Objective To explore the pharmacokinetic characteristics of sufentanil in individuals with normal body mass index(BMI),overweight BMI,and different CYP3A4/5 enzyme genotypes.Methods The patients receiving laparoscopic surgery under general anesthesia in the First Affiliated Hospital of Army Medical University from November 2020 to September 2021 were prospectively recruited in this study.Before the operation,the oral swabs were collected from all the patients for genotyping using the human CYP3A4/5 gene kit.Based on the potential impact of combination of their polymorphisms on sufentanil metabolism and the proportion of different genotype combinations of CYP3A4/5 enzymes,the patients were divided into groups I(3A4 homozygous mutation or 3A4 heterozygous mutation+3A5 homozygous mutation),II(3A4 heterozygous mutation+3A5 heterozygous mutation),and III(3A4 wild type or 3A4 heterozygous mutation+3A5 wild type).According to their BMI,they were also assigned into a normal body weight group(18.5～24.0 kg/m2)and an overweight group(24～<28 kg/m2),and the differences in drug metabolism parameters were statistically analyze between the 2 groups.After routine general anesthesia induction(sufentanil 0.5 μg/kg),venous blood samples were collected to detect the changes in its concentration using high performance liquid chromatography-mass spectrometry(HPLC-MS).The pharmacokinetic data of sufentanil were calculated between the normal BMI group and overweight group in all participants and between the 2 body weight groups among those with different genotype combinations.Results Among the 90 participants completing the blood drug concentration test,8 patients had their blood samples contaminated(including 1 case with an anesthesia duration of<2 h),and 3 were excluded due to low weight or overweight.Eventually,79 participants were included in the pharmacokinetic analysis on the normal body weight group and the overweight group.Compared with the normal body weight group,the central compartment volume of distribution in the overweight group was significantly reduced(P<0.05),while no obvious differences were observed between the 2 groups in terms of peripheral compartment volume of distribution,total clearance rate,peripheral compartment clearance rate,distribution half-life,clearance half-life,and area under the blood concentration-time curve.In group Ⅰ(n=26),the overweight patients(n=13)had significantly reduced central compartment volume of distribution,peripheral compartment volume of distribution,and peripheral compartment clearance rate when compared with the normal body weight patients(n=13)(P<0.05),while no differences were observed in other pharmacokinetic parameters.In groups Ⅱ(n=25)and Ⅲ(n=28),the overweight patients and normal body weight patients had no statistical differences in all pharmacokinetic parameters.Conclusion Among the patients with the same genotype combination of CYP3A4/5 mutations,there was no difference in the metabolism of sufentanil between the overweight and normal weight patients.Additionally,in the population of 3A4 homozygous mutation or 3A4 heterozygous mutation+3A5 homozygous mutation,the overweight patients have smaller peripheral distribution range of sufentanil,and weakened metabolic process.

Objective:To establish a cutoff level of anti-Müllerian hormone(AMH)which could help with the di-agnosis of polycystic ovary syndrome(PCOS)in adults,and to analyze the risk factors of metabolic syndrome(MS).Methods:A retrospectively analyzed 426 PCOS patients(PCOS group)and 205 healthy controls aged 20-39 years from the Health Checkup Center of the Gynecological Endocrine Center,Hunan Maternal and Child Health Hospital from January 2021 to December 2023.AMH diagnostic validity was estimated by receiver operating characteristic(ROC)curves.Patients were subgrouped into PCOS combined metabolic syndrome group(MS-PCOS)and the uncomplicated MS group(UMS-PCOS)according to metabolic status.Multivariate Logistic regression analysis was performed to determine the risk factors for MS in PCOS patients.Results:The serum AMH level was higher in PCOS group than that in the control group(8.42±3.71 ng/ml vs.2.99±0.94 ng/ml,P＜0.001).AMH cutoff for the diagnosis of PCOS was determined as≥4.87 ng/ml on ROC analysis,and the area under the curve is 0.981 with 92.7％sensitivity and 94.6％specificity.The prevalence of MS was 18.3％(78 ca-ses)in PCOS group.Subgroup analysis showed that MS-PCOS patients had higher waist circumference,BMI,fasting glucose,dyslipidemia,hypertension(BP＞130/85 mmHg),and hormone related index androgen level,but lower AMH vs.UMS-PCOS.Multivariate Logistic regression analysis identified insulin resistance(OR 39.17,95％CI 9.33-164.48),BMI ≥24 kg/m2(OR 3.72,95％CI 1.86-7.45),and hyperandrogenism(OR 2.56,95％CI 1.34-4.89)as independent risk factors of MS.AMH was negatively associated with MS,a single-unit increase in AMH was associated with an 17％decrease in odds of MS(OR 0.83,95％CI 0.73-0.95,P=0.006).Conclusions:Serum AMH levels were significantly higher in adult PCOS patients,with an optimal diagnostic threshold of 4.87 ng/ml.Hyperandrogenism and low AMH levels may predict a higher risk of MS,in addition to metabolism-related factors.

Objective:To establish a cutoff level of anti-Müllerian hormone(AMH)which could help with the di-agnosis of polycystic ovary syndrome(PCOS)in adults,and to analyze the risk factors of metabolic syndrome(MS).Methods:A retrospectively analyzed 426 PCOS patients(PCOS group)and 205 healthy controls aged 20-39 years from the Health Checkup Center of the Gynecological Endocrine Center,Hunan Maternal and Child Health Hospital from January 2021 to December 2023.AMH diagnostic validity was estimated by receiver operating characteristic(ROC)curves.Patients were subgrouped into PCOS combined metabolic syndrome group(MS-PCOS)and the uncomplicated MS group(UMS-PCOS)according to metabolic status.Multivariate Logistic regression analysis was performed to determine the risk factors for MS in PCOS patients.Results:The serum AMH level was higher in PCOS group than that in the control group(8.42±3.71 ng/ml vs.2.99±0.94 ng/ml,P＜0.001).AMH cutoff for the diagnosis of PCOS was determined as≥4.87 ng/ml on ROC analysis,and the area under the curve is 0.981 with 92.7％sensitivity and 94.6％specificity.The prevalence of MS was 18.3％(78 ca-ses)in PCOS group.Subgroup analysis showed that MS-PCOS patients had higher waist circumference,BMI,fasting glucose,dyslipidemia,hypertension(BP＞130/85 mmHg),and hormone related index androgen level,but lower AMH vs.UMS-PCOS.Multivariate Logistic regression analysis identified insulin resistance(OR 39.17,95％CI 9.33-164.48),BMI ≥24 kg/m2(OR 3.72,95％CI 1.86-7.45),and hyperandrogenism(OR 2.56,95％CI 1.34-4.89)as independent risk factors of MS.AMH was negatively associated with MS,a single-unit increase in AMH was associated with an 17％decrease in odds of MS(OR 0.83,95％CI 0.73-0.95,P=0.006).Conclusions:Serum AMH levels were significantly higher in adult PCOS patients,with an optimal diagnostic threshold of 4.87 ng/ml.Hyperandrogenism and low AMH levels may predict a higher risk of MS,in addition to metabolism-related factors.

The dynamic balance between bone formation and bone resorption is a critical process of bone remodeling.The imbalance of bone formation and bone resorption is closely associated with the occurrence and development of various bone-related diseases.Under both physiological and pathological conditions,non-coding RNAs(ncRNAs)play a crucial regulatory role in protein expression through either inhibiting mRNAs translation or promoting mRNAs degradation.Circular RNAs(circRNAs)are a type of non-linear ncRNAs that can resist the degradation of RNA exonucleases.There is accumulating evidence suggesting that circRNAs and microRNAs(miRNAs)serve as critical regulators of bone remodeling through their direct or indirect regulation of the expression of osteogenesis-related genes.Additionally,recent studies have revealed the involvement of the circRNAs-miRNAs regulatory network in the process by which mesenchymal stem cells(MSCs)differentiate towards the osteoblasts(OB)lineage and the process by which bone marrow-derived macrophages(BMDM)differentiate towards osteoclasts(OC).The circRNA-miRNA network plays an important regulatory role in the osteoblastic-osteoclastic balance of bone remodeling.Therefore,a thorough understanding of the circRNA-miRNA regulatory mechanisms will contribute to a better understanding of the regulatory mechanisms of the balance between osteoblastic and osteoclastic activities in the process of bone remodeling and the diagnosis and treatment of related diseases.Herein,we reviewed the functions of circRNA and microRNA.We also reviewed their roles in and the mechanisms of the circRNA-miRNA regulatory network in the process of bone remodeling.This review provides references and ideas for further research on the regulation of bone remodeling and the prevention and treatment of bone-related diseases.

Human adipose-derived stem cells (hASCs) are a promising cell type for bone tissue regeneration. Circular RNAs (circRNAs) have been shown to play a critical role in regulating various cell differentiation and involve in mesenchymal stem cell osteogenesis. However, how circRNAs regulate hASCs in osteogenesis is still unclear. Herein, we found circ_0003204 was significantly downregulated during osteogenic differentiation of hASCs. Knockdown of circ_0003204 by siRNA or overexpression by lentivirus confirmed circ_0003204 could negatively regulate the osteogenic differentiation of hASCs. We performed dual-luciferase reporting assay and rescue experiments to verify circ_0003204 regulated osteogenic differentiation via sponging miR-370-3p. We predicted and confirmed that miR-370-3p had targets in the 3'-UTR of HDAC4 mRNA. The following rescue experiments indicated that circ_0003204 regulated the osteogenic differentiation of hASCs via miR-370-3p/HDAC4 axis. Subsequent in vivo experiments showed the silencing of circ_0003204 increased the bone formation and promoted the expression of osteogenic-related proteins in a mouse bone defect model, while overexpression of circ_0003204 inhibited bone defect repair. Our findings indicated that circ_0003204 might be a promising target to promote the efficacy of hASCs in repairing bone defects.
Adipose Tissue/metabolism* ; Animals ; Cell Differentiation/genetics* ; Cells, Cultured ; Histone Deacetylases/metabolism* ; Humans ; Mice ; MicroRNAs/metabolism* ; Osteogenesis/genetics* ; RNA, Circular/metabolism* ; Repressor Proteins/metabolism* ; Signal Transduction ; Stem Cells/metabolism*

Objective:To analyze the clinical characteristics of patients with nocardiosis, so as to improve the diagnosis and treatment of nocardia infection in the future.Methods:From May 2016 to October 2020, 24 patients with nocardiosis in Xiangya Hospital, Central South University were enrolled, and their clinical data including clinical features, laboratory examinations, imaging findings, diagnosis and treatment process, and outcome were retrospectively analyzed.Results:Among the 24 patients with nocardiosis, 18 cases (75.0%) were males, and the median age was 54.5 years.Twenty-three patients had underlying diseases, of which the most common disease was antineutrophil cytoplasmic antibody-related vasculitis (16.7%(4/24)). Of nine species of Nocardia identified from the 24 patients, Nocardia farcinica was the most common species (seven cases). The lesion sites were mainly lungs (70.8%(17/24)), skin and soft tissues (42.0%(10/24)), brain (25.0%(6/24)) and blood system (17.0%(4/24)). There were 12 cases (50.0%) of patients with more than two lesion sites. The clinical manifestations, imaging examinations and laboratory tests of the 24 patients were not specific. The diagnosis depended on the etiology. Nineteen patients received trimethoprim-sulfamethoxazole-based combination therapy, and two were discontinued due to adverse reactions of sulfa drugs. After treatment, 19 cases (79.2%) were improved and five cases (20.8%) died. Conclusions:Patients with nocardiosis often have atypical clinical manifestations, and multiple organs are easily affected.Early and accurate identification and rapid and effective anti-biotic therapy are the keys to improve the overall prognosis of these patients.

OBJECTIVE:To establish a method for simultaneous determination of residual methylparaben,ethylparaben,nipa-sol and benzalkonium chloride in marketed eye drops. METHODS:HPLC method was adopted. The determination was performed on Hypersil GOLD C18 column with mobile phase consisted of 0.005 mol/L ammonium acetate(10 mL triehtylamine in 1 L solu-tion,pH adjusted to 5.0±0.5 with glacial acetic acid)-acetonitrile(45:55,V/V)at the flow rate of 1.0 mL/min. The detection wave-length was 262 nm(methylparaben,ethylparaben,nipasol)and 214 nm(benzalkonium chloride),respectively. The column tem-perature was 30 ℃ and sample size was 20 μL. RESULTS:The linear range were 1.2350-15.4380 μg/mL for methylparaben(r=0.9999),1.3170-16.3836 μ g/mL for ethylparaben (r=0.9997),1.2072-15.0894 μ g/mL for nipasol (r=0.9996) and 17.776-222.0 μg/mL for benzalkonium chloride(r=0.9999),respectively. Limits of quantitation were 2.0,2.0,2.0,1.11 μg,re-spectively;limits of determination were 0.375,0.375,0.375,0.333 μg,respectively. RSDs of precision,stability and reproducibili-ty tests were all lower than 2.0%. The average recoveries were 98.14%-102.48%(RSD=1.6%,n=9),98.79%-102.42%(RSD=1.3%,n=9),98.19%-102.49%(RSD=1.5%,n=9)and 98.76%-100.53%(RSD=0.6%,n=9),respectively. CONCLUSIONS:The method is accurate,reproducible,simple and suitable for the determination of residual methylparaben,ethylparaben,nipasol and benzalkonium chloride in marketed eye drops.

Objective To investigate the clinical value of hysteroscopy in helping identify the etiology of intrauterine fluid collection in postmenopausal women.Methods One hundred and thirtyfour postmenopausal women with intrauterine fluid collection,detected via transvaginal sonography,were admitted in the department of gynecology and obstetrics at Beijing ChaoYang Hospital from May 2014 to October 2016.All clinical data were retrospectively analyzed.All patients underwent hysteroscopy and biopsy of the endometrium.The results of hysteroscopy were analyzed against biopsy results.Results Hysteroscopy revealed that 116 of the 134 cases had uterine fluid(86.6％).Based on hysteroscopy results,44 (37.9 ％) had endometritis,10 (8.6 ％) had atrophic endometritis,5 (4.3 ％) had endometrial carcinoma,and 7(6.0 ％)had endometrial polyps,but the case numbers and rates for these conditions from microscopic examination were39 (33.6％),15 (12.9％),6 (5.2％),and 7 (6.0 ％),respectively.When microscopic examination was used as the gold standard,the sensitivities and specificities of hysteroscopy in the diagnosis of endometritis,atrophic endometritis,endometrial cancer and endometrial polyps were 82.0 ％ and 85.7 ％,66.7 ％ and 97.1％,80.0 ％ and 98.2 ％,and 100.0％ and 97.3％,respectively.The diagnostic coincidence rates were 84.6％,93.2％,94.4％ and 97.4％,respectively.Conclusions Postmenopausal women with intrauterine fluid collection shown on transvaginal sonography should receive hysteroscopy for early detection of endometrial lesions.

Objective: To evaluate the significance of bacteria detection with filter paper method on diagnosis of diabetic foot wound infection. Methods: Eighteen patients with diabetic foot ulcer conforming to the study criteria were hospitalized in Liyuan Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from July 2014 to July 2015. Diabetic foot ulcer wounds were classified according to the University of Texas diabetic foot classification (hereinafter referred to as Texas grade) system, and general condition of patients with wounds in different Texas grade was compared. Exudate and tissue of wounds were obtained, and filter paper method and biopsy method were adopted to detect the bacteria of wounds of patients respectively. Filter paper method was regarded as the evaluation method, and biopsy method was regarded as the control method. The relevance, difference, and consistency of the detection results of two methods were tested. Sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of filter paper method in bacteria detection were calculated. Receiver operating characteristic (ROC) curve was drawn based on the specificity and sensitivity of filter paper method in bacteria detection of 18 patients to predict the detection effect of the method. Data were processed with one-way analysis of variance and Fisher's exact test. In patients tested positive for bacteria by biopsy method, the correlation between bacteria number detected by biopsy method and that by filter paper method was analyzed with Pearson correlation analysis. Results: (1) There were no statistically significant differences among patients with wounds in Texas grade 1, 2, and 3 in age, duration of diabetes, duration of wound, wound area, ankle brachial index, glycosylated hemoglobin, fasting blood sugar, blood platelet count, erythrocyte sedimentation rate, C-reactive protein, aspartate aminotransferase, serum creatinine, and urea nitrogen (with F values from 0.029 to 2.916, P values above 0.05), while there were statistically significant differences among patients with wounds in Texas grade 1, 2, and 3 in white blood cell count and alanine aminotransferase (with F values 4.688 and 6.833 respectively, P<0.05 or P<0.01). (2) According to the results of biopsy method, 6 patients were tested negative for bacteria, and 12 patients were tested positive for bacteria, among which 10 patients were with bacterial number above 1×10⁵/g, and 2 patients with bacterial number below 1×10⁵/g. According to the results of filter paper method, 8 patients were tested negative for bacteria, and 10 patients were tested positive for bacteria, among which 7 patients were with bacterial number above 1×10⁵/g, and 3 patients with bacterial number below 1×10⁵/g. There were 7 patients tested positive for bacteria both by biopsy method and filter paper method, 8 patients tested negative for bacteria both by biopsy method and filter paper method, and 3 patients tested positive for bacteria by biopsy method but negative by filter paper method. Patients tested negative for bacteria by biopsy method did not tested positive for bacteria by filter paper method. There was directional association between the detection results of two methods (P=0.004), i. e. if result of biopsy method was positive, result of filter paper method could also be positive. There was no obvious difference in the detection results of two methods (P=0.250). The consistency between the detection results of two methods was ordinary (Kappa=0.68, P=0.002). (3) The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of filter paper method in bacteria detection were 70%, 100%, 1.00, 0.73, and 83.3%, respectively. Total area under ROC curve of bacteria detection by filter paper method in 18 patients was 0.919 (with 95% confidence interval 0-1.000, P=0.030). (4) There were 13 strains of bacteria detected by biopsy method, with 5 strains of Acinetobacter baumannii, 5 strains of Staphylococcus aureus, 1 strain of Pseudomonas aeruginosa, 1 strain of Streptococcus bovis, and 1 strain of bird Enterococcus. There were 11 strains of bacteria detected by filter paper method, with 5 strains of Acinetobacter baumannii, 3 strains of Staphylococcus aureus, 1 strain of Pseudomonas aeruginosa, 1 strain of Streptococcus bovis, and 1 strain of bird Enterococcus. Except for Staphylococcus aureus, the sensitivity and specificity of filter paper method in the detection of the other 4 bacteria were all 100%. The consistency between filter paper method and biopsy method in detecting Acinetobacter baumannii was good (Kappa=1.00, P<0.01), while that in detecting Staphylococcus aureus was ordinary (Kappa=0.68, P<0.05). (5) There was no obvious correlation between the bacteria number of wounds detected by filter paper method and that by biopsy method (r=0.257, P=0.419). There was obvious correlation between the bacteria numbers detected by two methods in wounds with Texas grade 1 and 2 (with r values as 0.999, P values as 0.001). There was no obvious correlation between the bacteria numbers detected by two methods in wounds with Texas grade 3 (r=-0.053, P=0.947). Conclusions The detection result of filter paper method is in accordance with that of biopsy method in the determination of bacterial infection, and it is of great importance in the diagnosis of local infection of diabetic foot wound.

OBJECTIVE To explore downstream regulatory pathway of microRNA-21 (miR-21) in colon cancer cells (RKO) through detecting miR-21 and its target PDCD4, and the influence of miR-21 regulation on the sensitivity of RKO cells to 5-fluorouracil (5-FU). METHODS 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay was used to determine the effect of 5-FU on the viability of RKO cells with knockout of miR-21 or high expression of PDCD4. Real-time was used to determine the expression of PDCD4, ABCC5 and CD44 in RKO cell after knockout of miR-21. RESULTS MTT assay reveals that the IC50 of 5-FU in RKO-WT cells (52.82 ± 0.06 umol/L) was about 67% higher than in miR-21 knockout cells (32.23 ± 0.05 umol/L) (P < 0.05), and the apoptosis ratio elevated after knockout of miR-21. High expression of PDCD4, a target gene of miR-21, can negatively regulate the expression of ABC transporter ABCC5 and the stem cell marker CD44. CONCLUSION MiR-21 can mediate the drug resistance to 5-FU by inhibiting its target PDCD4, which can regulate the expression of ABCC5 and CD44 genes.
ATP Binding Cassette Transporter, Sub-Family G, Member 5 ; ATP-Binding Cassette Transporters ; genetics ; Antimetabolites, Antineoplastic ; pharmacology ; Apoptosis Regulatory Proteins ; physiology ; Cell Line, Tumor ; Colonic Neoplasms ; drug therapy ; pathology ; Drug Resistance, Neoplasm ; Fluorouracil ; pharmacology ; Humans ; Hyaluronan Receptors ; genetics ; Lipoproteins ; genetics ; MicroRNAs ; physiology ; RNA-Binding Proteins ; physiology