Objective To explore the pathogenic roles of miR-21, estrogen (E2), and estrogen receptor (ER) in adenomyosis. Methods We examined the expression levels of miR-21 in specimens of adenomyotic tissue and benign cervical lesions using qRT-PCR. In primary cultures of cells isolated from the adenomyosis lesions, the effect of ICI82780 (an ER inhibitor) on miR-21 expression levels prior to E2 activation or after E2 deprivation were examined with qRT-PCR. We further assessed the effects of a miR-21 mimic or an inhibitor on proliferation, apoptosis, migration and autophagy of the cells. Results The expression level of miR-21 was significantly higher in adenomyosis tissues than in normal myometrium (P<0.05). In the cells isolated from adenomyosis lesions, miR-21 expression level was significantly higher in E2 activation group than in ER inhibition+E2 activation group and the control group (P<0.05);miR-21 expression level was significantly lower in cells in E2 deprivation+ER inhibition group than in E2 deprivation group and the control group (P<0.05). The adenomyosis cells transfected with miR-21 inhibitor showed inhibited proliferation and migration, expansion of mitochondrial endoplasmic reticulum, increased lysosomes, presence of autophagosomes, and increased cell apoptosis, while transfection of the cells with the miR-21 mimic produced the opposite effects. Conclusion MiR-21 plays an important role in promoting proliferation, migration, and anti-apoptosis in adenomyosis cells by altering the cell ultrastructure, which may contribute to early pathogenesis of the disease. In addition to binding with E2, ER can also regulate miR-21 through other pathways to participate in the pathogenesis of adenomyosis, thus having a stronger regulatory effect on miR-21 than E2.

Objective To explore the pathogenic roles of miR-21, estrogen (E2), and estrogen receptor (ER) in adenomyosis. Methods We examined the expression levels of miR-21 in specimens of adenomyotic tissue and benign cervical lesions using qRT-PCR. In primary cultures of cells isolated from the adenomyosis lesions, the effect of ICI82780 (an ER inhibitor) on miR-21 expression levels prior to E2 activation or after E2 deprivation were examined with qRT-PCR. We further assessed the effects of a miR-21 mimic or an inhibitor on proliferation, apoptosis, migration and autophagy of the cells. Results The expression level of miR-21 was significantly higher in adenomyosis tissues than in normal myometrium (P<0.05). In the cells isolated from adenomyosis lesions, miR-21 expression level was significantly higher in E2 activation group than in ER inhibition+E2 activation group and the control group (P<0.05);miR-21 expression level was significantly lower in cells in E2 deprivation+ER inhibition group than in E2 deprivation group and the control group (P<0.05). The adenomyosis cells transfected with miR-21 inhibitor showed inhibited proliferation and migration, expansion of mitochondrial endoplasmic reticulum, increased lysosomes, presence of autophagosomes, and increased cell apoptosis, while transfection of the cells with the miR-21 mimic produced the opposite effects. Conclusion MiR-21 plays an important role in promoting proliferation, migration, and anti-apoptosis in adenomyosis cells by altering the cell ultrastructure, which may contribute to early pathogenesis of the disease. In addition to binding with E2, ER can also regulate miR-21 through other pathways to participate in the pathogenesis of adenomyosis, thus having a stronger regulatory effect on miR-21 than E2.

Objective:To investigate the ultrasonographic features and genetic etiology of complex talipes equinovarus (TE) in fetuses.Methods:This retrospective study enrolled 95 cases of complex TE (TE complicated by other abnormalities) who were diagnosed by prenatal ultrasound in the Third Affiliated Hospital of Zhengzhou University from March 2018 to December 2022. Chromosome karyotype analysis and/or chromosomal microarray analysis (CMA) [or copy number variation-sequencing (CNV-seq)] were performed on all cases for prenatal genetic diagnosis and those with normal results were further tested by whole exome sequencing (WES). Prenatal ultrasonographic and genetic features of complex TE in fetuses were summarized. Complicated abnormalities in the fetuses were classified into nine categories according to the involved system or site and based on each category these subjects were divided into with or without the corresponding complicated abnormalities groups. Besides, these cases were also divided into single-system and multi-system abnormality groups based on the number of involved systems or sites of complicated abnormalities. The detection rates of WES abnormality (pathogenic or likely pathogenic variants) and the overall detection rate of genetic abnormality [karyotype abnormality detected by chromosome karyotype analysis, pathogenic or likely pathogenic copy number variations (CNVs) detected by CMA (or CNV-seq), and pathogenic or likely pathogenic variation detected by WES] were compared between different groups using Chi-square test or Fisher's exact test. Results:Abnormal chromosome karyotypes were identified in 10 (24.4%) of 41 cases receiving chromosome karyotype analysis, pathogenic and likely pathogenic CNVs were found in seven (7.6%) of 92 cases by CMA (or CNV-seq). WES was performed on 37 cases with negative results of chromosomal karyotype analysis and CMA (or CNV-seq) and the detection rate of pathogenic and likely pathogenic variants was 43.2% (16/37). The detection rate of WES abnormality was higher in the fetuses with musculoskeletal abnormalities than in those without the abnormalities [71.4% (15/21) vs. 1/16, Fisher's exact test, P<0.001], while in those with other postural abnormalities was higher than that in the group without other postural abnormalities [12/16 vs. 19.0% (4/21), Fisher's exact test, P=0.001]. The genetic causes of complex TE were identified in 34.7% (33/95) of the fetuses by the sequential genetic diagnosis using chromosome karyotype analysis, CMA (or CNV-seq), and WES. The overall detection rate of genetic abnormality was higher in the group with multi-system abnormality than in the group with single-system abnormality [48.9% (22/45) vs. 22.0% (11/50), χ2=7.55, P=0.006], in the group with musculoskeletal system abnormalities and without [46.8% (22/47) vs. 22.9% (11/48), χ2=5.98, P=0.014], and in the group with other postural abnormality and without [47.2% (17/36) vs. 27.1% (16/59), χ2=3.99, P=0.046]. Nine cases that were considered isolated TE on initial ultrasound were corrected to a complex diagnosis on subsequent ultrasound examinations. Of all the involved system or site, the neurologic abnormalities were the most diverse (13 kinds) and had a diversity of ultrasound presentations. Conclusions:Genetic diagnosis should be performed when prenatal ultrasound suggests fetal complex TE. WES is conducive to improving the prenatal detection rate of monogenic diseases, especially in fetuses complicated by musculoskeletal abnormalities. Isolated TE fetuses require serial ultrasound examinations to correct the diagnosis in time and genetic testing should be performed if necessary. Additional attention should be paid to the TE fetus for comorbid neurologic abnormalities at the time of ultrasonography to rule out TE as an intrauterine harbinger of neuromuscular disease.

Objective:To observe the lipid metabolism characteristics of type 2 diabetes mellitus (T2DM) patients with different levels of blood glucose control and preliminarily analyze their relationship with diabetic retinopathy (DR).Methods:A retrospective clinical study. From January 2019 to January 2024, 232 T2DM patients who underwent fundus examination in Department of Ophthalmology of Yichang Central People’s Hospital were included in the study. Based on the glycated hemoglobin A1c (HbA1c) test results, patients were divided into blood glucose standard group and blood glucose non standard group, with 100 and 132 cases respectively. Based on the results of fundus fluorescein angiography, patients were divided into non DR (NDR) group and DR group, with 89 and 143 cases, respectively. 100 healthy individuals who underwent physical examinations during the same period were selected as the control group. The thickness of peripapillary retinal nerve fiber layer (pRNFL) around the optic disc, the blood flow density of radial peripapillary capillaries (RPC) around the optic disc, and the thickness of ganglion cell complex (GCC) in the upper and lower parts of the optic disc and macular area were measured by optical coherence tomography angiography instrument. Fully automated biochemical analyzer was used to detect serum levels of total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and HbA1c. After adjusting for confounding factors, multiple linear regression model was used to analyze the correlation between HbA1c and blood lipids. Multiple logistic regression analysis was conducted to investigate the correlation between TG, HDL-C, and the occurrence of DR.Results:Compared with the control group, both the blood glucose standard group and the blood glucose non standard group had higher levels of HbA1c ( F=8.115), TC ( F=4.373), TG ( F=20.220), and LDL-C ( F=12.271), and lower levels of HDL-C ( F=6.349), with statistically significant differences ( P<0.05). Compared with the blood glucose standard group, patients in the blood glucose non standard group had higher levels of serum HbA1c ( t=3.531), TC ( t=2.561), TG ( t=6.418), LDL-C ( t=7.880), and lower levels of HDL-C ( t=5.152), with statistically significant differences ( P<0.05). Correlation analysis showed that HbA1c was positively correlated with TC, TG, and LDL-C ( P<0.05), and negatively correlated with HDL-C ( P<0.05). Multiple logistic regression analysis showed that TG, LDL-C, and HDL-C were independent risk factors for the occurrence of DR ( Ptrend<0.05). Compared with the NDR group, the DR group had thinner GCC and pRNFL thickness in the upper part of the optic disc, and lower overall and RPC blood flow density in the upper part of the optic disc, with statistically significant differences ( t=4.964, 2.406, 2.685, 2.404; P<0.05). Correlation analysis results showed that TG, LDL-C, HDL-C, HbA1c were correlated with GCC thickness, pRNFL thickness, and RPC blood flow density ( P<0.05). Conclusions:The higher the blood glucose level in T2DM patients, the more likely they are to experience dyslipidemia. TG, LDL-C, and HDL-C are independent risk factors for the occurrence of DR. Abnormal blood lipids and blood glucose levels in T2DM patients can affect retinal nerves, blood vessels, and function.

Objective To investigate the intervention effect and mechanism of LUO's Neiyi Prescription on the proliferation and angiogenesis of eutopic endometrium in rats with endometriosis(EMs)based on hypoxia-inducible factor 1A(HIF1A)/enhancer of zeste homolog 2(EZH2)/anthrax toxin receptor 2(ANTXR2)signaling pathway.Methods SD rats were randomly divided into sham operation group,model group,Danazol group(4.2 g·kg-1),low-dose LUO's Neiyi Prescription group and high-dose LUO's Neiyi Prescription group(15.74,31.48 g·kg-1),eight rats in each group.The rat model of EMs with qi stagnation and blood stasis syndrome was constructed by autologous endometrial transplantation combined with multi-factor intervention.Intragastric administration was given once a day for 28 consecutive days.The pathological changes of endometrial tissue were observed by HE staining.The expression levels of proliferating cell protein 67(Ki67)and platelet endothelial cell adhesion molecule 1(CD31)in endometrial tissue were detected by immunohistochemistry.The mRNA and protein expression levels of HIF1A,EZH2 and ANTXR2 in endometrial tissues were detected by qRT-PCR and Western Blot.The expression levels of YAP1,CD44 and β-catenin in endometrial tissues were detected by Western Blot.Results Compared with the sham operation group,the epithelial cells of the eutopic endometrium of the model group were thickened,the interstitial cells were arranged disorderly,and the inflammatory cells increased.The expression levels of Ki67 and CD31 in endometrial tissues were significantly increased(P＜0.01),the mRNA and protein expression levels of HIF1A and ANTXR2 were significantly increased(P＜0.01),while the mRNA and protein expression levels of EZH2 were significantly decreased(P＜0.01),and the protein expression levels of YAP1,CD44 and β-catenin were significantly increased(P＜0.01).Compared with the model group,the epithelial layer of the eutopic endometrial tissue of the rats in each administration group became thinner,the interstitial disorder and inflammatory infiltration were improved,and the levels of Ki67 and CD31 in the eutopic endometrial tissue were significantly decreased(P＜0.01).The mRNA and protein expression levels of HIF1A and ANTXR2 in the endometrium of rats in the Danazol group and the high-dose LUO's Neiyi Prescription group were significantly decreased(P＜0.05,P＜0.01),and the mRNA and protein expression levels of EZH2 were significantly increased(P＜0.05,P＜0.01).The protein expression levels of YAP1,CD44 and β-catenin in endometrial tissue of rats in each administration group were significantly decreased(P＜0.05,P＜0.01).Conclusion LUO's Neiyi Prescription can play a role in the treatment of EMs by inhibiting the proliferation and angiogenesis of eutopic endometrial cells,and its mechanism may be related to the inhibition of HIF1A/EZH2/ANTXR2 signaling pathway.

Objective: QL1604 is a highly selective, humanized monoclonal antibody against programmed death protein 1. We assessed the efficacy and safety of QL1604 plus chemotherapy as first-line treatment in patients with advanced cervical cancer. Methods: This was a multicenter, open-label, single-arm, phase II study. Patients with advanced cervical cancer and not previously treated with systemic chemotherapy were enrolled to receive QL1604 plus paclitaxel and cisplatin/carboplatin on day 1 of each 21-day cycle for up to 6 cycles, followed by QL1604 maintenance treatment. Results: Forty-six patients were enrolled and the median follow-up duration was 16.5 months. An 84.8% of patients had recurrent disease and 13.0% had stage IVB disease. The objective response rate (ORR) per Response Evaluation Criteria in Advanced Solid Tumors (RECIST) v1.1 was 58.7% (27/46). The immune ORR per immune RECIST was 60.9% (28/46).The median duration of response was 9.6 months (95% confidence interval [CI]=5.5–not estimable). The median progression-free survival was 8.1 months (95% CI=5.7–14.0). Fortyfive (97.8%) patients experienced treatment-related adverse events (TRAEs). The most common grade≥3 TRAEs (>30%) were neutrophil count decrease (50.0%), anemia (32.6%), and white blood cell count decrease (30.4%). Conclusion QL1604 plus paclitaxel-cisplatin/carboplatin showed promising antitumor activity and manageable safety profile as first-line treatment in patients with advanced cervical cancer. Programmed cell death protein 1 inhibitor plus chemotherapy may be a potential treatment option for the patient population who have contraindications or can’t tolerate bevacizumab, which needs to be further verified in phase III confirmatory study.

Objective: QL1604 is a highly selective, humanized monoclonal antibody against programmed death protein 1. We assessed the efficacy and safety of QL1604 plus chemotherapy as first-line treatment in patients with advanced cervical cancer. Methods: This was a multicenter, open-label, single-arm, phase II study. Patients with advanced cervical cancer and not previously treated with systemic chemotherapy were enrolled to receive QL1604 plus paclitaxel and cisplatin/carboplatin on day 1 of each 21-day cycle for up to 6 cycles, followed by QL1604 maintenance treatment. Results: Forty-six patients were enrolled and the median follow-up duration was 16.5 months. An 84.8% of patients had recurrent disease and 13.0% had stage IVB disease. The objective response rate (ORR) per Response Evaluation Criteria in Advanced Solid Tumors (RECIST) v1.1 was 58.7% (27/46). The immune ORR per immune RECIST was 60.9% (28/46).The median duration of response was 9.6 months (95% confidence interval [CI]=5.5–not estimable). The median progression-free survival was 8.1 months (95% CI=5.7–14.0). Fortyfive (97.8%) patients experienced treatment-related adverse events (TRAEs). The most common grade≥3 TRAEs (>30%) were neutrophil count decrease (50.0%), anemia (32.6%), and white blood cell count decrease (30.4%). Conclusion QL1604 plus paclitaxel-cisplatin/carboplatin showed promising antitumor activity and manageable safety profile as first-line treatment in patients with advanced cervical cancer. Programmed cell death protein 1 inhibitor plus chemotherapy may be a potential treatment option for the patient population who have contraindications or can’t tolerate bevacizumab, which needs to be further verified in phase III confirmatory study.

Objective: QL1604 is a highly selective, humanized monoclonal antibody against programmed death protein 1. We assessed the efficacy and safety of QL1604 plus chemotherapy as first-line treatment in patients with advanced cervical cancer. Methods: This was a multicenter, open-label, single-arm, phase II study. Patients with advanced cervical cancer and not previously treated with systemic chemotherapy were enrolled to receive QL1604 plus paclitaxel and cisplatin/carboplatin on day 1 of each 21-day cycle for up to 6 cycles, followed by QL1604 maintenance treatment. Results: Forty-six patients were enrolled and the median follow-up duration was 16.5 months. An 84.8% of patients had recurrent disease and 13.0% had stage IVB disease. The objective response rate (ORR) per Response Evaluation Criteria in Advanced Solid Tumors (RECIST) v1.1 was 58.7% (27/46). The immune ORR per immune RECIST was 60.9% (28/46).The median duration of response was 9.6 months (95% confidence interval [CI]=5.5–not estimable). The median progression-free survival was 8.1 months (95% CI=5.7–14.0). Fortyfive (97.8%) patients experienced treatment-related adverse events (TRAEs). The most common grade≥3 TRAEs (>30%) were neutrophil count decrease (50.0%), anemia (32.6%), and white blood cell count decrease (30.4%). Conclusion QL1604 plus paclitaxel-cisplatin/carboplatin showed promising antitumor activity and manageable safety profile as first-line treatment in patients with advanced cervical cancer. Programmed cell death protein 1 inhibitor plus chemotherapy may be a potential treatment option for the patient population who have contraindications or can’t tolerate bevacizumab, which needs to be further verified in phase III confirmatory study.

Aristolochic acid (AA), extracted from Aristolochiaceae plants, plays an essential role in traditional herbal medicines and is used for different diseases. However, AA has been found to be nephrotoxic and is known to cause aristolochic acid nephropathy (AAN).AA-induced acute kidney injury (AKI) is a syndrome in AAN with a high morbidity that manifests mitochondrial damage as a key part of its pathological progression. Melatonin primarily serves as a mitochondria-targeted antioxidant. However, its mitochondrial protective role in AA-induced AKI is barely reported. In this study, mice were administrated 2.5 mg/kg AA to induce AKI. Melatonin reduced the increase in Upro and Scr and attenuated the necrosis and atrophy of renal proximal tubules in mice exposed to AA. Melatonin suppressed ROS generation, MDA levels and iNOS expression and increased SOD activities in vivo and in vitro. Intriguingly, the in vivo study revealed that melatonin decreased mitochondrial fragmentation in renal proximal tubular cells and increased ATP levels in kidney tissues in response to AA. In vitro, melatonin restored the mitochondrial membrane potential (MMP) in NRK-52E and HK-2 cells and led to an elevation in ATP levels. Confocal immunofluorescence data showed that puncta containing Mito-tracker and GFP-LC3A/B were reduced, thereby impeding the mitophagy of tubular epithelial cells. Furthermore, melatonin decreased LC3A/B-II expression and increased p62 expression. The apoptosis of tubular epithelial cells induced by AA was decreased. Therefore, our findings revealed that melatonin could prevent AA-induced AKI by attenuating mitochondrial damage, which may provide a potential therapeutic method for renal AA toxicity.

Objective:To study the safety and treatment outcomes of portal vein embolization (PVE) combined with lenvatinib plus an anti-programmed death-1(PD-1) antibody to treat patients with initially unreasectable hepatocellular carcinoma (uHCC).Methods:This study retrospectively analyzed the data of six patients with uHCC who received first-line combined systemic therapy with lenvatinib plus an anti-PD-1 antibody, and then underwent pre-hepatectomy PVE at the Department of Liver Surgery at Zhongshan Hospital, Fudan University from May 2019 to November 2020. All enrolled patients were males, aged (54.6±6.2) (ranged 46 to 63) years. Tumor response and liver volume were evaluated by medical imagings once every 2 months (±2 weeks) and evaluated using the Response Evaluation Criteria in Solid Tumours (version 1.1). Patients were followed-up by outpatient interviews or by phone calls to record their survival and tumor outcome status.Results:Three of the six enrolled patients had Barcelona Clinic Liver Cancer stage A and three had stage B disease. One patient achieved a partial response and five patients had stable diseases. The mean ± s. d. future liver remnant (FLR) percentage was (29.0±8.9) % before PVE and the combination therapy, and was (41.3±10.8) % before the last evaluation for liver surgery ( t=10.79, P<0.001). Hepatectomy was carried out in five patients, and one patient who failed to develop significant FLR hypertrophy did not undergo hepatectomy. Grade B post-hepatectomy liver failure and major postoperative complications (i.e. pleural effusion requiring additional percutaneous drainage) occurred in one patient. After a median post-operative follow-up of 4.5 (range: 1.0-12.3) months, all five patients were alive and were tumor free. Conclusion:PVE followed by hepatectomy is feasible in a uHCC patients receiving systemic therapy with lenvatinib and an anti-PD-1 antibody.