Objective:To explore the efficacy of non-invasive prenatal testing (NIPT) of fetal free DNA in maternal peripheral blood in fetuses with increased nuchal translucency (NT).Methods:A retrospective analysis was conducted on 1 184 singleton pregnant women that underwent chromosomal microarray analysis (CMA) at Nanjing Drum Tower Hospital, Nanjing University Medical School from June 2014 to December 2022 due to fetal increased NT (≥3.0 mm). These subjects were categorized based on whether the increased NT was accompanied by other high-risk factors into isolated increased NT without advanced maternal age (further subdivided into 3.0 mm≤NT<3.5 mm, 3.5 mm≤NT<4.0 mm, and NT≥4.0 mm subgroups), isolated increased NT with advanced maternal age, increased NT with nasal bone abnormalities, increased NT with other soft markers, and increased NT with structural abnormalities groups. Assuming the sensitivity and specificity of NIPT and expanded NIPT at this center were both 100%, genomic abnormalities outside the detection range of NIPT or expanded NIPT were termed as residual risk of NIPT or expanded NIPT. Chi-square test and Bonferroni correction were used to compare the residual risks of NIPT and expanded NIPT among the three subgroups of isolated increased NT without advanced maternal age group. Results:(1) In the group of isolated increased NT without advanced maternal age: For the 3.0 mm≤NT<3.5 mm subgroup (329 cases), 19 abnormalities were detected by CMA [12 cases of chromosome aneuploidy, seven cases of pathogenic copy number variation (pCNV)], with residual risks of NIPT and expanded NIPT both at 2.1% (7/329). For the 3.5 mm≤NT<4.0 mm subgroup (173 cases), 29 abnormalities were detected by CMA (17 cases of chromosome aneuploidy, nine cases of pCNV, three cases of chromosome unbalanced translocation), with residual risks of NIPT at 8.1% (14/173) and expanded NIPT at 7.5% (13/173). For the NT≥4.0 mm subgroup (270 cases), CMA detected abnormalities in 70 cases (50 cases of chromosome aneuploidy, 16 cases of pCNV, three cases of unbalanced translocations, and one case of sex chromosome abnormality combined with pCNV). The residual risk of NIPT was 12.2% (33/270), and the residual risk of expanded NIPT was 7.0% (19/270). The residual risks of NIPT and expanded NIPT in the 3.0 mm≤NT<3.5 mm subgroup were lower than those in the 3.5 mm≤NT<4.0 mm and NT≥4.0 mm subgroups (Bonferroni correction, all P<0.017). (2) In the group of 92 cases with isolated increased NT and advanced maternal age, CMA detected abnormalities in 36 cases (29 cases of chromosome aneuploidy, five cases of pCNV, one case of trisomy 21 combined with sex chromosome abnormality, and one case of trisomy 18 combined with sex chromosome abnormality). The residual risk of NIPT was 7.6% (7/92), and that of expanded NIPT was 5.4% (5/92). (3) In the group of 49 cases with increased NT combined with nasal bone abnormalities, CMA detected abnormalities in 24 cases (23 cases of chromosome aneuploidy and one case of pCNV). The residual risks of NIPT and expanded NIPT were both 2.0% (1/49). (4) In the group of 26 cases with increased NT combined with other soft markers, CMA detected abnormalities in nine cases (six cases of chromosome aneuploidy, one case of pCNV, and two cases of chromosome unbalanced translocations). The residual risks of NIPT and expanded NIPT were both 11.5% (3/26). (5) In the group of 245 cases with increased NT combined with structural abnormalities, CMA detected abnormalities in 121 cases (107 cases of chromosome aneuploidy, seven cases of pCNV, four cases of chromosome unbalanced translocations, one case of trisomy 21 combined with trisomy 20, and two cases of trisomy 18 combined with sex chromosome abnormalities). The residual risk of NIPT was 16.7% (41/245), and that of expanded NIPT was 4.1% (10/245). Conclusions:For isolated NT≥3.5 mm or NT≥3.0 mm combined with other high-risk factors, chorionic villus sampling in early pregnancy can be recommended, advancing the timing of prenatal diagnosis from the second trimester to the first trimester. For fetuses with isolated 3.0 mm≤NT<3.5 mm, the 2.1% residual risk of chromosomal abnormalities should be fully informed during counseling, even if the risk of NIPT is low.

OBJECTIVE To explore the mechanism of improvement effect of Chanbao zhichuang suppository(CBZCS)on hemorrhoids in rats through network pharmacology and metabolomics.METHODS A hemorrhoid model was established by subcutaneous injection of rhododendron oil to induce anal swelling.SD rats were divided into blank group(NC group,0.32 g/kg vaseline),model group(Model group,0.32 g/kg vaseline),CBZCS low-,medium-,and high-dose groups(CBZCS-L,CBZCS-M,CBZCS-H groups,with dosages of 0.16,0.32,and 0.64 g/kg respectively),and Mayinglong musk hemorrhoids suppository group(Positive group,0.32 g/kg),with 9 rats in each group.Anal administration was performed at 6,12,24,48,and 72 hours after modeling.After the last administration,the pathological changes of the anal tissues in rats were observed,and the serum levels of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in rats were detected.Differential metabolite analysis and enrichment analysis were conducted by metabolomics methods,and the target proteins of CBZCS in treating hemorrhoids were obtained by network pharmacology.The core metabolic pathways were screened by interaction and enrichment analysis of differential metabolites and proteins,and the core proteins were experimentally verified.RESULTS Compared with the NC group,the anal tissues of the Model group showed obvious lesions,and the levels of IL-6 and TNF-α in the serum were significantly increased(P＜0.05);compared with the Model group,the pathological damage of the anal tissues in the treatment groups was alleviated to varying degrees,and serum levels of IL-6 in CBZCS-H group,CBZCS-M group,and Positive group as well as serum levels of TNF-α in CBZCS-H group were significantly reduced(P＜0.05).The metabolomics results showed that 34 differential metabolites were screened from the anal tissues of rats,and 22 of them showed a return after CBZCS administration.The differential metabolites mainly enriched in arachidonic acid metabolism,histidine metabolism,and glycerophospholipid metabolism.Through the network pharmacology,138 intersection genes of CBZCS against hemorrhoids were determined.The analysis results showed that differential metabolites and target proteins were mainly enriched in the arachidonic acid metabolism pathway,and the regulation of this pathway might be related to cyclooxygenase-2(COX-2),Myc proto-oncogene protein(c-MYC),cytochrome P450 1B1(CYP1B1),interleukin-1β(IL-1β),and IL-6 protein expression.The experimental verification results showed that the expression levels of key proteins(COX-2,c-MYC,CYP1B1,IL-6,IL-1β)in the anal tissues of the Model group were significantly higher than those in the NC group(P＜0.05),and the levels of the above proteins in the anal tissues of CBZCS-H group and Positive group were significantly lower than those in the Model group(P＜0.05).CONCLUSIONS The mechanism of CBZCS in treating hemorrhoids may be to inhibit the expression of COX-2,c-MYC and CYP1B1 proteins,thereby inhibiting arachidonic acid metabolism and reducing the release of inflammatory factors IL-6 and IL-1β.

Objective:To construct a preoperative prehabilitation program for prostate cancer based on evidence-based and Delphi method, so as to provide theoretical basis for medical staff to carry out preoperative prehabilitation research for prostate cancer patients.Methods:From June 2023 to March 2024, the first draft of the prehabilitation plan for prostate cancer before surgery was formed through evidence summary, semi-structured interviews and expert meetings. The experts in related fields were selected for two rounds of Delphi expert consultation. The items were revised according to the expert consultation opinions to establish the final plan.Results:Totally 16 experts were included, aged (43.19 ± 7.57) years. Five were males and 11 were females. The response rates of the two rounds of expert consultation were both 16/16, and the authority coefficients of the two rounds expert consultation were both 0.85. The Kendall coordination coefficients of the importance and feasibility of the items in the second round of consultation were 0.213 and 0.224, both P<0.05. In the second round of consultation, the value of importance assignment of items at all levels was 4.19-4.94 points, and the full score rate was 43.75%-93.75%. The final scheme included 5 first-level items, 13 second-level items, and 34 third-level items. Conclusions:The preoperative prehabilitation program for prostate cancer is scientific, importanceand applicable, which provides a theoretical basis for clinical preoperative prehabilitation for prostate cancer patients.

Background::B-cell maturation antigen (BCMA)-directed chimeric antigen receptor T (CAR-T) therapy yield remarkable responses in patients with relapsed/refractory multiple myeloma (R/RMM). Circulating tumor DNA (ctDNA) reportedly exhibits distinct advantages in addressing the challenges posed by tumor heterogeneity in the distribution and genetic variations in R/RMM.Methods::Herein, the ctDNA of 108 peripheral blood plasma samples from patients with R/RMM at the First Affiliated Hospital, School of Medicine, Zhejiang University was thoroughly investigated before administration of anti-BCMA CAR-T therapy to establish its predictive potential. Flow cytometry is used primarily to detect subgroups of T cells or CAR-T cells.Results::In this study, several tumor and T cell effector-mediated factors were considered to be related to treatment failure by an integrat analysis, including higher percentages of multiple myeloma (MM) cells in the bone marrow ( P = 0.0125), lower percentages of CAR-T cells in the peripheral blood at peak ( P = 0.0375), and higher percentages of CD8 + T cells ( P = 0.0340). Furthermore, there is a substantial correlation between high ctDNA level (>143 ng/mL) and shorter progression-free survival (PFS) ( P = 0.007). Multivariate Cox regression analysis showed that high levels of ctDNA (>143 ng/mL), MM-driven high-risk mutations (including IGLL5 [ P = 0.004], IRF4 [ P = 0.024], and CREBBP [ P = 0.041]), number of multisite mutations, and resistance-related mutation ( ERBB4, P = 0.040) were independent risk factors for PFS. Conclusion::Finally, a ctDNA-based risk model was built based on the above independent risk factors, which serves as an adjunct non-invasive measure of substantial tumor burden and a prognostic genetic feature that can assist in predicting the response to anti-BCMA CAR-T therapy.

Purpose: Erectile dysfunction (ED) is a common male sexual dysfunction. Gut microbiota plays an important role in various diseases. To investigate the effects and mechanisms of intestinal flora dysregulation induced by high-fat diet (HFD) on erectile function. Materials and Methods: Male Sprague–Dawley rats aged 8 weeks were randomly divided into the normal diet (ND) and HFD groups. After 24 weeks, a measurement of erectile function was performed. We performed 16S rRNA sequencing of stool samples. Then, we established fecal microbiota transplantation (FMT) rat models by transplanting fecal microbiota from rats of ND group and HFD group to two new groups of rats respectively. After 24 weeks, erectile function of the rats was evaluated and 16S rRNA sequencing was performed, and serum samples were collected for the untargeted metabolomics detection. Results: The erectile function of rats and the species diversity of intestinal microbiota in the HFD group was significantly lower, and the characteristics of the intestinal microbiota community structure were also significantly different between the two groups. The erectile function of rats in the HFD-FMT group was significantly lower than that of rats in the ND-FMT group. The characteristics of the intestinal microbiota community structure were significantly different. In the HFD-FMT group, 27 metabolites were significantly different and they were mainly involved in the several inflammation-related pathways. Conclusions Intestinal microbiota disorders induced by HFD can damage the intestinal barrier of rats, change the serum metabolic profile, induce low-grade inflammation and apoptosis in the corpus cavernosum of the penis, and lead to ED.

Purpose: Erectile dysfunction (ED) is a common male sexual dysfunction. Gut microbiota plays an important role in various diseases. To investigate the effects and mechanisms of intestinal flora dysregulation induced by high-fat diet (HFD) on erectile function. Materials and Methods: Male Sprague–Dawley rats aged 8 weeks were randomly divided into the normal diet (ND) and HFD groups. After 24 weeks, a measurement of erectile function was performed. We performed 16S rRNA sequencing of stool samples. Then, we established fecal microbiota transplantation (FMT) rat models by transplanting fecal microbiota from rats of ND group and HFD group to two new groups of rats respectively. After 24 weeks, erectile function of the rats was evaluated and 16S rRNA sequencing was performed, and serum samples were collected for the untargeted metabolomics detection. Results: The erectile function of rats and the species diversity of intestinal microbiota in the HFD group was significantly lower, and the characteristics of the intestinal microbiota community structure were also significantly different between the two groups. The erectile function of rats in the HFD-FMT group was significantly lower than that of rats in the ND-FMT group. The characteristics of the intestinal microbiota community structure were significantly different. In the HFD-FMT group, 27 metabolites were significantly different and they were mainly involved in the several inflammation-related pathways. Conclusions Intestinal microbiota disorders induced by HFD can damage the intestinal barrier of rats, change the serum metabolic profile, induce low-grade inflammation and apoptosis in the corpus cavernosum of the penis, and lead to ED.

Objective To investigate the protective mechanism of ergothioneine against oxidative stress and inflammatory damage in vascular dementia(VaD)neurons through activation of the nuclear factor erythroid 2-related factor 2/heme oxygenase-1(Nrf2/HO-1)signaling pathway.Methods A total of 15 VaD patients(VaD group)admitted in our hospital from December 2023 to April 2024 were prospectively recruited,and another 15 healthy individuals taking physical examination during same period were subjected and served as health group.Then,plasma ergothioneine levels in the two groups were quantified by liquid chromatography-tandem mass spectrometry(LC-MS/MS).In in vitro experiments,hippocampal neurons were isolated from neonatal rats,and then divided into control,oxygen-glucose deprivation model,ergothioneine,and ergothioneine+ML385 groups.The expression levels of key molecules of Nrf2/HO-1 signaling pathway,oxidative stress markers,inflammatory cytokines,and apoptosis-related proteins were detected with RT-qPCR and Western blotting.Results The VaD patients exhibited significantly reduced plasma ergothioneine level(694.71±106.87 vs 1268.84±152.87)and down-regulated mRNA levels of Nrf2(0.49±0.14 vs 1.00±0.17),HO-1(0.35±0.23 vs 1.00±0.14)and NQO1(0.39±0.16 vs 1.00±0.12)than the health group(P＜0.05,P＜0.01).Compared with the control group,the model group showed significant increases in inflammatory factor levels.Ergothioneine treatment resulted in significant decreases in malondialdehyde and reactive oxygen species contents,and neuronal apoptosis rate.The ergothioneine+ML385 group showed a significant increase in hippocampal neuronal apoptotic rate,when compared with the ergothionein group.Conclusion Ergothioneine protects VaD neurons against oxidative stress and inflammatory damage by activating the Nrf2/HO-1 signaling pathway.

Objective To explore the application effect of carbon dioxide ablative fractional la-ser(CO2-AFL)in photodynamic therapy(PDT)for condylomata acuminata(CA).Methods A to-tal of 80 patients with CA from January 2022 to December 2023 were randomly divided into two groups,with 40 cases in each group.Control group was treated with PDT mediated by 5-aminolevulin-ic acid(5-ALA)as a photosensitizer(5-ALA-PDT),while study group was treated with CO2-AFL combined with 5-ALA-PDT.Both groups were treated for 4 weeks.Clinical efficacy,recurrence,and adverse reactions were compared between the two groups;additionally,serum inflammatory factors,immune function related indicators,local human papillomavirus(HPV)load,and the Dermatology Life Quality Index(DLQI)scores before treatment and 4 weeks after treatment were compared be-tween the two groups.Results The total effective rate was 97.50％ in the study group,which was significantly higher than the 80.00％ in the control group(P＜0.05).Four weeks after treatment,the levels of interleukin-2(IL-2)and tumor necrosis factor-α(TNF-α)decreased significantly in both groups when compared with before treatment,while the levels of interferon-γ(IFN-γ),interleukin-4(IL-4),interleukin-10(IL-10),and the ratio of T helper 1 to T helper 2 cells(Th1/Th2)increased significantly when compared with before treatment(P＜0.05);moreover,the levels of IL-2 and TNF-α in the study group were significantly lower than those in the control group,while the levels of IFN-γ,IL-4,IL-10,and Th1/Th2 in the study group were significantly higher than thosein the control group(P＜0.05).Four weeks after treatment,CD8+level significantly decreased in both groups when compared with before treatment,while CD3+,CD4+,and CD4+/CD8+levels increased sig-nificantly when compared with before treatment(P＜0.05);additionally,the levels of CD3+,CD4+and CD4+/CD8+in the study group were significantly higher than those in the control group,while CD8+level was significantly lower in the study group(P＜0.05).Four weeks after treat-ment,the local HPV load and DLQI score significantly decreased in both groups when compared with those before treatment,and these indexes in the study group were significantly lower than those in the control group(P＜0.05).The incidence of adverse reactions was 10.00％ in the study group,which showed no significant difference when compared to 5.00％ in the control group(P＞0.05).The recurrence rate within 6 months was 5.00％ in the study group,which was significantly lower than the 22.50％ in the control group(P＜0.05).Conclusion Application of CO2-AFL in 5-ALA-PDT for CA is effective,which helps to eliminate latent HPV viruses,improve serum levels of inflammatory factors,promote the recovery of immune function,and improve the quality of life of CA patients,with a low recurrence rate.

The quality of traditional Chinese medicine (TCM) prescriptions (TCMPs) is critical to clinical efficacy; however, evaluating their consistency and identifying sources of variability remain challenging. This study proposes an integrated strategy to assess the quality of 100 widely sold TCMPs. A "one-for-all" chromatographic method was employed to analyze 645 sample batches. This large-scale data collection enabled statistical evaluations, such as hierarchical cluster analysis (HCA) and similarity heatmap, to identify quality inconsistencies. The introduction of a TCM-specific mass spectrometry (MS) database allowed for rapid, automated annotation of chemicals across 100 prescriptions and facilitated the tracing of raw material sources. Results indicate that 19% of prescriptions exhibited chemical inconsistencies, which are associated with high market value, low pricing, and substantial price disparities. The MS database allowed rapid annotation of 761 and 673 compounds in positive and negative modes, respectively, in 100 TCMPs, with 73 prescriptions reported for the first time. The tracing efforts succeeded in identifying >40% of the raw material sources for 51 prescriptions. P93 (Yinianjin (YNJ)) is a case in which the chromatographic profiles from three manufacturers displayed inconsistencies. Analysis using the database traced divergent peaks to Rhei Radix et R hizoma (RRER). Verification with self-prepared samples confirmed that manufacturers utilized three distinct botanical sources. This integrated strategy provides a scalable framework for quality control in TCMPs.

Objective To investigate the distribution and carriage of Bartonella in rodents and plateau pikas (Ochotona curzoniae) in Xining City, Hai Prefecture, and Menyuan County in Qinghai Province, providing basic data for the prevention and control of Bartonella infections and scientific evidence for the control natural zoonotic diseases in the local area. Methods In 2023, rodents were captured using rodent traps across Xining City, Hai Prefecture, and Menyuan County in Qinghai Province. Bartonella DNA was extracted and amplified from collected liver, spleen, and kidney tissue samples of plateau pikas. Statistical analysis was performed using SPSS 28.0 software, with the chi-square test and Fisher's exact probability method employed to analyze Bartonella positivity rates across different rodent species and ecological environments. Sequence analysis was also conducted on the Bartonella-positive strains from Menyuan County. Results A total of 489 rodents and plateau pikas were captured across the three regions: 206 in Xining City, 200 in Haixi Prefecture, and 83 in Menyuan County. The Bartonella positivity rates were 40.29% (83/206), 2.00% (4/200), and 12.05% (10/83) respectively. A statistically significant difference in Bartonella positivity was observed between Menyuan County and Haixi Prefecture (χ2=10.55, P<0.01). In Menyuan County, 74 Mus musculus linnaeus were captured, 9 Ochotona were captured, 10 Mus musculus linnaeus were Bartonella positive, Ochotona was not detected, the difference was statistically significant by Fisher's exact probability test (P<0.05). There were 14 and 69 rodents captured on grassland and stone land, respectively. 10 rodents captured on stone land were positive for Bartonella, but no rodents were detected on grassland, and the difference was statistically significant by Fisher's exact probability test (P<0.05). Genomic sequence analysis showed that the 10 Bartonella strains from Menyuan County clustered into three genotypes:Bartonella sp.AR 15-3, Bartonella grahamii as4aup, and Bartonella grahamii subsp. shimonis.Conclusion Natural Bartonella infections exist in rodents and plateau pikas across Xining City, Haixi Prefecture, and Menyuan County in Qinghai Province. Furthermore, gene types related to human pathogenicity pose a risk of human infection. Therefore, the monitoring of rodent-borne pathogens in these areas should be strengthened.