Originating but free from chromosomal DNA, extrachromosomal circular DNAs (eccDNAs) are organized in circular form and have long been found in unicellular and multicellular eukaryotes. Their biogenesis and function are poorly understood as they are characterized by sequence homology with linear DNA, for which few detection methods are available. Recent advances in high-throughput sequencing technologies have revealed that eccDNAs play crucial roles in tumor formation, evolution, and drug resistance as well as aging, genomic diversity, and other biological processes, bringing it back to the research hotspot. Several mechanisms of eccDNA formation have been proposed, including the breakage-fusion-bridge (BFB) and translocation-deletion-amplification models. Gynecologic tumors and disorders of embryonic and fetal development are major threats to human reproductive health. The roles of eccDNAs in these pathological processes have been partially elucidated since the first discovery of eccDNA in pig sperm and the double minutes in ovarian cancer ascites. The present review summarized the research history, biogenesis, and currently available detection and analytical methods for eccDNAs and clarified their functions in gynecologic tumors and reproduction. We also proposed the application of eccDNAs as drug targets and liquid biopsy markers for prenatal diagnosis and the early detection, prognosis, and treatment of gynecologic tumors. This review lays theoretical foundations for future investigations into the complex regulatory networks of eccDNAs in vital physiological and pathological processes.
Male ; Female ; Animals ; Humans ; Swine ; DNA, Circular/genetics* ; Genital Neoplasms, Female ; Semen ; DNA ; Reproduction

OBJECTIVE To investigate the monitoring of tacrolimus blood concentration in patients with nephrotic syndrome （NS），and to establish a prediction model for tacrolimus blood concentration. METHODS Data from 509 concentration monitoring sessions of 166 NS patients using tacrolimus were collected from January 1， 2020 to August 31， 2023 in Zhongshan Hospital Affiliated to Xiamen University. The relationship of efficacy and adverse drug reaction（ADR） with blood concentration was analyzed. A multilayer perceptron （MLP） prediction model was established by using the blood concentration monitoring data of 302 times from 109 NS patients with genetic information， and then verified. RESULTS In terms of efficacy， the median blood concentration of tacrolimus in the non-remission group was 2.20 ng/mL， which was significantly lower than that in the partial remission group （4.00 ng/mL， P＜0.001） and the complete remission group （3.60 ng/mL， P＝0.002）. In terms of ADR， the median blood concentration of tacrolimus in the ADR group was 5.01 ng/mL， which was significantly higher than that in the non-ADR group （3.37 ng/mL） （P＝0.001）. According to the subgroup analysis of the receiver operating characteristic curve， when the blood concentration of tacrolimus was ≥6.65 ng/mL， patients were more likely to develop elevated blood creatinine [area under the curve （AUC） was 0.764， P＜0.001）； when the blood concentration of tacrolimus was ≥6.55 ng/mL， patients were more likely to develop blood glucose （AUC＝0.615， P＝ 0.005）. The established MLP prediction model has a loss function of 0.9， with an average absolute error of 0.279 5 ng/mL between the predicted and measured values. The determination coefficient of the validation scatter plot was 0.984， indicating an excellent predictive performance of the model. CONCLUSION Tacrolimus blood concentration has an impact on both efficacy and ADR in NS patients. The use of the MLP model for predicting blood concentration exhibits high accuracy with minimal error between predicted and measured values. The model can be used as an important tool in clinical individualized medication regimens.

OBJECTIVE To establish an ion chromatography method for the simultaneous determination of chloride， sulfate and bicarbonate ions in Polyethylene glycol electrolyte powder （Ⅲ）. METHODS The chromatographic column was a Dionex IonpacTM AS11-HC anion analysis column， with a Dionex IonPacTM AG11-HC guard column. The mobile phase was 10 mmol/L potassium hydroxide at an isocratic elution flow rate of 1.2 mL/min. The detector was a conductivity detector， and the suppressor was a Dionex AERS with a suppressor current of 30 mA. The column temperature was maintained at 30 ° C， and the injection volume was 10 μL. Chloride and sulfate contents were calculated by external standard method， while bicarbonate content was determined by double logarithmic fitting standard curve method. RESULTS Under these chromatographic conditions， chloride， sulfate and bicarbonate ions were effectively separated with linear ranges of 0.055 to 0.219 mg/mL （r＝0.999 9）， 0.155 to 0.618 mg/mL （r＝1.000 0）， and 0.065 to 0.121 mg/mL （r＝0.999 9）， respectively. The recoveries were 98.06% to 101.34%， 97.37% to 101.25%， and 97.16% to 99.81%， respectively， with RSDs of 1.1%， 1.3% and 1.0% （n＝9）. The RSDs for the evaluation of precision， accuracy， stability and ruggedness were all less than 2%. CONCLUSIONS The established ion chromatography is simple， rapid， accurate， precise and durable， can simultaneously determine the contents of chloride， sulfate and bicarbonate ions in Polyethylene glycol electrolyte powder （Ⅲ）， which is suitable for its quality control.

Purpose To investigate the expression status of GNL3 in gastric cancer,and to explore the role of GNL3 in tumor proliferation,invasion and migration.Methods The ex-pression of GNL3 mRNA in gastric cancer tissues was analyzed by searching database.The expression of GNL3 protein in 51 gastric cancer tissues and 51 adjacent non-tumor tissues was de-tected by immunohistochemistry(IHC)SP method.The correla-tion between GNL3 protein expression and gastric cancer clinical pathological features was analyzed by x2 test.The expression of GNL3 in gastric cancer cells was silenced by transfection of sh-RNA,and the silencing efficiency was verified by Western blot.The effect of silencing GNL3 on the proliferation of gastric cancer cells was examined by CCK-8,colony formation and EdU stai-ning.In addition,wound-healing assay and Transwell assay were performed to detect the effect of GNL3 silencing on cell invasion and migration.Results SangerBox and UALCAN database re-trieval showed that the expression of GNL3 mRNA was signifi-cantly increased in gastric cancer tissues(P＜0.01).IHC stai-ning showed that the positive expression rate of GNL3 protein in gastric cancer tissues was 96.1％,and the high expression rate was 78.4％,which was significantly higher than that in adjacent non-tumor tissues(74.5％,51.0％,P＜0.01).Moreover,the high expression of GNL3 was significantly correlated with lymph node metastasis in gastric cancer patients(x2=4.933,P=0.026).CCK-8,colony formation and EdU staining showed that GNL3 silencing inhibited the proliferation of gastric cancer cell SGC-7901.The wound-healing and Transwell assay showed that GNL3 silencing inhibited the migration and invasion of gastric cancer cell.Conclusion The GNL3 protein is highly expressed in gastric cancer tissues,and closely related to the proliferation,migration and invasion of gastric cancer cells.

From January 1, 2013, to March 1, 2024, nine patients with hematological malignancies complicated by Gilbert’s syndrome in Peking University People’s Hospital underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT). The patients comprised seven male and two female cases, with a median age of 38 (13-60) years old. Among them, three cases were acute myeloid leukemia, three cases were acute lymphocytic leukemia, two cases were myelodysplastic syndrome, and one case was chronic myelomonocytic leukemia. None of the patients had viral hepatitis. Of the nine cases, seven cases received the Bu-Cy+ATG regimen, while the other two cases received the TBI-Cy+ATG regimen (Bu, busulfan; Cy, cyclophosphamide; ATG, antithymocyte immunoglobulin; and TBI, total body irradiation). All patients achieved neutrophil engraftment, and eight received platelet engraftment. The median total bilirubin level was 45.4 (22.5-71.2) μmol/L before transplantation and 22.0 (18.0-37.2) μmol/L on -1d of preconditioning. The total bilirubin level on +20d after the transplantation of eight patients decreased compared with the baseline level before transplantation. Moreover, one patient had a transient increase in the total bilirubin level on +5d after transplantation, which was considered to be attributed to the toxicity of Bu. No patients were complicated by hepatic veno-occlusive disease. The median follow-up time was 739 (42-2 491) days. During the follow-up period, one patient died of recurrence, and the remaining eight patients had disease-free survival events.

Objective: To explore the abnormal changes in neuroelectric activity in the primary visual cortex of rats deprived of vision in one eye and to investigate the regulatory effect of acupuncture in the sensitive period on the abnormal coding and conduction of electrical signals of rats' optic neurons.Methods: Sixty 14-day-old Sprague-Dawley rats were randomly divided into a blank group, a model group, an early-stage acupuncture group, a middle-stage acupuncture group, and a late-stage acupuncture group, with 12 rats in each group. Rats in every group except the blank group received right eyelid suturing to create a monocular deprivation model in the sensitive period of visual development (from the day rats open their eyes to the 45th day after their birth). Rats in the three acupuncture groups started to undergo acupuncture respectively on the 3rd, 12th, and 21st days after the model replication was done, with each group receiving nine-day treatment. The activity level of the neuroelectrical signal of the primary visual cortex in each group, including the latency and amplitude of P100 wave, average discharge frequency and amplitude of neurons, the power spectral density (PSD), and interspike interval (ISI), were measured by neuroelectric evaluation technology after the acupuncture treatment was finished. Results: Compared with the blank group, the latency of P100 wave in the visual center of vision-deprived eyes was significantly prolonged, and the amplitude was significantly reduced (P<0.05); the average discharge frequency and amplitude of the neurons in the visual cortex also decreased significantly (P<0.05); PSD decreased and ISI was prolonged significantly (P<0.05). Compared with the model group, the abnormal electrical activity of optic neurons in the three acupuncture groups ameliorated, the latency of P100 shortened, and the amplitude of P100 increased (P<0.05), the discharge frequency and amplitude increased significantly (P<0.05), the PSD reduced, and the ISI shortened (P<0.05). In addition, among the three acupuncture groups, the early-stage acupuncture group had the best effect on various indicators. Conclusion: Abnormal electrophysiological activity is significant in the visual center of vision-deprived rats, and acupuncture treatment in the sensitive period of visual development can enhance the bioelectrical activity of visual nerve cells, improve the efficiency of optic nerve conduction, and regulate the inhibition and retardation of visual response caused by visual deprivation.

Objective:To investigate the prevalence and associated risk factors of pre-hypertension and hypertension in young and middle-aged population in Nanjing.Methods:Subjects of the study were those who underwent physical examination in the physical examination center of Nanjing Drum Tower Hospital from 2009 to 2016. The prevalence and risk factors of pre-hypertension and hypertension in young (aged 18-44 years old) and middle-aged people (aged 45-59 years old) were analyzed.Results:A total of 142 857 participants aged 18-59 years old were analyzed. Among them, 64 220 cases in the pre-hypertension group and 13 912 cases in the hypertension group. The prevalence of hypertension was 9.74% (12.51% in males and 5.82% in females). The prevalence of pre-hypertension was 44.95% (53.31% in males and 33.15% in females). In the middle-aged group, the prevalence of pre-hypertension and hypertension were 51.68% and 15.13%, respectively, which was higher than that in the young group (37.95% and 4.13%, respectively). The prevalence of pre-hypertension and hypertension in 2013-2016 was 45.37% and 10.65%, respectively, which was higher than that in 2009-2012(44.52% and 8.78%). In addition, the prevalence of abnormal blood glucose metabolism, abnormal blood lipid metabolism and abnormal glucose and lipid metabolism in the pre-hypertension group was higher than that in the normal blood pressure group, but lower than that in the hypertension group ( P<0.001). A logistic regression analysis indicated that age, overweight or obesity, hyperglycemia, hypertriglyceridemia and hypercholesterolemia were risk factors of pre-hypertension in male. Age, overweight or obesity, hyperglycemia, hypertriglyceridemia, hypercholesterolemia and hyper-low density cholesterolemia were associated with hypertension in male and with pre-hypertension and hypertension in female. Conclusions:Middle age, overweight/obesity, elevated fasting plasma glucose, elevated triglyceride and elevated total cholesterol were risk factors of pre-hypertension and hypertension in both men and women. Intervention on the related risk factors should be conducted as early as possible.

Objective:To explore the expression of circular RNA circ-MYBL2 in prostate cancer tissue and the molecular mechanism of its influence on the occurrence and metastasis of prostate cancer.Methods:From February 2017 to April 2021, 45 cases of prostate cancer tissues and paracancerous tissues from patients with prostate cancer in the Department of Urology, Jingmen No.2 People′s Hospital were selected. quantitative real-time fluorescence polymerase chain reaction (qRT-PCR) was used to detect the difference in expression of circ-MYBL2 in prostate cancer tissues and adjacent tissues, and the difference in expression of circ-MYBL2 in prostate cancer cell lines and immortalized prostate duct epithelial cells. Cell lines with low circ-MYBL2 expression were respectively transfected with circ-MYBL2 plasmid (circ-MYBL2 group) or negative control plasmid (control group). qRT-PCR was used to detect the transfection efficiency of circ-MYBL2 plasmid. CCK-8 method and cell scratch test were used to detect the effect of circ-MYBL2 on cell proliferation and migration. The starBase v2.0 software was used to predict the miRNA bound by circ-MYBL2 and the target gene of miRNA. The dual luciferase reporter gene experiment was used to verify the regulatory relationship between circ-MYBL2 and miRNA. qRT-PCR was used to detect the influence of circ-MYBL2 on miRNA expression and the influence of miRNA on target gene mRNA expression. Western blotting was used to detect the expression of target gene protein and Wnt/β-catenin signaling pathway proteins. The measurement data were expressed as mean±standard deviation ( Mean± SD), the comparison between the means of multiple samples used one-way analysis of variance, and the comparison between the means of two samples used the t-test. Results:The expression of circ-MYBL2 of DU-145 cells in prostate cancer tissue was significantly lower than that in adjacent tissues ( P<0.01). The expression of circ-MYBL2 in prostate cancer cell lines was significantly lower than that of prostate ductal epithelial cells ( P<0.01), and the expression of DU-145 cells was the lowest ( P<0.01). Compared with the control group, the expression of circ-MYBL2 of DU-145 cells in the circ-MYBL2 group increased significantly ( P<0.01), and circ-MYBL2 reduced the proliferation activity ( P<0.05) and migration ability ( P<0.01) of DU-145 cells. circ-MYBL2 acted as a sponge to adsorb miR-324-3p, and miR-324-3p complementarily bound to the suppressor of SUFU gene. circ-MYBL2 inhibited the expression of miR-324-3p ( P<0.01), SUFU gene expression was increased ( P<0.01), and Wnt/β-catenin signal pathway transduction was inhibited. Conclusion:circ-MYBL2 promotes the expression of SUFU gene by adsorbing miR-324-3p, inhibits the Wnt/β-catenin signaling pathway, thereby reducing the proliferation activity and migration ability of prostate cancer cells.

Objective:To observe the effects of miR-146a on human retinal endothelial cell (HREC) under high glucose condition.Methods:Total of 57 cases diagnosed as diabetic mellitus and 40 cases with diabetic retinopathy (DR) in Wuxi People's Hospital Affiliated to Nanjing Medical University from October to December 2013.Forty-one healthy volunteers were enrolled and served as control group.The clinical data and venous blood samples of subjects were collected.HRECs were cultured in normal glucose (5.5 mmol/L) or high glucose medium (30 mmol/L). Real-time PCR was used to detect the expression of miR-146a.The cultured HRECs were transfected with miR-146a mimic, mimic negative control, inhibitor and inhibitor negative control by lipofectamine2000, respectively.The expression of miR-146a and intercellular cell adhesion molecule-1 (ICAM-1) mRNA was examined by real-time PCR and the expression of nuclear factor-кB (NF-кB) p65 and NF-кB p65 Ser536 was detected by Western blot assay. Results:The relative expression of miR-146a mRNA in the diabetic mellitus group and DR group was 0.36±0.08 and 0.27±0.08, respectively, which were significantly lower than 1.00±0.16 in the control group (both at P<0.01). The expression of miR-146a mRNA was 0.37±0.11 in the high glucose group, which was lower than 1.00±0.18 in the normal control group ( t=5.57, P<0.01). The relative expression of miR-146a mRNA in the miR-146a mimic group was 2 540.00±105.00, which was significantly higher than 61.00±17.90 in the miR-146a mimic control group; The relative expression of miR-146a mRNA in the miR-146a inhibitor group was 0.04±0.01, which was significantly lower than 0.88±0.04 in the miR-146a inhibitor control group ( t=23.23, 17.12; both at P<0.01). The relative expression of ICAM-1 mRNA in the miR-146a mimic group was 0.35±0.12, which was significantly lower than 1.00±0.13 in the miR-146a mimic control group; The relative expression of ICAM-1 mRNA in the miR-146a inhibitor group was 2.74±0.48, which was significantly higher than 1.00±0.16 in the miR-146a inhibitor control group ( t=3.58, 3.37; both at P<0.05). The relative expression of NF-кB p65 Ser536 in the miR-146a mimic group was 0.43±0.03, which was significantly lower than 1.07±0.09 in the miR-146a mimic control group ( t=6.74, P<0.01). The relative expression of NF-кB p65 Ser536 in the miR-146a inhibitor group was 2.08±0.12, which was significantly higher than 1.00±0.01 in the miR-146a inhibitor control group ( t=8.76; P<0.01). Conclusions:miR-146a can reduce inflammation of HREC in high glucose condition through inhibiting ICAM-1 expression and NF-кB phosphorylation.

Objective:To establish autoverification rules for coagulation tests in multicenter cooperative units, in order to reduce workload for manual review of suspected results and shorten turnaround time (TAT) of test reports, while ensure the accuracy of results.Methods:A total of 14 394 blood samples were collected from fourteen hospitals during December 2019 to March 2020. These samples included: Rules Establishment Group 11 230 cases, including 1 182 cases for Delta check rules; Rules Validation Group 3 164 cases, including 487cases for Delta check; Clinical Application Trial Group 77 269 cases. Samples were analyzed for coagulation tests using Sysmex CS series automatic coagulation analyzers, and the clinical information, instrument parameters, test results, clinical diagnosis, medication history of anticoagulant and other relative results such as HCT, TG, TBIL, DBIL were summarized; on the basis of historical data, the 2.5 and 97.5 percentile of all data arranged from low to high were initially accumulated; on the basis of clinical suggestions, critical values and specific drug use as well as relative guidelines, autoverification rules and limits were established.The rules were then input into middleware, in which Stage I/Stage II validation was done. Positive coincidence, negative coincidence, false negative, false positive, autoverification pass rate, passing accuracy (coincidence of autoverification and manual verification) were calculated. Autoverification rules underwent trial application in coagulation results reports.Results:(1) The autoverification algorisms involve 33 rules regarding PT/INR, APTT, FBG, D-dimer, FDP,Delta check, reaction curve and sample abnormalities; (2)Autoverification Establishment Group showed autoverification pass rate was 68.42% (7 684/11 230), the false negative rate was 0%(0/11230), coincidence of autoverification and manual verification was 98.51%(11 063/11 230), in which positive coincidence and negative coincidence were respectively 30.09% (3 379/11 230) and 68.42%(7 684/11 230); Autoverification Validation Group showed autoverification pass rate was 60.37%(1 910/3 164), the false negative rate was 0%(0/11 230), coincidence of autoverification and manual verification was 97.79%(3 094/3 164), in which positive coincidence and negative coincidence were respectively 37.42%(1 184/3 164) and 60.37%(1 910/3 164); (3) Trialed implementation of these autoverification rules on 77 269 coagulation samples showed that the average TAT shortened by 8.5 min-83.1 min.Conclusions:This study established 33 autoverification rules in coagulation tests. Validation showedthese rules could ensure test quality while shortening TAT and lighten manual workload.