Objective:To investigate the effect of DNA methylation of laminin α3 (LAMA3) on the prognosis of platinum-resistant epithelial ovarian cancer (EOC) and its possible mechanism.Methods:(1) The relationship between DNA methylation of LAMA3 and platinum resistance in EOC was evaluated by bioinformatics. (2) A total of 67 EOC patients treated at Guangxi Medical University Cancer Hospital from January 2000 to December 2012 were selected to detect the levels of LAMA3 DNA methylation in EOC tissues using pyrophosphate sequencing technology to explore its diagnostic efficacy for platinum resistance and prognosis in EOC patients. Furthermore, its impact on chemotherapy efficacy and prognosis of platinum resistant EOC patients were also analyzed.Results:(1) Ten proteins highly interacting with LAMA3 were screened from the Gene Interaction Retrieval Platform (STRING) database, including laminin β (LAMB) 3, laminin γ (LAMC) 3, integrin α (ITGA) 6, intestine protein β4 (ITGB4), ITGA3, LAMC1,LAMB2, dystrophin associated glycoprotein 1 (DAG1), LAMB1 and cytochrome P450c17α (COL17A1) protein; kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis showed that LAMA3 and its related interacting proteins participate in the regulation of malignant tumor occurrence and development through signaling pathways such as apoptosis, cell cycle, DNA damage response, epithelial mesenchymal transition (EMT), androgen receptor (AR), estrogen receptor (ER), phosphatidylinositol 3 kinase (PI3K)/protein kinase B (Akt), RAS/mitogen activated protein kinase (MAPK), receptor tyrosine kinase (RTK), tuberous sclerosis protein complex (TSC)/mammalian target of rapamycin (mTOR), and their expression levels were related to the sensitivity of chemotherapy drugs such as cisplatin in EOC. (2) Our clinical data analysis found that the LAMA3 DNA methylation level in EOC tissue of the platinum-sensitive group (35 cases) was 71% (25/35), which was higher than 69% (22/32) in the platinum-resistant group (32 cases), with statistically insignificant difference ( χ2=0.057, P=0.811). The area under the curve (AUC) of LAMA3 DNA methylation level for assessing platinum resistance in EOC was 0.601, and the AUC for predicting EOC patient prognosis was 0.686. The chemotherapy efficacy of EOC patients with high methylation of LAMA3 DNA was worse than that of patients with low methylation, 50% (12/24) vs 15/15, with statistically significant difference ( χ2=10.833, P=0.001). The level of LAMA3 DNA methylation had a significant impact on the progression free survival and overall survival of EOC patients (both P<0.05). Conclusion:The level of LAMA3 DNA methylation has certain diagnostic and predictive value for platinum resistance and prognosis in EOC patients, which may be closely related to the regulatory mechanism, platinum resistance and prognosis of EOC.

Oral potentially malignant disorders (OPMDs) are precursors of oral squamous cell carcinoma (OSCC). Deregulated cellular energy metabolism is a critical hallmark of cancer cells. Peroxisome proliferator-activated receptor-gamma coactivator-1 alpha (PGC1α) plays vital role in mitochondrial energy metabolism. However, the molecular mechanism of PGC1α on OPMDs progression is less unclear. Therefore, we investigated the effects of knockdown PGC1α on human dysplastic oral keratinocytes (DOKs) comprehensively, including cell proliferation, cell cycle, apoptosis, xenograft tumor, mitochondrial DNA (mtDNA), mitochondrial electron transport chain complexes (ETC), reactive oxygen species (ROS), oxygen consumption rate (OCR), extracellular acidification rate (ECAR), and glucose uptake. We found that knockdown PGC1α significantly inhibited the proliferation of DOKs in vitro and tumor growth in vivo, induced S-phase arrest, and suppressed PI3K/Akt signaling pathway without affecting cell apoptosis. Mechanistically, downregulated of PGC1α decreased mtDNA, ETC, and OCR, while enhancing ROS, glucose uptake, ECAR, and glycolysis by regulating lactate dehydrogenase A (LDHA). Moreover, SR18292 (an inhibitor of PGC1α) induced oxidative phosphorylation dysfunction of DOKs and declined DOK xenograft tumor progression. Thus, our work suggests that PGC1α plays a crucial role in cell proliferation by reprograming energy metabolism and interfering with energy metabolism, acting as a potential therapeutic target for OPMDs.
Humans ; Carcinoma, Squamous Cell/metabolism* ; Cell Proliferation ; DNA, Mitochondrial ; Energy Metabolism ; Glucose ; Mouth Neoplasms/metabolism* ; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha/metabolism* ; Phosphatidylinositol 3-Kinases ; Reactive Oxygen Species

Objective:To comprehensively evaluate the tricuspid valve, right heart anatomical characteristics and related dynamic parameters in patients with different degrees of functional tricuspid regurgitation (FTR) using four-dimensional auto tricuspid valve quantitative(4D Auto TVQ), four-dimensional auto right ventricle quantitative(4D Auto RVQ), and four-dimensional auto left atrium quantitative(4D Auto LAQ), and to investigate the structural and functional changes of the tricuspid valve and right heart in them.Methods:Sixty-three patients with FTR diagnosed by echocardiography at the First Affiliated Hospital of Guangxi Medical University from February to July 2022 were prospectively selected as the case group, including 30 patients with mild FTR and 33 patients with moderate or above FTR, and 30 healthy subjects were selected as the control group. Transthoracic echocardiography was used for two-dimensional and three-dimensional image acquisition of the heart. The tricuspid regurgitation volume, left ventricular ejection fraction (LVEF), right ventricular global strain (RVGS) were measured by 2D images, and pulmonary artery systolic pressure (PASP) were measured from the tricuspid regurgitation pressure difference. The 3D images were imported into EchoPAC 204 to obtain the tricuspid valve, right heart structure and related dynamic parameters. The annulus area (AA), annulus perimeter(AP), spherical index (SI), annulus area change fraction (AC), coaptation point height (CPH), and tenting volume (TV) were measured by 4D Auto TVQ. The right atrial maximum volume (RAVmax) and right atrial minimum volume (RAVmin) were measured by 4D Auto LAQ. Right ventricular end-diastolic volume (RVEDV), right ventricular end-systolic volume (RVESV), right ventricular fractional area change (RVFAC) and tricuspid annular plane systolic excursion (TAPSE) were measured by 4D Auto RVQ. After standardizing the dimension parameters with body surface area (BSA), the differences in the above parameters were compared between the three groups, the correlation between regurgitant volume and each parameter was compared by correlation analysis, and the independent factors of increased tricuspid regurgitant volume were investigated by univariate and multivariate linear regression analysis.Results:There were statistically significant differences in PASP, AA/BSA, AP/BSA, AC, TV, RAVmax/BSA, RAVmin/BSA, RVFAC, RVGS, and TAPSE between the three groups (all P<0.05). There were statistically significant differences in LVEF, CPH, RVEDV/BSA, and RVESV/BSA in the moderate and above FTR group compared with the control and mild FTR groups (all P<0.05). Correlation analysis showed that RAVmin was the most highly correlated with tricuspid regurgitant volume ( r=0.875, P<0.001) and TV and end-systolic annulus area(ESAA) were highly correlated with tricuspid regurgitant volume ( r=0.747, 0.683; both P<0.001) in patients with FTR. Multifactorial linear regression showed that RAVmin, TV and regurgitant volume were independently positively correlated (β=0.721, 0.205; both P<0.05). Conclusions:The four quantification technique can provide valid structural and functional information by quantifying the tricuspid valve as well as the right heart in patients with FTR, and RAVmin and TV are independent correlates of increased tricuspid regurgitant volume.

Objective To establish a UHPLC method for the determination of zolpidem tartrate tablets after radiation, and to investigate the effect of different radiation doses on the content of zolpidem tartrate tablets. Methods Ultra high performance liquid chromatography was used. The content of zolpidem tartrate tablets irradiated by γ-ray was determined. Using C₁₈ column, acetonitrile methanol-0.05 mol/L phosphoric acid solution (the pH value as 5.5 with triethylamine) (18∶26∶56) was used as the mobile phase. The flow rate was 0.7 ml/min, and the detection wavelength was 254 nm. Results The method validation showed good linearity in the concentration range of 5-80 μg/ml (r=0.999 6); The average recovery was 98.2%, RSD was 1.72%, and the repeatability was 0.87%. The contents of zolpidem tartrate were 105.1%, 106.4%, 102.7% and 105.4% under 0, 8, 25 and 80 kGy radiation. Conclusion UHPLC has accurate results with short analysis cycle in this study. It is suitable for the determination of zolpidem tartrate tablets after radiation. The content of zolpidem tartrate tablets remained basically unchanged after radiation.

Objective To analyze the electrophysiological characteristics of Charcot-Marie-Tooth (CMT) disease 1A,1X,2A and myelin protein zero (MPZ)-related CMT in Chinese patients.Methods Baseline electrophysiological data from 36 CMT1A patients,78 CMT1X patients,31 CMT2A patients and 10 MPZ-related CMT patients in the Third Xiangya Hospital and Xiangya Hospital of Central South University during 2004-2018 were analyzed.Electrophysiological recordings were taken from the upper limbs (median nerve,ulnar nerve) and lower limbs (tibial nerve,peroneal nerve).Demyelination in different nerve segments was assessed by measurement of distal motor latency,motor nerve conduction velocity (MNCV),sensory nerve conduction velocity and F-wave latency,and calculation of conduction block,terminal latency index (TLI) and modified F ratio (MFR);Axonal degeneration was assessed by measuring compound motor action potential (CMAP) and sensory nerve action potential.The relationship between the gender,age at onset,duration,Overall Neuropathy Limitation Scale (ONLS) score and indexes of peripheral nerve electrophysiology was statistically analyzed.Results The peripheral nerves of CMT1A patients were characterized by uniform demyelination and axonal degeneration.MNCV ((21.39± 6.72) m/s) and CMAP amplitude (2.40 (3.50) mY) of median nerve of CMT1A patients were decreased.The peripheral nerves of CMT1X patients were also characterized by uniform demyelination and axonal degeneration.MNCV (35.20 (6.77) m/s) and CMAP amplitude (2.60 (3.79) mY) of median nerve of CMT1X patients were decreased.CMT2A patients showed axonal degeneration of the peripheral nerves and CMAP amplitude ((4.75 ±2.38) mV) of median nerve of CMT2A patients was decreased.The electrophysiological data in MPZ-related CMT patients demonstrated variability.The TLI and MFR for the median and ulnar nerves in these four subtypes were normal.MNCV (r=0.423,P=0.025) of median nerve in CMT1A patients was positively correlated with age at onset.MNCV (r=0.782,P=-0.013) of median nerve in MPZ-related CMT patients was positively correlated with age at onset.CMAP amplitude (r=0.652,P＜0.01) of median nerve in CMT2A patients was positively correlated with age at onset.Demyelination and axonal degeneration in male CMT1X patients were relatively more severe than those in female patients,and MNCV (Z=-3.300,P＜0.01) and CMAP amplitude (Z=-3.960,P＜0.01) of median nerve,MNCV (Z=-2.56,P=0.011) and CMAP amplitude (Z=-2.311,P=0.048) of ulnar nerve of male patients were lower than those of female patients.The ONLS score of CMT1A (r=-0.494,P＜0.01),CMT1X (r=-0.596,P＜0.01) and CMT2A patients (r=-0.494,P=0.012) was inversely associated with CMAP amplitude.Conclusions The electrophysiological characteristics of CMT1A,CMT1X,CMT2A and MPZ-related CMT are different.Electrophysiological examinations are the basis of clinical classification and could provide guidance for further genetic testing and diagnosis.CMAP amplitude may serve as an objective index to assess the severity of functional disability in CMT patients.

Objective To observe the changes of renal tubular injury and the extent of interstitial fibrosis in the C57BL/6 mouse models of chronic kidney disease(CKD),and provide experimental animal evidence for study of the pro-gression of acute kidney injury(AKI)to chronic kidney disease as well as its mechanisms. Methods Twenty-four 8-week-old male C57BL/6 mice were randomly and equally divided into control group, low-dose, medium-dose, and high-dose cisplatin groups,6 mice in each group. Mice in the cisplatin groups were administrated with 5,7 or 10 mg/kg cispla-tin by intraperitoneal injection once a week for 4 weeks. Plasma creatinine and 24-hour urinary protein were detected to as-sess the renal function. The mice were sacrificed, and plasma and kidney samples were collected for subsequent tests. Pathological changes were observed using periodic acid-Schiff(PAS)staining. To evaluate renal tubules injury, the ex-pression of kidney injury molecule 1(KIM-1)was examined by immunohistochemistry and the level of urinary N-Acetyl-β-D-glucosaminidase was detected with a commercial kit. The infiltration of CD3-positive T cells and F4/80-positive macro-phages was observed by immunohistochemistry(IHC)and immunofluorescence. The expression of collagen I and α-smooth muscle actin(α-SMA)were tested by immunohistochemistry to assess the renal fibrosis, while total kidney collagen was detected by Picrosirius red staining. Results In contrast to the normal control group,the kidney injury became more seri-ous in the cisplatin-treated mice as cisplatin concentration increased. Particularly,significant kidney damage was observed in the high-dose cisplatin group. Compared with the control group,the plasma creatinine and 24-hour urinary protein were significantly increased in the high-dose cisplatin group(P<0.05 and P<0.001)indicating impaired renal function. Mor-phologically,numerous clear vacuoles and necrosis were present in renal tubule epithelial cells in the high-dose cisplatin group. The expression of KIM-1 was markedly up-regulated and the level of urinary NAG was elevated. Infiltration of CD3-positive T cells and F4/80-positive macrophages was enhanced in the mice of high-dose cisplatin group. Data from immuno-histochemistry and picrosirius red staining showed that mice of the high-dose cisplatin group developed renal fibrosis evi-denced by markedly up-regulated expression of collagen I and α-SMA. Conclusions Repeated administration of 10 mg /kg cisplatin for 4 weeks can induce chronic renal insufficiency in mice,which may serve as a novel model for the research on underlying mechanisms of progression from acute kidney injury to chronic kidney disease.

Objective To investigate clinical effects and prognostic factors of transabdominal laparoscopic-assisted and open radical resection for Siewert type Ⅱ and Ⅲ adenocarcinoma of the esophagogastric junction (AEG).Methods The retrospective cohort and case-control studies were conducted.The clinicopathologieal data of 84 patients with Siewert type Ⅱ and Ⅲ AEG who were admitted to the First Affiliated Hospital of Xiamen University from January 2014 to January 2017 were collected.Among 84 patients,42 undergoing transabdominal laparoscopic-assisted radical gastectomy (LARG) were allocated into LARG group and 42 undergoing transabdominal open radical gastectomy (ORG) were allocated into ORG group.Observation indicators:(1) comparison of intraoperative and postoperative recovery situations between groups;(2)comparison of follow-up and survival situations between groups;(3) prognostic analysis of patients with Siewert type Ⅱ and Ⅲ AEG undergoing transabdominal radical resection.Follow-up using outpatient examination and telephone interview was performed to detect postoperative survival situations,tumor recurrence and metastasis of patients up to January 2018.Measurement data with normal distribution were representde as-x±s,and comparison between groups was analyzed using the t test.Measurement data with skewed distribution were described as M (range).Comparison of count data were analyzed using the ehi-square test or Fisher exact probability.Ordinal data were compared between groups using the Mann-Whitney U nonparametric test.The survival rate and curve were respectively calculated and drawn by the Kaplan-Meier method,and Log-rank test was used for survival and univariate analyses.Multivariate analysis was done by COX proportional hazard model.Results (1) Comparison of intraoperative and postoperative recovery situations between groups:patients with Siewert type Ⅱ and Ⅲ AEG in the LARG and ORG group underwent successful transabdominal radical resection,without conversion to thoracotomy.All patients in the LARG group underwent esophagojejunostomy with circular stapler device,38 and 4 patients in the ORG group underwent esophagojejunostomy with circular stapler and linear cut stapler respectively.Operation time,volume of intraoperative blood loss,length of incision,time of postoperative analgesia,cases with anastomotic bleeding,anastomotic leakage,abdominal bleeding,incisional infection,pulmonary infection,abdominal infection and reflux esophagitis of grade Ⅰ ～ Ⅱ postoperative complications and duration of postoperative stay were respectively (261±50)minutes,(119±111)mL,(7.8±1.6)cm,(2.1±1.3)days,1,1,0,0,1,0,0,(12.8 ± 1.9) days in LARG group and (216 ± 52) minutes,(230± 178) mL,(17.3± 1.8) cm,(3.4±1.2)days,2,0,2,2,2,1,2,(18.4±15.3)days in ORG group,with statistically significances between groups (t =2.357,2.960,2.195,2.013,x2 =5.486,t =2.125,P＜0.05).All patients with complications were improved by symptomatic treatment.(2) Comparison of follow-up and survival situations between groups:81 of 84patients including 41 in LARG group and 40 in ORG group were followed up for 6-48 months,with a median time of 29 months.The postoperative 2-year overall and tumor-free survival rates were respectively 85.1％ and 82.1％ of 41 patients in LARG group and 83.1％ and 79.3％ of 40 patients in ORG group,with no statistically significance between groups (x2 =0.013,0.049,P＞0.05).(3) Prognostic analysis of patients with Siewert type Ⅱ and Ⅲ AEG undergoing transabdominal radical resection:results of univariate analysis showed that tumor diameter,tumor TNM staging,tumor T staging,tumor N staging and postoperative adjuvant chemotherapy were related factors affecting prognosis of patients with Siewert type Ⅱ and Ⅲ AEG undergoing transabdominal radical resection (x2 =8.349,14.376,9.732,17.250,8.012,P＜0.05).Results of multivariate analysis showed that tumor TNM staging and postoperative adjuvant chemotherapy were independent factors affecting prognosis of patients with Siewert type Ⅱ and Ⅲ AEG undergoing transabdominal radical resection (risk ratio =4.305,0.031,95％ confidence interval:1.858-9.977,0.004-0.246,P＜0.05).Conclusions Transabdominal laparoscopicassisted radical resection for AEG is safe and feasible,with advantage of minimally invasiveness,having equivalent long-term effects compared to open surgery.Tumor TNM staging and postoperative chemotherapy are independent factors affecting prognosis of patients with Siewert type Ⅱ and Ⅲ AEG undergoing transabdominal radical resection.

UNLABELLEDObjective To explore the effects of sperm DNA integrity rate, acrosome integrity rate and acrosome reaction rate on the outcomes of rescue intracytoplasmic sperm injection (ICSI).

METHODSThis retrospective analysis was conducted among 97 infertile couples receiving rescue ICSI due to failure of in vitro fertilization procedures in our Reproductive Medicine Center. Of these 97 women, 41 had clinical pregnancy and 56 did not, and the effects of sperm DNA integrity rate (estimated by DNA fragmentation index, DFI), acrosome integrity rate and acrosome reaction rate on rescue ICSI outcomes were analyzed.

RESULTSNo significant difference was found in paternal age, testosterone value, testicular volume, FSH, female patient' age or the number of eggs retrieved between the two groups (P>0.05), but the infertility years was significantly shorter in the pregnancy group than in the non-pregnancy group (P<0.05). The fertilization rate and cleavage rate were similar between the two groups (P>0.05), but the good embryo rate was significantly higher in the pregnancy group (P<0.05). The sperm DNA integrity or acrosome reaction rate did not differ significantly between the two groups (P>0.05), but the acrosome integrity rate was significantly higher in the pregnancy group (P<0.05). The sperm DNA integrity rate, acrosome integrity or acrosome reaction rate were not correlated with the fertilization rate, cleavage rate or good embryo rate (P>0.05). The pregnancy rate, twin and single fetus rates were 42.3%, 10.3% and 32.0% in this cohort after recue ICSI, respectively.

CONCLUSIONRescue ICSI is an effective treatment after failed in vitro fertilization procedure, and sperm acrosome integrity rate is associated with the outcome of rescue ICSI.

Acrosome ; pathology ; Acrosome Reaction ; DNA Fragmentation ; Female ; Fertilization ; Fertilization in Vitro ; Humans ; Infertility ; Male ; Pregnancy ; Pregnancy Rate ; Retrospective Studies ; Sperm Injections, Intracytoplasmic

The dysfunction of subthalamic nucleus is the main cause of Parkinson's disease. Local field potentials in human subthalamic nucleus contain rich physiological information. The present study aimed to quantify the oscillatory and dynamic characteristics of local field potentials of subthalamic nucleus, and their modulation by the medication therapy for Parkinson's disease. The subthalamic nucleus local field potentials were recorded from patients with Parkinson's disease at the states of on and off medication. The oscillatory features were characterised with the power spectral analysis. Furthermore, the dynamic features were characterised with time-frequency analysis and the coefficient of variation measure of the time-variant power at each frequency. There was a dominant peak at low beta-band with medication off. The medication significantly suppressed the low beta component and increased the theta component. The amplitude fluctuation of neural oscillations was measured by the coefficient of variation. The coefficient of variation in 4-7 Hz and 60-66 Hz was increased by medication. These effects proved that medication had significant modulation to subthalamic nucleus neural oscillatory synchronization and dynamic features. The subthalamic nucleus neural activities tend towards stable state under medication. The findings would provide quantitative biomarkers for studying the mechanisms of Parkinson's disease and clinical treatments of medication or deep brain stimulation.
Antiparkinson Agents ; therapeutic use ; Beta Rhythm ; Electrodes ; Evoked Potentials ; Humans ; Oscillometry ; Parkinson Disease ; drug therapy ; physiopathology ; Subthalamic Nucleus ; physiopathology ; Theta Rhythm

Objective To evaluate correlations between CT features and clinical activity index of Crohn’s disease(CD).Methods The CTE datasets from 37 patients with moderate active,severe active and remission stages CD were retrospectively analyzed.To ob-serve imaging features of intestinal wall,parenteral and complications.And analyze whether there was an association between imaging findings and clinical activity.Results A total of 147 bowel segments in 37 cases were involved.Wall thickening (intestinal wall≥4 mm)were observed in 121 segments.An average thickness was (12.1±3.2)mm.There are 42 bowel segments of trilaminar mural stratifica-tion,71 of the double-layed stratification and 34 of non-stratification.Enlarged mesenteric lymph nodes were observed in 20 patients. Comb sign was observed in 27 patients.Higher density of surrounding fat appeared in 22 cases.The other 37 cases inluced 4 cases with abdominal abscess,7 cases with fistula,1 6 cases with intestinal obstruction or stenosis and 1 7 cases of perianal disease.The in-testinal stratification,wall thickening,comb sign,mesenteric fat density increased,abdominal abscess,intestinal stenosis and obstruc-tion were closely associated with clinical activity.Conclusion CTE images could provide information about intestinal wall,parenteral changes and complications of Crohn’s disease,and effectively help identifying remission and active stages of Crohn’s disease.