Objective To evaluate the accuracy of using HIrisPlex-S system for eye color,hair color,and skin color prediction in forensic DNA phenotyping(FDP).Methods A total of 49 unrelated individuals were enrolled.First,based on the collected facial photographs and questionnaire responses,the actual phenotypic traits,including eye color,hair color,and skin color,of the participants were manually assessed and categorized.Subsequently,peripheral blood samples were collected from the participants,and DNA was extracted.Genotyping of the HIrisPlex-S system was performed using the SNaPshot method combined with capillary electrophoresis.Single nucleotide polymorphism(SNP)sites were analyzed using the online HIrisPlex-S prediction platform(https://HIrisPlex.erasmusmc.nl/)to predict the aforementioned phenotypic traits.Two different methods were used to interpret the prediction results.In Method 1,the phenotypic trait with the highest probability value were directly selected as the predicted phenotype.In Method 2,the prediction guidelines proposed by Manfred Kayser's team was applied.The sensitivity and specificity of the predictions were calculated by comparing the prediction results with the manually interpreted actual phenotypic data,and the differences between the two interpretation methods in these performance metrics were compared.Additionally,two real case samples were included for phenotypic inference analysis to validate the system's effectiveness in practical applications.Results Among the participants,approximately 55%had"brown"eyes,while"intermediate"eye color was less common;approximately 45%had"black"hair;and approximately 39%had"pale"skin.After optimizing the concentration of single base extension(SBE)primers,the peak balance of most SNP loci in the HIrisPlex-S system improved,though inter-laboratory reproducibility remained suboptimal.In eye color prediction,the sensitivity of Method 1(0.875 0,0.000 0,and 1.000 0)was slightly higher than that of Method 2(0.812 5,0.000 0,and 0.925 9),while the specificity of Method 1(0.848 5,1.000 0,and 0.818 2)was slightly lower than that of Method 2(0.909 1,1.000 0,and 0.909 1).For hair color prediction,the two methods showed identical sensitivity(0.772 7,0.615 4,and 0.857 1),but Method 2 showed slightly better specificity(1.000 0,0.944 4,and 0.714 2)than Method 1(1.000 0,0.916 7,and 0.742 9).In skin color prediction,both the sensitivity(0.000 0,0.789 5,1.000 0,0.333 3,and 0.875 0)and specificity(1.000 0,0.966 7,0.833 3,0.976 7,and 0.878 0)of Method 2 were slightly higher than those of Method 1(sensitivity:0.000 0,0.578 9,0.923 1,0.166 7,and 0.750 0;specificity:1.000 0,0.966 7,0.694 4,0.953 5,and 0.878 0).However,overall,there was no statistically significant difference in sensitivity or specificity between the two methods(P>0.05).Additionally,the phenotypic predictions for the two real case samples were consistent with genetic ancestry analysis.Conclusion The HIrisPlex-S system can be used to predict the eye and hair colors in population samples,but its accuracy in skin color prediction needs further improvement.

In recent years, the incidence of infertility has been steadily increasing. Obesity, recognized as a chronic inflammatory condition, can not only lead to various chronic diseases such as hypertension and diabetes, but also adversely affects female reproductive health,including rare ovulation, anovulation, menstrual irregularities, and even infertility. The reproductive outcomes of overweight/obese women are not optimistic even among people undergoing assisted reproductive technology (ART).However, the mechanisms of overweight and obesity on ART outcomes remain poorly understood. This review aims to summarize current evidence on the impact of overweight and obesity on ART outcomes in women and explore the potential mechanisms.

In recent years, the incidence of infertility has been steadily increasing. Obesity, recognized as a chronic inflammatory condition, can not only lead to various chronic diseases such as hypertension and diabetes, but also adversely affects female reproductive health,including rare ovulation, anovulation, menstrual irregularities, and even infertility. The reproductive outcomes of overweight/obese women are not optimistic even among people undergoing assisted reproductive technology (ART).However, the mechanisms of overweight and obesity on ART outcomes remain poorly understood. This review aims to summarize current evidence on the impact of overweight and obesity on ART outcomes in women and explore the potential mechanisms.

AIM:To investigate the therapeutic effects and potential mechanism of pachymic acid(PA)on li-popolysaccharide(LPS)-induced acute kidney injury(AKI)in mice.METHODS:(1)Genes related to AKI were screened using the DAVID database.Core genes were identified by intersecting related genes and analyzed using Cyto-scape software.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses were performed through the DAVID database for the cross-targets.Molecular docking and activity assays were conducted on the primary core targets.(2)A total of 100 C57BL/6J mice were randomly divided into five groups:normal control(NC),model(LPS),solvent control(LPS+DMSO),and treatment groups(LPS+PA-10 and LPS+PA-20),with 20 mice in each group.The LPS-AKI model was established by intraperitoneal injection of 18 mg/kg LPS.The treatment groups received 10 mg/kg and 20 mg/kg PA,respectively,and the solvent control group was administered an equivalent dose of DMSO.Mice were euthanized 24 h after injection.Serum was collected for biochemical analysis,and Western blot was used to detect neutro-phil gelatinase-associated lipocalin(NGAL),kidney injury molecule-1(KIM-1),caspase-3,cleaved caspase-3,interleu-kin-1β(IL-1β),and monocyte chemoattractant protein-1(MCP-1)protein expression.RT-qPCR was employed to detect inflammatory factor mRNA levels.Molecular docking was used to simulate the optimal binding site of PA to caspase-3.En-zyme activity assays were performed to assess caspase protein activity,and renal lesions were observed via hematoxylin and eosin(HE)staining.Apoptosis was detected by TUNEL staining.RESULTS:(1)Thirty-one potential targets of PA against AKI were identified through network pharmacology.GO and KEGG enrichment analyses indicated that these tar-gets were primarily involved in immune response,inflammatory processes,apoptosis and survival,angiogenesis and hemo-dynamics,oxidative stress,and endoplasmic reticulum stress.Key targets included CASP3(caspase-3),PTGS2,BCL2,CCL2,and CYP219.(2)PA treatment improved renal function and reduced tubular epithelial injury.It significantly de-creased NGAL,KIM-1,and cleaved caspase-3 protein levels,as well as inflammatory factors TNF-α,IL-1β,and MCP-1 mRNA and protein expression.PA also reduced apoptosis of renal tubular epithelial cells.Enzyme activity assays and mo-lecular docking revealed that PA exerted its anti-apoptotic effect by directly binding to caspase-3,thereby inhibiting its ac-tivation by caspase-8.CONCLUSION:PA demonstrated a therapeutic effect in LPS-AKI,potentially through the inhibi-tion of inflammatory factor synthesis and release,as well as the inhibition of caspase-3 activation by caspase-8,reducing apoptosis in renal tubular epithelial cells.

Objective:To establish a flow cytometric assay for myeloid-derived suppressor cells (MDSC) in human peripheral whole blood and reference intervals for healthy adults in Shanghai region.Method:A whole blood eight-color and twelve-parameter flow cytometric assay was designed, utilizing fluorescently labeled antibodies against CD45, CD3, CD19, CD123, CD56, CD16, HLA-DR, CD33, CD11b, CD14, CD15 and CD20.A total of 246 healthy participants who met the health standards from the health check-ups conducted at the Tongji University Affiliated Shanghai East Hospital between May 8 to December 2, 2024 were enrolled. Peripheral venous whole blood was collected using EDTA-K 2 anticoagulant vacuum tubes for MDSC detection. A single-platform flow cytometry based relative count technique was used to quantify the percentage of each MDSC subpopulation. Kolmogorov Smirnov (K-S) test was used to test the distribution of specimens. Mann-Whitney U test and Kruskal-Wallis (K-W) test were used to evaluate whether reference intervals should be established separately based on gender or age. According to the clinical significance of MDSC, bilateral reference intervals were taken. Non parametric methods were used to take the percentile P2.5 and P97.5 to represent the rank of the lower and upper reference limits, respectively. Results:The results showed that a gating strategy was designed to exclude granulocytes, lymphocyte lineage cells, and natural killer cells. The K-S test results showed that the MDSC in each group of healthy individuals were distributed in a skewed manner. The U test showed significant gender differences ( P0.05) in the distribution of total myeloid-derived suppressor cells (T-MDSC) and polymorphonuclear myeloid-derived suppressor cells (PMN-MDSC). The K-W test showed no significant differences in MDSC among different age groups (21-30 years old, 30-40 years old, 40-50 years old, and 50-60 years old). T-MDSC reference interval is 0.056%-0.485%, PMN-MDSC reference interval is 0.035%-0.406%, Monocytic myeloid-derived suppressor cells (M-MDSC) reference interval is 0.000%-0.221%, early-stage myeloid-derived suppressor cells (E-MDSC) reference interval is 0.004%-0.125%. Reference interval verification was conducted on 20 healthy individuals, with a pass rate of 100%. Conclusion:A whole blood eight-color and twelve-parameter flow cytometric assay was established in this experiment. Based on the flow cytometry single platform method, reference intervals for healthy adults in Shanghai region were established.

AIM:To investigate the therapeutic effects and potential mechanism of pachymic acid(PA)on li-popolysaccharide(LPS)-induced acute kidney injury(AKI)in mice.METHODS:(1)Genes related to AKI were screened using the DAVID database.Core genes were identified by intersecting related genes and analyzed using Cyto-scape software.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analyses were performed through the DAVID database for the cross-targets.Molecular docking and activity assays were conducted on the primary core targets.(2)A total of 100 C57BL/6J mice were randomly divided into five groups:normal control(NC),model(LPS),solvent control(LPS+DMSO),and treatment groups(LPS+PA-10 and LPS+PA-20),with 20 mice in each group.The LPS-AKI model was established by intraperitoneal injection of 18 mg/kg LPS.The treatment groups received 10 mg/kg and 20 mg/kg PA,respectively,and the solvent control group was administered an equivalent dose of DMSO.Mice were euthanized 24 h after injection.Serum was collected for biochemical analysis,and Western blot was used to detect neutro-phil gelatinase-associated lipocalin(NGAL),kidney injury molecule-1(KIM-1),caspase-3,cleaved caspase-3,interleu-kin-1β(IL-1β),and monocyte chemoattractant protein-1(MCP-1)protein expression.RT-qPCR was employed to detect inflammatory factor mRNA levels.Molecular docking was used to simulate the optimal binding site of PA to caspase-3.En-zyme activity assays were performed to assess caspase protein activity,and renal lesions were observed via hematoxylin and eosin(HE)staining.Apoptosis was detected by TUNEL staining.RESULTS:(1)Thirty-one potential targets of PA against AKI were identified through network pharmacology.GO and KEGG enrichment analyses indicated that these tar-gets were primarily involved in immune response,inflammatory processes,apoptosis and survival,angiogenesis and hemo-dynamics,oxidative stress,and endoplasmic reticulum stress.Key targets included CASP3(caspase-3),PTGS2,BCL2,CCL2,and CYP219.(2)PA treatment improved renal function and reduced tubular epithelial injury.It significantly de-creased NGAL,KIM-1,and cleaved caspase-3 protein levels,as well as inflammatory factors TNF-α,IL-1β,and MCP-1 mRNA and protein expression.PA also reduced apoptosis of renal tubular epithelial cells.Enzyme activity assays and mo-lecular docking revealed that PA exerted its anti-apoptotic effect by directly binding to caspase-3,thereby inhibiting its ac-tivation by caspase-8.CONCLUSION:PA demonstrated a therapeutic effect in LPS-AKI,potentially through the inhibi-tion of inflammatory factor synthesis and release,as well as the inhibition of caspase-3 activation by caspase-8,reducing apoptosis in renal tubular epithelial cells.

Objective:To establish a flow cytometric assay for myeloid-derived suppressor cells (MDSC) in human peripheral whole blood and reference intervals for healthy adults in Shanghai region.Method:A whole blood eight-color and twelve-parameter flow cytometric assay was designed, utilizing fluorescently labeled antibodies against CD45, CD3, CD19, CD123, CD56, CD16, HLA-DR, CD33, CD11b, CD14, CD15 and CD20.A total of 246 healthy participants who met the health standards from the health check-ups conducted at the Tongji University Affiliated Shanghai East Hospital between May 8 to December 2, 2024 were enrolled. Peripheral venous whole blood was collected using EDTA-K 2 anticoagulant vacuum tubes for MDSC detection. A single-platform flow cytometry based relative count technique was used to quantify the percentage of each MDSC subpopulation. Kolmogorov Smirnov (K-S) test was used to test the distribution of specimens. Mann-Whitney U test and Kruskal-Wallis (K-W) test were used to evaluate whether reference intervals should be established separately based on gender or age. According to the clinical significance of MDSC, bilateral reference intervals were taken. Non parametric methods were used to take the percentile P2.5 and P97.5 to represent the rank of the lower and upper reference limits, respectively. Results:The results showed that a gating strategy was designed to exclude granulocytes, lymphocyte lineage cells, and natural killer cells. The K-S test results showed that the MDSC in each group of healthy individuals were distributed in a skewed manner. The U test showed significant gender differences ( P0.05) in the distribution of total myeloid-derived suppressor cells (T-MDSC) and polymorphonuclear myeloid-derived suppressor cells (PMN-MDSC). The K-W test showed no significant differences in MDSC among different age groups (21-30 years old, 30-40 years old, 40-50 years old, and 50-60 years old). T-MDSC reference interval is 0.056%-0.485%, PMN-MDSC reference interval is 0.035%-0.406%, Monocytic myeloid-derived suppressor cells (M-MDSC) reference interval is 0.000%-0.221%, early-stage myeloid-derived suppressor cells (E-MDSC) reference interval is 0.004%-0.125%. Reference interval verification was conducted on 20 healthy individuals, with a pass rate of 100%. Conclusion:A whole blood eight-color and twelve-parameter flow cytometric assay was established in this experiment. Based on the flow cytometry single platform method, reference intervals for healthy adults in Shanghai region were established.

Acute high altitude disease (AHAD) is a general term for a series of clinical reactions that occur when the body fails to adapt to the low-pressure hypoxic environment of high altitudes. Mild cases can cause symptoms such as headache, nausea and vomiting, while more severe cases can lead to life-threatening conditions such as pulmonary edema, cerebral edema and other critical conditions that can be fatal. With the increasing demand for high altitudes deployment, understanding the common preventive measures of AHAD can reduce its morbidity or mortality to a certain extent, which is of great benefit to those who reside temporarily at high altitudes. In recent years, as people's health awareness has improved, there has been a growing attention towards non-pharmacological methods of disease prevention. At the same time, non-pharmacological therapy has significant therapeutic effects in preventing and treating high-altitude diseases, which has attracted the attention of researchers in this field. This review summarizes the major non-pharmacological preventive components of modern medicine and outlines the current non-pharmacological approaches to AHAD from the perspective of traditional Chinese medicine, intending to serve clinical purposes and improve the onset and prognosis of AHAD.

Objective The characteristics of supramolecular"imprinting template"of volatile oil of Curcuma kwangsiensis and Curcuma phaeocaulis were analyzed and studied based on the supramolecular"qixi"theory of Chinese materia medica combined with chemometrics.Methods The volatile oil of C.kwangsiensis and C.phaeocaulis were extracted by steam distillation,and the fingerprint and composition information of each batch were obtained by GC-MS.Total statistical moment method was used to compare the imprinting characteristics of the"imprinting template"of C.kwangsiensis and C.phaeocaulis.The core index(CI)of each batch of essential oil of C.kwangsiensis and C.phaeocaulis was calculated,and the topological characteristics of types of"imprinting template"of C.kwangsiensis and C.phaeocaulis were compared by chemometrics.Results There was no significant difference in the extraction rate of volatile oil between C.kwangsiensis and C.phaeocaulis.The average values of total zero order moment(AUCT)were(1.907±0.177)×108,(1.979±0.413)×108 μV·s,respectively,showing that there was no significant difference in the total content of volatile oil between the two groups.The mean values of the total first order moment(MCRTT)were(30.969±0.962)and(33.198±0.409)min.The average value of total second order moment(VCRTT)was(56.176±11.368)and(43.891±4.113)min2,respectively,indicating that there were significant differences in the content ratio and species of volatile oils between the two groups.The similarity of total statistical moments of C.kwangsiensis and C.phaeocaulis was mostly lower than the defined value,indicating that the chemical composition and composition ratio of the volatile oil were different.Principal component analysis and orthogonal partial least squares discriminant analysis could obviously divide C.kwangsiensis and C.phaeocaulis into two categories.Through the analysis of P value and VIP value,the CI values of Xvp 4th order,Xvpc 5th order,Xvpc 6th order,Xvpc 7th order,Xvc 3rd order,Xvpc 4th order were the main difference values of C.kwangsiensis and C.phaeocaulis.Conclusion Through the characterization of"imprinting property"and"topological characteristics"of the supramolecular"imprinting template"and combining with chemometric analysis,it is possible to successfully distinguish C.kwangsiensis and C.phaeocaulis,and find the different CI values between two"imprinting templates".

Purpose: Male overweight and obesity could affect sperm quality and reproductive health. However, the impact of body mass index (BMI) on assisted reproductive technology (ART) outcomes in oligospermia and/or asthenospermia patients is yet lacking. This study aims to assess the impact of paternal BMI on ART and neonatal outcomes among oligozoospermia and/or asthenospermia patients undergoing in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI). Materials and Methods: In this study, 2,075 couples undergoing their first fresh embryo transfer between January 2015 and June 2022 were recruited. Following the World Health Organization’s (WHO’s) categories, couples were stratified into three cohorts based on paternal BMI: normal weight (18.5–24.9 kg/m2), overweight (25.0–29.9 kg/m2), and obese (≥30.0 kg/m2). Modified Poisson regression models were used to assess the associations of paternal BMI with fertilization, in vitro embryonic development, and pregnancy outcomes. Logistic regression models were performed to investigate the associations of paternal BMI with pregnancy loss and neonatal outcomes. Furthermore, stratified analyses were performed based on fertilization methods, male infertility cause, and maternal BMI. Results: Higher paternal BMI is associated with a lower likelihood of achieving normal fertilized (p-trend=0.002), Day 3 transferable (p-trend=0.007), and high-quality embryos (p-trend=0.046) in IVF cycles, rather than in ICSI cycles. Paternal BMI of oligospermia or asthenospermia was negatively correlated with day 3 transferable (p-trend=0.013 and 0.030) and high-quality embryos (p-trend=0.024 and 0.027). Moreover, for neonatal outcomes, paternal BMI was positively associated with macrosomia (p-trend=0.019), large for gestational age (LGA) (p-trend=0.031), and very LGA (p-trend=0.045). Conclusions Our data suggested that higher paternal BMI was associated with fetal overgrowth, reduced fertilization, and embryonic development potential. Among males with oligospermia and/or asthenospermia, the impact of overweight and obesity on the choice of fertilization method and the long-term effects on their offspring need to be further investigated.