BACKGROUND: Temozolomide (TMZ) resistance is a significant challenge in treating glioblastoma (GBM). Collagen remodeling has been shown to be a critical factor for therapy resistance in other cancers. This study aimed to investigate the mechanism of TMZ chemoresistance by GBM cells reprogramming collagens. METHODS: Key extracellular matrix components, including collagens, were examined in paired primary and recurrent GBM samples as well as in TMZ-treated spontaneous and grafted GBM murine models. Human GBM cell lines (U251, TS667) and mouse primary GBM cells were used for in vitro studies. RNA-sequencing analysis, chromatin immunoprecipitation, immunoprecipitation-mass spectrometry, and co-immunoprecipitation assays were conducted to explore the mechanisms involved in collagen accumulation. A series of in vitro and in vivo experiments were designed to assess the role of the collagen regulators prolyl 4-hydroxylase subunit alpha 1 (P4HA1) and yes-associated protein (YAP) in sensitizing GBM cells to TMZ. RESULTS: This study revealed that TMZ exposure significantly elevated collagen type I (COL I) expression in both GBM patients and murine models. Collagen accumulation sustained GBM cell survival under TMZ-induced stress, contributing to enhanced TMZ resistance. Mechanistically, P4HA1 directly binded to and hydroxylated YAP, preventing ubiquitination-mediated YAP degradation. Stabilized YAP robustly drove collagen type I alpha 1 ( COL1A1) transcription, leading to increased collagen deposition. Disruption of the P4HA1-YAP axis effectively reduced COL I deposition, sensitized GBM cells to TMZ, and significantly improved mouse survival. CONCLUSION P4HA1 maintained YAP-mediated COL1A1 transcription, leading to collagen accumulation and promoting chemoresistance in GBM.
Temozolomide ; Humans ; Glioblastoma/drug therapy* ; Animals ; Mice ; Cell Line, Tumor ; Drug Resistance, Neoplasm/genetics* ; YAP-Signaling Proteins ; Hydroxylation ; Dacarbazine/pharmacology* ; Adaptor Proteins, Signal Transducing/metabolism* ; Transcription Factors/metabolism* ; Collagen/biosynthesis* ; Collagen Type I/metabolism* ; Prolyl Hydroxylases/metabolism* ; Antineoplastic Agents, Alkylating/therapeutic use*

Objective:To investigate the effect of the embryo cryopreservation duration on pregnancy and obstetric outcome.Methods:A retrospective cohort study of 2 662 frozen-thawed embyro tranfer (FET) cycles was conducted in the Reproductive Medicine Center, Department of Obstetrics and Gynecology, Tianjin Medical University General Hospital from January 2016 to December 2020. According to embryo cryopreservation duration, the patients were divided into group A (≤1 year, n=2 115), group B (>1 years and ≤3 years, n=319), group C (>3 years and ≤6 years, n=174), and group D (>6 years, n=54). We used the propensity score matching (PSM) to match the baseline data of oocyte retrieval age of the other three groups according to group D at a ratio of 1∶3. Clinical and obstetric outcomes were compared among the four groups. Multiple logistic regression analysis was used to analyze the effect of oocyte retrieval age, embryo transfer age, the duration of embryo cryopreservation, endometrial preparation scheme, endometrial thickness, the number of transferred embryos and the number of high-quality embryos on pregnancy and live birth outcome. Results:1) Before PSM, there were significant differences in the maternal age at oocyte retrieval and embryo transfer and duration of embryo cryopreservation among the four groups(all P<0.001). 2) After PSM, the baseline characteristics of oocyte retrieval age reached a balance among the four groups. There were no statistical differences in the number of embryos transfer, the number of high-quality embryos, the transferred embryo stage, the endometrial regimen among the groups (all P>0.05). The clinical pregnancy rate [37.04% (20/54)] and the live birth rate [33.33% (18/54)] in group D were lower than those in group A [51.57% (82/159), 40.88% (65/159)], group B [50.00% (65/130), 40.77% (53/130)] and group C [49.59% (61/123), 39.02% (48/123)], but the difference was not statistically significant between the four groups ( P=0.310, P=0.781). There were no statistical differences among the four groups in the ratio of male to female newborns, gestational age, birth weight, preterm delivery rate, low birth weight rate, macrosomia rate, birth defects, and premature repture of membranes (all P>0.05). 3) Multiple logistic regression analysis showed that the number of high-quality embryos transferred affected the clinical pregnancy outcome (before PSM, OR=2.614, 95% CI: 2.168-3.151, P<0.001; after PSM, OR=1.984, 95% CI: 1.406-2.800, P<0.001) and live birth (before PSM, OR=2.708, 95% CI: 2.198-3.336, P<0.001; after PSM, OR=2.122, 95% CI: 1.474-3.053, P<0.001). The duration of embryo cryopreservation does not affect the clinical outcome and live birth (all P>0.05). Conclusion:The duration of embryo cryopreservation does not affect the clinical outcome and live birth, but large sample data are still needed to support this conclusion in the future.

Objective:To investigate the effect of the embryo cryopreservation duration on pregnancy and obstetric outcome.Methods:A retrospective cohort study of 2 662 frozen-thawed embyro tranfer (FET) cycles was conducted in the Reproductive Medicine Center, Department of Obstetrics and Gynecology, Tianjin Medical University General Hospital from January 2016 to December 2020. According to embryo cryopreservation duration, the patients were divided into group A (≤1 year, n=2 115), group B (>1 years and ≤3 years, n=319), group C (>3 years and ≤6 years, n=174), and group D (>6 years, n=54). We used the propensity score matching (PSM) to match the baseline data of oocyte retrieval age of the other three groups according to group D at a ratio of 1∶3. Clinical and obstetric outcomes were compared among the four groups. Multiple logistic regression analysis was used to analyze the effect of oocyte retrieval age, embryo transfer age, the duration of embryo cryopreservation, endometrial preparation scheme, endometrial thickness, the number of transferred embryos and the number of high-quality embryos on pregnancy and live birth outcome. Results:1) Before PSM, there were significant differences in the maternal age at oocyte retrieval and embryo transfer and duration of embryo cryopreservation among the four groups(all P<0.001). 2) After PSM, the baseline characteristics of oocyte retrieval age reached a balance among the four groups. There were no statistical differences in the number of embryos transfer, the number of high-quality embryos, the transferred embryo stage, the endometrial regimen among the groups (all P>0.05). The clinical pregnancy rate [37.04% (20/54)] and the live birth rate [33.33% (18/54)] in group D were lower than those in group A [51.57% (82/159), 40.88% (65/159)], group B [50.00% (65/130), 40.77% (53/130)] and group C [49.59% (61/123), 39.02% (48/123)], but the difference was not statistically significant between the four groups ( P=0.310, P=0.781). There were no statistical differences among the four groups in the ratio of male to female newborns, gestational age, birth weight, preterm delivery rate, low birth weight rate, macrosomia rate, birth defects, and premature repture of membranes (all P>0.05). 3) Multiple logistic regression analysis showed that the number of high-quality embryos transferred affected the clinical pregnancy outcome (before PSM, OR=2.614, 95% CI: 2.168-3.151, P<0.001; after PSM, OR=1.984, 95% CI: 1.406-2.800, P<0.001) and live birth (before PSM, OR=2.708, 95% CI: 2.198-3.336, P<0.001; after PSM, OR=2.122, 95% CI: 1.474-3.053, P<0.001). The duration of embryo cryopreservation does not affect the clinical outcome and live birth (all P>0.05). Conclusion:The duration of embryo cryopreservation does not affect the clinical outcome and live birth, but large sample data are still needed to support this conclusion in the future.

Studies have suggested that the nucleus accumbens (NAc) is implicated in the pathophysiology of major depression; however, the regulatory strategy that targets the NAc to achieve an exclusive and outstanding anti-depression benefit has not been elucidated. Here, we identified a specific reduction of cyclic adenosine monophosphate (cAMP) in the subset of dopamine D1 receptor medium spiny neurons (D1-MSNs) in the NAc that promoted stress susceptibility, while the stimulation of cAMP production in NAc D1-MSNs efficiently rescued depression-like behaviors. Ketamine treatment enhanced cAMP both in D1-MSNs and dopamine D2 receptor medium spiny neurons (D2-MSNs) of depressed mice, however, the rapid antidepressant effect of ketamine solely depended on elevating cAMP in NAc D1-MSNs. We discovered that a higher dose of crocin markedly increased cAMP in the NAc and consistently relieved depression 24 h after oral administration, but not a lower dose. The fast onset property of crocin was verified through multicenter studies. Moreover, crocin specifically targeted at D1-MSN cAMP signaling in the NAc to relieve depression and had no effect on D2-MSN. These findings characterize a new strategy to achieve an exclusive and outstanding anti-depression benefit by elevating cAMP in D1-MSNs in the NAc, and provide a potential rapid antidepressant drug candidate, crocin.

Objective:To investigate mRNA and N6-methyladenosine (m6A) changes in retinal pigment epithelium (RPE) cells treated with recombinant human methyl-CpG binding protein 2 (MeCP2) and the mechanisms.Methods:The passaged ARPE-19 cells were divided into normal control and MeCP2 groups after adhesion culture.Cells in the normal control group were continuously cultured in normal culture medium, and the cells in the MeCP2 group were cultured in culture medium containing a final concentration of 20 ng/ml of recombinant human MeCP2 protein for 72 hours.Transcriptomic sequencing (RNA-seq) and methylated RNA immunoprecipitation sequencing (MeRIP-seq) were used to extract and analyze total RNA.Differentially methylated genes (DMGs) and differentially expressed genes (DEGs) were screened using the edgeR software package based on P<0.05.The biological function of differential genes was determined by gene ontology (GO) enrichment analysis, and the pathway enrichment analysis was performed by Kyoto Encyclopedia of Genes and Genomes (KEGG).Intersection of genes between DEGs and DMGs were screened, and real-time fluorescence quantitative PCR was used to determine the mRNA expression levels of differential genes. Results:A total of 100 DEGs and 7 441 DMGs genes were screened.According to enrichment analysis, the DEGs were enriched to extracellular matrix (ECM)-receptor interaction, cell division, phosphatidylinositol 3 kinase/protein kinase B (PI3K/AKT) signaling pathway and so on.The DMGs were associated with microtubule cytoskeleton, angiogenesis, epidermal growth factor receptor (ErbB) signaling pathway, advanced glycation end-products (AGEs) -glycation end-products receptor (RAGE) signaling pathway, mammalian target of rapamycin (mTOR) signaling pathway, Notch signaling pathway and transforming growth factors-β (TGF-β) signaling pathway and so on.There were 24 up-regulated and 76 down-regulated DEGs.Five DMGs had hypermethylation peaks, and 7 439 DMGs had hypomethylation peaks.After annotation of peaks, 7 626 genes in the normal control group and 8 006 genes in the MeCP2 group had m6A methylation, with 7 360 intersecting genes between the two groups.The m6A methylation in the normal control group and MeCP2 group was concentrated in the CDS, intron and 3'-untranslated region (3'UTR) regions of the transcript, with the methylation ratio of 23.62%/22.27%, 48.53%/48.35% and 23.66%/25.28%, respectively.Joint analysis showed that CSPG5 and RBP1 genes related to the epithelial-mesenchymal transition (EMT) had lower amount of mRNA and m6A.Fluorescence quantitative PCR results showed that the relative mRNA expression levels of GSPG5, RBP1 and ZNF484 in MeCP2 group were significantly lower than those in normal control group ( t=7.885, 7.613, 7.345; all at P<0.01). Conclusions:The regulatory mechanism of MeCP2 on EMT in RPE cells is related to m6A methylation modification. CSPG5 and RBP1 genes may be the target genes of m6A methylation and participate in the EMT regulated by MeCP2.

Objective To evaluate the economic value of fluticasone furoate/umeclidinium/vilanterol(FF/UMEC/VI)powder for inhalation in the treatment of symptomatic chronic obstructive pulmonary disease patients with acute exacerbation risk from the perspective of the Chinese health system.Methods Based on subgroup analysis of the China cohort in the IMPACT trial,a four-state lifetime Markov model was established with a 3-month cycle.The model simulation period was 11 years.Clinical efficacy,health benefits,and cost data were obtained through published literature.The health outcomes included quality adjusted life year(QALY).Using 3 times Chinas per capita gross domestic product(GDP)in 2023 as the willingness-to-pay threshold,the cost-utility analysis method was used for analysis the economic viability of FF/UMEC/VI.The scenario analysis,one-way sensitivity analysis and probability sensitivity analysis were used to verify the robustness of the results.Results Compared with fluticasone furoate/vilanterol(FF/VI),FF/UMEC/VI in the treatment of symptomatic chronic obstructive pulmonary disease patients with acute exacerbation risk saved costs 8 118.66 yuan and obtained an additional 0.000 06 QALYs,giving it an economic advantage.Compared with umeclidinium/vilanterol(UMEC/VI),FF/UMEC/VI treatment paid 2 784.41 yuan more and received 0.000 45 QALYs less,making UMEC/VI more cost-utility.The scenario analysis results further confirmed the robustness of the model.The sensitivity analysis results showed that when the drug cost of FF/UMEC/VI per cycle decreases to 637.29 yuan,FF/UMEC/VI had economic benefits under a willingness-to-pay threshold of 3 times China's per capita GDP in 2023.Conclusion For patients with symptomatic chronic obstructive pulmonary disease at risk of acute exacerbation,FF/UMEC/VI is more cost-utility than FF/VI.Compared with UMEC/VI,FF/UMEC/VI has economic viability after price reducing.

Recurrent hydatidiform moles refer to patients with at least two molar pregnancies. Mutations in NLRP7 and KHDC3L genes have been implicated in this disease. A pedigree with a history of recurrent hydatidiform mole who visited Tianjin Medical University General Hospital Reproductive Medicine Center, was selected as the study subject. Clinical data of the family member were collected, peripheral blood samples were taken from each member. Whole exon sequencing was carried out for the proband. Candidate genes were validated by Sanger sequencing of her family members. The whole exome sequencing showed that the proband had a homozygous mutation of c.2282G>A (p.Cys761Tyr) in the NLRP7 gene, the Sanger sequencing results were consistent with the results. Sanger sequencing results verified that the parents and her sisters carried heterozygous mutations. The proband obtained a clinical pregnancy through egg donation and intracytoplasmic sperm injection. Therefore, homozygous mutation of NLRP7 gene c.2282G>A (p.Cys761Tyr) is the genetic cause of recurrent hydatidiform mole. The discovery of this mutation broadens the spectrum of NLRP7 gene pathogenic variants and provide fertility guidance for such patients.

Objective:To explore the parameters of controlled ovarian stimulation (COS) on the occurrence of de novo chromosomal abnormalities in preimplantation genetic testing for structural rearrangements (PGT-SR) cycles. Methods:A retrospective analysis control study was performed on 70 PGT-SR cycles and 39 preimplantation genetic testing for monogenic (PGT-M) embryos in the Department of Obstetrics and Gynecology, Reproductive Medicine Center of Tianjin Medical University General Hospital from January 2023 to August 2024. The correlation between de novo chromosomal abnormalities and ovarian stimulation protocol, duration and total dosage of gonadotropin (Gn) used, estradiol and progesterone levels on human chorionic gonadotropin (hCG) trigger day in COS were analyzed. Results:1) Biopsies were performed on 341 blastocysts in the PGT-SR group and 196 blastocysts in the PGT-M group. There was a significant difference in aneuploid rate of blastocyst between PGT-SR and PGT-M groups [62.1% (205/330) vs. 30.2% (58/192), P<0.001]. There was no significant difference in the incidence of de novo chromosomal abnormalities, between PGT-SR and PGT-M groups ( P>0.05). 2) After adjusting for couples' age and blastocyst grade, there were no significant differences in the incidence of de novo chromosomal abnormalities among different ovarian stimulation protocols, different duration and total dosage of Gn used, different estradiol and progesterone levels on hCG trigger day (all P>0.05). 3) There were no significant differences the incidence of de novo whole, fragment and complex chromosomal abnormalities among different ovarian stimulation protocols, different duration and total dosage of Gn used, different estradiol and progesterone levels on hCG trigger day (all P>0.05). Conclusion:Different ovarian stimulation protocols, duration and total dosage of Gn used, estrogen and progesterone levels on hCG trigger day don't affect the whole de novo chromosomal abnormalities, de novo whole, fragment, complex chromosomal abnormalities in patients with structural chromosomal abnormalities.

A retrospective analysis was conducted on a case of duplicate right kidney and ureter with ectopic opening in the vagina discovered before embryo transfer, and literature review was conducted. The patient planned to undergo a third embryo transfer. Prior to the transfer, laparoscopic bilateral salpingectomy was performed due to "right fallopian tube hydrosalpinx". About a week after the surgery, there was a continuous increase in vaginal discharge with an odor for more than 50 d. Repeated antibiotic treatments in the outpatient department were ineffective. Vaginal ultrasound examination revealed a cyst in the right adnexal area. After multiple gynecological examinations, vaginal ultrasound, urological ultrasound, whole abdominal CT, magnetic resonance urography, etc., the diagnosis was right kidney and ureteral malformation with ectopic opening of the ureter in the vagina. Upon reviewing the preoperative images of the patient undergoing salpingectomy, it was found that the dilated right repeated ureter was misdiagnosed as hydrosalpinx. Women with adnexal cysts accompanied by continuous vaginal discharge should consider the possibility of repeated ureters and ectopic ureteral opening, and should pay attention to distinguishing it from hydrosalpinx.

Objective:To investigate the effect of triglyceride-glucose index (TyG) on the severity of depression in patients with depression.Methods:The clinical data of 653 patients with depression admitted to Hebei General Hospital from January to December 2021 were retrospectively analyzed. The patients were categorized into mild-moderate depression group ( n=324) and severe depression group ( n=329) according to the 17-item Hamilton depression scale (HAMD-17) score. Patients' demographics and data on fasting plasma glucose (FPG), triglyceride (TG), total cholesterol, high-density lipoprotein, and low-density lipoprotein were extracted from the electronic medical records, and TyG was calculated. Statistical analyses were performed by SPSS 25.0 software.The Mann-Whitney U test was used to compare the differences between the two groups. Spearman correlation analysis and binary Logistic regression analysis were used to investigate the relationship between TyG level and depression severity. Results:(1) In the severe depression group, FPG( 956.9 (118.9) mg/L vs 865.0 (1 44.2) mg/L), TG(1 275.8 (1 027.8) mg/L vs 983.5 (766.4) mg/L), total cholesterol (2 007.0 (477.6) mg/L vs 1 730.5 (547.2) mg/L), high-density lipoprotein (514.3 (168.2) mg/L vs 464.0 (138.2) mg/L), low-density lipoprotein (1 264.5 (326.8) mg/L vs 1 088.6 (412.8) mg/L), and TyG (8.72 (0.77) vs 8.37 (0.78)) were higher than those in the mild-moderate depression group, and the differences were statistically significant (all P<0.05).(2) Both TyG level and HAMA score were positively correlated with HAMD-17 score ( r=0.231, 0.586, both P<0.05); (3) The results of binary logistic regression analysis showed that TyG ( B=1.116, OR=3.054, 95% CI=2.058-4.531, P<0.05) was a risk factor for the severity of depression in patients with depression. Conclusion:The higher the TyG, the more severe the severity of depression in patients. Insulin resistance may affect the severity of depression in patients with depression.