Objective To explore the role and mechanism of warm malate ringer's solution(MR)in resuscitation of the lethal triad caused by severe trauma.Methods A rat model of severe trauma was established in SPF-grade SD rats(half male and half female,weighing 200～220 g)using combined multiple injuries and hemorrhagic shock,and the rats were randomly divided into 8 groups(n=8):Sham group,only arterial and venous catheterization;Trauma(Tra)groups with different time points(10,30,60,90,120,180 min)and a Trauma group that were observed without any treatment for 180 min after model establishment.The changes of activated clotting time(ACT),reaction time(R),maximum amplitude(MA),and rate of blood clot formation(Angle)at different time points were detected by using thromboelastography,and tail bleeding,core body temperature and arterial blood gas parameters,were also observed and detected.The plasma von Willebrand Factor(vWF)level,mitochondrial respiratory control ratio in pulmonary venous endothelium,and expression levels of vascular endothelial cadherin(VE-Cadherin),peroxisome proliferator activating receptor gamma coactivator 1α(PGC1α),dynamin-related protein 1(Drp1),p-Drp1,and mitofusin 2(Mfn2)were detected to evaluate the vascular endothelial injury and mitochondrial dysfunction.Another group of SD rats were randomly divided into severe trauma group(no treatment for 180 min after injury),and MR solution at room temperature and at 37 ℃ groups.MR solution at room temperature or at 37 ℃ was given to the rats using a medical blood transfusion apparatus at 60 min post-trauma.Above indicators were observed and detected to investigate the resuscitation effect of the MR solution.Results Compared with the Sham group,the severely traumatic rats at 180 min after injury had significantly prolonged ACT and R values(P＜0.05),shortened MA and decreased Angle values(P＜0.05),extended tail bleeding time(P＜0.05),lower partial pressure of carbon dioxide(PCO2)and HCO3-and base excess(BE)levels(P＜0.05),and continuously increasing K+(P＜0.05)and decreasing Na+(P＜0.05)and Ca2+levels(P＜0.05).Additionally,plasma vWF level(P＜0.05)and protein levels of VE-cadherin,PGC1α and Mfn2 in pulmonary vein endothelium were significantly reduced(P＜0.05),the expression of p-Drp1 was enhanced and the mitochondrial respiration control rate was declined in the rats at 180 min after injury(P＜0.05).MR solution resuscitation shortened tail bleeding time(P＜0.05),increased core body temperature(P＜0.05),elevated plasma vWF level(P＜0.05),increased protein levels of VE-cadherin,PGC1α and Mfn2(P＜0.05),and decreased that of p-Drp1 protein expression(P＜0.05)when compared with the rats at 180 min after severe traumatic injury.The above effects were more significant in the rats infused with the solution at 37 ℃ than those at room temperature.Conclusion Warm MR solution significantly improves the lethal triad in rats after severe trauma,which may be associated with its improving mitochondrial function and attenuating vascular endothelial damage.

Objective:To explore the clinical characteristics of neonatal necrotizing enterocolitis（NEC）and analyze the risk factors for early-onset NEC.Methods:A total of 220 children with NEC admitted to the Department of Pediatrics，Tianjin Medical University General Hospital from January 1st，2018 to February 29th，2024 were retrospectively selected as the research objects. According to the time of onset，the early-onset group（ n=120）and the late-onset group（ n=100）were established，and the clinical characteristics of the two groups were compared. Another 150 cases of normal healthy newborns born in this hospital in the same period were selected as the control group，and the clinical data of the control group were collected. The clinical characteristics of the early-onset group and the control group were compared，and the risk factors of early-onset NEC were analyzed by multivariate Logistic regression. Results:Compared with the late-onset group，the early-onset group had fever［50.0%（60/120）vs. 40%（40/100）， χ2=7.333， P=0.007］，apnea［39.17%（47/120）vs. 28%（28/100）， χ2=7.568， P=0.006］，no rise in body temperature［56.67%（68/120）vs. 39%（39/100）， χ2=6.815， P=0.009］，abdominal distension［25%（30/120）vs. 40%（40/100）， χ2=13.200， P<0.001］，vomiting［30.83%（37/120）vs. 45%（45/100）， χ2=12.797， P<0.001］was significantly different（all P<0.05）；Multivariate Logistic regression analysis：weight<1 500 g（ OR=5.871，95% CI：3.153～9.673， P<0.001），gestational age<30 weeks（ OR=4.256，95% CI：2.641～7.896， P=0.007），hemodynamically significant patent ductus arteriosus（hs-PDA）（ OR=3.113，95% CI：1.865～5.133， P=0.033），severe anemia（ OR=3.057，95% CI：2.165～4.802， P=0.001），feeding intolerance（ OR=4.215，95% CI：1.579～10.802， P=0.005），amniotic fluid pollution（ OR=2.452，95% CI：1.579～3.111， P<0.001）were the independent risk factors for early-onset NEC（all P<0.05）. Conclusion:Weight<1 500 g，gestational age<30 weeks，hs-PDA，severe anemia，feeding intolerance，and amniotic fluid contamination are independent risk factors for early-onset NEC. In clinical practice，more attention should be paid to these factors for disease prevention，early identification，and timely intervention in newborns to reduce the occurrence of NEC.

Background::Total human immunodeficiency virus (HIV) DNA and integrated HIV DNA are widely used markers of HIV persistence. Droplet digital polymerase chain reaction (ddPCR) can be used for absolute quantification without needing a standard curve. Here, we developed duplex ddPCR assays to detect and quantify total HIV DNA and integrated HIV DNA.Methods::The limit of detection, dynamic ranges, sensitivity, and reproducibility were evaluated by plasmid constructs containing both the HIV long terminal repeat (LTR) and human CD3 gene (for total HIV DNA) and ACH-2 cells (for integrated HIV DNA). Forty-two cases on stable suppressive antiretroviral therapy (ART) were assayed in total HIV DNA and integrated HIV DNA. Correlation coefficient analysis was performed on the data related to DNA copies and cluster of differentiation 4 positive (CD4 +) T-cell counts, CD8 + T-cell counts and CD4/CD8 T-cell ratio, respectively. The assay linear dynamic range and lower limit of detection (LLOD) were also assessed. Results::The assay could detect the presence of HIV-1 copies 100% at concentrations of 6.3 copies/reaction, and the estimated LLOD of the ddPCR assay was 4.4 HIV DNA copies/reaction (95% confidence intervals [CI]: 3.6-6.5 copies/reaction) with linearity over a 5-log 10-unit range in total HIV DNA assay. For the integrated HIV DNA assay, the LLOD was 8.0 copies/reaction (95% CI: 5.8-16.6 copies/reaction) with linearity over a 3-log 10-unit range. Total HIV DNA in CD4 + T cells was positively associated with integrated HIV DNA ( r = 0.76, P <0.0001). Meanwhile, both total HIV DNA and integrated HIV DNA in CD4 + T cells were inversely correlated with the ratio of CD4/CD8 but positively correlated with the CD8 + T-cell counts. Conclusions::This ddPCR assay can quantify total HIV DNA and integrated HIV DNA efficiently with robustness and sensitivity. It can be readily adapted for measuring HIV DNA with non-B clades, and it could be beneficial for testing in clinical trials.

Objective To investigate the factors influencing the pregnancy outcomes during frozen-thawed embryo transfer(FET)cycles in patients with polycystic ovary syndrome(PCOS).Methods A retrospective analysis was conducted on patients'data from 882 FET cycles.According to the pregnancy outcome,the patients were divided into non-implantation group(Group A),abortion group(Group B1)and live birth group(Group B2).Clinical data and laboratory parameters were compared among the three groups,and ordered Logistic regression analysis was used to study the factors influencing pregnancy outcomes after FET.Patients were also divided into four groups(C1-C4)based on the number of high-quality embryos obtained(0-3,4-6,7-10,≥11),and their clinical data and laboratory parameters were compared.Results The clinical pregnancy rate,live birth rate,and miscar-riage rate in the 882 treatment cycles were 71.09%(627/882),61.68%(544/882),and 13.24%(83/627),respectively.Single-factor analysis showed significant differences in body mass index(BMI),infertility type,hu-man chorionic gonadotropin(hCG)day estradiol(E2)level,number of retrieved oocytes,and number of high-quality embryos among Groups A,B1,and B2(P<0.05).Further multiple Logistic regression analysis revealed that BMI(OR=1.046,95%CI:1.001-1.093,P=0.044)and a history of previous pregnancy(OR=1.417,95%CI:1.030-1.950,P=0.032)were independent risk factors for successful FET in PCOS patients,while an in-creased number of high-quality embryos was an independent protective factor for successful pregnancy.Based on the results of Group B2,compared to Group A,OR=0.920,95%CI:0.880-0.962,P=0.000;compared to Group B1,OR=0.923,95%CI:0.862-0.988,P=0.022.Compared with the other three groups(C1-C3),the total amount of gonadotropin(Gn)in the C4 group was the lowest and the number of oocytes obtained was the high-est(P<0.05).Multiple comparisons showed that Group C4 had lower BMI,follicle-stimulating hormone(FSH),very low-density lipoprotein(vLDL)levels,a higher luteinizing hormone and follicle-stimulating hormone(LH/FSH)ratio compared to Group C1(P<0.05).Group C4 had lower fasting insulin(FINS)and homeostasis model assessment of insulin resistance(HOMA-IR)levels compared to Group C3,and higher high-density lipoprotein-cholesterol(HDL-C)and apolipoprotein A1(Apo A1)levels compared to Groups C2 and C3(P<0.05).Con-clusion BMI,the history of previous pregnancy and the number of high-quality embryos were both independent factors for predicting pregnancy outcomes in PCOS patients undergoing FET cycles.Patients with a higher number of high-quality embryos have a higher clinical pregnancy rate during FET cycles.

Objective To investigate the role of member septin family(SEPT12)in human spermatogenesis and its influence on sperm motility and sperm ultrastructure.Methods Whole exome sequencing(WES)was performed on peripheral blood DNA extracted from 375 patients with asthenoteratozoospermia,and a patient with idiopathic in-fertility carrying compound heterozygous mutation of SEPT12 was screened out.Sanger sequencing was performed to verify the mutation,and co-segregation analysis was performed in the family.The morphological abnormalities of sperm were analyzed by hematoxylin-eosin(HE)staining and scanning electron microscopy(SEM),and the ultra-structural defects of sperm were analyzed by transmission electron microscopy(TEM).Then the effects of the muta-tion on the level and position of the protein and the changes of the location and level of the defect structure markers were analyzed by Western blot and immune-fluorescence(IF).Results The compound heterozygous mutations c.C332A(p.Ti111K)and c.406_416 del TGCTCGTATTG(p.q136 VFS*39)in the SEPT12 gene were screened and identified in a patient with asthenoteratozoospermia.The mutations were verified by Sanger sequencing,which was consistent with the co-segregation genetic pattern of the family.The mutations resulted in loss of protein expres-sion,decreased sperm motility and sperm morphological deformities,mainly including short tail,curly tail and ir-regular sperm head.The ultrastructure of sperm showed that the annulus between the mid-piece and the principle-piece was missing,the acrosome membrane of sperm head fell off and the nucleus contained vacuoles.In the mid-piece of sperm flagella,the arrangement of mitochondrial sheath was disordered,most of flagella axoneme central pair was absent,microtubules doublet was missing or disordered,and some radical spoke was absent.By Western blot and IF,the marker proteins of related structural components were detected,and the results showed that the level of SEPT4 protein decreased,SEPT6 protein unchanged,acrosomal related proteins ACTL7A and ACROSIN protein missing,and the expression levels of mitochondrial and axoneme related proteins TOMM20,SPAG6 and RSPH3 protein significantly decreased.Conclusion The deletion of SEPT12 protein caused by SEPT12 gene mu-tation leads to the deletion of the annulus between the mid-piece and the principle-piece,and the abnormal assem-bly of sperm acrosome,mitochondrial sheath and flagella.

BACKGROUND:In vitro fertilization-embryo transfer is commonly used to solve infertility,but its success rate is not high,the more common reasons are poor endometrial receptivity,poor egg quality,etc.The follicular fluid melatonin can inhibit the aging of the ovary,to a certain extent,can promote the development of embryos,improve the probability of conception,but whether there is a correlation between the two is not known. OBJECTIVE:To explore the correlation between follicular fluid melatonin level and pregnancy rate of single-cycle in vitro fertilization-embryo transfer women. METHODS:A total of 112 female patients who received in vitro fertilization-embryo transfer treatment in the First Affiliated Hospital of Anhui Medical University from December 2020 to April 2021 were selected as the study subjects.They were divided into quartile array(Q1-Q5)according to the follicular fluid melatonin level from low to high.Among them,the melatonin level of group Q1 was<6.99 ng/L(n=18),that of group Q2 was 7.00-9.99 ng/L(n=26),that of group Q3 was 10.00-11.99 ng/L(n=27),and that of group Q4 was 12.00-13.99 ng/L(n=18);and melatonin levels in group Q5 were 14.00-19.99 ng/L(n=23).Clinical data characteristics of the five groups were compared.Multi-factor Logistic regression was used to analyze the correlation between follicular fluid melatonin level and pregnancy rate of women with single-cycle in vitro fertilization-embryo transfer and embryo transfer.A restricted cubic spline Logistic regression model was established to analyze the dose-response relationship,and the model was evaluated by clinical decision curve. RESULTS AND CONCLUSION:(1)Compared with the study population with the lowest melatonin quintile(Q1),with the increase of melatonin level(Q2-Q5),the levels of egg harvest and pregnancy success were gradually increased,and the body mass index was gradually decreased,and the differences were significant(P<0.05).(2)Multivariate Logistic regression analysis showed that after adjusting for confounding factors such as global mass index,number of eggs retrieved,luteinizing hormone,estradiol,progesterone and other confounding factors,follicular fluid melatonin level was still independently correlated with pregnancy rate of single-cycle in vitro fertilization-embryo transfer women(OR=1.538,95%CI:1.032-1.837,P<0.05),and there was significant difference in trend test of follicular fluid melatonin level from low to high quintile array(Ptrend<0.05).(3)The sensitivity test analysis showed that E value was 2.117.Subgroup analysis showed that the study population with higher levels of luteinizing hormone in follicular fluid had a more significant association between follicular fluid melatonin and pregnancy rate in single-cycle in vitro fertilization-embryo transfer women(P interaction=0.008).(4)The results of restricted cubic spline model analysis showed that there was a nonlinear dose-response relationship between follicular fluid melatonin level and pregnancy rate of single-cycle in vitro fertilization-embryo transfer women(P<0.05),and there was an overall positive correlation between follicular fluid melatonin level and pregnancy rate of single-cycle in vitro fertilization-embryo transfer women.(5)The results of clinical decision curve analysis showed that the follicular fluid melatonin level had important clinical value in predicting the pregnancy rate of single-cycle in vitro fertilization-embryo transfer women.(6)Follicular fluid melatonin level is closely related to the pregnancy rate of single-cycle in vitro fertilization-embryo transfer women,and with the decrease of follicular fluid melatonin level,the pregnancy rate of single-cycle in vitro fertilization-embryo transfer women also decreases.

Objective:To explore the developmental characteristics of different cognitive styles on verbal and visuospatial working memory of primary school students.Methods:The Mosaic graph test was used to test the cog-nitive style of students in grades 2,4 and 6.The three-factor mixed experiment of 2(cognitive style:field independ-ent,field dependent)× 3(grade:2,4,6)× 2(task type:letter task,spatial task)was carried out.The 2-back task was used to measure the working memory characteristics of students with different cognitive styles,and the repeated measurement ANOVA was performed on the accuracy of letter and spatial tasks of 175 effective subjects.Results:The accuracy of letter task and spatial task of field independent primary school students were higher than that of field-dependent primary school students(P＜0.001).The accuracy rate of letter task for field independent primary school students was higher than that of space task(P＜0.001).The accuracy of two kinds of tasks of field inde-pendent students in grade 6 was better than that of field independent students in grade 2 and grade 4(P＜0.05).Conclusion:There are differences in the development of verbal and visuospatial working memory of primary school students with different cognitive styles,which may also be related to task type and age.

Objective To observe the ameliorative effect of inhibiting acetyl-CoA carboxylase 2(ACC2)on cardiac dysfunction in septic mice and investigate its underlying mechanism.Methods Mouse model of sepsis was established by cecal ligation and perforation.A total of 24 male C57BL/6 mice(aged 8 weeks,weighing 20～25 g)were divided into sham operation group,sepsis group and ND-630+sepsis group.The cardiac specific ACC2 knockout(ACC2△CM)mice were constructed by Cre-LoxP recombinase system,and ACC2flox/flox Myh6-Cre-(ACC2fl/fl)mice were used as control.Several genetically engineered mice(8 weeks old,20～25 g,male)were divided into ACC2fl/fl+sham operation group,ACC2fl/fl+sepsis group,ACC2△CM+sepsis group,ACC2△CM+Mal-CoA+sepsis group,and ACC2△CM+sham operation group according to the random number table method.The contractile function and myofilament calcium sensitivity of cardiomyocytes were measured by a cell microtensiometer.Western blotting was used to detect the expression level of ACC2,and ELISA was employed to measure the level of malonyl-CoA(Mal-CoA)in myocardial tissue.The survival of the mice in 36 h after sepsis was observed.Results Compared with the sham operation group,the contraction amplitude of cardiomyocytes in sepsis group was decreased markedly,and the calcium sensitivity decreased significantly as well(P＜0.05).Based on the ACC2fl/fl+sham operation group,the ACC2△CM+sepsis group showed significant improvement in myocardial contraction compared with the ACC2fl/fl+sepsis group,with the contraction amplitude of cardiomyocytes recovered by 48.1％,and significantly restored calcium sensitivity(P＜0.05).ACC2△CM+Mal-CoA+sepsis group showed a notable decrease in myocardial cell contraction amplitude and a significant decrease in calcium sensitivity when compared to the ACC2△CM+sepsis group(P＜0.05).Compared with the sepsis group,the contraction amplitude of cardiomyocytes in the ND-630+sepsis group increased significantly,and the calcium sensitivity of myofilament also increased(P＜0.05).The expression level of ACC2 protein in the myocardial tissue of mice in the sepsis group increased compared to the sham operation group(P＜0.05).The level of myocardial Mal-CoA in the sepsis group was higher than that in the sham operation group(P＜0.05);Mal-CoA level in the myocardium of ACC2△CM+sepsis group mice was lower than that of sepsis group(P＜0.05).The 36-hour survival rate of the ACC2△CM+sepsis group.mice was 37.5％higher than that of the ACC2fl/fl+sepsis group mice.(P＜0.05).Conclusion Inhibition of ACC2 exerts a protective effect on myocardial contractility and calcium sensitivity in septic mice by reducing Mal-CoA.

Objective:To explore the causal associations of two blood pressure phenotype and four lipid fractions with type 2 diabetes mellitus(T2DM) in European and East Asian populations using Two-Sample Mendelian randomization analysis.Methods:Blood pressure phenotype, lipid fractions and T2DM genetic loci from two ethnics were matched and combined according to single nucleotide polymorphisms(SNPs) numbering. With SNPs closely related to the exposure phenotype as instrumental variables, the inverse variance weighting method was used to analyze the causal effects of blood pressure phenotype and lipid fractions on T2DM in different ethnic groups. The sensitivity analysis was conducted using MR-Egger regression model, Weighted Median method, MR-PRESSO, MR-robust Adjusted Profile Score, and leave-one-out method.Results:Among European populations, systolic blood pressure( OR=1.40, 95% CI 1.23-1.59, P<0.001) and diastolic blood pressure( OR =1.24, 95% CI 1.08-1.42, P=0.002)were associated with increased risk of T2DM while high density lipoprotein-cholesterol( OR=0.68, 95% CI 0.62-0.76, P<0.001) reduced the risk of T2DM. In East Asian ethnicity, elevated diastolic blood pressure( OR=0.75, 95% CI 0.59-0.95, P=0.007) reduced the risk of T2DM. Sensitivity analysis confirmed the results. Conclusion:There are differences in the effects of blood pressure phenotype and lipid composition on T2DM in different ethnic groups, which may be related to population heterogeneity and exposure sensitivity. It should be taken into consideration in extrapolation.

Objective: To explore the characteristics of overweight children s attention bias to food cues, so as to provide a theoretical basis for the behavior and life intervention of overweight children. Methods: From September to October 2017, 28 normal weight children and 28 overweight children in grades 3 to 4 of a primary school in Shenyang were screened by body mass index (BMI). The point detection paradigm was used to evaluate the attention bias of overweight children to food cues. Results: There was no significant difference between overweight children and control children in the score of attention bias to food cues [ F (1,52)=0.01, P> 0.05 , ηp 2 <0.01]; there was significant difference between overweight children and normal weight children in the score of facilitated attention to food cues[ F (1,52)=5.94, P<0.05, ηp 2 =0.10], normal weight children had higher score of facilitated attention to food cues than that of overweight children; there was significant difference between overweight children and normal weight children in the score of difficulty in disengaging to food cues[ F (1,52)=7.74, P<0.05, ηp 2 =0.13], overweight children had higher score of difficulty in disengaging to food cues than that of normal weight children. Conclusion Compared with normal weight children, overweight children didn t have higher attention bias to food cues; in the specific indicators of attention bias to food cues, normal weight children are more alert to food cues, while overweight children are more difficult to remove attention from food cues.