1.Improvement of neutral protease activity of Bacillus amyloliquefaciens LX-6 by combined ribosome engineering and medium optimization and its application in soybean meal fermentation.
Yifan ZHU ; Xinyi HUANG ; Tao HAN ; Jiteng WANG ; Xiaoping YU ; Zheng MA
Journal of Zhejiang University. Science. B 2025;26(8):805-812
Soybean meal (SBM) prepared by soybean crushing is the most popular protein source in the poultry and livestock industries (Cai et al., 2015) due to its economic manufacture, high protein content, and good nutritional value. Despite these benefits, SBM contains various antigen proteins such as glycinin and β-conglycinin, which account for approximately 70% of the total proteins of the SBM and reduce digestibility and damage intestinal function (Peng et al., 2018). Treating SBM with proteases (neutrase, alcalase, and trypsin) or fermentation can eliminate these antigen proteins (Contesini et al., 2018). Because of its safety and rapid growth cycle, Bacillus strains are considered ideal for the fermentation industry (Yao et al., 2021). SBM fermented by Bacillus yields products with high nutritional value and low levels of antinutritional factors (ANFs), stimulating research in this area (Yuan et al., 2017). Kumari et al. (2023) demonstrated that fermentation with Bacillus species effectively degrades antigen proteins and increases crude protein content. The degradation of antigen proteins relies on protease hydrolysis. Low protease production is the major obstacle hindering the widespread use of microbial fermentation techniques.
Bacillus amyloliquefaciens/metabolism*
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Fermentation
;
Glycine max/metabolism*
;
Soybean Proteins/metabolism*
;
Peptide Hydrolases/metabolism*
;
Ribosomes/metabolism*
;
Globulins
;
Antigens, Plant
;
Seed Storage Proteins
2.Intranodal injection of neoantigen-bearing engineered Lactococcus lactis triggers epitope spreading and systemic tumor regressions.
Junmeng ZHU ; Yi SUN ; Xiaoping QIAN ; Lin LI ; Fangcen LIU ; Xiaonan WANG ; Yaohua KE ; Jie SHAO ; Lijing ZHU ; Lifeng WANG ; Qin LIU ; Baorui LIU
Acta Pharmaceutica Sinica B 2025;15(4):2217-2236
Probiotics are natural systems bridging synthetic biology, physical biotechnology, and immunology, initiating innate and adaptive anti-tumor immune activity. We previously constructed an all-in-one engineered food-grade probiotic Lactococcus lactis (FOLactis) which could boost the crosstalk among different immune cells such as dendritic cells (DCs), natural killer cells, and T cells. Herein, considering the limited clinical efficacy of naked personalized neoantigen peptide vaccines, we decorate FOLactis with tumor antigens by employing a Plug-and-Display system comprising membrane-inserted peptides. Intranodal injection of FOLactis coated with neoantigen peptides (Ag-FOLactis) induces robust DCs presentation and neoantigen-specific cellular immunity. Notably, Ag-FOLactis not only triggers a 45-fold rise in the quantity of locally reactive neoantigen-specific T cells but also induces epitope spreading in both subcutaneous and metastatic tumor-bearing models, leading to potent inhibition of tumor growth. These findings imply that Ag-FOLactis represents a powerful platform to rapidly and easily display antigens, facilitating the development of a bio-activated platform for personalized therapy.
3.Chinese version of the Needs and Provision Complexity Scale and its reliability and validity in stroke patients
Mengke SU ; Shufan CHEN ; Zining GUO ; Xiaoli YU ; Xiaoping ZHU
Chinese Journal of Modern Nursing 2025;31(35):4784-4790
Objective:To translate the Needs and Provision Complexity Scale (NPCS) into Chinese and validate its reliability and validity among stroke patients.Methods:The Chinese version of NPCS was developed through forward translation, back-translation, cultural adaptation, and pre-survey using the modified Brislin translation model. Convenience sampling was used to select 330 stroke patients who visited Shanghai Tenth People's Hospital between May and July 2024 for questionnaire surveys, thereby validating the reliability and validity of the scale. A total of 330 questionnaires were distributed, with 316 valid responses collected for the need version and 305 valid responses collected for the provision version.Results:The Chinese version of NPCS was divided into a need version and a provision version, each comprising two dimensions and 15 items. The content validity index at the item level of the need and provision version of the scale ranged from 0.833 to 1.000, with an average content validity index of 0.943 for both. The Cronbach's α coefficients of the scale were 0.916 and 0.918, and the Cronbach's α coefficients of each dimension were from 0.882 to 0.919, and the test-retest reliability coefficients were 0.913 and 0.897, respectively. Exploratory factor analysis yielded two common factors of personal health care and social support, with cumulative variance contribution rates of 71.437% and 70.266%, respectively. The Chinese version of the scale demonstrated good model fit.Conclusions:The Chinese version of NPCS has good reliability and validity, and can be used for investigating the provision and need of rehabilitation and social support services for stroke patients.
4.Application of Wrist-Ankle Acupuncture in Postoperative Analgesia of Mixed Hemorrhoids
Haiyan ZHU ; Xiaoping SHU ; Weixia CHAI
Journal of Zhejiang Chinese Medical University 2025;49(4):470-474
[Objective]To explore the therapeutic effect of wrist-ankle acupuncture in postoperative analgesia of mixed hemorrhoids.[Methods]Eighty patients with mixed hemorrhoid after surgery under lumbar anesthesia were randomly divided into experimental group and control group,with 40 cases in each group.The control group received routine postoperative treatment and nonsteroidal anti-inflammatory drugs(NSAIDs),100 mg Tramadol was intramuscularly injected when the visual analogue score(VAS)≥7.The experimental group received wrist-ankle acupuncture treatment firstly when the VAS≥7,if the VAS was still≥7 after half an hour,the 100 mg Tramadol was intramuscularly injected.Compare the VAS scores of two groups before the first intervention and 0.5 hour after the intervention,as well as at 6,12,18,24,36 and 48 hours after surgery.Compare the frequency of Tramadol use at 48 hours after surgery and the changes in prostaglandin E2(PGE2)and 5-hydroxytryptamine(5-HT)levels before and 0.5 hour after the first intervention.[Results]There was no significant statistical difference in VAS scores between experimental group and control group before the first intervention(P>0.05).The VAS scores of both groups 0.5 hour after the first intervention were lower than before the intervention in the same group(P<0.05).The VAS score of experimental group patients after the first intervention for 0.5 hour was higher than that of control group(P<0.05).The VAS scores of experimental group at 6,12,18,24,36 and 48h after surgery were lower than those of control group at the same time point(P<0.05).The frequency of using Tramadol in experimental group was lower than that in control group 48 hours after surgery(P<0.05).There was no significant statistical difference in PGE2 and 5-HT levels between experimental group and control group before intervention(P>0.05).The PGE2 and 5-HT levels in experimental group and control group decreased after intervention for 0.5 hour compared to before intervention in the same group(P<0.05).The PGE2 and 5-HT levels in experimental group were higher than those in control group after 0.5 hour of intervention(P<0.05).[Conclusion]Wrist-ankle acupuncture therapy can effectively alleviate postoperative pain in patients with mixed hemorrhoids,reduce the use of postoperative analgesics,and its analgesic mechanism may be related to downregulating the serum levels of PGE2 and 5-HT,which has certain clinical application value..
5.Correlations of expression levels of microRNA-147,Ki-67 and carcinoembryonic antigen with lymph node metastasis in invasive pulmonary adenocarcinoma
Journal of Clinical Medicine in Practice 2025;29(14):7-12
Objective To explore correlations of expression levels of microRNA-147(miR-147),Ki-67 and carcinoembryonic antigen(CEA)with lymph node metastasis of invasive lung adenocarci-noma.Methods A total of 160 patients with invasive pulmonary adenocarcinoma were selected as study subjects.Based on presence of lymph node metastasis,patients were divided into lymph node metastasis group(n=74)and non-lymph node metastasis group(n=86).Immunohistochemistry was employed to detect the expression levels of miR-147 and Ki-67 in cancer tissues.The serum CEA level was measured using the chemiluminescence method.The correlations of the expression levels of miR-147,Ki-67 and serum CEA with the clinicopathological characteristics of patients were analyzed.The value of miR-147,Ki-67 and CEA in predicting lymph node metastasis was evaluated using the receiver operating characteristic(ROC)curve.Results The expression level of miR-147 in the lymph node metastasis group was significantly lower than that in the non-lymph node metastasis group,while the expression levels of Ki-67 and serum CEA were significantly higher in the lymph node me-tastasis group than in the non-lymph node metastasis group(P<0.05).Patients with a tumor diameter≥3 cm or TNM stage Ⅲ to Ⅳ had significantly lower miR-147 expression levels in cancer tissues compared to those with a tumor diameter<3 cm or TNM stage Ⅰ to Ⅱ(P<0.05).The Ki-67 ex-pression level in cancer tissues was significantly higher in patients with a tumor diameter ≥3 cm than in those with a tumor diameter<3 cm(P<0.05).The serum CEA level was significantly higher in patients with TNM stage Ⅲ to Ⅳ than in those with TNM stage Ⅰ to Ⅱ(P<0.05).The expres-sion level of miR-147 was negatively correlated with Ki-67,serum CEA and lymph node metastasis(P<0.05);Ki-67 and serum CEA were positively correlated with lymph node metastasis(P<0.05);Ki-67 was positively correlated with serum CEA(P<0.05).The areas under the ROC curves for miR-147,Ki-67 and CEA in predicting lymph node metastasis were 0.785,0.835 and 0.798,respectively(P<0.05).Conclusion The expression levels of miR-147 and Ki-67 in cancer tissues and the serum CEA level are correlated with lymph node metastasis in invasive pulmo-nary adenocarcinoma.
6.A single-center retrospective study on pacemaker lead-related tricuspid regurgitation
Danqing YU ; Qifeng ZHU ; Yan LIN ; Xiaoping LIN ; Xianbao LIU ; Jian'an WANG
Chinese Journal of Emergency Medicine 2025;34(10):1410-1417
Objective:This study aimed to retrospectively analyze the incidence and influencing factors of tricuspid regurgitation (TR) in the short term after pacemaker lead implantation at the Second Affiliated Hospital of Zhejiang University School of Medicine, so as to provide evidence for understanding pacemaker lead-related TR.Methods:Consecutive patients who underwent single- or dual-chamber permanent pacemaker implantation between June 2019 and December 2023 in the Department of Cardiology were enrolled. General clinical data and relevant parameters were collected. Changes in TR severity before and shortly after the procedure were assessed using echocardiography. TR progression was defined as an increase by one grade or more, and TR improvement as a decrease by one grade or more. Logistic regression analysis was employed to identify factors associated with TR progression.Results:A total of 219 patients were included (128 males, 91 females), with a mean age of 69.7 ± 11.2 years. The median follow-up time was 99 (26, 199) days. TR remained unchanged in 114 patients (52.1%), improved in 46 (21.0%)—including 36 (16.4%) with one-grade reduction, 9 (4.1%) with two-grade reduction, and 1 (0.5%) with three-grade reduction—and progressed in 59 patients (26.9%). Among those with progression, 51 (23.3%) had mild-to-moderate TR worsening by one grade, and 8 (3.7%) had moderate or worse TR worsening by at least two grades. Notably, one case involved lead perforation of the leaflet and two cases had lead impingement. Compared with the non-progression group ( n = 114), pacemaker indication (AV block vs. sick sinus syndrome), baseline left atrial diameter, pulmonary artery systolic pressure (PASP), and the severity of mitral regurgitation (MR) and TR were significantly associated with TR progression or improvement (all P < 0.05). Ordinal logistic regression analysis identified preoperative TR severity [ OR=10.57 (3.77–29.68), P < 0.001] and pacemaker indication [ OR=0.452 (0.222–0.918), P = 0.028] as independent predictors of postoperative TR progression. Patients with AV block were more likely to receive left bundle branch pacing ( P < 0.001), which may contribute to their lower risk of TR. Conclusions:Short-term progression of TR after pacemaker implantation is relatively common, although severe TR remains rare. Preoperative TR severity and pacemaker indication are independent predictors of short-term TR progression. The use of physiological pacing modalities may help reduce the incidence of TR following pacemaker lead implantation.
7.Identification of HMA gene family and response to cadmium stress in Ophiopogon japonicas.
Zhihui WANG ; Erli NIU ; Yuanliang GAO ; Qian ZHU ; Zihong YE ; Xiaoping YU ; Qian ZHAO ; Jun HUANG
Chinese Journal of Biotechnology 2025;41(2):771-790
Soil cadmium (Cd) pollution is one of the major environmental problems globally. Ophiopogon japonicus, a multifunctional plant extensively used in traditional Chinese medicine, has demonstrated potential in environmental remediation. This study investigated the Cd accumulation pattern of O. japonicus under cadmium stress and identified the heavy metal ATPase (HMA) family members in this plant. Our results demonstrated that O. japonicus exhibited a Cd enrichment factor (EF) of 2.75, demonstrating strong potential for soil Cd pollution remediation. Nine heavy metal ATPase (HMA) members of P1B-ATPases were successfully identified from the transcriptome data of O. japonicus, with OjHMA1-OjHMA6 classified as the Zn/Co/Cd/Pb-ATPases and OjHMA7-OjHMA9 as the Cu/Ag-ATPases. The expression levels of OjHMA1, OjHMA2, OjHMA3, and OjHMA7 were significantly up-regulated under Cd stress, highlighting their crucial roles in cadmium ion absorption and transport. The topological analysis revealed that these proteins possessed characteristic transmembrane (TM) segments of the family, along with functional A, P, and N domains involved in regulating ion absorption and release. Metal ion-binding sites (M4, M5, and M6) existed on the TM segments. Based on the number of transmembrane domains and the residues at metal ion-binding sites, the plant HMA family members were categorized into three subgroups: P1B-1 ATPases, P1B-2 ATPases, and P1B-4 ATPases. Specifically, the P1B-1 ATPase subgroup included the motifs TM4(CPC), TM5(YN[X]4P), and TM6(M[XX]SS); the P1B-2 ATPase subgroup featured the motifs TM4(CPC), TM5(K), and TM6(DKTGT); the P1B-4 ATPase subgroup contained the motifs TM4(SPC) and TM6(HE[X]GT), all of which were critical for protein functions. Molecular docking results revealed the importance of conserved sequences such as CPC/SPC, DKTGT, and HE[X]GT in metal ion coordination and stabilization. These findings provide potential molecular targets for enhancing Cd uptake and tolerance of O. japonicus by genetic engineering and lay a theoretical foundation for developing new cultivars with high Cd accumulation capacity.
Cadmium/metabolism*
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Adenosine Triphosphatases/metabolism*
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Ophiopogon/drug effects*
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Soil Pollutants/toxicity*
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Plant Proteins/metabolism*
;
Stress, Physiological
;
Multigene Family
;
Gene Expression Regulation, Plant
8.The safety and efficacy of boron neutron capture therapy (BNCT) in the treatment of recurrent malignant tumors
Junqiang HONG ; Xiaoyi LIN ; Youqun LAI ; Ye CAO ; Xiangquan KONG ; Yuanhao LIU ; Shuiying LUO ; Zhicheng XIONG ; Mei GONG ; Yalai LIN ; Qiaoyun CHEN ; Mingang YING ; Li HUO ; Xiaohua ZHU ; Xiaoping SUN ; Yiqiao DENG ; Diyun SHU ; Haige ZHANG ; Cheng HUANG ; Jianji PAN
Chinese Journal of Radiation Oncology 2025;34(10):985-992
Objective:To evaluate the safety and efficacy of accelerator-based boron neutron capture therapy (AB-BNCT) in the treatment of recurrent and refractory malignant tumors.Methods:The data of 14 patients admitted to Xiamen Humanity Hospital from September 2022 to April 2023 were prospectively collected, including 7 patients with primary brain malignancies and 7 patients with locally recurrent inoperable head and neck malignancies. All patients received intravenous infusion of boron drug (NBB-001, p-dihydroxyborylphe nylalanine, a patented freeze-dried formulation) at a total nominal dosage of 500 mg/kg (11 patients) or 750 mg/kg (3 patients), and were irradiated with neutrons (operating with NeuPex system). Adverse events after treatment were recorded and assessed. The primary efficacy endpoint was the 90 d objective response rate (ORR), while the secondary endpoints included progression-free survival (PFS) and complete response rate (CRR). Data were compiled and analyzed by SAS 9.4 software. The rate and 95% CI were calculated using Clopper-Pearson method. Results:The median dose delivered to 80% of the target volume (D 80%) was 16.80 GyE (range: 8.93-23.79 GyE). The most common adverse reactions were hyperamylasemia, alopecia, and hyperprolactinemia. Five patients experienced 8 cases of grade 3 or above adverse events, including 1 case of grade 4 acute kidney injury and 7 cases of grade 3 adverse events. All adverse events were recovered after observation or treatment. At 90 d after treatment, the ORR of all patients was 9/14 (64%, 95% CI: 35%-87%), disease control rate (DCR) was 10/14 (71%, 95% CI: 42%-92%), CRR was 2/14 (14%, 95% CI: 2%-42%); and the best overall response during the entire course included an ORR of 10/14 (71% ,95% CI: 42%-92%), DCR of 13/14 (93%, 95% CI: 66%-100%), and CRR of 3/14 (21% ,95% CI: 5%-51%). The 1-year survival rate for head and neck malignancies was 71.4%, and the 2-year survival rate was 42.8%. The 1-year survival rate for recurrent brain malignancies was 42.8%. Conclusion:AB-BNCT demonstrates favorable safety and promising efficacy in treating primary brain malignancies and recurrent/refractory head and neck malignancies, representing a potential therapeutic option.
9.Guideline for Adult Weight Management in China
Weiqing WANG ; Qin WAN ; Jianhua MA ; Guang WANG ; Yufan WANG ; Guixia WANG ; Yongquan SHI ; Tingjun YE ; Xiaoguang SHI ; Jian KUANG ; Bo FENG ; Xiuyan FENG ; Guang NING ; Yiming MU ; Hongyu KUANG ; Xiaoping XING ; Chunli PIAO ; Xingbo CHENG ; Zhifeng CHENG ; Yufang BI ; Yan BI ; Wenshan LYU ; Dalong ZHU ; Cuiyan ZHU ; Wei ZHU ; Fei HUA ; Fei XIANG ; Shuang YAN ; Zilin SUN ; Yadong SUN ; Liqin SUN ; Luying SUN ; Li YAN ; Yanbing LI ; Hong LI ; Shu LI ; Ling LI ; Yiming LI ; Chenzhong LI ; Hua YANG ; Jinkui YANG ; Ling YANG ; Ying YANG ; Tao YANG ; Xiao YANG ; Xinhua XIAO ; Dan WU ; Jinsong KUANG ; Lanjie HE ; Wei GU ; Jie SHEN ; Yongfeng SONG ; Qiao ZHANG ; Hong ZHANG ; Yuwei ZHANG ; Junqing ZHANG ; Xianfeng ZHANG ; Miao ZHANG ; Yifei ZHANG ; Yingli LU ; Hong CHEN ; Li CHEN ; Bing CHEN ; Shihong CHEN ; Guiyan CHEN ; Haibing CHEN ; Lei CHEN ; Yanyan CHEN ; Genben CHEN ; Yikun ZHOU ; Xianghai ZHOU ; Qiang ZHOU ; Jiaqiang ZHOU ; Hongting ZHENG ; Zhongyan SHAN ; Jiajun ZHAO ; Dong ZHAO ; Ji HU ; Jiang HU ; Xinguo HOU ; Bimin SHI ; Tianpei HONG ; Mingxia YUAN ; Weibo XIA ; Xuejiang GU ; Yong XU ; Shuguang PANG ; Tianshu GAO ; Zuhua GAO ; Xiaohui GUO ; Hongyi CAO ; Mingfeng CAO ; Xiaopei CAO ; Jing MA ; Bin LU ; Zhen LIANG ; Jun LIANG ; Min LONG ; Yongde PENG ; Jin LU ; Hongyun LU ; Yan LU ; Chunping ZENG ; Binhong WEN ; Xueyong LOU ; Qingbo GUAN ; Lin LIAO ; Xin LIAO ; Ping XIONG ; Yaoming XUE
Chinese Journal of Endocrinology and Metabolism 2025;41(11):891-907
Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.
10.Association and mechanism between genetic variants in binding region of pancreatic and duodenum homeobox-1 and susceptibility to pancreatic cancer
Wen TIAN ; Yizhuo LIU ; Bin LI ; Jianbo TIAN ; Xiaoping MIAO ; Ying ZHU
Chinese Journal of Preventive Medicine 2025;59(5):581-587
Objective:To systematically investigate the association and regulatory mechanism between genetic variants in the binding region of pancreatic and duodenal homeobox 1 (PDX1) and pancreatic cancer susceptibility in the Chinese population.Methods:Chromatin immunoprecipitation sequencing (ChIP-seq) was performed using the human pancreatic cancer cell line BxPC-3 to identify and annotate genetic variants within the PDX1 binding region. A two-center case-control study was conducted, and logistic regression models were employed to analyze the association between PDX1-related variants and pancreatic cancer susceptibility. Functional experiments were performed to elucidate the molecular mechanisms of these genetic variants.Results:ChIP-seq analysis identified 1 608 PDX1 binding regions. SNPs within these regions were significantly enriched in susceptible areas of pancreatic cancer ( P<0.001). The common variant rs154659, located within the most significant PDX1 binding peak, was further investigated. The multivariate logistic regression model showed that compared with individuals with TT genotype, individuals with CC genotype had a reduced risk of pancreatic cancer by 29.2% ( OR=0.708, 95% CI: 0.589-0.850). Functional studies demonstrated that the rs154659[C] allele displayed higher relative luciferase activity than the rs154659[T] allele. Knockdown of PDX1 significantly attenuated the relative luciferase differences between the two alleles. Conclusion:Genetic variants in the PDX1 binding region are associated with pancreatic cancer risk. The rs154659 modulates pancreatic cancer susceptibility by specifically altering PDX1 binding activity.

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