Sucrose non-fermenting 1-related protein kinase 1 (SnRK1) is one of the highly conserved Ca²⁺ non-dependent serine/threonine protein kinases, playing a crucial role in regulating the stress responses, growth, and development of plants. SnRK1 is a three-subunit complex, and it is involved in responding to the signaling transduction induced by low-energy/low-sugar conditions. SnRK1 responds biotic and abiotic stress conditions (such as salt, drought, low/high temperatures, and diseases) through phosphorylation of key metabolic enzymes and regulatory proteins, regulation of transcription, and interactions with other proteins. Furthermore, SnRK1 is not only involved in hormone signaling pathways mediated by abscisic acid (ABA), jasmonic acid (JA) and salicylic acid (SA), but also regulates plant autophagy by inhibiting the activity of target of rapamycin (TOR). In this review, we summarized the current results of research on the discovery, structure, and classification of plant SnRK1 and its roles in the stress responses, growth, and development of plants. Furthermore, this article proposes the directions of future research. This review provides good genetic resources and a theoretical basis for the genetic improvement and biological breeding for enhancing the stress tolerance of crops.
Stress, Physiological/physiology* ; Protein Serine-Threonine Kinases/metabolism* ; Plant Development/genetics* ; Signal Transduction ; Gene Expression Regulation, Plant ; Plant Proteins/physiology* ; Plants/metabolism* ; Arabidopsis Proteins/physiology* ; Plant Growth Regulators/metabolism*

Abstract In recent years, the prevalence of overweight and obesity among children and adolescents has risen rapidly, posing a serious threat to their physical and mental health. To provide scientific, systematic, and standardized weight management guidance for overweight and obese children and adolescents, the study focuses on the core concept of healthy lifestyle intervention, integrates multidisciplinary expert opinions and research findings,and proposes a comprehensive multidisciplinary intervention framework covering scientific exercise intervention, precise nutrition and diet, optimized sleep management, and standardized psychological support. It calls for the establishment of a multi agent collaborative management mechanism led by the government, implemented by families, fostered by schools, initiated by individuals, optimized by communities, reinforced by healthcare, and coordinated by multiple stakeholders. Emphasizing a child and adolescent centered approach, the consensus advocates for comprehensive, multi level, and personalized guidance strategies to promote the internalization and maintenance of a healthy lifestyle. It serves as a reference and provides recommendations for the effective prevention and control of overweight and obesity, and enhancing the health level of children and adolescents.

Objective To investigate the mechanism of Yes-associated protein 1(YAP1)ameliorating aristolochic acid 1(AAI)-induced liver injury in mice based on untargeted metabolomics techniques.Methods There were 83-week-old male hepatocyte-specific Yap1 gene knockout mice(genotyped as Yap1Flox/Flox,Albumin-Cre,aka.Yap1LKO)were randomly selected as the Yap1LKO+AAI group,and 8 Yap1Flox control mice as the Yap1Flox+AAI group.Both groups were injected intraperitoneally with AAI at a dose of 2.5 mg·kg-1·d-1 for 14 consecutive days.Genotypes were identified by tail PCR;serum alanine transaminase(ALT)and aspartate transaminase(AST)activities were determined by microplate assay;histopathological changes of liver tissue were observed by HE staining;and the protein expression of YAP1 in liver tissue was determined by immunohistochemistry.The untargeted metabolomics approach was used to analyze the liver tissue differential metabolites,and the samples were analyzed by ultra performance liquid chromatography-quadrupole-electrostatic field orbit trap high-resolution mass spectrometry,and the differential metabolites were screened by principal component analysis(PCA),Partial least square-discriminant analysis(PLS-DA),and orthogonal partial least squares-discriminant analysis(OPLS-DA);using HMDB database and METLIN database to identify metabolites,and the pathway enrichment of differential metabolites was analyzed by KEGG database.Results(1)After 14 days of AAI induction,the increase of body mass in Yap1LKO mice was lower than that in Yap1Flox mice,but there was no statistical significance(P＞0.05).On day 14,compared with the Yap1Flox+AAI group,the serum ALT and AST enzyme activities in the Yap1LKO+AAI group of mice were significantly increased(P＜0.05),and the histopathological damage of the liver was significantly aggravated.The livers of the Yap1Flox mice had a positive protein expression of YAP1,whereas the Yap1LKO mice did not have a positive protein expression of YAP1.(2)A total of 139 differential metabolites with significant changes(VIP＞1 and P＜0.05)were screened by metabonomic analysis;compared with Yap1LKO+ AAI group,62 liver metabolites in Yap1Flox+AAI group were up-regulated,including choline,taurine,hypotaurine,α-linolenic acid,eleostearic acid,chenodeoxycholic acid and so on.Seventy-seven metabolites were down-regulated including glycerophosphocholine,L-phosphatidylcholine,L-glutamine,L-serine,L-glutathione,5-methionine,phenylalanine,glucose 6-phosphate,lactic acid,uric acid glycosides,etc..KEGG-enriched pathways were mainly choline metabolism,glycerophospholipid metabolism,insulin resistance,glutathione metabolism,etc..Conclusion Hepatocyte-specific Yap1 gene knockout exacerbated AAI-induced liver injury in mice,and YAP1 was involved in the regulation of choline metabolism and glycerophospholipid metabolism through the up-regulation of unsaturated fatty acids,such as choline and taurine,which ameliorated AAI-induced liver injury in mice.

Objective To investigate the effect of plasma microRNA(miR)-93-5p on ovarian granulosa cell proliferation in patients with polycystic ovary syndrome(PCOS)and its possible mechanism.Methods A total of 120 PCOS patients of childbearing period who were treated at Jiaozuo People's Hospital from January 2022 to January 2023 were selected as the PCOS group,and 18 healthy women of childbearing period who were physically examined at the same hospital during the same period were selected as the control group.The age,body weight and height of the subjects in the two groups were recorded,and the body mass index(BMI)was calculated.Fasting venous blood was collected from the subjects during the follicular phase of the natural menstrual cycle or the progesterone-induced withdrawal bleeding phase,and serum levels of luteinizing hormone(LH),follicle-stimulating hormone(FSH),total testosterone(TT),anti-Mullerian hormone(AMH),aspartate aminotransferase(AST),alanine aminotransferase(ALT),fasting insulin(FINS),and fasting blood glucose(FBG)were measured by using the electrochemical method;and the homeostasis model assessment of insulin resistance(HOMA-IR)was calculated.Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the expression level of miR-93-5p in plasma of patients in the two groups.Human ovarian granulosa cells(KGN cells)in logarithmic growth phase were cultured in 6-well plates with 3 × 105 cells per well,and they were randomly divided into the miR-93-5p mimics group,LY294002+miR-93-5p group,AZD5363+miR-93-5p group,negative control(NC)group,and blank control group.The KGN cells in the miR-93-5p group were transfected with miR-93-5p mimics;the KGN cells in the LY294002+miR-93-5p group were transfected with miR-93-5p mimics and treated with 50 mmol·L-1 LY294002 one hour before transfection;the KGN cells in the AZD5363+miR-93-5p group were transfected with miR-93-5p mimics and treated with 50 μmol·L-1 AZD5363 one hour before transfection;the KGN cells in the NC group were transfected with the negative control plasmids;and the KGN cells in the blank control group were not treated at all.The expression of miR-93-5p in the KGN cells in each group was detected by qRT-PCR,and the proliferation of the KGN cells in each group was detected by cell counting kit-8.Results There was no significant differences in age,FSH,ALT,and AST levels of patients between the PCOS group and the blank control group(P＞0.05).The BMI,TT,AMH,LH,FINS,FBG,and HOMA-IR of patients in the PCOS group were significantly higher than those in the blank control group(P＜0.05).The relative expression of miR-93-5p in plasma of patients in the PCOS group was significantly higher than that in the blank control group(t=-5.549,P＜0.001).miR-93-5p was moderately positively correlated with TT,FINS and HOMA-IR(r=0.434,0.622,0.586;P＜0.001)and was mildly positively correlated with FBG and LH(r=0.398,0.398;P＜0.001).The receiver operating characteristic curve showed that the optimal cut-off value for plasma miR-93-5p in diagnosing PCOS was 1.380,the area under the curve was 0.906(95％confidence interval:0.839-0.973,P＜0.001),the sensitivity was 0.858,the specificity was 0.833,and the Youden index was 0.691.The relative expression of miR-93-5p in the KGN cells in the miR-93-5p mimics group,LY294002+miR-93-5p group and AZD5363+miR-93-5p group was significantly higher than that in the blank control group and NC group(P＜0.05).After 24,48 and 72 hours of culture,the proliferation of the KGN cells in the miR-93-5p mimics group,LY294002+miR-93-5p group and AZD5363+miR-93-5p group was significantly higher than that in the blank control group and the NC group(P＜0.05);the proliferation of the KGN cells in the LY294002+miR-93-5p group and AZD5363+miR-93-5p group was significantly lower than that in the miR-93-5p mimics group(P＜0.05).Conclusion miR-93-5p in plasma is overexpressed in PCOS patients,and it may be involved in the occurrence and development of PCOS by mediating the proliferation of ovarian granulosa cells through the phosphoinositide 3 kinase/protein kinase B signaling pathway.The miR-93-5p level in plasma has a certain diagnostic value for PCOS.

Objective To explore the expression level of microRNA(miR)-320a-3p in peripheral blood in patients with polycystic ovary syndrome(PCOS)and its possible mechanism for regulating PCOS.Methods A total of 149 PCOS patients admitted to the Jiaozuo People's Hospital from July 2021 to July 2022 were selected as the research subjects(PCOS group),and 18 healthy volunteers with a regular menstrual cycle(28-35 d),no clinical or biochemical manifestations of hyperandrogenism,and no polycystic ovary changes detected by ultrasonography were selected as the control group.Clinical data of the subjects in the two groups were collected and compared.The expression level of miR-320a-3p in plasma of subjects in the two groups was detected by using the real-time quantitative polymerase chain reaction.The correlation between plasma miR-320a-3p expression and clinical indexes was evaluated by using the Pearson partial correlation analysis.The predictive value of miR-320a-3p for PCOS was analyzed by using the receiver operating characteristic curve.The relationship between miR-320a-3p and androgen receptor was evaluated by using the dual luciferase reporter gene assay.Results There was no significant difference in age,hip circumference,follicle stimulating hormone(FSH),prolactin(PRL),and high-density lipoprotein cholesterol(HDL-C)between the two groups(P＞0.05).The body mass index,waist circumference,total testosterone(TT),anti-Miillerian hormone(AMH),luteinizing hormone(LH),fasting insulin(FINS),fasting blood glucose(FBG),2-hour postprandial blood glucose(2 h BG),homeostasis model assessment of insulin resistance(HOMA-IR),total cholesterol,triglycerides(TG),and low-density lipoprotein cholesterol(LDL-C)of patients in the PCOS group were significantly higher than those in the control group,while the level of estradiol(E2)was significantly lower than that in the control group(P＜0.05).The relative expression level of miR-320a-3p in plasma of patients in the PCOS group was significantly lower than that in the control group(P＜0.05).The expression level of miR-320a-3p was moderately negatively correlated with TT(r=-0.594,P＜0.05)and slightly negatively correlated with waist circumference,FINS,2 h BG,HOMA-IR,and LDL-C(r=-0.293,-0.208,-0.227,-0.208,-0.208;P＜0.05),and showed no significant correlation with hip circumference,AMH,FSH,E2,PRL,FBG,TG,and HDL-C(r=-0.079,0.020,-0.042,0.089,0.005,-0.141,-0.116,0.059;P＞0.05).The area under the curve of miR-320a-3p for predicting PCOS was 0.968(95％confidence interval:0.922-1.000,P＜0.05),with a cut-off value of 0.515,a sensitivity of 0.917,a specificity of 0.789,and a Jordon index of 0.706.miR-320a-3p negatively regulated androgen receptors.Conclusion miR-320a-3p is abnormally low expressed in peripheral blood of PCOS patients and may participate in the occurrence and development of PCOS by negatively regulating androgen receptors to increase TT levels.It has high predictive value for diagnosing PCOS.

Objective:To analyze and determine the clinical and genetic characteristics of children with Helsmoortel-Van der Aa syndrome(HVDAS).Methods:Clinical data of 6 children with HVDAS treated at the First Hospital of Peking University from November 2018 to October 2022 and their family members were collected and analyzed retrospectively.Whole exome sequencing was performed on children and their family members to identify the genetic variants.Genotype and phenotype correlation was analyzed.Results:(1) Clinical analysis results: among the 6 children, there were 5 boys and 1 girl, and their age at diagnosis ranged from 11 months and 17 days to 12 years and 9 months.Six patients all presented with developmental delays/intellectual disabilities; (2) Genetic analysis results: 6 de novo ADNP variants were discovered in 6 children, including 1 initial codon deletion variant c. 1_2del, 2 nonsense variants c. 1175dup, p.(Tyr392*) and c. 2213C>G, p.(Ser738*), and 3 frameshift variants c. 2632dup, p.(Ser878Lysfs*3), c.1695_1696insATGGTATGTATGTATGTATG, p.(Val566Metfs*8) and c. 2120_2123del, p.(Asn707Serfs*8).All variants were classified as pathogenic variants by the American College of Medical Genetics and Genomics.Except the c. 2213C>G, p.(Ser738*), the other 5 variants are all novel variants that have not been reported before. Conclusions:All of the 6 cases of HVDAS showed typical clinical manifestations, and expanded the phenotype spectrum of microcephaly and tall stature.Six de novo mutations were discovered, expanding the ADNP mutation spectrum and providing accurate genetic counseling and prenatal genetic diagnosis of the disease.

Objective:To investigate the effectiveness and feasibility of whole exome sequencing (WES), as a molecular diagnosis technique, for adult patients with genetic diseases.Methods:The present retrospective analysis included 445 adult patients (ages 18-80 years) with suspected genetic diseases who underwent whole exome sequencing (WES) from August 2021 to December 2022. The pathogenicity classification of each variant was assessed in accordance with the recommendations developed by the American Society of Medical Genetics and Genomics.Results:The overall positive rate of WES among adult patients with suspected genetic diseases was 28.08% (125/445). The highest positive rate was observed in the age group of 41-50 years (34.33%, 23/67). Among the diagnosed genetic diseases, those affecting the cardiovascular system (63.16%, 84/133), nervous system (18.05%, 24/133), and endocrine system (13.53%, 18/133) ranked as the top three. The most common genetic diseases identified through WES in adult patients were hypertrophic cardiomyopathy (18.80%, 25/133), dilated cardiomyopathy (16.54%, 22/133), Marfan syndrome (15.04%, 20/133), epilepsy (9.02%, 12/133), and familial hypercholesterolemia (4.51%, 6/133). The main causative genes identified included FBN1 (14.29%, 19/133), MYBPC3 (9.02%, 12/133), MYH7 (9.02%, 12/133), LDLR (3.76%, 5/133), TTN (3.76%, 5/133), and TNNI3 (3.01%, 4/133).Conclusion:Applying the WES technique in clinical practice can improve the diagnostic rate of adult genetic diseases, especially in adult patients with suspected genetic conditions involving the cardiovascular system, nervous system, and endocrine system.

Clinical research coordinator (CRC) can improve the efficiency of clinical research, ensure the accuracy and reliability of the research process, as well as the standardized implementation of research projects and the safety of subjects. However, the CRCs dispatched by the site management organization have problems such as high liquidity, uneven quality, low work initiative, and high management costs. Since 2004, a research hospital in South China had started to employ CRCs by itself, exploring and forming three self-employed CRC management modes: hospital employ-hospital manage, researcher employ hospital manage, and researcher employ-third-party labor dispatch agency manage. The CRCs under hospital employ-hospital manage mode were recruited and selected by the hospital. The hospital established a labor relationship with CRC, and the hospital′s personnel department was responsible for salary distribution and promotion management of professional titles/positions. The relevant labor costs were borne by the hospital′s funds, and the salary standards were also formulated by the hospital. This type of CRC could participate in the promotion of health technology series titles. The CRCs under researcher employ-hospital manage mode were recruited and selected by the researcher. The hospital established a labor relationship with them. The relevant labor costs were borne by the research funding, and the researchers determined the specific salary standards for CRCs based on the salary guidance plan formulated by the hospital. These kind of CRCs could not be promoted to professional titles in hospitals. The CRCs under researcher employ-third-party labor dispatch agency manage mode were recruited and selected by researchers. The third-party labor dispatch agency established an employment relationship with them and provided personnel services. The relevant labor costs were borne by the research funding, and the researchers formulated their own salary standards. Such CRCs could not be promoted to professional titles in hospitals. As of July 2023, the number of self-employed CRCs in the hospital had reached 185, with 75.8% having a bachelor′s degree or above and an average work experience of 4.58 years. Among them, there were 26 people in the hospital hiring-hospital management mode, 45 people in the researcher hiring-hospital management mode, and 114 people in the researcher hiring-third-party labor dispatch agency management mode. These measures had fully released management efficiency, effectively met manpower needs, promoted rapid growth in the number of clinical research projects, and reduced management costs through third-party labor dispatch mode, which could provide references for CRC management in clinical trial institutions of China.

Objective To correctly identify the blood group of ABO and study its molecular biological characteristics.Methods Blood samples from blood donors with discrepancies in forward and reverse typing using the microplate method were subjected to both saline tube agglutination test for serological identification and polymerase chain reaction-sequence specific primers(PCR-SSP)for genotyping.Additionally,direct sequencing of exons 1 to 7 of the ABO gene was performed on some donor samples to analyze their blood phenotype,genotype and gene sequence.Results For 256 samples showing discrepancy between forward and reverse typing in microwell method,119 were identified as normal ABO blood types,90 were weakened ABO antibodies and 47 were ABO subtypes by serology tube test.According to the PCR-SSP genotyping test,233 of 256(91.02%)were consistent with serological phenotype and genotype,17 of 256(6.64%)were inconsistent,and 6 samples can′t read genotype based on the kit result typing table.A total of 17 genotypes were identified in 250 samples as AO1 in 56,AO2 in 58,AA in 50,BO1 in 31,BO2 in 17,BB in 8,O1O1 in 2,O1O2 in 7,AB in 13,AO4 in 1,A205O2 in 1,A205A in 1,A201A in 1,O1O4 in 1,O2O2 in 1,A201B in 1 and A205B in 1.Sequencing of exons 1 to 7 of the ABO gene was carried out in 78 samples,and 29 ABO alleles were detected,seven of which were common alleles(?A101,?A102,?A104,?B101,?O01,?O02,?O04),22 of which were rare alleles(?A201,?A205,?Ax01,?Ax03,?Ax13,?Ax19,?Ax22,?Ael10,?B305,?Bel03,?Bel06/?Bx02,?Bw07,?Bw12,?Bw17,?Bw37,?O05,?O26,?O61,?B(A)04,?B(A)07,?cisAB01 and ?cisAB01var).In addition,six rare allele mutation sites were identified(c.101A＞G;c.103_106delG;c.146_147insGC;c.259G＞T;c.322C＞T;c.932T＞C).Conclusion The identification of ambiguous ABO blood group requires the combination of serological testing and molecular biological examination to correctly identify the blood type and ensure the safety of clinical blood transfusion.

Objective To express recombinant Burkholderia pseudomallei(B.pseudomallei)type Ⅲ secretion system BipD protein,prepare its polyclonal antibodies and verify their immunological traits.Methods The recombinant pET-28a-BipD plasmid was generated,and the pET-28a-BipD-carried E.coli BL21(DE3)bacteria were induced with isopropyl-β-d-thiogalactoside(IPTG)to express recombinant BipD(rBipD)protein.The rBipD was obtained by affinity chromatography using His Trap column,then mixed with Fredrick's adjuvant to immunize BALB/c mice by intraperitoneal injection in order to obtain anti-rBipD polyclonal antibodies.The immunoreactivity of rBipD was detected by Western blot assay using rabbit anti-melioidosis serum and the serum from melioidosis patients.The immunogenicity of rBipD was evaluated using Western blotting and immunofluorescence staining.Finally,rBipD was used to establish an indirect ELISA to detect serum antibodies of clinical melioidosis patients.Results The recombinant plasmid pET-28a-BipD was successfully constructed and transformed into E.coli BL21(DE3)to induce rBipD expression with IPTG treatment.The obtained rBipD had a relative molecular weight of 36×103 and a purity of 95.4％,and had good immunogenicity and immunoreactivity.It could induce the production of specific antibodies after immunizing mice,and mouse polyclonal antibodies against rBipD were prepared with the titer of 1∶512 000.rBipD of 5.0 μg/mL produced specific immune response with the serum of melioidosis patients,but had no specific reaction with the serum of tuberculosis patients,with statistical difference(P＜0.01).Conclusion rBipD with immunological activity is successfully prepared and purified,and its polyclonal antibodies are also developed,which provide a good tool for clinical immunological diagnosis and study of immune mechanism of B.pseudomallei infection.