Microalgae possess high photosynthetic efficiency, robust adaptability, and substantial biomass, serving as excellent biological resources for large-scale cultivation. Malic enzyme (ME), a ubiquitous metabolic enzyme in living organisms, catalyzes the decarboxylation of malate to produce pyruvate, CO₂, and NAD(P)H, playing a role in multiple metabolic pathways including energy metabolism, photosynthesis, respiration, and biosynthesis. In this study, we identified the Scenedesmus quadricauda malic enzyme gene (SqME) and its biological functions, aiming to provide excellent target genes for the genetic improvement of higher plants. Based on the RNA-seq data from S. quadricauda under the biofilm cultivation mode with high CO₂ and light energy transfer efficiency and small water use, a highly expressed gene (SqME) functionally annotated as ME was cloned. The physicochemical properties of the SqME-encoded protein were systematically analyzed by bioinformatics tools. The subcellular localization of SqME was determined via transient transformation in Nicotiana benthamiana leaves. The biological functions of SqME were identified via genetic transformation in Nicotiana tabacum, and the potential of SqME in the genetic improvement of higher plants was evaluated. The ORF of SqME was 1 770 bp, encoding 590 amino acid residues, and the encoded protein was located in chloroplasts. SqME was a NADP-ME, with the typical structural characteristics of ME. The ME activity in the transgenic N. tabacum plant was 1.8 folds of that in the wild-type control. Heterologous expression of SqME increased the content of chlorophyll a, chlorophyll b, and total chlorophyll by 20.9%, 26.9%, and 25.2%, respectively, compared with the control. The transgenic tobacco leaves showed an increase of 54.0% in the fluorescence parameter NPQ and a decrease of 30.1% in Fo compared with the control. Moreover, the biomass, total lipids, and soluble sugars in the transgenic tobacco leaves enhanced by 20.5%, 25.7%, and 9.5%, respectively. On the contrary, the starch and protein content in the transgenic tobacco leaves decreased by 22.4% and 12.2%, respectively. Collectively, the SqME-encoded protein exhibited a strong enzymatic activity. Heterologous expressing of SqME could significantly enhance photosynthetic protection, photosynthesis, and biomass accumulation in the host. Additionally, SqME can facilitate carbon metabolism remodeling in the host, driving more carbon flux towards lipid synthesis. Therefore, SqME can be applied in the genetic improvement of higher plants for enhancing photosynthetic carbon fixation and lipid accumulation. These findings provide scientific references for mining of functional genes from S. quadricauda and application of these genes in the genetic engineering of higher plants.
Nicotiana/genetics* ; Photosynthesis/physiology* ; Malate Dehydrogenase/biosynthesis* ; Plant Leaves/genetics* ; Scenedesmus/enzymology* ; Carbon Cycle/genetics* ; Lipid Metabolism/genetics* ; Plants, Genetically Modified/metabolism*

Massive Open Online Courses （MOOCs） in medical ethics play a significant role in cultivating the humanistic quality of medical students. Through quantitative analysis， this paper compared the commonalities and differences in “medical ethics” MOOC courses in different types of colleges and universities， as well as proposed new paths for optimizing the cultivation of medical students’ humanistic quality. By searching and analyzing course data from “medical ethics” offered on mainstream MOOC platforms in China from 2013 to 2024， this paper explored new paths for the integration of MOOCs and humanistic quality from the perspectives of course content， faculty construction， student participation， and comparison between traditional Chinese medicine （TCM） universities and comprehensive universities. This study found that medical MOOCs focused on theoretical foundations and medical professional ethics and spirit， medical equity and social responsibility， as well as end-of-life care and medical humanistic care， forming a multidimensional curriculum system. Faculty teams reflected interdisciplinary characteristics， gathering teachers from disciplines such as medicine， philosophy， and education. Differences were observed between TCM universities and comprehensive universities. The former focused more on the origins and inheritance of humanistic care in TCM culture and ancient Chinese medical classics， while the latter emphasized more on Western ethical spirit. This paper can provide valuable insights for the diversified development of medical humanistic quality education， contribute to the integration of TCM ethics with modern medical ethics， provide references for online medical ethics education， implement new medical concepts， promote interdisciplinary integration of medicine and the humanities， and assists the reform and development of medical education.

Objective:To investigate the therapeutic effect of Brucea javanica oil emulsion combined with chemotherapy for advanced lung squamous cell carcinoma.Methods:A retrospective cohort study was conducted. The clinical data from 120 patients with lung squamous cell carcinoma at stage Ⅳ in Shanxi Province Cancer Hospital between January 2019 and December 2020 were retrospectively analyzed. According to the treatment regimens, all patients were divided into Brucea javanica oil emulsion combined with chemotherapy group (the observation group, 60 cases) and chemotherapy alone group (the control group, 60 cases). Immune function, efficacy, and chemotherapy-related adverse effects were compared between the 2 groups before and after treatment.Results:Among 120 patients, there were 77 males and 43 females, with the age of (67±7) years. There were no statistically differences in basic information between the 2 groups (all P > 0.05). The proportion of CD4 + cells and the ratio of CD4 + to CD8 + in the observation group after treatment increased compared to those before treatment, while the proportion of CD8 + decreased (all P < 0.001). In the control group, the proportions of CD3 +, CD4 +, NK cells decreased after treatment (all P < 0.001). There were no statistically significant differences in the T cell subsets between the 2 groups before treatment (all P > 0.05); after treatment, the proportions of CD3 +, CD4 +, NK cells and the ratio of CD4 + to CD8 + in the observation group were all higher than those in the control group, and the proportion of CD8 + cells was lower than that in the control group (all P < 0.001). The disease control rate (DCR) in the observation group was higher than that in the control group [73.3% (44/60) vs. 55.0% (33/60)], and the difference was statistically significant ( χ2 = 4.39, P = 0.036). The median progression-free survivals (PFS) time was 7.833 months (95% CI: 6.927-8.739 months), 5.433 months (95% CI: 3.878-6.988 months), and the difference was statistically significant ( χ2 = 4.84, P = 0.028). The incidence of leukopenia was 38.3% (23/60), 60.0% (36/60), respectively in the observation group and the control group ( χ2 = 5.64, P = 0.018); the incidence of thrombocytopenia was 21.7% (13/60), 38.3% (23/60), respectively in the observation group and the control group ( χ2 = 3.97, P = 0.046). Conclusions:Brucea javanica oil emulsion combined with chemotherapy can improve immune function, increase DCR, and reduce myelosuppression of patients with advanced lung squamous cell carcinoma.

Objective To evaluate the application value of nucleic acid mass spectrometry in detecting ERG11 gene mutations associated with azole resistance in Candida tropicalis(C.tropicalis),thereby providing a scientific basis for the rational clinical use of azole antifungal agents.Methods Clinical isolates of C.tropicalis were obtained from the Affiliated Hospital of Xuzhou Medical University between July 2023 and November 2024.For each isolate,specific ERG11 mutations(A395T and C461T)were analyzed using Sanger sequencing and nucleic acid mass spectrometry.Sanger sequencing was used as the reference standard to evaluate the performance of mass spectrometry in mutation detection.Results The established nucleic acid mass spectrometry method effectively identified four genotypes of ERG11(A395T and C461T),with distinct spectral peaks for each mutation and no cross-interference observed.Among 88 clinical isolates,the sensitivity,specificity,positive predictive value(PPV),and negative predictive value(NPV)for the A395T mutation were 86.7%,100%,100%,and 97.3%,respectively.For the C461T mutation,the corresponding values were 81.3%,100%,100%,and 96.0%,respectively.Kappa statistics demonstrated a high level of agreement between mass spectrometry and sequencing results.Conclusions Nucleic acid mass spectrometry exhibits high sensitivity and specificity in the detection of ERG11 mutations,enabling rapid,accurate,and adaptable high-throughput analysis.This makes it a highly effective method for identifying mutations associated with fungal resistance.

Objective:To develop a Knowledge-Attitude-Practice (KAP) Scale for Dietary Management During Hemodialysis and to test its reliability and validity.Methods:Based on the KAP theoretical framework, an initial version of the scale was developed through a literature review and expert consultations. A convenience sampling method was used to recruit hemodialysis patients from four hospitals in Suzhou in March 2024. Questionnaire item analysis and reliability and validity tests were conducted.Results:A total of 460 questionnaires were distributed and 438 valid responses were collected, with an effective response rate of 95.22%. The final scale included three dimensions (knowledge, attitude, and practice) with 34 items. Content validity at the scale level was 0.910, and the item level ranged from 0.800 to 1.000. Exploratory factor analysis extracted three common factors, with a cumulative variance contribution rate of 74.520%. Confirmatory factor analysis showed a good model fit. The total Cronbach's α coefficient of the scale was 0.971, and the Cronbach's αcoefficients for the three dimensions were 0.963, 0.933, and 0.934, respectively. The test-retest reliability coefficient was 0.839.Conclusions:The Knowledge-Attitude-Practice Scale for Dietary Management During Hemodialysis demonstrates good reliability and validity, making it a valuable tool for assessing the KAP level of dietary management in hemodialysis patients.

Objective To evaluate the application value of nucleic acid mass spectrometry in detecting ERG11 gene mutations associated with azole resistance in Candida tropicalis(C.tropicalis),thereby providing a scientific basis for the rational clinical use of azole antifungal agents.Methods Clinical isolates of C.tropicalis were obtained from the Affiliated Hospital of Xuzhou Medical University between July 2023 and November 2024.For each isolate,specific ERG11 mutations(A395T and C461T)were analyzed using Sanger sequencing and nucleic acid mass spectrometry.Sanger sequencing was used as the reference standard to evaluate the performance of mass spectrometry in mutation detection.Results The established nucleic acid mass spectrometry method effectively identified four genotypes of ERG11(A395T and C461T),with distinct spectral peaks for each mutation and no cross-interference observed.Among 88 clinical isolates,the sensitivity,specificity,positive predictive value(PPV),and negative predictive value(NPV)for the A395T mutation were 86.7%,100%,100%,and 97.3%,respectively.For the C461T mutation,the corresponding values were 81.3%,100%,100%,and 96.0%,respectively.Kappa statistics demonstrated a high level of agreement between mass spectrometry and sequencing results.Conclusions Nucleic acid mass spectrometry exhibits high sensitivity and specificity in the detection of ERG11 mutations,enabling rapid,accurate,and adaptable high-throughput analysis.This makes it a highly effective method for identifying mutations associated with fungal resistance.

Objective:To develop a Knowledge-Attitude-Practice (KAP) Scale for Dietary Management During Hemodialysis and to test its reliability and validity.Methods:Based on the KAP theoretical framework, an initial version of the scale was developed through a literature review and expert consultations. A convenience sampling method was used to recruit hemodialysis patients from four hospitals in Suzhou in March 2024. Questionnaire item analysis and reliability and validity tests were conducted.Results:A total of 460 questionnaires were distributed and 438 valid responses were collected, with an effective response rate of 95.22%. The final scale included three dimensions (knowledge, attitude, and practice) with 34 items. Content validity at the scale level was 0.910, and the item level ranged from 0.800 to 1.000. Exploratory factor analysis extracted three common factors, with a cumulative variance contribution rate of 74.520%. Confirmatory factor analysis showed a good model fit. The total Cronbach's α coefficient of the scale was 0.971, and the Cronbach's αcoefficients for the three dimensions were 0.963, 0.933, and 0.934, respectively. The test-retest reliability coefficient was 0.839.Conclusions:The Knowledge-Attitude-Practice Scale for Dietary Management During Hemodialysis demonstrates good reliability and validity, making it a valuable tool for assessing the KAP level of dietary management in hemodialysis patients.

Objective:To investigate the therapeutic effect of Brucea javanica oil emulsion combined with chemotherapy for advanced lung squamous cell carcinoma.Methods:A retrospective cohort study was conducted. The clinical data from 120 patients with lung squamous cell carcinoma at stage Ⅳ in Shanxi Province Cancer Hospital between January 2019 and December 2020 were retrospectively analyzed. According to the treatment regimens, all patients were divided into Brucea javanica oil emulsion combined with chemotherapy group (the observation group, 60 cases) and chemotherapy alone group (the control group, 60 cases). Immune function, efficacy, and chemotherapy-related adverse effects were compared between the 2 groups before and after treatment.Results:Among 120 patients, there were 77 males and 43 females, with the age of (67±7) years. There were no statistically differences in basic information between the 2 groups (all P > 0.05). The proportion of CD4 + cells and the ratio of CD4 + to CD8 + in the observation group after treatment increased compared to those before treatment, while the proportion of CD8 + decreased (all P < 0.001). In the control group, the proportions of CD3 +, CD4 +, NK cells decreased after treatment (all P < 0.001). There were no statistically significant differences in the T cell subsets between the 2 groups before treatment (all P > 0.05); after treatment, the proportions of CD3 +, CD4 +, NK cells and the ratio of CD4 + to CD8 + in the observation group were all higher than those in the control group, and the proportion of CD8 + cells was lower than that in the control group (all P < 0.001). The disease control rate (DCR) in the observation group was higher than that in the control group [73.3% (44/60) vs. 55.0% (33/60)], and the difference was statistically significant ( χ2 = 4.39, P = 0.036). The median progression-free survivals (PFS) time was 7.833 months (95% CI: 6.927-8.739 months), 5.433 months (95% CI: 3.878-6.988 months), and the difference was statistically significant ( χ2 = 4.84, P = 0.028). The incidence of leukopenia was 38.3% (23/60), 60.0% (36/60), respectively in the observation group and the control group ( χ2 = 5.64, P = 0.018); the incidence of thrombocytopenia was 21.7% (13/60), 38.3% (23/60), respectively in the observation group and the control group ( χ2 = 3.97, P = 0.046). Conclusions:Brucea javanica oil emulsion combined with chemotherapy can improve immune function, increase DCR, and reduce myelosuppression of patients with advanced lung squamous cell carcinoma.

A 49-year-old female examinee, employed at a paper company in Guigang, Guangxi (engaged in the production of napkins), visited the Qintang District People′s Hospital in Guigang on June 11, 2023, to apply for a health certificate. Salmonella was isolated during a physical examination screening for enteric pathogens ( Salmonella, Shigella), and repeated use of empirical antibacterial drugs failed to eradicate the pathogen. A reference laboratory identified the two isolates as S. Kentucky ST198 by performing serotyping, drug resistance phenotyping, and whole genome sequencing. Based on the regional serotype database of Salmonella in China, a phylogenetic tree of S. Kentucky ST198 resistance genotypes was constructed, defining both isolated strains as high-level drug-resistant ST198.2-1 subclade clones. Following a multidisciplinary consultation and based on antimicrobial susceptibility test results, the treatment was adjusted to amoxicillin/clavulanate and azithromycin. On August 19, three consecutive negative test results confirmed the eradication of Salmonella Kentucky.

Objective To explore the relationship between serum iron(SI)metabolism and glucose and lipid metabolism in patients with type 2 diabetes mellitus(T2DM).Methods A total of 170 T2DM patients hospitalized in the Department of Endocrinology,Lanzhou University First Hospital from 2019 to 2021 were included.During the same period,30 healthy individuals from physical examination center were selected as the normal control(NC)group.Based on HbA1c control,T2DM patients were divided into subgroups with good blood glucose control(H1,HbA1c＜7%,n=39),poor glucose control(H2,7%＜HbA1c＜9%,n=63)and very poor glucose control(H3,HbA1c＞9%,n=68).According to the level of blood lipids,T2DM patients were divided into subgroups with normal blood lipids(L1,n=36)and high blood lipids(L2,n=134).Results Compared with NC group,age,SBP,DBP,BMI,serum ferritin(SF),FPG,FIns,HOMA-IR,TyG,TG,LDL-C and SUA increased inT2DM group(P＜0.05),while SI,TF,total iron binding capacity(TIBC),DI,HDL-C and eGFR decreased(P＜0.05).The levels of TF and TIBC in H3 subgroup were lower than those in H1 subgroup(P＜0.05).LDL-C of L2 subgroup was higher than that of L1 subgroup(P＜0.05),while HDL-C was lower than that of L1 subgroup(P＜0.05).Pearson correlation analysis showed that SF was positively correlated with HbA1c,TyG,TG and SUA(P＜0.05),and negatively correlated with HDL-C and eGFR(P＜0.05).TF was positively correlated with HDL-C and eGFR(P＜0.05),but negatively correlated with age,SBP,DBP,TyG and SUA(P＜0.05).Multiple linear regression analysis showed that SF and FPG were influencing factors for HbA1c in T2DM patients.Conclusion SI metabolism is closely related to glucose and lipid metabolism in T2DM patients.