ObjectiveTo establish a quality evaluation method for Zhuye Shigao granules（Zhuye Shigaotang） based on fingerprint and determination of index components， and to investigate the therapeutic effect of Zhuye Shigao granules on mice with cold-dampness pestilence attacking lung syndrome. MethodsThe fingerprint of Zhuye Shigao granules was established by high performance liquid chromatography（HPLC）， and the methods for determination of total calcium， orientin， isoorientin， ginsenosides Rg₁， Re and Rb₁ and other 2 index components were established. Fifty ICR mice were randomly divided into the blank group， model group， Zhuye Shigao granules low， medium and high dose groups（9.3， 18.6， 37.2 g·kg^-1·d^-1）， with 10 mice in each group. In addition to the blank group， the model mice with cold-dampness pestilence attacking lung syndrome was prepared by nasal drip of lipopolysaccharide combined with cold-dampness environment. Each administration group was given the corresponding liquid by gavage according to the dose， while the blank group and model group were given the same volume of normal saline by gavage. Then， the body temperature and organ index of mice in each group were measured， hematoxylin-eosin（HE） staining was used to investigate the lung tissue injury of mice in each group， and enzyme-linked immunosorbent assay（ELISA） was used to detect the changes of tumor necrosis factor-α（TNF-α）， interleukin（IL）-lβ， IL-6， IL-10 levels in serum and lung tissue， as well as immunoglobulin（Ig）A and IgM levels in serum. ResultsThe fingerprint similarity of 10 batches of Zhuye Shigao granules was>0.950， and 20 common peaks were calibrated. Seven of them were identified， including peak 11（isoorientin）， peak 12（orientin）， peak 14（apioside liquiritin）， peak 15（liquiritin）， peak 17（apioside isoliquiritin）， peak 19（isoliquiritin） and peak 20（liquiritigenin）. The results of quantitative analysis showed that the content range of each index component in 10 batches of Zhuye Shigao granules was as follows：Total calcium of 9.978-11.294 mg·g^-1， isoorientin of 0.033-0.041 mg·g^-1， orientin of 0.046-0.055 mg·g^-1， ginsenoside Rg₁+ginsenoside Re of 0.748-0.762 mg·g^-1， ginsenoside Rb₁ of 0.151-0.197 mg·g^-1， liquiritin of 1.106-1.366 mg·g^-1， glycyrrhizic acid of 0.904-1.182 mg·g^-1_. Compared with the blank group， the body temperature of mice in the model group was significantly increased， the organ indexes of liver， lung and spleen were significantly decreased， the organ index of thymus was significantly increased， HE staining of lung tissue showed infiltration of inflammatory cells， a small amount of serous exudation was observed in the alveoli， and lung tissue was damaged. After the intervention of Zhuye Shigao granules， the pathological changes were improved compared with the model group. The expression levels of IL-1β， IL-6 and TNF-α were significantly increased， the expression level of IL-10 was significantly decreased in serum and lung tissue. The levels of IgA and IgM in serum were significantly decreased（P<0.01）. Compared with the model group， the body temperature， the organ indexes and immune factor levels in serum and lung tissue of mice in the Zhuye Shigao granules medium and high dose groups were significantly reduced（P<0.05， P<0.01）. ConclusionIn this study， the quality evaluation of Zhuye Shigao granules was carried out based on fingerprint combined with determination of index components， and the fingerprint of four herbs（Lophatheri Herba， Ophiopogonis Radix， Pinelliae Rhizoma and Glycyrrhizae Radix et Rhizoma） in this formula and the determination of 8 index components were established. The therapeutic effect of Zhuye Shigao granules on mice with cold-dampness pestilence attacking lung syndrome may be related to inhibiting inflammatory response and mediating immune regulation.

Objective An HPLC-MS/MS method was established for the simultaneous determination of 7 components in Qingkailing Oral Liquid.Methods The assay was performed on a Waters ACQUITY UPLC BEH C18 column(2.1 mm×10 mm,1.7 μm)and the sample was eluted with a gradient mobile phase containing 10 mmol·L-1 of ammonium acetate and 0.1%of formic acid in water(A)-methanol(B).The mass spectrometry was carried out by electrospray ionization(ESI)with positive/negative ions in multiple reaction monitoring(MRM)mode for quantitative analysis.Results The linear ranges of adenine,chlorogenic acid,caffeic acid,geniposide,baicalin,hyodeoxycholic acid and cholic acid were 0.100 4-3.213,0.784 5-8.982,0.998-3.194,0.622 5-19.92,25.05-300.6,2.513-30.15 and 7.775-93.30 μg·mL-1(r≥0.999 0).The average recoveries(n=6)were 100.9%,98.74%,101.2%,100.2%,100.8%,99.97%and 98.94%with RSD of 1.58%,0.59%,1.78%,1.25%,0.65%,1.69%and 1.07%.The contents of the above mentioned 7 components in 15 tested samples were in the ranges of 0.12-0.18,0.19-0.24,0.06-0.09,0.34-0.37,4.54-4.85,0.49-0.67 and 1.82-2.19 mg·mL-1.The contents of 7 components in tested sample from different manufacturers were closed.Conclusion The method has shown good sensitivity,accuracy,and repeatability.The study can provide reference and data support for the quality control and subsequent research of Qingkailing Oral Liquid.

Objective:Care bundles for critically ill patients with early enteral nutrition up to goal was constructed. Its purpose was to improve early enteral nutrition, prognosis and provide reference basis for improving the rate of standard of early enteral nutrition in critically ill patients.Methods:By conducting systematic searching of domestic and foreign Chinese and English databases, related guide websites, relevant documents on early enteral nutrition in critically ill patients up to goal, which were obtained, evaluated, extracted, summarized and graded. After discussion by the research group, the first draft was prepared. Delphi method was used to conduct two rounds of expert correspondence, and the final draft of the proposal was established through the reliability analysis of correspondence results.Results:Twenty experts participated finally, and their opinions tended to be consistent after two rounds of expert inquiry. The authority coefficients were 0.92 and 0.91 respectively. The variation coefficients of the importance and operability of the two rounds of correspondence items were 0.05-0.20 and 0.05-0.21, 0.00-0.17 and 0.00-0.20 respectively. The Kendall concordance coefficients for the importance and operability of the two rounds of correspondence items were 0.16 and 0.13, 0.27 and 0.18 respectively. The differences were statistically significant ( χ2 values were 117.01-228.43, all P<0.05). Finally, the final draft of bundle of care for early enteral nutrition up to goal in critically ill patients was established which included three aspects related to evaluation, implementation, and effectiveness monitoring, besides care bundle included 12 intervention perspectives and 29 specific intervention measures. Conclusions:Based on evidence-based and delphi method constructing care bundles for critically ill patients with early enteral nutrition up to goal was scientific, reliable and practical which could provide theoretical and practical guidance for bundled nursing interventions to meet early enteral nutrition standards in critically ill patients.

Seven novel linear polyketides,talaketides A-G(1-7),were isolated from the rice media cultures of the mangrove sed-iment-derived fungus Talaromyces sp.SCSIO 41027.Among these,talaketides A-E(1-5)represented unprecedented unsaturated lin-ear polyketides with an epoxy ring structure.The structures,including absolute configurations of these compounds,were elucidated through detailed analyses of nuclear magnetic resonance(NMR)and high-resolution mass spectrometry(HR-MS)data,as well as elec-tronic custom distributors(ECD)calculations.In the cytotoxicity screening against prostate cancer cell lines,talaketide E(5)demon-strated a dose-dependent inhibitory effect on prostate cancer PC-3 cell lines,with an IC50 value of 14.44 μmol·L-1.Moreover,com-pound 5 significantly inhibited the cloning formation of PC-3 cell lines and arrested the cell cycle in S-phase,ultimately inducing ap-optosis.These findings indicate that compound 5 may serve as a promising lead compound for the development of a potential treat-ment for prostate cancer.

ObjectiveTo compare the regulatory effects of five plant polysaccharides on immune function of cyclophosphamide （CTX）-induced immunosuppressed mice by network Meta-analysis， to provide evidence for the clinical application of polysaccharides and the development of effective polysaccharides and oligosaccharides. MethodSeven databases including PubMed， Embase and Web of Science were searched， and studies that met the inclusion criteria were selected. The methodological quality of the included studies was evaluated using the risk of bias tool of Systematic Review Centre for Laboratory Animal Experimentation （SYRCLE）， and the data were analyzed using RStudio and StataSE 17. ResultA total of 62 randomized controlled trials （RCTs） were included， involving 1 512 mice and five plant polysaccharides： Astragalus polysaccharide （APS）， lentinan （LNT）， Ganoderma lucidum polysaccharide （GLP）， Poria cocos polysaccharide （PCC）， and Codonopsis pilosula polysaccharide （CPP）. The network Meta-analysis showed that APS ranked first in increasing spleen index （mean deviation （MD）=3.54， 95% confidence interval （CI） ［2.10， 5.96］）， thymus index （MD=1.98， 95%CI ［1.55， 2.54］） and T helper cells （CD4⁺）/T suppressor cells （CD8⁺） （MD=1.63， 95%CI ［1.13， 2.37］）， while CPP ranked first in up-regulating the number of peripheral blood leukocytes （MD=24.16， 95%CI ［8.21， 71.12］）， macrophage phagocytosis index （MD=2.52， 95%CI ［1.32， 4.82］） and immunoglobulin M （IgM） content （MD=1.79， 95%CI ［1.12， 2.85］）. ConclusionAll the five plant polysaccharides can regulate the immune function of immunosuppressed mice. Among them， APS has advantages in elevating spleen index， thymus index and CD4⁺/CD8⁺， while CPP focuses on increasing the number of peripheral blood leukocytes， macrophage phagocytosis index and IgM content. Due to limited number and quality of included studies， the conclusions needs to be further verified with large samples and high-quality studies.

L-asparaginase (L-ASN) is widely applied in the treatment of malignant tumor and low-acrylamide food production, however, the low expression level hampers its application. Heterologous expression is an effective strategy to increase the expression level of target enzymes, and Bacillus is generally used as the host for efficient production of enzymes. In this study, the expression level of L-asparaginase in Bacillus was enhanced through optimization of expression element and host. Firstly, five signal peptides (SP_SacC, SP_AmyL, SP_AprE, SP_YwbN and SP_WapA) were screened, among which SP_SacC showed the best performance, reaching an activity of 157.61 U/mL. Subsequently, four strong promoters (P43, P_ykzA-P43, P_Ubay and P_bacA) from Bacillus were screened, and tandem promoter P_ykzA-P43 showed the highest yield of L-asparaginase, which was 52.94% higher than that of control strain. Finally, three Bacillus expression hosts (B. licheniformis Δ0F3 and BL10, B. subtilis WB800) were investigated, and the maximum L-asparaginase activity, 438.3 U/mL, was reached by B. licheniformis BL10, which was an 81.83% increase compared with that of the control. This is also the highest level of L-asparaginase in shake flask reported to date. Taken together, this study constructed a B. licheniformis strain BL10/P_ykzA-P43-SP_SacC-ansZ capable of efficiently producing L-asparaginase, which laid the foundation for industrial production of L-asparaginase.
Bacillus licheniformis/metabolism* ; Asparaginase/genetics* ; Bacillus/genetics* ; Protein Sorting Signals ; Promoter Regions, Genetic/genetics* ; Bacillus subtilis/genetics* ; Bacterial Proteins

Objective:To explore the selection criteria of the donor for islet transplantation of Chinese people by analyzing the correlation between pancreas characteristics and success rate of islets isolation.Methods:Data from 113 cases of human islet isolation were collected. According to the result of islet isolation, the donors were divided into two groups, the success group(IEQ≥250 000, purification≥30%, and viability≥80%), and the failure group(IEQ<250 000, or purification<30%, or viability<80%). The modified Ricordi method was used to digest pancreas tissue, and the continuous density gradient method was performed to purify islets. The islets were identified by staining with the Dithizone(DTZ), the islets were analyzed for cell viability and purity.Results:The donor age in success group was significantly younger than failure group in the range of age eligible for this study( t=2.479, P=0.015). Pearson correlation showed that donor age was positively corelated with islet yield( r=-0.214, P=0.047). There was more fat on the pancreas surface in the successful islet isolation group( z=-2.007, P=0.045). The digestibility( t=2.133, P=0.035) and recovery rate( t=5.912, P=0.001) were elevated in success group. Conclusion:The pancreases from younger donors could obtain the higher-yielding islet, the pancreas with more surface fat or with higher weight was associated with islet isolation success in the scope covered by the inclusion criteria of this study.

Objective:To investigate the prognosis and influencing factors in critically ill surgical patients of different feeding intolerance trajectories.Methods:The retrospective cohort study was conducted. The clinical data of 354 critically ill surgical patients who were admitted to 69 medical centers in the Chinese Critical Care Nutrition Trials Group -NEED database from March 2018 to July 2019 were selected. There were 247 males and 107 females, aged 58(46,68)years. According to the trajectory model of feeding intolerance change, 354 patients were divided into 3 categories as feeding intolerance, decreased feeding intolerance, continuous feeding intolerance, including 164, 49, 141 cases respectively. Observation indicators: (1) general situations of patients of different feeding intolerance trajectories; (2) treatment of patients of different feeding intolerance trajectories; (3) survival of patients of different feeding intolerance trajectories; (4) analysis of pro-gnostic factors in critically ill surgical patients. Measurement data of normal distribution were expressed as Mean± SD, and one-way analysis of variance was used for comparison between groups. Measurement data of skewed distribution were expressed as M( Q1, Q3), and Kruskal-Wallis rank sum test was used for comparison between groups. Count data were expressed as absolute numbers or percentages, and chi-square test was used for comparison between groups. Ordinal data were compared using the Kruskal-Wallis rank sum test. Bonferroni correction was used for pairwise comparison. Group-based trajectory model was constructed according to Traj plug-in in Stata17.0 statistical software, and the optimal trajectory model was evaluated by Bayesian information criterion and average posterior probability parameter. The Kaplan-Meier method was used to draw the survival curve and calculate the survival rate, and Log-Rank test was used for survival analyses. Univariate and multivariate analyses were conducted using the COX proportional hazard regression model. Results:(1) General situations of patients of different feeding intolerance trajectories. Of 354 critically ill surgical patients, 257 cases underwent enteral nutrition and 97 cases underwent enteral plus parenteral nutrition. The acute physiological and chronic health score (APACHEII) was 17(13,21), and the sequential organ failure score (SOFA) was 6(5,8). The modified Critical Illness Nutritional risk score (mNUTRIC) was 4 (2,5), the number of complications was 2(1,3). There were 293, 55 and 6 patients with grade Ⅰ, grade Ⅱ and grade Ⅲ acute gastrointestinal injury (AGI), and there were 224, 17 and 61 patients who were treated with mechanical ventilation, continuous renal replacement therapy and vasoactive drugs, respectively. The incidence of feeding intolerance in 354 patients increased first and then decreased, reaching a peak of 25.42%(90/354) on the third day and 53.67%(190/354) within 7 days. Of 354 critically ill surgical patients, cases with no feeding intolerance, decreased feeding intolerance, continuous feeding intolerance had the APACHE Ⅱ as 16(12,20), 17(14,25), 18(13,22), mNUTRIC as 3(2,5), 4(3,6), 4(3,5), the number of complications as 2(1,2), 2(2,3), 2(2,3). There were 152, 27, 114 cases with grade Ⅰ AGI, 12, 22, 27 cases with grade Ⅱ-Ⅲ AGI, 95, 39, 90 cases with mechanical ventilation. There were significant differences in the above indicators among the three groups ( H=6.14, 13.11, 28.05, χ2=37.96, 7.65, P< 0.05). Further analysis showed that compared with patients with no feeding intolerance, patients with decreased feeding intolerance and continuous feeding intolerance had the higher number of complications and grade of AGI ( Z=60.32, 54.69, χ2=39.72, 9.52, P<0.05), patients with decreased feeding intolerance had the higher mNUTRIC scores and ratio of mechanical ventilation ( Z=53.41, χ2=7.59, P<0.05). (2) Treatment of patients of different feeding intolerance trajectories. Cases with prokinetic drugs use and post-pyloric feeding were 36, 13 of patients with no feeding intolerance, 25 and 10 of patients with decreased feeding intolerance, 46 and 19 of patients with continuous feeding intolerance, respectively, showing significant differences in the above indicators among the three groups ( χ2=15.76, 6.20, P<0.05). Further analysis showed that compared with patients with no feeding intolerance, patients with decreased feeding intolerance had higher ratio of prokinetic drugs use and ratio of post-pyloric feeding ( χ2=15.60, 6.10, P<0.05). (3) Survival of patients of different feeding intolerance trajectories. The 28-day overall survival rates of patients with no feeding intolerance, decreased feeding intolerance, and continued feeding intolerance were 96.96%, 95.92%, and 87.94%, respectively, showing a significant difference ( χ2=10.39, P<0.05). Further analysis showed a significant difference between patents with no feeding intolerance and patients with continuous feeding intolerance ( χ2=9.19, P<0.05). (4) Analysis of prognostic factors in critically ill surgical patients. Multivariate analysis showed that continuous feeding intolerance was an independent risk factor for 28-day death in critically ill surgical patients ( hazard ratio=3.92, 95% confidence interval as 1.43-10.79, P<0.05). Conclusion:For surgical critically ill patients, patients with continuous feeding intolerance have a higher 28-day mortality than patients with no feeding intolerance, and the continuous feeding intolerance is an independent risk factor for 28-day death in critically ill surgical patients.

Objective To evaluate the impact of hepatobiliary surgery in a tertiary public hospital on its core medical operation indicators after the implementation of subspecialty reform.Methods The Department of Hepatobiliary Sur-gery,which implemented the subspecialty reform,was the experimental group,and six surgical departments with-out subspecialties were the control group.Seven indicators in the dimensions of medical quality,medical efficiency,and rational use of medication were collected from September to December 2022,and the difference between the experimental group and the control group was analyzed by the differences-in-differences method using the Septem-ber data before the implementation of sub-specialty reform as the baseline value.Results After the subspecialty re-form,the changes of the indicators of the percentage of discharged patients for surgery,intensity of antimicrobial drug use and average hospital stay in hepatobiliary surgery were better than those of the control group;the percentage of discharged patients for minimally invasive surgery and the percentage of medical service revenue were inferior to those of the control group;the intergroup differences of the two indicators of the percentage of discharged patients for level IV surgery and the case-mix index were not significant.Conclusion By setting up similar control departments in the same period and excluding the influence of external confounding factors on the study results,overall,the core medical operation indexes of hepatobiliary surgery were slightly better than those of the control group,but the advantage was not obvious,which might be related to the short observation time after the subspecialty reform,and its long-term effects need to be continuously followed up and analyzed.

The fungus Xylaria sp. KYJ-15 was isolated from Illigera celebica. Based on the one strain many compounds (OSMAC) strategy, the strain was fermented on potato and rice solid media, respectively. As a result, two novel steroids, xylarsteroids A (1) and B (2), which are the first examples of C₂₈-steroid with an unusual β- and γ-lactone ring, respectively, along with two new dihydroisocoumarin glycosides, xylarglycosides A (3) and B (4), were identified. Their structures were elucidated by spectroscopic methods, X-ray diffraction and electronic circular dichroism (ECD) experiments. All isolated compounds were evaluated for cytotoxicity, DPPH radical scavenging activity, acetylcholinesterase inhibitory and antimicrobial effect. Compound 1 exhibited potent AChE inhibitory activity with an IC₅₀ value of 2.61 ± 0.05 μmol·L^-1. The β-lactone ring unit of 1 is critical for its AChE inhibitory activity. The finding was further confirmed through exploring the interaction of 1 with AChE by molecular docking. In addition, both compounds 1 and 2 exhibited obvious antibacterial activity against Bacillus subtilis with a minimum inhibitory concentration (MIC) of 2 μg·mL^-1. Compounds 3 and 4 exhibited antibacterial activities against Staphylococcus aureus with MICs of 4 and 2 μg·mL^-1, respectively, which also exhibited DPPH radical scavenging activity comparable to the positive control with IC₅₀ values of 9.2 ± 0.03 and 13.3 ± 0.01 μmol·L^-1, respectively.
Humans ; Acetylcholinesterase ; Molecular Docking Simulation ; Anti-Bacterial Agents ; Glycosides ; Lactones ; Pain