influence of different injection time of carbon nanoparticle tracer on the acquisition of lymph nodes in adenocarcinoma of esophagogastric junc-tion (AEG) treated by neoadjuvant chemoradiotherapy (nCRT) combined with surgical resection.Methods:The prospective randomized controlled study was conducted. The clinicopathological data of 120 AEG patients who were treated by nCRT combined with surgical resection in the Fourth Hospital of Hebei Medical University from March 2020 to March 2021 were selected. Based on random number table, patients were allocated into two groups. Patients undergoing endoscopic injection of carbon nanoparticle tracer 24 hours before nCRT were allocated into the experiment group, and patients undergoing endoscopic injection of carbon nanoparticle tracer 24 hours before surgical resection were allocated into the control group. All patients received the same plan of nCRT combined with D 2 radical gastrectomy. Observation indicators: (1) grouping situations of the enrolled patients; (2) surgical and postoperative pathological situations; (3) postoperative complications and treatment. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was analyzed using the independent sample t test. Measurement date with skewed distribution were represented as M( Q1, Q3), and comparison between groups was conducted using the Mann-Whitney U test. Count data were described as absolute numbers, and comparison between groups was analyzed using the chi-square test. Comparison of ordinal data was analyzed using the non-parameter rank sum test. Results:(1) Grouping situations of the enrolled patients. A total of 120 patients were selected for eligibility. There were 85 males and 35 females, aged (60±9)years. There were 60 patients in the experiment group and 60 patients in the control group, respectively. (2) Surgical and postoperative pathological situations. Patients in the two groups underwent D 2 radical gastrectomy successfully, with R 0 resection. The number of lymph nodes harvest, the number of lymph nodes stained, the number of metastatic lymph nodes stained, the number of micro lymph nodes, the number of inferior mediastinal lymph nodes, the number of inferior mediastinal lymph nodes stained, cases in postoperative pathological stage N0, stage N1, stage N2, stage N3a were 40.6±13.9,20.1±7.7, 1.0(0,3.0), 8.1±2.8, 3.7±1.3, 2.0(1.0,2.0), 18, 13, 23, 6 in patients of the experiment group, respectively. The above indicators were 30.4±8.3, 12.7±3.5, 0(0,1.0), 6.2±2.0, 2.4±1.2, 1.0(0,1.0), 23, 21, 15, 1 in patients of the control group, respectively. There were significant differences in the above indicators between the two groups ( t=-5.01, 6.85, Z=-3.78, t=-4.04, -5.57, Z=-5.48, -2.12, P<0.05). (3) Postoperative complications and treatment. There were 5 cases of the experiment group and 7 cases of the control group with postoperative complications, showing no significant difference between the two groups ( χ2=0.37, P>0.05). The patients with postoperative complications were improved after symptomatic treatment. Conclusion:Compared with injection of carbon nanoparticle tracer 24 hours before surgical resection, injection of carbon nanoparticle tracer 24 hours before nCRT can improve the acquisition of lymph nodes in AEG treated by nCRT combined with surgical resection.

Objective:To analyze the core elements′ index design of physical examination reports and pilot data.Methods:We searched and analyzed the core element indexes of the physical examination report in databases such as CNKI and Wanfang through the literature analysis method, forming the initial core element indicator set. Special working meetings were organized to discuss revisions and improvements. The core elements′ index set was formed using a mixed research method combined with the quantitative analysis results of pilot data and expert opinions. Six hundred and thirteen cases from five hospitals were included in this study, including Anzhen Hospital, Peking University Third Hospital, Beijing Physical Examination Center, Beijing Tongren Hospital, China-Japan friendship hospital.Results:The subjects were (49.5±17.6) years old, including 302 males (49.3%) and 311 females (50.7%). The physical examination conclusion indicates that the number of health problems is between five and eight, with no significant difference between men and women. The health history data analysis of different sexes showed that hypertension in men was significantly higher than in women (17.9% vs. 10.6%, P=0.010). The lifestyle and eating habits survey showed that men had significantly higher incidences of smoking and drinking than women (24.8% vs. 2.2%, P?0.001; 9.9% vs. 1.3%, P?0.001). The exercise habits and sleep survey showed that the proportion of men who did not exercise was lower than that of women (13.2% vs. 22.8%, P=0.022). There was no statistically significant difference in sleep duration between the sexes. Conclusions:The core elements of health examination reports can standardize the collection of physical examination data and improve its sharing and application. This can allow the reports to better assist the decisions based upon physical examination report data.

Objective:To explore the clinico-pathological characteristics and risk factors affecting prognosis in elderly patients with gastric cancer.Methods:A retrospective study was used to retrospectively analyze 2386 patients with gastric cancer undergoing radical surgery in Surgery Department of the Fourth Hospital of Hebei Medical University from 1 January 2012 to 1 January 2015.Patients aged 70 years and older were screened so as to analyze clinical characteristics and influencing factors for the prognosis.Results:A total of 2386 patients with gastric cancer were divided into the elderly group aged 70 years and older(342 of 2386 cases, 14.3%). There were statistically significant differences between the two groups in gender, number of concomitant diseases, NRS2002 score, PG-SGA score, tumor location, tumor diameter, histological type, Borrmann classification, tumor invasion depth staging(pT), lymph node metastasis staging(pN), the anatomic extent of tumor staging(TNM, pTNM), and Lauren classifications( P<0.05). The 981 of 2386 cases(41.4%)had postoperative complications, accompanied by 413 cases(17.3%)of surgery-related complications and 568 cases(24.0%)of non-surgery-related complications.A multivariate logistic analysis showed that the number of preoperative co-existing diseases ≥ 2 was an independent influencing factor for postoperative complications in elderly gastric cancer patients( HR=4.478, 95% CI: 1.121-7.918, P=0.006). The 5-year OS and DSS was 21.10% and 62.73% in the ≥70 years gastric cancer group, and was 54.1% and 70.0% in the <70 years gastric cancer group, respectively.The difference in the 5-year OS between the two groups was statistically significant( P<0.05), while the difference in the 5-year DSS between the two groups was not statistically significant( P>0.05). Multivariate analysis by the Cox proportional hazard model showed that the independent risk factors for the prognosis of elderly patients with gastric cancer included the low-undifferentiated histological type of the tumor( P=0.004), the depth of tumor invasion pT stage of pT4a-pT4b( P=0.007), lymph node metastasis( P=0.034), tumor pTNM stage ⅢA-ⅢC( P=0.002)and vascular tumor thrombus( P=0.034). Conclusions:Elderly patients with gastric cancer have many preoperative co-existing diseases, which increases the risk of postoperative non-surgical complications.Therefore, we should focus on the peri-operative management of their comorbid diseases so as to improve the safety and efficacy of surgery.The advanced age is not the independent risk factors for the prognosis.