Objective To construct 5 machine-learning models and compare their performance in predicting the associations between pre-pregnancy socio-psycho-behavioral exposures of both spouses and preconception outcomes.Methods Based on Chongqing Preconception Reproductive Health and Birth Outcome Cohort of volunteers recruited from Chongqing Health Center for Women and Children during January 2019 and March 2022,5 447 couples were recruited and surveyed through interviewer-interview for the demographic and social-psychological-behavioral data of both spouses(221 variables).According to the inclusion and exclusion criteria,4 097 couples were finally included,and randomly assigned into a training set(n=2 867 spouses)and a validation set(n=1 230 spouses)at a ratio of 7∶3.Feature analysis and collinear screening were applied to select the potential exposure factors.In consideration of difficulty to carry out semen parameters analysis in primary healthcare institutions,feature Set 1 including sperm parameters and feature Set 2 excluding semen parameters were constructed by including or excluding sperm quality simultaneously in the training set and the validation set.Five algorithms,that is,Logistic Regression,Naive Bayes,Random Forest,Gradient Boosting Machine,and Support Vector Machine,were used to construct preconception outcome prediction models,and the parameters of each model were optimized using random search combined with grid search.The predictive performance of each model was compared using precision,recall,F1 score,area under the receiver operating characteristic curve(AUC),and calibration curve.The optimal model was then selected by comparing the changes in the predictive ability of the questionnaire data for fertility outcomes with or without semen parameters.Results There were 24 variables screened out in feature Set 1,and 16 variables in feature Set 2.In feature Set 1,the gradient boosting machine performed better,with a relatively higher AUC value(0.651)and better F1 score(0.61).The logistic regression model performed stably(AUC value=0.647)and was suitable as the reference model.The random forest(AUC value=0.641),Naive Bayes(AUC value=0.641),and support vector machine(AUC value=0.634)performed second-best.By utilizing the gradient boosting machine,comparable results were found between the predictions from feature sets with or without semen parameters,as in feature Set 1,the AUC value of its validation set was 0.651(95%CI:0.629～0.681),the prediction accuracy was 0.63,the recall rate was 0.65,and the average precision value F1 was 0.61;and in feature Set 2,the AUC value of its validation set was 0.649(95%CI:0.624～0.663),and both the calibration curves were close to the ideal curve.The prediction results indicated that in feature Set 1,the features highly negatively correlated with preconception outcomes were female age,male age,and no pregnancy within 1 year without contraception,while the features highly positively correlated with preconception outcomes were female pregnancy history,total sperm vitality,and use of contraceptive measures before enrollment.Conclusion Among the 5 machine-learning algorithms performed in this cohort data,the gradient boosting machine shows slightly better performance.There are 24 factors being associated with preconception outcomes in both spouses,and the performance of the simplified model excluding semen parameters is not significantly declined.It is feasible to use machine-learning methods to predict human preconception outcomes through social-psychological-behavioral questionnaires.

Objective To analyze the detrimental effects of exposure to environmental noise alone and combined with carbon monoxide(CO)on spermatogenesis in male rats,investigate the underlying mechanisms involved in such damage,and evaluate the protective role of pterostilbene(PTE)against these adverse effects.Methods Sixty male SD rats(6～8 weeks old,weighing 200±10 g)were randomly divided normal control group(standard housing),sham-exposure control group(restraint stress only),noise exposure(85 dB),CO exposure(460 mg/m3),combined exposure(simultaneous exposure),and PTE intervention(80 mg/kg pretreatment),with 10 animals in each group.The rats were exposed daily for 2 h via a nose-only inhalation exposure system within a multifactorial environmental simulation chamber for 60 consecutive days.Sperm count and viability were measured after exposure.Histopathological changes of testicular tissues were observed with HE staining.qRT-PCR was used to measure stage-specific mRNA levels in germ cells.Serum sex hormone levels and adenosine triphosphate(ATP)concentrations in testes and sperm were detected.Transmission electron microscopy(TEM)was applied to observe the ultrastructural damage in the spermatocytes.Additionally,transcriptome sequencing was performed on testicular tissue,followed by bioinformatics analysis.Results Compared with the negative control group,the combined exposure group exhibited significant reductions in sperm viability and count(P<0.05),and developmental arrest of immature germ cells in the testicular tissue,with obviously less spermatogonia,spermatocytes,and round/elongated spermatids(P<0.01).Additionally,significantly reduced levels of reproductive-related hormones,such as gonadotropin-releasing hormone,follicle-stimulating hormone,luteinizing hormone and testosterone,and ATP levels in testes and sperm were observed in the mice after combined exposure(P<0.01),accompanied by mitochondrial rupture and cristae disruption in spermatocytes.Conversely,the PTE intervention group showed marked alleviation of these impairments,with parameters recovering almost to normal levels.Transcriptome sequencing identified biological processes related to reproductive development and ATP-dependent pathways as potential contributors to testicular injury induced by noise and CO exposure,with key genes including Nppa,Adm,Gnrh1,Ptafr,Atp13a5,Atp8b1,and LOC102555469.Conclusion Noise and CO exposure induce spermatogenic damage in rats,which may be related with energy metabolism and hormonal regulation,while PTE demonstrates significant protective effects against such reproductive impairments.

Objective To investigate the transgenerational effects of paternal PM2.5 exposure on offspring growth and development,and to preliminarily elucidate the role of sperm DNA methylation modifications in mediating these effects.Methods Eight-week-old male C57BL/6 mice were randomly divided into filtered air(FA),unfiltered air(UA),and concentrated PM2.5(CAP)groups,with 10 animals in each group.The exposure was conducted from November 2019 to April 2020,and then,these male mice were mated with unexposed females to generate F1 offspring,which were bred successively to produce F2 and F3 generations.All the offspring were living in PM2.5-free environment.The birth body weight,birth number,and sex ratio of the offspring were recorded,body weight growth was monitored,and organ coefficients of the heart,liver,lung,and brain were calculated.Whole-genome methylation sequencing was performed on the sperm DNA of the CAP group,FA group,and their F1 generation offspring to screen for differentially methylated regions,and the genes and pathways associated with these regions were analyzed.Results When compared with the F1～F3 offspring of the FA group,the CAP group had significantly reduced birth body weight in the F1 generation(P<0.05),no statistical differences were observed in the birth body weight in the F2 and F3 generations(P>0.05),or either in the sex ratio and birth number among the F1,F2 and F3 generations.Compared with the FA group offspring,the F1～F3 offspring of CAP group exhibited delayed body weight gain,especially in the males(P<0.05),the CAP-F1 male generation had obviously elevated liver organ coefficient(P<0.01),but no statistical changes were observed in the heart,lung,or brain coefficients among the F1～F3 generations.Between the FA group and the CAP group,37 997 differentially methylated regions were detected,with a reduction of approximately 50%in the number of differentially methylated regions in the F1 generation.Differentially methylated genes in F0 and F1 sperm were potentially related to developmental processes,including imprinting genes(Gnas,Igf2)and metabolic genes(Ppard,Rps6kb1).Conclusion Paternal exposure to PM2.5 leads to reduced birth weight and intergenerational growth retardation in offspring.Its impact on phenotypic effects is gradually weakened during intergenerational transmission.Changes in the methylation of development-related genes in sperm may be one of the mechanisms mediating this intergenerational effect.

This study aims to describe the population and regional distribution characteristics of smoking behavior among adult twins in the China Twin Registry (CNTR), as well as the concordance rates for smoking behavior in monozygotic and dizygotic twins, and estimate the heritability. The study population included adult twins in CNTR who had smoking questionnaire data. A random-effects regression model was used to describe the distribution of smoking behavior among different subgroups based on various characteristics. The concordance of smoking behavior between different zygosity groups was calculated, and heritability was estimated. A total of 28 444 twin pairs were included in this study, with an average age of (36.6±12.0) years. Among male twins, 41.2% were current smokers, while only 1.2% of females smoked. Higher smoking rates were observed among male smokers in the 50-59 age group ( z=23.0, P<0.001), northern regions ( z=2.9, P<0.01), rural areas ( z=-5.2, P<0.001), those who were divorced/widowed ( z=3.8, P<0.001), and first-born twins ( z=-4.3, P<0.001), while lower smoking rates were found in those with higher education ( z=-16.1, P<0.001) and unmarried individuals ( z=-16.0, P<0.001). The smoking concordance rate for male monozygotic twins was 69.6%, significantly higher than the 57.3% concordance rate for dizygotic twins ( χ 2=105.0, P<0.05). The heritability of smoking behavior in male twins was estimated at 28.9% (95% CI: 24.3%-33.4%). Stratified analyses showed differences in heritability across regions and age groups: the heritability in northern regions was 32.6% (95% CI: 27.3%-38.0%), higher than the 21.0% (95% CI: 12.4%-29.5%) observed in southern regions; the highest heritability of 35.1% (95% CI: 26.3%-43.9%) was found in the 18-29 age group, with heritability decreasing with age. In conclusion, the smoking rate and influencing factors in the twin population are similar to those in the general population, with unique characteristics, such as higher smoking rates in first-born twins. Genetic factors have a significant impact on smoking behavior.

Objective To explore the correlation between triglyceride-glucose(TyG)index and hyperuricemia in men with normal fasting blood glucose(FPG)levels.Methods A total of 309 men with normal FPG who participated in a health examination at the Ninth Medical Center of the People's Liberation Army General Hospital in April 2024 were enrolled in this study.All the subjects were divided into the normal uric acid(NUA,n=218)group and the hyperuricemia(HUA,n=91)group according to serum uric acid(SUA)levels.Results Scr,TG,weight,SBP,DBP,BMI,ALT,AST,γ-GGT,and TyG index were higher in the HUA group than in the NUA group(P<0.05).Pearson and Spearman correlation analysis showed that SUA were positively correlated with Scr,eGFR,TG,weight,SBP,DBP,BMI,ALT,AST,γ-GGT and TyG(P<0.05),and negatively correlated with HDL-C(P<0.05).Logistic regression analysis showed that after adjusting for confounding factors,TyG index remained an important influencing factor for HUA.ROC curve analysis showed that the area under the ROC curve of TyG index predicting hyperuricemia in men with normal FPG was 0.665,with an cutoff value of 8.45.Conclusions TyG index in men with normal FPG are influencing factors for hyperuricemia,indicating that hyperuricemia has a close association with insulin resistance,and is an important component of metabolic syndrome.

Purpose To investigate the changes of free water(FW)values in the key basal ganglia of dopamine system in the brain of early Parkinson's disease(PD)by free water imaging,and to reveal the potential relationship between the changes and early PD.Materials and Methods From February to December 2023,62 patients with early PD and 27 healthy controls were enrolled in the Parkinson's Disease Progression Markers Project database,and 25 age-and sex-matched healthy controls were self-recruited from Henan Provincial People's Hospital.All subjects underwent clinical scale assessment and MRI data collection.Free water imaging was used to calculate the FW values of key nuclei in dopamine system,and the difference of FW values between PD group and healthy controls group,the correlation between FW values of PD group and clinical scale scores were analyzed.Results The FW values of compact part of substantia nigra(SNc)and ventral tegmental area(VTA)in PD group were significantly higher than those in healthy controls group(Z=2.458-3.914,all P<0.05).Regression analysis showed that FW values of SNc were correlated with the movement disorder society-sponsored revision unified Parkinson′s disease rating scale(MDS-UPDRS)Ⅲ scores and state-trait anxiety inventory(STAI)scores on both sides(r2=0.223,P<0.001;r2=0.125,P=0.018;r2=0.151,P=0.003;r2=0.128,P=0.017),the FW value of VTA on the left was significantly correlated with MDSUPDRS Ⅲ score(r2=0.143,P=0.004)and the FW value of the right VTA was significantly correlated with the STAI score(r2=0.125,P=0.019).Conclusion The FW values of SNc and VTA in PD patients are significantly increased in the early stage,and are correlated with motor and non-motor symptoms in PD patients.FW values may reflect brain degeneration,and this change may be related to the decline of motor and non-motor function in patients.

Objective:To explore whether changes in the deltoid muscle tone of the affected side before and after arthroscopic rotator cuff repair (ARCR) are correlated with shoulder joint pain and shoulder joint function.Methods:A total of 55 patients (22 males and 33 females, aged 60.44±9.54 years with a range of 41 to 78 years, BMI 25.86±4.25 kg/m 2 with a range of 18.82 to 42.32 kg/m 2, symptom duration 5.94±7.27 months with a range of 0.3 to 36.0 months) diagnosed with rotator cuff tear and underwent ARCR at the First Affiliated Hospital of Tsinghua University between December 2019 and June 2023 were included. All the patients suffered from unilateral symptom (23 left side and 32 right side), the size of rotator cuff tear was 16.47±5.98 mm (range, 6.92-36.38 mm). Following the operation, postoperative rehabilitation exercises were arranged with a minimum follow-up duration of six months. The muscle tone of the deltoid was measured bilaterally using a muscle tone tester before the operation and at 1, 3, 6 months after operation. The area under curve (AUC) of the pressure-displacement curve was used to reflect the muscle tone, with larger AUC values represent less muscle tone. Visual analogue scale (VAS) and Constant-Murley score were evaluated and recorded at the same time points. Results:At 1, 3, and 6 months postoperatively, the VAS scores were all significantly reduced compared to preoperative values, with the Constant-Murley scores and the AUC values of the pressure-displacement of the affected-side deltoid significantly increased ( P<0.05). The postoperative AUC values of the pressure-displacement of the affected side deltoid muscle were significantly lower than those of the contralateral side ( P<0.05). There was no significant correlation between the changes in deltoid muscle pressure-displacement AUC and age, gender, disease duration, or BMI ( P>0.05), while a significant positive correlation was observed between the AUC and the size of the rotator cuff tear ( r=0.369, P=0.006). There was a positive correlation between the change in VAS and the change in deltoid muscle pressure-displacement AUC ( t=0.424, P<0.001), and the unary linear regression equation was: change in VAS=4.314+0.837×(change in deltoid muscle pressure-displacement AUC). There was a positive correlation between the changes in Constant-Murley shoulder score and the changes in deltoid AUC ( t=5.336, P<0.001), and the unary linear regression equation was: change in Constant-Murley score=24.838+6.097×(change in deltoid muscle pressure-displacement AUC). Conclusions:ARCR can reduce the affected side deltoid muscle tone, relieve shoulder pain and improve shoulder joint function. The changes in deltoid muscle tone are positively correlated with the degree of shoulder pain and function.

Objective To explore the biological functions and mechanisms of PIWI-interacting RNA(piRNA)in cervical cancer(CC).Methods RT-qPCR was used to detect the expression level of piRNA-2732 in CC tissue,CaSki cells and End1/E6E7 cells.EpiQuik m6A RNA methylation quantification kit was used to detect the methylation level of m6A RNA in CaSki cells.The expression levels of methyltransferases(METTL3,METTL14 and WTAP)and demethylases(FTO,ALKBH5)mRNA in CaSki cells were detected by RT-qPCR.After culturing CaSki cells to logarithmic growth stage,they were divided into six groups:piRNA-2732 mimic negative control group(mi-NC group),piRNA-2732 mimic group(mi-2732 group),piRNA-2732 inhibitor negative control group(in-NC group),piRNA-2732 inhibitor group(in-2732 group),piRNA-2732 mimic+METTL3 knockdown control group(mi-2732+si-NC group),and piRNA-2732 mimic+METTL3 knockdown group(mi-2732+si-METTL3 group).The viability of CaSki cells was detected by CCK8 assay.Colony formation assay was used to detect the proliferation ability of CaSki cells.Transwell experiment was used to detect the migration and invasion ability of CaSki cells.RT-qPCR and Western blot were used to detect the expression of methyltransferase like protein 3(METTL3).Transfected METTL3 wild-type(METTL3-WT)and METTL3 mutant(METTL3-MUT)in the mi-NC group,mi-2732 group,in-NC group,and in-2732 group respectively,and detected the effect of piRNA-2732 on METTL3 through dual luciferase reporter gene assay.Results Compared with the adjacent tissues,the expression of piRNA-2732(3.84±1.08 vs 1.32±0.53)was significantly higher in the cancer tissues of CC patients,and the difference was statistically significant(t=5.115,P<0.001).Compared with end1/E6E7 cells,the expression of piRNA-2732(1.00±0.13 vs 1.67±0.16)in CaSki cells was significantly higher,and the difference was statistically significant(t=5.632,P<0.01).Compared with mi-NC group,mi-2732 group promoted the viability,proliferation,migration and invasion of CaSki cells,and the differences were statistically significant(t=4.410～11.040,all P<0.01).Compared with mi-NC group,mi-2732 group increased m6A RNA methylation level and METTL3 mRNA and protein,the differences were statistically significant(t=6.176,9.211,12.550,all P<0.05).The results of dual luciferase reporter gene testing showed that compared with the mi-NC+METTL3-WT group,the relative luciferase activity of mi-2732+METTL3-WT group was significantly increased(t=11.850).Compared with mi-2732+METTL3-WT group,the relative luciferase activity of mi-2732+METTL3-MUT group was significantly lower(t=12.740),and the difference was statistically significant(all P<0.000 1).Compared with in NC+METTL3-WT group,the relative luciferase activity of in-2732+METTL3-WT group was significantly lower(t=7.828),compared with in-2732+METTL3-WT group,the relative luciferase activity of CaSki cells in in-2732+METTL3-MUT group was significantly increased(t=8.146),and the difference was statistically significant(all P<0.001).Compared with mi-2732+si-NC group,the expression level of m6A in mi-2732+si-METTL3 group was significantly lower,and the difference was statistically significant(t=7.630,P<0.01).Compared with mi-2732+si-NC group,the proliferation ability,colony number,cell migration and invasion ability of CaSki cells in mi-2732+si-METTL3 group were significantly decreased,and the differences were statistically significant(t=3.695～4.891,all P<0.001).Conclusion piRNA-2732 is overexpressed in CC tissues and cells,and piRNA-2732 promotes tumor development in CC through METTL3 mediated m6A methylation.

Objective:To explore whether changes in the deltoid muscle tone of the affected side before and after arthroscopic rotator cuff repair (ARCR) are correlated with shoulder joint pain and shoulder joint function.Methods:A total of 55 patients (22 males and 33 females, aged 60.44±9.54 years with a range of 41 to 78 years, BMI 25.86±4.25 kg/m 2 with a range of 18.82 to 42.32 kg/m 2, symptom duration 5.94±7.27 months with a range of 0.3 to 36.0 months) diagnosed with rotator cuff tear and underwent ARCR at the First Affiliated Hospital of Tsinghua University between December 2019 and June 2023 were included. All the patients suffered from unilateral symptom (23 left side and 32 right side), the size of rotator cuff tear was 16.47±5.98 mm (range, 6.92-36.38 mm). Following the operation, postoperative rehabilitation exercises were arranged with a minimum follow-up duration of six months. The muscle tone of the deltoid was measured bilaterally using a muscle tone tester before the operation and at 1, 3, 6 months after operation. The area under curve (AUC) of the pressure-displacement curve was used to reflect the muscle tone, with larger AUC values represent less muscle tone. Visual analogue scale (VAS) and Constant-Murley score were evaluated and recorded at the same time points. Results:At 1, 3, and 6 months postoperatively, the VAS scores were all significantly reduced compared to preoperative values, with the Constant-Murley scores and the AUC values of the pressure-displacement of the affected-side deltoid significantly increased ( P<0.05). The postoperative AUC values of the pressure-displacement of the affected side deltoid muscle were significantly lower than those of the contralateral side ( P<0.05). There was no significant correlation between the changes in deltoid muscle pressure-displacement AUC and age, gender, disease duration, or BMI ( P>0.05), while a significant positive correlation was observed between the AUC and the size of the rotator cuff tear ( r=0.369, P=0.006). There was a positive correlation between the change in VAS and the change in deltoid muscle pressure-displacement AUC ( t=0.424, P<0.001), and the unary linear regression equation was: change in VAS=4.314+0.837×(change in deltoid muscle pressure-displacement AUC). There was a positive correlation between the changes in Constant-Murley shoulder score and the changes in deltoid AUC ( t=5.336, P<0.001), and the unary linear regression equation was: change in Constant-Murley score=24.838+6.097×(change in deltoid muscle pressure-displacement AUC). Conclusions:ARCR can reduce the affected side deltoid muscle tone, relieve shoulder pain and improve shoulder joint function. The changes in deltoid muscle tone are positively correlated with the degree of shoulder pain and function.

BACKGROUND:Small diameter artificial vessels are urgently needed to treat coronary artery and peripheral artery diseases in clinical practice.At present,vascular tissue engineering has become the main method for preparing small diameter artificial vessels.Selecting suitable biomaterials and cell sources is the key factor for successful construction of small diameter tissue engineered vessels. OBJECTIVE:To observe the effect of four kinds of electrospinning membrane materials on proliferation,adhesion and differentiation of bone marrow mesenchymal stem cells into vascular smooth muscle cells. METHODS:Bone marrow mesenchymal stem cells were isolated and extracted from SD rats.The bone marrow mesenchymal stem cells were inoculated separately on polycaprolactone(PCL),polycaprolactone-hyaluronic acid(PCL-HA),polycaprolactone-silk-filament proteins(PCL-SF),and polycaprolactone-gelatin(PCL-GEL)electrospinning membrane materials.After 1,3,and 7 days of culture,the cell arrangement on the material was observed under scanning electron microscope.The proliferation and adhesion of the material were observed by phalloidin staining.The mRNA expressions of CD90,Meflin,and transforming growth factor β were detected by qRT-PCR.After 7 days of induced differentiation into vascular smooth muscle cells,the mRNA expression ofɑ-smooth muscle actin on the material was detected by qRT-PCR. RESULTS AND CONCLUSION:(1)Bone marrow mesenchymal stem cells were arranged along the fibers of the four kinds of electrospinning membranes under scanning electron microscopy.(2)Phalloidin staining showed the regular distribution of bone marrow mesenchymal stem cells on the four kinds of electrospinning membranes and parallel distribution along the fiber direction.Moreover,PCL-HA,PCL-SF,and PCL-GEL electrospinning membranes were more conducive to the proliferation and adhesion of bone marrow mesenchymal stem cells than PCL electrospinning membranes.Compared with PCL-HA and PCL-GEL electrospinning membranes,PCL-SF electrospinning membranes were more conducive to the proliferation and adhesion of bone marrow mesenchymal stem cells.(3)qRT-PCR showed that the four kinds of electrospun membrane materials could maintain the mRNA expression of CD90 and Meflin in bone marrow mesenchymal stem cells,but there was no significant difference between groups(P>0.05).The mRNA expression of transforming growth factor β in PCL-HA,PCL-SF,and PCL-GEL groups was higher than that in PCL group on days 1 and 7(P<0.05),and the mRNA expression of transforming growth factor β in PCL-SF group was higher than that in the other three groups on days 3 and 7(P<0.05).The mRNA expression of transforming growth factor β in PCL-HA group was higher than that in PCL-GEL group on day 7(P<0.05).(4)qRT-PCR showed that the mRNA expression of ɑ-smooth muscle actin in PCL-SF group was higher than that in the other three groups(P<0.05),and that in PCL-HA group was higher than that in PCL group(P<0.05).(5)The results indicate that compared with PCL,PCL-HA and PCL-GEL electrospinning membranes,PCL-SF electrospinning membranes combined with bone marrow mesenchymal stem cells are more suitable for the preparation of small diameter tissue engineered vessels.