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
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Glycine max/metabolism*
;
Soybean Proteins/metabolism*
;
Peptide Hydrolases/metabolism*
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Ribosomes/metabolism*
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Globulins
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Antigens, Plant
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Seed Storage Proteins
2.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*
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Stress, Physiological
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Multigene Family
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Gene Expression Regulation, Plant
3.Expert consensus on clinical application of 177Lu-prostate specific membrane antigen radio-ligand therapy in prostate cancer
Guobing LIU ; Weihai ZHUO ; Yushen GU ; Zhi YANG ; Yue CHEN ; Wei FAN ; Jianming GUO ; Jian TAN ; Xiaohua ZHU ; Li HUO ; Xiaoli LAN ; Biao LI ; Weibing MIAO ; Shaoli SONG ; Hao XU ; Rong TIAN ; Quanyong LUO ; Feng WANG ; Xuemei WANG ; Aimin YANG ; Dong DAI ; Zhiyong DENG ; Jinhua ZHAO ; Xiaoliang CHEN ; Yan FAN ; Zairong GAO ; Xingmin HAN ; Ningyi JIANG ; Anren KUANG ; Yansong LIN ; Fugeng LIU ; Cen LOU ; Xinhui SU ; Lijun TANG ; Hui WANG ; Xinlu WANG ; Fuzhou YANG ; Hui YANG ; Xinming ZHAO ; Bo YANG ; Xiaodong HUANG ; Jiliang CHEN ; Sijin LI ; Jing WANG ; Yaming LI ; Hongcheng SHI
Chinese Journal of Clinical Medicine 2024;31(5):844-850,封3
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.
4.Atrial fibrillation diagnosis algorithm based on improved convolutional neural network.
Yu PU ; Junjiang ZHU ; Detao ZHANG ; Tianhong YAN
Journal of Biomedical Engineering 2021;38(4):686-694
Atrial fibrillation (AF) is a common arrhythmia, which can lead to thrombosis and increase the risk of a stroke or even death. In order to meet the need for a low false-negative rate (FNR) of the screening test in clinical application, a convolutional neural network with a low false-negative rate (LFNR-CNN) was proposed. Regularization coefficients were added to the cross-entropy loss function which could make the cost of positive and negative samples different, and the penalty for false negatives could be increased during network training. The inter-patient clinical database of 21 077 patients (CD-21077) collected from the large general hospital was used to verify the effectiveness of the proposed method. For the convolutional neural network (CNN) with the same structure, the improved loss function could reduce the FNR from 2.22% to 0.97% compared with the traditional cross-entropy loss function. The selected regularization coefficient could increase the sensitivity (SE) from 97.78% to 98.35%, and the accuracy (ACC) was 96.62%, which was an increase from 96.49%. The proposed algorithm can reduce the FNR without losing ACC, and reduce the possibility of missed diagnosis to avoid missing the best treatment period. Meanwhile, it provides a universal loss function for the clinical auxiliary diagnosis of other diseases.
Algorithms
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Atrial Fibrillation/diagnosis*
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Electrocardiography
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Humans
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Neural Networks, Computer
;
Stroke
5.BRICS report of 2018-2019: the distribution and antimicrobial resistance profile of clinical isolates from blood culture in China
Yunbo CHEN ; Jinru JI ; Chaoqun YING ; Peipei WANG ; Zhiying LIU ; Qing YANG ; Haishen KONG ; Hui DING ; Yongyun LIU ; Haifeng MAO ; Ying HUANG ; Zhenghai YANG ; Yuanyuan DAI ; Guolin LIAO ; Lisha ZHU ; Liping ZHANG ; Yanhong LI ; Hongyun XU ; Junmin CAO ; Baohua ZHANG ; Liang GUO ; Haixin DONG ; Shuyan HU ; Sijin MAN ; Lu WANG ; Zhixiang LIAO ; Rong XU ; Dan LIU ; Yan JIN ; Yizheng ZHOU ; Yiqun LIAO ; Fenghong CHEN ; Beiqing GU ; Jiliang WANG ; Jinhua LIANG ; Lin ZHENG ; Aiyun LI ; Jilu SHEN ; Yinqiao DONG ; Lixia ZHANG ; Hongxia HU ; Bo QUAN ; Wencheng ZHU ; Kunpeng LIANG ; Qiang LIU ; Shifu WANG ; Xiaoping YAN ; Jiangbang KANG ; Xiusan XIA ; Lan MA ; Li SUN ; Liang LUAN ; Jianzhong WANG ; Zhuo LI ; Dengyan QIAO ; Lin ZHANG ; Lanjuan LI ; Yonghong XIAO
Chinese Journal of Clinical Infectious Diseases 2021;14(1):32-45
Objective:To investigate the distribution and antimicrobial resistance profile of clinical bacteria isolated from blood culture in China.Methods:The clinical bacterial strains isolated from blood culture from member hospitals of Blood Bacterial Resistant Investigation Collaborative System (BRICS) were collected during January 2018 to December 2019. Antibiotic susceptibility tests were conducted with agar dilution or broth dilution methods recommended by US Clinical and Laboratory Standards Institute (CLSI). WHONET 5.6 was used to analyze data.Results:During the study period, 14 778 bacterial strains were collected from 50 hospitals, of which 4 117 (27.9%) were Gram-positive bacteria and 10 661(72.1%) were Gram-negative bacteria. The top 10 bacterial species were Escherichia coli (37.2%), Klebsiella pneumoniae (17.0%), Staphylococcus aureus (9.7%), coagulase-negative Staphylococci (8.7%), Pseudomonas aeruginosa (3.7%), Enterococcus faecium (3.4%), Acinetobacter baumannii(3.4%), Enterobacter cloacae (2.9%), Streptococci(2.8%) and Enterococcus faecalis (2.3%). The the prevalence of methicillin-resistant S. aureus (MRSA) and methicillin-resistant coagulase-negative Staphylococcus were 27.4% (394/1 438) and 70.4% (905/1 285), respectively. No glycopeptide-resistant Staphylococcus was detected. More than 95% of S. aureus were sensitive to amikacin, rifampicin and SMZco. The resistance rate of E. faecium to vancomycin was 0.4% (2/504), and no vancomycin-resistant E. faecalis was detected. The ESBLs-producing rates in no carbapenem-resistance E. coli, carbapenem sensitive K. pneumoniae and Proteus were 50.4% (2 731/5 415), 24.6% (493/2001) and 35.2% (31/88), respectively. The prevalence of carbapenem-resistance in E. coli and K. pneumoniae were 1.5% (85/5 500), 20.6% (518/2 519), respectively. 8.3% (27/325) of carbapenem-resistance K. pneumoniae was resistant to ceftazidime/avibactam combination. The resistance rates of A. baumannii to polymyxin and tigecycline were 2.8% (14/501) and 3.4% (17/501) respectively, and that of P. aeruginosa to carbapenem were 18.9% (103/546). Conclusions:The surveillance results from 2018 to 2019 showed that the main pathogens of bloodstream infection in China were gram-negative bacteria, while E. coli was the most common pathogen, and ESBLs-producing strains were in majority; the MRSA incidence is getting lower in China; carbapenem-resistant E. coli keeps at a low level, while carbapenem-resistant K. pneumoniae is on the rise obviously.
6.Surveillance of drug resistance of clinically isolated fungi strains from 46 hospitals in Shandong Province
Linna KOU ; Chunyan ZHANG ; Zheng LI ; Bin JI ; Sijin MAN ; Maoli YI ; Renzhe LI ; Mingyan SUN ; Yuanqi ZHU ; Jiliang WANG ; Mingju HAO ; Chengjie GUO ; Jing LI ; Wenwen YU ; Zhongtao GAI ; Shifu WANG
Chinese Journal of Clinical Infectious Diseases 2021;14(1):46-53,80
Objective:To analyze the distribution of clinically isolated fungal strains and their resistance to common antifungal drugs in Shandong province.Methods:Through the Shandong Children’s Bacterial & Fungal Drug Resistance Surveillance and Research Collaborative Network, a total of 1 030 fungi were collected in 46 hospitals of Shandong province from January 1 to December 31, 2018. The source and type of strains were analyzed, and antifungal drug sensitivity tests were performed by using the micro-dilution method. Whonet 5.6 and SPSS 22.0 were applied to analyze the data.Results:The overall main strains were Candida albicans (38.74%, 399/1 030), Candida tropicalis (16.99%, 175/1 030) and Candida parapsilosis (16.41%, 169/1 030); the main fungi strains in child patients were C. albicans (52.50%, 63/120), C. parapsilosis (12.50%, 15/120) and C. tropicalis (9.17%, 11/120); the main fungi strains in adult patients were C. albicans (36.37%, 331/910), C. tropicalis (17.03%, 155/910) and C. parapsilosis (15.27%, 139/910). The isolation rate of main Candida strains from January to March and August to December was much higher than that of other months. The drug resistance rates of C. albicans to fluconazole and voriconazole were 7.14% and 7.43%, respectively, and the drug resistance rates to itraconazole were 50.44%. The resistance rates of C. tropicalis to fluconazole, voriconazole and itraconazole were 29.05%, 23.29% and 48.65%, respectively. The sensitivity rates of C. parapsilosi to fluconazole, voriconazole and itraconazole were 93.06%, 93.75% and 94.44%, respectively. Candida glabrata showed a dose-dependent sensitivity rate of 2.33% to fluconazole. Analysis of 244 blood fungi strains showed that non-candida albicans bacteremia accounted for 70.08%. In the pathogen spectrum covering 92.22%, fluconazole was sensitive to 64.65% of the pathogens, voriconazole was 68.88%, and amphotericin B was 88.75%. After quantification, the effective rates of fluconazole, voriconazole and amphotericin B in the clinical treatment of fungal bacteremia were 70.10%, 74.69% and 96.23%, respectively. Among them, the sensitivity rate of voriconazole to C. tropicalis was lower than that of fluconazole. Conclusions:Candida is the main clinical fungus isolates in hospitals of Shandong province. The resistance rate of C. tropicalis to azole antifungal drugs is on the rise, and the sensitivity of other Candida species to clinically used antifungal drugs is basically stable.
7.Screening of tomato cultivars in cadmium-polluted areas and study on their antioxidant capacity.
Jiamin YANG ; Hexigeduleng BAO ; Jiayue WAN ; Yanfei DING ; Feijuan WANG ; Cheng ZHU
Chinese Journal of Biotechnology 2021;37(1):242-252
To screen the available tomato pollution-safe cultivar varieties and reduce the potential food safety risks in Cd-polluted areas, the differences of Cd accumulation in different tomato (Solanum lycopersicum) varieties in southern China were studied by soil culture and hydroponic experiments. Firstly, the high and low accumulation varieties were selected from 25 tomato varieties under 2.94 mg/kg Cd stress by soil culture test, and then the responses of high and low accumulation tomato varieties to Cd stress were determined by hydroponic experiments. The results of soil culture test show that under 2.94 mg/kg Cd stress, there were significant differences in plant height, total biomass and yield among 25 tomato cultivars, and the Cd contents of fruits of all 25 tomato cultivars exceeded the highest limit value (0.05 mg/kg) of CAC (Codex alimentarius commission). Through cluster analysis, 7, 4 and 14 varieties accumulating relatively high, medium, and low concentrations of Cd in the fruits were screened, among which the highest, the lowest, and the average Cd contents in the fruits were 3.06 mg/kg DW, 1.47 mg/kg DW, and 2.21 mg/kg DW, respectively. The results of hydroponic experiment show that under the same concentration of Cd stress, Qiantangxuri F1, a high Cd accumulating variety, absorbed Cd faster, accumulated more Cd, used shorter oxidative stress response time and had stronger tolerance to Cd than Zhefen 3053, a low Cd accumulating variety. The typical high and low Cd accumulating varieties can provide a reference for agricultural production in heavy metal polluted areas and the development of molecular-assisted breeding methods of PSC. At present, cultivating low Cd accumulating PSC varieties and dynamic monitoring of Cd contents in tomato fruits are feasible methods in medium and light Cd-polluted areas.
Antioxidants
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Cadmium/toxicity*
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China
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Lycopersicon esculentum
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Plant Breeding
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Plant Roots/chemistry*
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Soil Pollutants/toxicity*
8.Evaluation of hepatic fibrosis in chronic hepatitis B with parameters obtained from multi-models of diffusion-weighted MRI
Fangfang FU ; Meiyun WANG ; Dapeng SHI ; Shaocheng ZHU ; Jiliang ZHANG ; Yan BAI ; Shewei DOU
Chinese Journal of Radiology 2018;52(2):113-118
Objective To explore the value of various diffusion parameters obtained from monoexponential, biexponential and stretched exponential diffusion-weighted imaging models in assessing hepatic fibrosis in chronic hepatitis B(CHB). Methods A total of 52 patients who were diagnosed hepatitis B by the markers of hepatitis and were confirmed by liver biopsy pathology were prospectively recruited between June 2014 and May 2016 in People's Hospital of Henan Province. Concomitantly, 30 healthy volunteers who had no history of hepatitis B and liver dysfunction were enrolled in the control group. All patients underwent multi-b values DWI on a 3.0 T MRI unit. ADC was calculated by using the monoexponential model. True diffusion coefficient(Dt),pseudo diffusion coefficient(Dp)and fraction of perfusion(f)were calculated by using the biexponential model.Distributed diffusion coefficient(DDC)and water molecular diffusion heterogeneity index(α)were calculated by using the stretched exponential model. Liver biopsy specimens were staged according to the degree of hepatic fibrosis (S0 to S4). The Kruskal-Wallis test was employed for the comparison of each parameter among the control group and the fibrosis stage groups. The Mann-Whitney U test was adopted to compare each parameter between fibrosis stage≤S1and≥S2,between≤S2 and≥S3.Spearman rank correlation coefficients were obtained to assess the correlation of the parameters with the fibrosis stages.ROC analysis was used to evaluate the performance of various parameters in predicting stage≥S2 and≥S3.Results The hepatic fibrosis stage distributions were as follows:1 cases with S0,9 cases with S1,22 cases with S2,11 cases with S3,9 cases with S4.ADC,Dt,f and DDC values all showed significant difference among the control group and groups S1,S2,S3,S4(all P<0.05), while Dp and α values showed no significant difference(P>0.05). Dt, DDC and ADC showed a moderate negative correlation with the fibrosis stage(r=-0.630,-0.603 and-0.464,respectively,all P<0.01),and f showed a mild negative correlation with the fibrosis stage(r=-0.379,P<0.05),while Dp and α values showed no correlation with the fibrosis stages(all P>0.05).The ADC, Dt, f and DDC values all showed significant difference between stage≥S2 and≤S1,between stage≥S3 and≤S2(all P<0.05),and the parameter values of the former were greater than those of the latter. While Dp and α values showed no significant difference among both groups(all P>0.05).The AUCs of ADC,Dt,f and DDC values for detecting fibrosis stage ≥S2 were 0.738,0.835,0.740 and 0.831, and the AUCs of ADC, Dt, f and DDC values for detecting fibrosis stage≥S3 were 0.716, 0.811, 0.672 and 0.798. Conclusion The Dt derived from biexponential and DDC derived from stretched exponential DWI could be useful for the staging of hepatic fibrosis in CHB.
9.Research on improving memory impairment of blue lavender volatile oil.
Li-Yun ZHU ; Yong-Sheng GAO ; Lin-Zhen SONG ; Su-Fang LI ; Jun-Qing QIAN
China Journal of Chinese Materia Medica 2017;42(24):4819-4826
In order to study the potential application value of lavender volatile oil (LVO), the chemical composition of the volatile oil of lavender was analyzed by GC-MS, and the mouse model of Alzheimer's disease (AD) was established. Additionally, the antioxidant enzymes activity of T-SOD, GSH-PX, CAT and MDA content were studied. Experimental results showed that 55 kinds of chemical constituents including terpene, terpene alcohol and ester compounds from LVO were identified, and the content of linalool and linalyl acetate was the highest, accounting for 49.71% of the total volatile oil. The ability of mouse platform memory was improved significantly. The levels of GSH-PX, CAT and T-SOD of mouse brain tissue in the treatment group were significantly higher than those in the model group (P<0.05). The level of MDA reached the maximum value in the model group, while there was no notable difference between the levels of MDA in the drug group and the normal group. The result indicated the significant oxidative activity of LVO, the possibility of induced oxidative stress reduction in neurons, and the reversal effect of memory acquired disorder.
10.Effect of compound flavonoids on atherosclerosis in ApoE -/- mice
Xiaoqing ZHU ; Jie YANG ; Juan SHAO ; Chao LIU ; Jiliang WU
Chinese Pharmacological Bulletin 2017;33(2):180-184
Aim To investigate the protective effect of compound total flavonoids on atherosclerosis in ApoE -/- knockout mice.Methods Seven-week old C57BL/6 mice considered of the normal group (n =1 5 );seven-week old ApoE -/- mice were fed with high-fat diet and were assigned randomly into 5 groups:model group,simvastatin group,the low com-pound flavonoids group,the middle compound fla-vonoids group and the high compound flavonoids group.After 1 6 weeks,mice serum and aortas were harvested.The formation of atherosclerotic plaque was analyzed by HE staining,The serum level of lipids pro-files and superoxide dismutase (SOD )were deter-rnined.The levels of IL-1 βand NF-κB in serum were detected by ELISA assay.Results Area of atheroscle-rotic lesion was significantly less in the compound fla-vones group than in model.The level of TC,TG,LDL-C,IL-1 β,NF-κB in serum of the compound flavonoids group were decreased significantly,while SOD and HDL-C increased significantly compared with the mod-el group,and the difference was significant (P <0.05).Conclusion The compound flavonoids have a good protective effect on early atherosclerosis in mice, which may be due to its alleviating effects on hyperlipi-demia and inflammation and oxidation.

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