Objective To explore the application value of anhydrous ethanol as an alternative to methanol in the preparation of chromosomal specimens from peripheral blood lymphocytes, and to establish a set of quantitative analytical methods for objectively evaluating the effectiveness of specimen preparation. Methods Residual blood samples from routine laboratory slide preparation were used for lymphocyte culture. The standard slide preparation method was employed. The fixative in the control group was methanol and glacial acetic acid (3∶1). Four experimental groups were set up based on the ratio of anhydrous ethanol to glacial acetic acid in the fixative (volume ratios of 3∶1, 5∶1, 7∶1, and 9∶1 for experimental groups 1, 2, 3, and 4, respectively). A chromosomal analysis was conducted using an automated chromosome scanning/image analysis system to evaluate the morphology and dispersion of metaphase chromosomes in both control and experimental groups. Comparisons were made between the control and experimental groups regarding the dic + r aberration rate, ace aberration rate, chromosomal aberration rate, chromosome dispersion index, chromosome overlapping ratio, and dispersion index/overlapping ratio. Results Microscopic evaluation revealed that the preparation quality of experimental groups 1 and 2 was comparable to the control group. No statistically significant differences were observed in dic + r aberration rate between each of the experimental groups and the control (P > 0.05). All experimental groups except group 4 showed no significant differences in ace aberration rate and chromosome aberration rate compared with the control group (P > 0.05). Experimental groups 1 and 2 showed no significant differences in chromosome dispersion index, overlapping ratio, and dispersion index/overlapping ratio compared with the control group (P > 0.05). Conclusion A mixture of anhydrous ethanol and glacial acetic acid at a 5∶1 ratio is recommended for use as a fixative in the preparation of chromosomal specimens from peripheral blood lymphocytes. A quantitative index system for assessing the quality of chromosomal specimens was established, enabling objective evaluation of slide preparation effectiveness.

Objective·To evaluate the quality of clinical practice guidelines of obstructive sleep apnea(OSA)published worldwide.Methods·The guidelines of OSA were retrieved in PubMed,Embase,China National Knowledge Infrastructure(CNKI),Wanfang Data,SinoMed,MedSci,The Cochrane Library,and the websites such as Medlive,U.S.Preventive Services Task Force(USPSTF),National Institute for Health and Care Excellence(NICE),New Zealand Guidelines Group(NZGG),Scottish Intercollegiate Guidelines Network(SIGN),and Guidelines International Network(GIN)from establishment to December 2022.Two reviewers screened the literature and extracted the main information independently,using the Appraisal of Guidelines for Research and Evaluation Ⅱ(AGREE Ⅱ)and Reporting Items for Practice Guidelines in Healthcare(RIGHT)to evaluate the quality of the included OSA guidelines.Subgroup analysis was performed according to the publication regions of guidelines.The inter-evaluator consistency test was also performed and the results were expressed as the intra-class correlation coefficient(ICC).All the included guidelines were read entirely and the clinical questions they raised were summarized.Results·A total of 35 OSA guidelines were included.The ICC value of 0.975 showed a good inter-evaluator agreement.The results of AGREE Ⅱ showed that the average score of all guidelines was(63.60±16.45)%,with a minimum of 23.40%and a maximum of 91.67%.In the six domains,the scores of"Rigor of development"[(56.07±25.89)%]and"Applicability"[(53.57±15.52)%]were relative low.The average reporting rate of RIGHT of all the included guidelines was(67.84±20.03)%,with a minimum of 14.29%and a maximum of 94.29%,and the three domains with the lowest reporting rates were"Review and quality assurance"[(31.40±45.51)%],"Funding and conflict of interest declaration and management"[(56.43±33.95)%]and"Other aspects"[(56.19±36.85)%].Subgroup analysis showed that guidelines in Asian had a lower score in"Rigor of development"and a lower overall score of AGREE Ⅱ than the guidelines in America and Europe(both P<0.05),and the reporting rates in the domains of"Evidence"and"Other information"of RIGHT of the Asian guidelines were also lower than those in the guidelines in America and Europe(P<0.05).These guidelines focused on 42 clinical questions which were classified to 3 aspects,i.e.screening and diagnosis,treatment and long-term management of OSA.Conclusion·The quality of current global OSA guidelines varies a lot,and they need to be strengthened in terms of rigor of development,applicability,review and quality assurance,funding and conflict of interest declaration and management,especially those in Asia.

Objective:To compare the recurrence of common bile duct stones (CBDS) in patients with gallstones and concurrent CBDS treated by two surgical approaches: synchronous laparoscopic cholecystectomy (LC) combined with laparoscopic common bile duct exploration (LCBDE) (LC+LCBDE) and synchronous LC combined with intraoperative endoscopic sphincterotomy (IO-EST) (LC+IO-EST).Methods:From Apr 2013 to Apr 2020, the clinical data of 903 patients with gallstones with CBDS who were admitted to the Clinical Medical College, Yangzhou University were retrospectively analyzed.Results:Based on the chosen surgical method, we categorized 389 cases into group A (LC+LCBDE) and 514 cases into group B (LC+IO-EST). Our findings revealed that group A had a significantly lower rate of CBDS recurrence and re-recurrent CBDS compared to group B (4.4% vs. 8.4%, P=0.024; 0.8% vs. 3.1%, P=0.010). Moreover, Logistic regression analysis after inverse probability of treatment weighting, revealed that the surgical approach implemented in group A was an independent protective factor of recurrent CBDS and second recurrence of CBDS ( OR=0.482, 95% CI: 0.365-0.637, P<0.001; OR=0.118, 95% CI:0.080-0.173, P<0.001). Conclusion:LC+LCBDE is an optimal treatment option to LC+IO-EST for patients with gallstones combined with CBDS and common bile duct ≥8 mm.

Objective To observe the effects of Levo-tetrahydropalmatine(l-THP)on the expression,regression and relapse of conditioned place preference(CPP)in ketamine induced rats,and to detect the content of dopamine(DA)in the striatum(caudate putamen,CPu)of the rat brain at different time points.Methods Ketamine addiction rat model was established by CPP.The effects of l-THP on the expression,regression and relapse of ketamine induced rat CPP were investigated using CPP score as the index.The content of DA in CPu of rats was determined by ultra-performance liquid chromatography coupled to tandem mass spectrometry(UPLC-MS/MS)after ketamine administration and l-THP intervention at 30 min,60 min,90 min,120 min and 150 min.Results It indicated that 1-THP could decrease the expression of CPP in ketamine induced rats,promote the process of CPP resolution and inhibit the process of relapse.In addition,l-THP combined with ketamine administration significantly inhibited the ketamine-induced increase in DA content in the CPu of the rats.Conclusion The mechanism of l-THP inhibiting the reward effect of ketamine may be related to blocking DA receptors and reducing the release of DA neurotransmitters.l-THP has potential implications for the treatment of ketamine addiction.

OBJECTIVE To assess the efficacy of bioabsorbable steroid-releasing sinus stents for improving surgical outcomes and subjective symptoms when placed in the bilateral frontal sinus opening(FSO)following full functional endoscopic sinus surgery in patients with chronic rhinosinusitis with nasal polyps(CRSwNP).METHODS CRSwNP patients who had under full functional endoscopic sinus surgery with complete data of nasal endoscopy and sinus computed tomography data were identified and included in the study.The patients were divided into a control group consisting of patients receiving only full functional endoscopic sinus surgery(n=92)and a stent group consisting of patients receiving full functional endoscopic sinus surgery combined with placement of steroid implants in both FSO(n=38).The visual analogue scale(VAS)subjective symptom scores and surgical outcomes were compared preoperatively,and on postoperative day(PD30 and PD90)between the two groups.RESULTS Compared to baseline,the overall symptom VAS scores of patients after operation decreased significantly in both groups(P<0.05),and the degree of improvement of overall symptoms in the stent group was significantly better than in the control group(P<0.05).On PD30,the proportion of patients requiring postoperative interventions for bilateral FSO was reduced by 42.3%in the stent group,and was significantly lower than in the control group(P<0.05).Compared to the control group,the proportion of patients needing postoperative intervention in both ethmoid sinus on the stent group decreased by 17.7%(P>0.05).The results at PD90 were consistent with those at PD30.CONCLUSION Full functional endoscopic sinus surgery in combination with bilateral frontal sinus stent implantation is better than full functional endoscopic sinus surgery alone.

Objective To observe the value of dual-energy CT(DECT)radiomics combined with clinical and CT features for predicting differentiation degree of gastric adenocarcinoma(GAC).Methods Totally 254 patients with GAC were prospectively analyzed and divided into high-grade group(low differentiation GAC,n=88)and low-grade group(middle-high differentiation GAC,n=166)according to pathological results.The patients were also divided into training set(n=203,including 70 high-grade and 133 low-grade GAC)and verification set(n=51,including 18 high-grade and 33 low-grade GAC)at the ratio of 8∶2.Radiomics features were extracted and screened based on venous phase single-level(40,70,100 and 140 keV)DECT,and a multi-energy radiomics model was constructed to predict GAC classification.Univariate analysis and multivariate logistic regression were used to analyze clinical and CT features as well as DECT parameters in training set to construct a clinic-CT model.Then a combined model was constructed through combining clinic-CT model with radiomics model.The predictive efficacy of the models were evaluated,and the calibration degree of the combined model was assessed.Results The area under the curve(AUC)of clinic-CT model,multi-energy radiomics model and combined model was 0.74,0.75 and 0.78 in training set,and 0.73,0.77 and 0.78 in verification set,respectively.Except for AUC of combined model was higher than that of clinic-CT model in training set(P＜0.05),no significant difference of AUC was found among models in training set nor verification set(all P＞0.05).The calibration degree of combined model was good in both training set and verification set(both P＞0.05).Conclusion DECT radiomics combined with clinical and CT features could effectively predict differentiation degree of GAC.

Objective:To design an improved lamina hook system and compare its biomechanical properties with traditional lamina hook system in fixation of lumbar spondylolysis.Methods:The thin layer CT data of the lumbosacral vertebrae of 20 healthy young male servicemen who underwent physical examination in the outpatient department of the 940th Hospital of Joint Logistics Support Force of PLA from January 2021 to August 2022 were collected. The age of the subjects was 20-30 years [(25.0±3.0)years]. A 3-dimensional model of the L 5 vertebral body was constructed using the 3-dimensional modeling software. The new improved lamina hook was designed according to the measurements including the thickness of the middle area, the longest longitudinal diameter, the curvature radius of the lower edge, the angle between the upper and lower tail ends, the thickness of the lower edge, and the longest diameter of the lower edge of the bilateral L 5 vertebral plates. One serviceman was selected from the aforementioned group to construct a linear finite element model of segments L 4-S using the 3-dimensional virtual software (normal model, model A), based on which, the L 5 bilateral spondylolysis model (model B), improved lamina hook model (model C) and traditional lamina hook models (model D) were designed. By constraining both sides of the sacrum and applying a longitudinal load of 400 N on the L 4 vertebral body, the upper 1/3 gravity of the body was simulated, and with a bending moment of 10 N·m along the X, Y, and Z directions, motions of forward flexion, backward extension, lateral bending, rotation, etc were simulated. The range of motion of segment L 4/5 and L 5/S 1 of model A was evaluated and compared with the findings of the previous researches to verify its effectiveness. The overall range of motion of models A, B, C, and D, the range of motion of segment L 4/5 and L 5/S 1, the maximum overall displacement, the maximum displacement and stress of the isthmus, the stress distribution and maximum stress of internal fixation of models C and D, and the stress distribution and maximum stress of the vertebral body of models C and D were compared. Results:(1) During forward flexion, backward extension, lateral bending and rotation, the range of motion of model A was 5.01°, 4.03°, 3.91° and 1.42° in segment L 4/5, and was 4.62°, 2.51°, 2.40° and 1.23° in segment L 5/S 1. (2) The overall range of motion, range of motion of segment L 4/5 and L 5/S 1 and maximum overall displacement of models A, C, and D were similar in axial compression, forward flexion, backward extension, left bending, and left rotation, while those of model B were significantly increased. (3) There was no significant difference in the maximum displacement of the isthmus of models A, C, and D under different motion modes, while the maximum displacement of model B in the isthmus was significantly larger than that of models A, C, and D, especially during rotation, increased by 295%, 277%, and 276% respectively. The maximum stress of the isthmus of model C was 0.938 MPa, 1.698 MPa, 0.410 MPa, 2.775 MPa, and 1.554 MPa respectively. The maximum stress in the isthmus of model D was 0.590 MPa, 1.297 MPa, 0.520 MPa, 3.088 MPa, and 2.072 MPa respectively. The maximum stress of the isthmus of models C and D was similar during axial compression and forward flexion, while the stress of the isthmus of model C was smaller than that of model D during backward extension, lateral bending, and rotation, decreased by 21.1%, 10.2%, and 25.0% respectively compared with model D. (4) The maximum stress of internal fixation in models C and D during forward flexion, backward extension, left bending, and left rotation was 135.220 MPa, 130.180 MPa, 200.940 MPa and 306.340 MPa respectively, and was 131.840 MPa, 112.280 MPa, 349.980 MPa and 370.140 MPa respectively. The maximum stress of internal fixation in the two models of internal fixation during forward flexion and backward extension was similar, while it was decreased by 42.6% and 17.2% in model C during left bending and left rotation, compared with model D. (5) The maximum stress of the vertebral body during forward flexion, backward extension, left bending, and left rotation was 79.787 MPa, 36.857 MPa, 37.943 MPa and 96.965 MPa respectively in model C, but was 80.104 MPa, 64.236 MPa, 196.010 MPa and 193.020 MPa respectively in model D. The maximum stress of models C and D was all distributed in the contact area with the internal fixation, and especially during backward extension, left bending, and left rotation, when it was reduced by 42.6%, 80.6%, and 49.8% of model C respectively, compared with that of model D. Conclusions:The improved laminar hook is more consistent with the Chinese anatomized structure of the lamina. Compared with the traditional lamina hook system, the improved lamina hook system can effectively reduce the displacement in all directions and range of motion of lumbar spondylolysis, therefor can significantly reduce the stress of internal fixation and vertebral body and has better biomechanical performance.

Objective:To investigate the effects of polyethylene glycol-poly (lactic-co-glycolic acid) (PEG-PLGA) co-loaded resveratrol nanoparticles on colon cancer cells through epithelial-mesenchymal transition (EMT) and phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt) signaling pathways.Methods:Human colon cancer HCT116 cells were randomly divided into the control group and the experimental group. The experimental group was supplemented with 50 μl of 10 μmol/L PEG-PLGA co-loaded resveratrol nanoparticle solution, and the control group was supplemented with the same volume of medium without adding any treatment substance. Cell viability was determined using the MTT assay, apoptosis rate was analyzed by flow cytometry, and cell migration ability was detected by a scratch test. Western Blot was used to analyze the expression of proteins of related signaling pathways such as apoptosis, EMT, and PI3K/Akt.Results:Compared with the control group, the survival rate of colon cancer cells in the experimental group was reduced and the apoptosis rate was increased ( P < 0.05). Compared with the control group, the scratch width of HCT116 cells in the experimental group was greater ( P < 0.001). Compared with the control group, the expression of B-cell lymphoma-2 (Bcl-2) protein in the experimental group was down-regulated ( P < 0.01), while the expression of Bcl-2 associated X protein (Bax) was up-regulated ( P < 0.05). Compared with the control group, the expression of N-cadherin protein in the experimental group was down-regulated, while the expression of E-cadherin was up-regulated ( P < 0.01). Compared with the control group, the levels of p-PI3K and p-Akt in the experimental group were down-regulated (both P < 0.01). Conclusions:PEG-PLGA co-loaded resveratrol nanoparticles can reduce the cell viability, proliferation, and migration of colon cancer cells, which may be related to the inhibition of EMT and PI3K/Akt signaling pathways.

BACKGROUND: Low-density computed tomography (LDCT) improved early lung cancer diagnosis but introduces an excess of false-positive pulmonary nodules data. Hence, accurate diagnosis of early-stage lung cancer remains challenging. The purpose of the study was to assess the feasibility of using circulating tumour cells (CTCs) to differentiate malignant from benign pulmonary nodules. METHODS: 122 patients with suspected malignant pulmonary nodules detected on chest CT in preparation for surgery were prospectively recruited. Peripheral blood samples were collected before surgery, and CTCs were identified upon isolation by size of epithelial tumour cells and morphological analysis. Laser capture microdissection, MALBAC amplification, and whole-exome sequencing were performed on 8 samples. The diagnostic efficacy of CTCs counting, and the genomic variation profile of benign and malignant CTCs samples were analysed. RESULTS: Using 2.5 cells/5 mL as the cut-off value, the area under the receiver operating characteristic curve was of 0.651 (95% confidence interval: 0.538-0.764), with a sensitivity and specificity of 0.526 and 0.800, respectively, and positive and negative predictive values of 91.1% and 30.3%, respectively. Distinct sequence variations differences in DNA damage repair-related and driver genes were observed in benign and malignant samples. TP53 mutations were identified in CTCs of four malignant cases; in particular, g.7578115T>C, g.7578645C>T, and g.7579472G>C were exclusively detected in all four malignant samples. CONCLUSIONS CTCs play an ancillary role in the diagnosis of pulmonary nodules. TP53 mutations in CTCs might be used to identify benign and malignant pulmonary nodules.
Humans ; Lung Neoplasms ; Exome Sequencing ; Multiple Pulmonary Nodules ; Carcinoma ; DNA Repair

Inflammatory bowel disease(IBD)is a non-specific and easily recurrent chronic inflammatory disease of the intestine,mainly including ulcerative colitis(UC)and Crohn's disease(CD).Its etiology and pathogenesis have not been fully elucidated,but it is currently believed to be related to environmental,genetic,immune response abnormalities,and other factors.At present,IBD has become a disease of great concern worldwide.According to the epidemiological data and data of inflammatory bowel disease in China,the incidence rate of IBD is on the rise year by year in China,but so far,there has not been a drug that can completely cure the disease.Recently,glucagon-like peptide(GLP)and its degradation enzyme inhibitor dipeptidyl peptidase-4(DPP-4)inhibitors have attracted widespread research and attention in the treatment of IBD.This article reviewed the roles and mechanisms of GLP and DPP-4 inhibitors in the treatment of IBD diseases.