BACKGROUND:Macrophage migration inhibitory factor(MIF)is a pleiotropic cytokine,which is secreted in different types of stem cells and can regulate the proliferation,differentiation and migration of various types of stem cells.Our previous research has confirmed that human embryonic stem cells secrete MIF and that its concentration in the culture medium is relatively stable.However,whether MIF is involved in the survival,proliferation and differentiation of human embryonic stem cells remains unclear. OBJECTIVE:To investigate the effects of MIF on survival,proliferation,and differentiation of human embryonic stem cells. METHODS:(1)Human embryonic stem cells H9 were cultured.The growth curve of cells was detected and plotted by CCK-8 assay.Enzyme-linked immunosorbent assay was used to determine the level of MIF in the medium.(2)To determine the effects of exogenous MIF on the survival and proliferation of human embryonic stem cells,different groups were established:the control group,which was cultured in stem cell medium without any modifications;the exogenous MIF group,which was treated with different concentrations(30,100,300 ng/mL)of MIF in the stem cell medium;the MIF inhibitor ISO-1 group,which was treated with different concentrations(2,7,21 μmol/L)of ISO-1 in the stem cell medium;and the MIF+ISO-1 group,which was treated with different concentrations of ISO-1 along with 100 ng/mL of MIF.Cell viability was assessed using the CCK-8 assay.(3)To further elucidate the effect of MIF gene on survival and proliferation of human embryonic stem cell,the MIF knockout H9 cell line was constructed by CRISPR-Cas 9 technology to observe the lineage establishment.(4)To determine the effect of high concentrations of MIF on human embryonic stem cell differentiation,100 ng/mL MIF and 100 ng/mL of CXCR4 neutralizing antibody were separately added to the normal stem cell culture medium.The expression levels of self-renewal factors(KLF4,c-MYC,NANOG,OCT4,and SOX2)and differentiation transcription factors(FOXA2,OTX2)were measured using real-time quantitative polymerase chain reaction,immunofluorescence staining,and western blot analysis. RESULTS AND CONCLUSION:(1)The logarithmic growth phase of H9 cells was between 3-6 days.Under normal growth conditions,human embryonic stem cells secreted MIF at a concentration of approximately 20 ng/mL,independent of cell quantity.(2)Compared to the control group,the addition of different concentrations of MIF had no effect on the proliferation of human embryonic stem cells(P>0.05).ISO-1 significantly inhibited the proliferation of human embryonic stem cells,with a stronger inhibition observed at higher concentrations of ISO-1(P<0.05).The addition of MIF in the presence of ISO-1 reduced the inhibitory effect of ISO-1(P<0.05).(3)Real-time quantitative polymerase chain reaction showed that knocking out 50%of the MIF gene resulted in a significant decrease in the growth vitality of human embryonic stem cells and failure to establish cell lines.(4)Adding 100 ng/mL exogenous MIF to the culture medium resulted in a decrease in the mRNA,protein,and fluorescence expression levels of the self-renewal transcription factor KLF4,while the mRNA,protein,and fluorescence expression levels of the differentiation factor FOXA2 increased.(5)When 100 ng/mL CXCR4 neutralizing antibody was added to the culture medium,the mRNA and protein expression levels of KLF4 increased,while the mRNA and protein expression levels of FOXA2 decreased,contrary to the expression trend observed in the MIF group.In conclusion,the endogenous secretion of MIF by human embryonic stem cells is essential for their survival.The addition of MIF to the culture medium does not promote the proliferation of human embryonic stem cells.However,it can lead to a decrease in the expression of the self-renewal factor KLF4 and an increase in the expression of the transcription factor FOXA2.This provides a clue for further investigation into the effects and mechanisms of MIF on the differentiation of human embryonic stem cells.The MIF-CXCR4 axis plays a regulatory role in this process.

ObjectiveTo develop the Evidence Grading Scale for Ancient classical prescriptions in Traditional Chinese medicine， assess its reliability and validity， and apply it in practice to provide multi-source evidence for clinical practice guidelines development. MethodsLiterature retrieval was conducted to extract and screen existing evaluation dimensions， then the initial items were summarized using thematic analysis. Experts in the clinical medicine， medical history and literature participated in the Delphi questionnaire survey to evaluate and refine the items. An expert consensus meeting was conducted to finalize the included items， refine the method for items evaluation and evidence grading. The evidence quality rating scale for ancient classical traditional Chinese medicine （TCM） prescriptions was then established and tested for reliability and validity. ResultsThrough literature review， extraction， screening and summarization， a total of 3 dimensions and 12 initial items were formed. Questionnaires were sent to 69 experts to evaluate the initial items， with a questionnaire response rate of 100% and an expert authority coefficient of 0.92. All 12 items were retained for they had importance scores above 4. The Evidence Grading Scale on Ancient classical prescriptions in Traditional Chinese medicine includes 3 dimensions with 12 items. The 3 dimensions includes ancient evidence， inheritance status， and modern application. Each dimension contains 4 items， and each item has a full score of 5 points. The evidence was rated as high-level， moderate-level， and low-level according to the final scores. The content validity index （CVI） of the 12 items was ＞0.9， the average CVI of the scale was 0.98， and the intraclass correlation coefficient （ICC） was 0.90. ConclusionThe Evidence Grading Scale on Ancient classical prescriptions in Traditional Chinese medicine has good reliability and validity， which is practical for use in the development of TCM clinical guidelines and can better support clinical decision-making.

Objective To predict the core targets and action pathways of Hedysari Radix based on UPLC-MS/MS and network pharmacology methods,and to verify the results of network pharmacology by molecular docking and molecular dynamics techniques.This article aims to investigate immune regulation mechanism of effective components absorbed into blood from Hedysari Radix.Methods Qualitative quantification of effective components absorbed into blood from Hedysari Radix were operated by using UPLC-MS/MS technique.The corresponding targets of effective components absorbed into blood from Hedysari Radix were screened by TCMSP and HERB databases.Targets of immune-related disease were obtained through DisGeNET,OMIM,TTD,and MalaCards databases.The network of"components absorbed into blood from Hedysari Radix-immune-related diseases"was then constructed.GO and KEGG enrichment analysis and mapped the PPI network were performed.Molecular docking and molecular dynamics techniques were applied for validation.Results A total of 8 prototype components absorbed into blood,synergistically acting on 101 targets,were identified by UPLC-MS/MS.They mediated 538 biological processes including immune response,positive regulation of gene expression,receptor binding,and cytokine activity.Meanuhile,116 signaling pathways,such as HIF-1,Toll-like receptor,JAK-STAT,T cell receptor,PI3K-Akt,and FoxO etc.were involved.The core targets were MAPK14,PTGS2,MMP9,PPARG,CCND1,etc..The results of molecular docking showed that formononetin and calycosin had strong docking binding activity with MAPK14.And molecular dynamics simulations further demonstrated that the binding between MAPK14 and formononetin or calycosin had good structural stability and binding affinity.Conclusion The results of serum pharmacochemistry,network pharmacology and molecular dynamics were verified to reveal the material basis and mechanism of Hedysari Radix in regulating immunity.The aim of this study is to provide scientific basis for its immunomodulatory mechanism.

Nonsteroidal anti-inflammatory drugs(NSAIDs)are widely used to treat osteoarthritis,rheumatoid arthritis,and acute and chronic painful diseases.Significant interindividual heterogeneity in the efficacy of NSAIDs has been found in patients,which can lead to treatment failure or life-threatening adverse drug reactions.This review will discuss the causes of the variability in the efficacy of NSAIDs in terms of CYP2C9 enzyme gene polymorphism,cyclooxygenase enzyme gene polymorphism,and differences in the intestinal microbiota,to provide a reference for the development of individualized dosing regimens for NSAIDs.

In order to achieve the purpose of rapid detection of duck astrovirus type 1(DAstV-1),specific primers were designed based on the conservative region of ORF1a which belonged to DAstV-1(WF1202 strain).A real-time fluorescent quantitative PCR(qPCR)detective method for DAstV-1 was established.Clinical samples were detected by the qPCR method and the positive samples were used for virus isolation and identification.Results showed that the detection limit of the established method was 4.64×103 copies/μL,which was 10 times higher than the normal RT-PCR method.In addition,no cross-reactions were found with other common infectious disease pathogens in poultry,indicating that the qPCR method had good specificity.What's more,the coef-ficient of variations(Cv)in intra-and inter-assays were 0.85％-2.85％and 0.21％-2.94％,re-spectively,both less than 3％,indicating that the qPCR method had a good repeatability.Using this method,35 tissue samples from different duck farms in 10 provinces from 2020 to 2022 were detected for DAstV-1.Results showed that the positive rate was 25.71％(9/35),and the coinci-dence rate was 94.29％when compared with the normal RT-PCR method.A positive sample ran-domly taken for the virus isolation through duck embryo passage,and the allantoic fluid was col-lected and then was verified by the qPCR method and inoculated with 1-day-old healthy ducklings for the animal regression experiment.The infected ducklings suffered from transient disease but did not die.The liver tissues were all positive with DAstV-1 when detected by qPCR.Meanwhile,autopsy showed that there were slight changes in the livers,and the histopathological observation showed that the liver cells were steatosis.These findings indicated that the isolated DAstV-1 strain had weak pathogenicity and might be a low virulent strain.To sum up,the qPCR detection method of DAstV-1 was successfully established in this work,and could provide technical support for clini-cal diagnosis,isolation and identification,and molecular epidemiology monitoring of DAstV-1.

Objective To observe and compare of the effects of esketamine and sufentanil induction for general anesthesia on postoperative analgesia in obese patients undergoing laparoscopic sleeve gastrectomy.Methods Patients scheduled for elective laparoscopic sleeve gastrectomy between March 2023 and March 2024,irrespective of gender,aged between 20 and 50 years,with a BMI ranging from 30.0 to 50.0 kg/m2 and ASA Ⅰ or Ⅱ classification,were randomly allocated into two groups:the esketamine group(Group E)and the sufentanil group(Group S),each consisting of 32 cases.During anesthesia induction,Group E received a dose of esketamine at 0.5 mg/kg while Group S received sufentanil at a dosage of 0.5 μg/kg;the remaining protocol remained unchanged.Heart rate(HR),mean arterial pressure(MAP),and SpO2 were recorded at various time points:upon arrival(T0),prior to anesthesia induction(T1),immediately after induction(T2),during intubation(T3),at the conclusion of anesthesia adminis-tration(T4),and during extubation(T5).Postoperative pain scores using the Numeric Rating Scale(NRS)were assessed at specific intervals following surgery:one hour post-surgery(P0),six hours post-surgery(P1),twelve hours post-surgery(P2),twenty-four hours post-surgery(P3)and forty-eight hours post-surgery(P4).Additionally,intraoperative remifentanil consumption as well as the number of presses on the analgesic pump within forty-eight hours after surgery in both patient groups were documented along with any occurrences of adverse reactions.Results During the surgery,there was no statistically significant difference in mean arterial pressure(MAP)and heart rate(HR)between the two patient groups at T0～T5(P>0.05).At T2,both groups exhibited lower MAP and HR compared to T0;specifically,group S had a MAP of(91.81±8.94)mmHg and HR of(81.75±13.37)beats/min,while group E had a MAP of(93.69±9.96)mmHg and HR of(80.38±13.2)beats/min,with group E showing values closer to baseline levels.At T3,both groups experienced a transient increase in MAP and HR(P<0.05);specifi-cally,group S had a MAP of(97.56±8.96)mmHg and HR of(86.47±13.84)beats/min,while group E had a MAP of(101.03±8.29)mmHg and(89.41±15.32)times/min,with S group closer to baseline values.There was no statistically significant difference in the amount of remifentanil used during surgery between group S and group E(P>0.05),which were(2071.88±717.63)μg and(2093.75±718.39)μg,respectively.Compared with the postoperative conditions of the two groups,the NRS scores of group E(0.41±0.61±1.870.75,2.47±0.62)at P0,P1 and P2 were lower than those in group S(0.88±0.71,2.47±0.72,2.97±0.54),and the difference was statisti-cally significant(P<0.05).The number of intravenous analgesia pump presses was significantly reduced in group E after surgery(P<0.05),with the postoperative analgesic pump compressions occurring 11.25±2.70 times in group S and 8.56±2.23 times in group E.The incidence of postoperative nausea and vomiting(PONV)and hypotension in Group E(21.88%,15.63%)was lower than that observed in group S(46.88%,37.50%),demonstrating statistical significance(P<0.05).Conclusion In comparison to sufentanil induction,the utilization of esketamine anesthesia induction is deemed safe for laparoscopic sleeve gastrectomy in obese patients,effectively mitigating postoperative acute pain and reducing the incidence of PONV.

Acute kidney injury(AKI)is a common perioperative complication,which is associated with an increased incidence rate of other postoperative complications and death risk of the patients.Remote ischemic preconditioning(RIPC)performs multiple transient ischemia/reperfusion cycles at remote sites far from the target organs to enhance the tolerance of target organs to oxidative stress.A large number of animal experiments have shown that RIPC can improve kidney function,alleviate AKI,and increase the survival rate of AKI patients.However,there is still controversy about the mechanism of RIPC in alleviating AKI in clinical studies.In this article,the research progress on the role and mechanism of RIPC in renal protection in recent years is reviewed,in order to provide a reference for further exploration of perioperative renal protection strategies and mechanisms.

Objective To understand the occurrence characteristics of occupational chemical incidents in China by media surveillance, to provide scientific evidences for formulating effective prevention and control strategies, and to provide new ideas for public surveillance and early warning of chemical incidents. Methods Occupational chemical incidents were collected and sorted through the Internet every day. The statistical analysis was conducted from time, region, industry, and causes of the collected incidents. Results A total of 99 occupational chemical incidents were collected in 2021, involving 356 peoples and 186 deaths. April to July was the high incidence period of events, mainly limited space poisoning and asphyxiation events. Majority of the incidents were general grade events, mainly occurring in small and medium enterprises. Guangdong Province had the most incident reporting areas. Incidents mainly occurred in chemical raw material and chemical product manufacturing industry as well as in water production and supply industry, mainly in manufacturing, cleaning and maintenance positions. The main types of poisons were asphyxiating gases and organic solvents, mainly involving hydrogen sulfide and benzene. The incidents could have been caused by lack of safety awareness, leakage of equipment, and lack of protective measures. Conclusion Occupational chemical incidents still occur frequently and cause serious harm. It is suggested that the relevant departments and employers should strengthen supervision and management, effectively manage and control operational risks, and increase supervision of key industries and positions, to ensure the health and safety of workers.

ObjectiveTo investigate the predictive indicators of early efficacy of Bushen Shengxue prescription combined with western medicine in the treatment of aplastic anemia， and provide prognosis indicators for the treatment of aplastic anemia （AA） with kidney-tonifying therapy in traditional Chinese medicine （TCM） combined with western medicine. MethodA total of 126 patients treated by Bushen Shengxue prescription combined with western medicine in 19 hospitals including Xiyuan Hospital of the China Academy of Chinese Medical Sciences from September 2018 to March 2021 were selected for a retrospective study. The therapy was proven to be effective after six months of treatment. According to the efficacy after 4 months of treatment， the patients were assigned into a 4-month effective group and a 4-month ineffective group. The age， sex， disease severity （including severe aplastic anemia and non-severe aplastic anemia）， course of disease， degree of bone marrow nucleated cell proliferation， baseline hemogram levels ［including white blood cell count （WBC）， absolute neutrophil count （ANC）， hemoglobin （HGB）， platelets （PLT）， and reticulocytes （RET）］， T lymphocytes subsets， and the expression levels of T-box transcription factor （T-bet） and GATA-binding protein-3 （GATA-3） were compared between the two groups before treatment. ResultThe proportions of patients within the age ranges of ［20， 40） and ［60， 80） were higher in the 4-month effective group （P<0.05）. The sex， disease severity， course of disease， and comorbidities had no significant differences between the two groups. The 4-month effective group had higher baseline levels of HGB， WBC， ANC， and PLT than the 4-month ineffective group （P<0.05）， and there was no significant difference in the RET level between the two groups before treatment. Binary Logistic regression analysis showed that the PLT level before treatment was an independent factor affecting the onset time， while other indicators did not affect the onset time. The receiver operating characteristic （ROC） curve was established to analyze the value of PLT level before treatment for predicting the onset time， and the area under the curve was 0.691. With the critical value of 40.5×10⁹/L， the sensitivity and specificity of the prediction that the therapy will take effect within 4 months were 0.569 and 0.893， respectively. The two groups of patients were graded according to age ｛（14， 20）， ［20， 40）， ［40， 60）， and ［60， 80）｝ and PLT level before treatment （PLT<40×10⁹/L， PLT≥40×10⁹/L）. The proportion of the patients with PLT≥40×10⁹/L before treatment in the 4-month effective group was significantly higher than that in the 4-month ineffective group （P<0.05）. The degree of bone marrow nucleated cell proliferation before treatment had no significant difference between the two groups. The level of total T lymphocytes in the 4-month effective patients was lower than that in the 4-month ineffective patients before treatment （P<0.05）. The levels of Th1 cells， Th2 cells， CD4⁺ T cells， and CD8⁺ T cells showed no significant differences between the two groups before treatment. The T-bet expression level in the 4-month effective group was higher than that in the 4-month ineffective group before treatment （P<0.05）， while the expression level of GATA-3 showed no significant difference between the two groups before treatment. ConclusionBushen Shengxue prescription combined with western medicine will achieve faster effect for the patients within the age ranges of ［20， 40） or ［40， 60）， with higher levels of HGB， WBC， ANC， and PLT （especially those with PLT≥40×10⁹/L）， lower level of total T lymphocytes， or higher T-bet expression level before treatment.

Proteins play a variety of functional roles in cellular activities and are indispensable for life. Understanding the functions of proteins is crucial in many fields such as medicine and drug development. In addition, the application of enzymes in green synthesis has been of great interest, but the high cost of obtaining specific functional enzymes as well as the variety of enzyme types and functions hamper their application. At present, the specific functions of proteins are mainly determined through tedious and time-consuming experimental characterization. With the rapid development of bioinformatics and sequencing technologies, the number of protein sequences that have been sequenced is much larger than those can be annotated, thus developing efficient methods for predicting protein functions becomes crucial. With the rapid development of computer technology, data-driven machine learning methods have become a promising solution to these challenges. This review provides an overview of protein function and its annotation methods as well as the development history and operation process of machine learning. In combination with the application of machine learning in the field of enzyme function prediction, we present an outlook on the future direction of efficient artificial intelligence-assisted protein function research.
Artificial Intelligence ; Machine Learning ; Proteins/genetics* ; Computational Biology/methods* ; Drug Development