AIM:To investigate the impact of total flavonoids of Pterocarya hupehensis Skan(PHSTF)on the migration,invasion,and ferroptosis of non-small-cell lung cancer A549 cells.METHODS:The A549 cells were divided into control group,low-,medium-and high-dose(100,150 and 200 μg/mL)PHSTF groups,ferroptosis inhibitor liprox-statin-1(Lip-1)group,and high-dose PHSTF combined with Lip-1 group,each cultured in corresponding media.Cell via-bility was assessed using the CCK-8 assay,while cell migration and invasion abilities were determined through scratch and Transwell assays.Cell lipid peroxidation levels were measured using the glutathione(GSH)assay kit.RT-qPCR was em-ployed to assess the mRNA expression of solute carrier family 7 member 11(SLC7A11)and glutathione peroxidase 4(GPX4),while Western blot was utilized to examine the protein expression of SLC7A11,GPX4,Kelch-like epichlorohy-drin-associated protein-1(Keap-1),nuclear factor E2-related factor 2(Nrf2)and heme oxygenase-1(HO-1).RE-SULTS:Compared with control group,PHSTF significantly diminished the viability of A549 cells in a time-and dose-de-pendent manner(P＜0.01),and the cell migration and invasion were also reduced(P＜0.01),along with a significant de-crease in GSH level(P＜0.01).Treatment with PHSTF inhibited the mRNA and protein expression levels of ferroptosis-re-lated proteins,including SLC7A11 and GPX4(P＜0.01),suppressed the protein expression of Nrf2 and HO-1(P＜0.01),and enhanced the expression of Keap-1(P＜0.01).The Lip-1 partially restored the decrease in cell viability in-duced by PHSTF(P＜0.01),significantly up-regulated the protein expression levels of SLC7A11,GPX4,Nrf2 and HO-1,and suppressed the protein expression of Keap-1(P＜0.01).CONCLUSION:Total flavonoids of Pterocarya hupehen-sis Skan can inhibit the migration and invasion of non-small-cell lung cancer A549 cells,and induce the cell ferroptosis by regulating the Keap-1/Nrf2/HO-1 pathway.

AIM:To investigate the mechanism by which wogonin(WOG)induces ferroptosis in collagen-in-duced arthritis rat fibroblast-like synoviocytes(rat CIA-FLS cells)through the nuclear factor E2-related factor 2(NRF2)/heme oxygenase-1(HO-1)signaling pathway.METHODS:Rat CIA-FLS cells were divided into:control group,low,medium,and high dose of(25,50 and 100 μmol/L)WOG group,ferroptosis inhibitor(LIP-1)group,LIP-1+high dose WOG group,HO-1 agonist cobalt protoporphyrin(COPP)group,and COPP+high dose WOG group.CCK-8 assay was used for cell viability.Crystal violet staining was used for for cell morphology.The levels of oxidative stress markers gluta-thione(GSH),malondialdehyde(MDA),and superoxide dismutase(SOD)were measured.DCFH-DA fluorescent probe was used to detect the intracellular reactive oxygen species(ROS)content as well as Western blot to detect the protein ex-pression levels of Kelch-like ECH-associated protein 1(KEAP-1),NRF2 and HO-1.RESULTS:Compared with the nor-mal control group,administration of WOG treatment resulted in a significant decrease in CIA-FLS cell viability(P<0.01),a significant increase in the level of oxidative stress(P<0.01),a significant increase in the content of ROS(P<0.01),a significant decrease in the level of expression of NRF2 and HO-1 proteins(P<0.01),and a significant increase in the level of KEAP-1(P<0.01)in the rat.Compared with the WOG group,the LIP-1-treated group showed a significant increase in cell viability(P<0.01),a significant decrease in the level of oxidative stress(P<0.01),and a significant de-crease in the content of ROS(P<0.01).Compared with the WOG group,the addition of COPP resulted in a significant in-crease in the protein expression levels of NRF2 and HO-1(P<0.01)and a significant decrease in KEAP-1 levels(P<0.01).CONCLUSION:WOG can induce ferroptosis in rat CIA-FLS cells by promoting oxidative stress through the NRF2/HO-1 signaling pathway.

Objective:To identify risk factors for infections by multidrug-resistant organisms(MDRO)in elderly patients admitted to the intensive care unit(ICU)based on an retrospective analysis of data from the Medical Information Mart for Intensive Care-Ⅳ v2.0(MIMIC-Ⅳ v2.0).Methods:Structured query language was used to extract basic information, disease severity scores, laboratory test results, medications, medical procedures, and outcome events of elderly patients from MIMIC-Ⅳ v2.0.Patients were divided into a non-MDRO group and an MDRO group based on whether they had MDRO infections.Univariate analysis was performed according to the type of variables.For significant variables identified from univariate analysis, logistic regression was used to calculate the odds ratio and the 95% confidence interval for MDRO infections.The Kaplan-Meier method was used to draw survival curves.The log-rank test was used to analyze the 28-day survival rate.Results:A total of 31 237 cases were enrolled, including 26 032 with non-MDRO infections and 5 205 with MDRO infections.The MDRO infection rate was 16.7%.MDRO were most frequently found in urine cultures, and the most common bacteria belonged to the genus Enterobacter.Multivariate logistic regression analysis showed that age( P=0.006), being male( P<0.001), coronary artery disease( P<0.001), brain disease( P=0.042), hemoglobin( P<0.001), albumin( P<0.001), and mechanical ventilation( P<0.001)were protective factors against MDRO infections.Diabetes( P<0.001), COPD( P<0.001), chronic kidney disease( P<0.001), white blood cell( P=0.001), SAPS Ⅱ( P<0.001), previous use of antibiotics( P<0.001), use of proton pump inhibitors( P<0.001), glucocorticoids( P<0.001), immunosuppressants( P<0.001)and sedatives( P<0.001), and continuous renal replacement therapy(CRRT)( P=0.015)were risk factors for MDRO infections.The 28-day mortality rates of the non-MDRO and MDRO groups were 16.3% and 19.1%, respectively.The log-rank test showed that the difference between two groups was statistically significant( P<0.001). Conclusions:The mortality rate among patients with MDRO infections is higher than among those with non-MDRO infections.Previous use of antibiotics, use of proton pump inhibitors, glucocorticoids, immunosuppressants and sedatives, mechanical ventilation, and CRRT are risk factors for MDRO infections.

Objective:To investigate the risk factors for postsurgical gastroparesis syndrome (PGS) after laparoscopic complete mesocolic excision (CME) for right colon cancer.Methods:The clinical data of 358 patients who underwent laparoscopic CME for right colon cancer were retrospectively analyzed. Univariate and multivariate logistics regression were used to analyze the independent risk factors for PGS.Results:PGS occurred in 19 patients (4.8%). Logistic regression analysis showed that preoperative anxiety score (PAS-7)≥14 ( OR=6.450, P=0.039), preoperative serum albumin<35 g/L ( OR=9.302, P=0.011), colon cancer at hepatic flexura ( OR=9.782, P=0.007), No.206 group lymph node dissection ( OR=8.317, P=0.004), and intra-abdominal infection ( OR=5.755, P=0.043) were independent risk factors for PGS. Conclusion:Patient's preoperative health status, tumor location, scope of lymph node dissection and postoperative intra-abdominal infection are all risk factors related to PGS after CME for right colon cancer.

Objective:To investigate the clinical efficacy of totally laparoscopic pylorus-preserving gastrectomy (TLPPG) with preservation of the first branch of the right gastroepiploic vein in early gastric cancer (EGC).Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 38 EGC patients who were admitted to the Subei Hospital Affiliated to Yangzhou University from July 2018 to May 2021 were collected. There were 18 males and 20 females, aged 60 (range, 39?73) years. All patients underwent TLPPG with preservation of the first branch of the right gastroepiploic vein.Observation indicators: (1) surgical and postoperative condi-tions; (2) postoperative histopathological examination. (3) follow-up. Follow-up was conducted using outpatient examination, WeChat interview and medical record review to detect the nutritional status, residual stomach function, cholecystolithiasis, tumor recurrence and metastasis and death of patients. Follow-up was up to July 2022. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers. Results:(1) Surgical and postoperative conditions. All 38 patients underwent TLPPG with preservation of the first branch of the right gastroepiploic vein successfully, without laparotomy conversion. The operation time, volume of intraoperative blood loss, time to postoperative first flatus, time to postoperative first liquid food intake and duration of postoperative hospital stay of the 38 patients were (180±28)minutes, (58±38)mL, (2.7±0.6)days, (3.4±0.7)days and (10.3±2.8)days, respectively. Of the 38 patients, there were 6 cases with postoperative complications ≥grade Ⅱ of Clavien-Dindo classification. (2) Postoperative histopatho-logical examination. The tumor diameter, distance from proximal resection margin to tumor and distance from distal resection margin to tumor of the 38 patients were (1.8±0.5)cm, (3.4±0.2)cm and (4.3±0.4)cm, respectively. Both of proximal and distal resection margin was negative. Numbers of lymph node examined and numbers of lymph node examined in the No.6 lymph node of the 38 patients were 23.3±3.9 and 3.4±1.1, respectively. There were 38 cases with pathological T1 stage including 23 cases of T1a stage and 15 cases of T1b stage. There were 36 cases with pathological N0 stage and 2 cases with pathological N1 stage. There were 36 cases with pathological ⅠA stage and 2 cases with pathological ⅠB stage of TNM staging. (3) Follow-up. All 38 patients were followed up for 18(range, 12?48)months. The hemoglobin, serum albumin and total serum protein of the 38 patients were (125.4±5.8)g/L, (42.4±2.3)g/L and (71.6±2.1)g/L, respectively, at postoperative 6 month. Endo-scopy was used to evaluate the function of residual stomach of patients at postoperative 12 month. There were 4 patients with moderate amount of food remaining in the residual stomach. No patient suffered reflux esophagitis, reflux gastritis and bile reflux. None of the 38 patients received post-operative chemotherapy, and there was no tumor recurrence and metastasis or death occured in patient.Conclusion:TLPPG with preservation of the first branch of the right gastroepiploic vein is safe and feasible for the treatment of EGC patients with tumor located at 1/3 of the middle segment of stomach.

Objective:To analyze the genetic etiology and prognosis in fetuses with increased nuchal translucency (NT) in order to assist in the clinical prenatal genetic counseling and diagnosis.Methods:This study retrospectively enrolled 1 658 cases of singleton pregnancy (<35 years old) receiving invasive prenatal diagnosis, including karyotype analysis and/or chromosome microarray analysis or copy number variation (CNV) sequencing, due to NT value ≥2.5 mm in the first trimester in Henan Provincial People's Hospital from August 2014 to December 2021. They were divided into different groups according to the thickness of NT (≥2.5-<3.0, ≥3.0-<3.5, ≥3.5-<4.5, ≥4.5-<5.5, ≥5.5-<6.5 and ≥6.5 mm groups) and abnormal ultrasound findings (isolated increased NT group, increased NT complicated by soft markers/non-severe structural abnormality group and increased NT complicated by severe structural abnormality group). The results of invasive prenatal diagnosis and pregnancy outcomes were compared between different groups using Chi-square test and trend Chi-square test. Results:The detection rates of numerical abnormalities of chromosomes were 15.8% (262/1 658) and 17.6% (252/1 431) when the NT thickness cut-off value were 2.5 mm or 3.0 mm, respectively. Overall, the detection rate of numerical abnormalities of chromosomes increased with thickness of NT ( χ2trend=180.75, P<0.001), ranging from 6.6% (44/671) in the NT≥2.5-<3.5 mm group to 45.6% (113/248) in the NT≥5.5 mm group. The incidence of pathogenic/likely pathogenic CNV(P/LP CNV) did not increased with NT thickness ( χ2trend=3.26, P=0.071), and the highest detection rate was observed in the NT≥4.5-<5.5 mm group (9.0%, 19/211). The detection rate of numerical abnormalities of chromosomes plus P/LP CNV in the isolated NT≥2.5-<3.0 mm group and NT≥3.0-<3.5 mm group were 5.3% (10/188) and 9.6% (36/375), respectively, however, the difference was not statistically significant ( χ2=3.06, P=0.080). The detection rates of numerical abnormalities of chromosomes plus P/LP CNV in the isolated NT≥3.5-<4.5 mm group and NT≥2.5-<3.0 mm complicated by soft markers/ non-severe structural abnormality group were 12.7% (52/410) and 24.1% (7/29), respectively, and the risk were 2.6 times (95% CI: 1.3-5.2) and 5.7 times (95% CI: 2.0-16.4) of the isolated NT≥2.5-<3.0 mm group, respectively. The pregnancy termination rate increased with the NT thickness ( χ2trend=304.42, P<0.001), ranging from 10.8% (23/212) in the NT≥2.5-<3.0 mm group to 90.7% (117/129) in the NT≥6.5 mm group. After exclusion of the pregnancies terminated due to numerical abnormalities of chromosomes and P/LP CNV, 87.6% (862/984) of the fetus with increased NT were born alive. Conclusions:The detection rate of numerical abnormalities of chromosomes increases with the thickness of NT. Invasive prenatal diagnosis is required for non-advance aged singleton pregnant women when fetuses present with isolated NT≥2.5 mm with or without soft markers/structural abnormalities.

Objective:To retrospectively analyze the contrast-enhanced ultrasound(CEUS) features of primary hepatic lymphoepithelioma-like carcinoma (LELC) and investigate the value of CEUS in the diagnosis of hepatic LELC.Methods:The images of CEUS of 12 cases with hepatic LELC were retrospectively analyzed. The perfusion patterns and time of enhancement were observed.Results:During the arterial phase, 11 lesions showed diffuse enhancement, while 1 lesion showed rim-like enhancement. The mean time of begin enhancement, time to peak, time to iso-enhancement and slightly hypo-enhancement were (17.92±5.81)s, (24.50±5.52)s, (29.55±6.25)s, (45.50±25.15)s, respectively. Compared with adjacent liver parenchyma, rapid enhancement was observed in 11 lesions and synchronous enhancement was observed in 1 lesion.As to time of peak enhancement, hyper-enhancement and iso-enhancement were observed in 11 lesions and 1 lesion, respectively. In portal phase, 8 lesions manifested slight hypo-enhancement, 3 lesions with marked hypo-enhancement and 1 lesion with iso-enhancement.And in delayed phase, 10 lesions showed marked hypo-enhancement and 1 lesion with slight hypo-enhancement. Ten lesions showed peripheral hyper-enhancement like a bright ring in the portal and delayed phase.Conclusions:CEUS is valuable for the diagnosis and differential diagnosis of hepatic LELC.

OBJECTIVE: To analyze the genomic variation characteristics of fetal with abnormal serological screening, and to further explore the value of copy number variation (CNV) detection technology in prenatal diagnosis of fetal with abnormal serological screening. METHODS: 7617 singleton pregnant women who underwent amniocentesis for prenatal diagnosis solely due to abnormal Down's serological screening were selected. According to the results of serological screening, the patients were divided into high risk group, borderline risk group and single abnormal multiple of median (MOM) group. CMA and CNV-Seq were used to detect the copy number variation of amniotic fluid cell genomic DNA and combined with amniotic fluid cell karyotype analysis for prenatal diagnosis. Outpatient revisit combined with telephone inquiry was used for postnatal follow-up. RESULTS: Among 7617 amniotic fluid samples, aneuploidy was detected in 138cases (1.81%) by CMA and CNV-Seq, 9 cases of aneuploid chimerism were detected by amniotic fluid cell karyotype analysis, and 203 cases of fetus carrying pathogenic and likely pathogenic CNV (P/LP CNV) were detected, the variant of uncertain significance (VUS) was detected in 437 cases (5.7%), the overall abnormal detection rate was 10.33%. The detection rate of aneuploidy by CMA and CNV-Seq in three group were 123 cases (2.9%), 13 cases (1.3%) and 2 cases (0.4%), respectively,and showing no significant difference (χ ²=7.469, P=0.024). The detection rate of pathogenic and likely pathogenic CNV in three group were 163cases (2.6%); 24 cases (2.6%) and 16 cases (3.3%), respectively, and showing no significant difference (χ ²=0.764, P=0.682). The CMA reported 2.9% (108/3729)P/LP CNV, and CNV-seq reported 2.4% (95/3888)P/LP CNV, both tests showed similar detective capabilities (χ ²=1.504, P=0.22).The most popular P/LP CNV in this cohort were Xp22.31 microdeletion, 16p13.11 microduplication /microdeletion, 22q11.21 microduplication /microdeletion. In fetuses with P/LP CNV CNV, 59 fetuses were terminated pregnancy, and 32 of 112 fetuses born had abnormal clinical manifestations. Non-medically necessary termination of pregnancy occurred in 11 fetuses carrying VUS CNV, 322 fetuses carrying VUS CNV were born, 4 of them presented abnormal clinical manifestations. CONCLUSION Compared with the traditional chromosome karyotype, CMA and CNV-Seq can improve the detection rate of pathogenic and likely pathogenic CNV. CMA and CNV-seq can be used for first tier diagnosis of pregnant women in the general population with abnormal Down's serological screening.
Amniotic Fluid ; Aneuploidy ; Chromosome Aberrations ; DNA Copy Number Variations ; Female ; Genomics ; Humans ; Pregnancy ; Pregnancy Trimester, Second ; Pregnant Women ; Prenatal Diagnosis/methods* ; Technology

OBJECTIVE: To carry out genetic testing and prenatal diagnosis for a family affected with Duchenne muscular dystrophy (DMD). METHODS: Multiplex ligation dependent probe amplification (MLPA) was used to detect potential deletion and duplication of the Dystrophin gene. Haplotype analysis was performed using five short tandem repeat polymorphism loci (3'-STR, 5'-STR, 45-STR, 49-STR, 50-STR of the DMD gene. RESULTS: A same deletional mutation (exons 51-55) of the DMD gene was detected in two brothers but not in their mother. The patients and fetus have inherited different haplotypes of the Dystrophin gene from their mother, suggesting that the fetus was unaffected. CONCLUSION The mother was very likely to harbor germline mosaicism for the Dystrophin gene variant. Genetic testing of peripheral blood samples cannot rule out germline mosaicism in the mother. Prenatal diagnosis should be provided for subsequent pregnancies in this family.
Dystrophin ; genetics ; Exons ; Female ; Gene Deletion ; Germ-Line Mutation ; Humans ; Male ; Mosaicism ; Muscular Dystrophy, Duchenne ; genetics ; Pregnancy ; Prenatal Diagnosis

OBJECTIVE: To provide genetic testing for two brothers with mental retardation and epilepsy. METHODS: Array comparative genomic hybridization (aCGH) was used to detect copy number variations in the two patients, their parents and maternal grandparents. Methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) was utilized to delineate the deleted region in the pedigree. RESULTS: A 138 kb deletion in 15q11.2 region was detected by aCGH in both patients, which encompassed part of the UBE3A gene. MS-MLPA has narrowed down the region to exons 8 to 14 of the UBE3A gene. The same deletion was also found in their mother and grandfather. CONCLUSION The pathogenesis of this rare form of recurrent Angelman syndrome may be attributed to the partial deletion of maternal UBE3A gene.
Angelman Syndrome ; Comparative Genomic Hybridization ; DNA Copy Number Variations ; Female ; Gene Deletion ; Humans ; Male ; Sequence Deletion ; Ubiquitin-Protein Ligases