Objective To evaluate the role and potential mechanism of apolipoprotein E(APOE)in drug resistance of colon cancer by bioinformatic tools and cellular experiments.Methods After downloading the microarray dataset GSE196900 from the GEO database,the online tool GEO2R was used to identify genes that were expressed differently in the drug-resistant and control groups.The differently expressed genes were then examined for Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment.The STRING database and Cytoscape software were used to build protein-protein interaction(PPI)networks and find hub genes.Hub genes'predictive significance in colon cancer was further assessed.Western blod and qRT-PCR were used to identify changes in APOE expression,whereas Transwell was used to identify changes in the colon cancer cells'capacity for invasion and migration.Results The analysis of GO and KEGG enrichment revealed that the differential genes derived from the GSE196900 dataset were primarily focused on receptor-ligand activity and cytokine-cytokine receptor interaction pathways.Using the CytoNCA plug-in in Cytoscape software,ten hub genes were obtained through PPI construction.Of these,the prognosis of the patients with colon cancer was negatively correlated with the expression of the APOE gene(P<0.05)and the overexpression of the APOE gene might significantly increase the migration and nvasivenessability of colon cancer cells(P<0.05).Conclusion The increased expression of APOE significantly promotes the migration and invasion ability of colon cancer cells,which may be one of the mechanisms by which APOE gene promotes tumor progression in the patients with colon cancer.

Objective:To explore independent predictors for poor outcome at 3 months in elderly patients with acute cerebral infarction(ACI)treated with intravenous thrombolysis(IVT), and to develop a nomogram-based predictive model.Methods:This was a retrospective cohort study.Clinical, laboratory and imaging data of 346 elderly patients with ACI treated with IVT from January 2016 to April 2021 in our hospital were collected.Poor outcome was defined as a modified Rankin Scale(mRS)score >2 at 3 months after the stroke.Logistic regression analysis was used to screen for independent factors predicting poor outcome in elderly ACI patients treated with IVT, and a corresponding nomogram model was developed using the R software.The ROC curve, calibration plots and decision curve analysis were used to evaluate discrimination, calibration and clinical application value of the nomogram model.Results:Among 346 candidates, 109 developed a poor outcome, representing a rate of 31.5%.Logistic regression analysis showed that symptomatic hemorrhagic transformation( OR=15.647, 95% CI: 8.913-27.454), stroke severity(moderate stroke, OR=3.322, 95% CI: 1.414-7.811; moderate-severe stroke, OR=8.169, 95% CI: 4.102-16.258; severe stroke, OR=9.653, 95% CI: 5.440-17.121), stroke-associated pneumonia( OR=2.239, 95% CI: 1.134-4.420), and heart failure( OR=2.758, 95% CI: 1.424-5.336)were independent predictors for poor outcome at 3 months in elderly ACI patients treated with intravenous thrombolysis(all P<0.05). With the area under curve(AUC-ROC)value at 0.85(95% CI: 0.80-0.89), the nomogram model, which was composed of the above four predictors, demonstrated good discrimination.On the calibration plot, the mean absolute error was 0.020, indicating that the model had good calibration.Decision curve analysis revealed that the model had good clinical application value. Conclusions:The nomogram model composed of symptomatic hemorrhagic transformation, stroke severity, stroke-associated pneumonia and heart failure may predict poor outcome at 3 months in elderly ACI patients treated with IVT, with high prediction accuracy and high clinical application value.

Objective:To evaluate the value of p16 INK4a detected by p16 INK4a immunostaining as a new generation of cervical cytology for primary screening and secondary screening in population-based cervical cancer screening, and in improving cytological diagnosis. Methods:Between 2016 and 2018, 5 747 non-pregnant women aged 25-65 years with sexual history were recruited and underwent cervical cancer screening via high-risk (HR)-HPV/liquid-based cytological test (LCT) test in Shenzhen and surrounding areas. All slides were immuno-stained using p16 INK4a technology, among them, 902 cases were offered p16 INK4a detection during primary screening, and the remaining 4 845 cases were called-back by the virtue of abnormal HR-HPV and LCT results for p16 INK4a staining. Participants with complete LCT examination, HR-HPV test, p16 INK4a staining and histopathological examination results were included in this study. The performance of p16 INK4a in primary and secondary screening, and in assisting cytology to detect high grade squamous intraepithelial lesion [HSIL, including cervical intraepithelial neoplasia (CIN) Ⅱ or Ⅲ] or worse [HSIL (CIN Ⅱ) + or HSIL (CIN Ⅲ) +] were analyzed. Results:(1) One-thousand and ninety-seven cases with complete data of p16 INK4a and histology were included. Pathological diagnosis: 995 cases of normal cervix, 37 cases of low grade squamous intraepithelial lesion (LSIL), 64 cases of HSIL and one case of cervical cancer were found. Among them, 65 cases of HSIL (CIN Ⅱ) + and 34 cases of HSIL (CIN Ⅲ) + were detected. The positive rate of p16 INK4a in HSIL (CIN Ⅱ) + was higher than that in CINⅠ or normal pathology (89.2% vs 10.2%; P<0.01). (2) p16 INK4a as primary screening for HSIL (CIN Ⅱ) + or HSIL (CIN Ⅲ) + was equally sensitive to primary HR-HPV screening (89.2% vs 95.4%, 94.1% vs 94.1%; P>0.05), but more specific than HR-HPV screening (89.8% vs 82.5%, 87.7% vs 80.2%; P<0.05). p16 INK4a was equally sensitive and similarly specific to cytology (≥LSIL; P>0.05). (3) The specificity of LCT adjunctive p16 INK4a for detecting HSIL (CIN Ⅱ) + or HSIL (CIN Ⅲ) + were higher than that of LCT alone or adjunctive HR-HPV ( P<0.01), while the sensitivity were similar ( P>0.05). (4) p16 INK4a staining as secondary screening: p16 INK4a was significantly more specific (94.1% vs 89.7%, 91.9% vs 87.4%; P<0.01) and comparably sensitive (84.6% vs 90.8%, 88.2% vs 91.2%; P>0.05) to cytology for triaging primary HR-HPV screening. HPV 16/18 to colposcopy and triage other HR-HPV with p16 INK4a was equally sensitive (88.2% vs 94.1%; P=0.500) and more specific (88.3% vs 83.0%; P<0.01) than HPV 16/18 to colposcopy and triage other HR-HPV with LCT≥ atypical squamous cells of undetermined significance (ASCUS), and the referral rate decreased (14.0% vs 19.4%; P=0.005). Conclusions:For primary screening, p16 INK4a is equally specific to cytology and equally sensitive to HR-HPV screening. p16 INK4a alone could be an efficient triage after primary HR-HPV screening. In addition, p16 INK4a immunostaining could be used as an ancillary tool to cervical cytological diagnosis, and improves its accuracy in cervical cancer screening.

Objective To investigate the impulsivity and aggressiveness characteristics of patients with hepatolenticular degeneration (HLD) and its relationship with brain structure.Methods The Chinese version of the Barratt impulsiveness scale,11 version (BIS-1 1-C) and Buss-Perry aggression questionnaire (BPAQ) were assessed in 78 patients with hepatolenticular degeneration(HLD group) and 86 normal adults (health control group).HLD patients were examined by 3.0T magnetic resonance imaging (MRI).The differences in impulsivity and aggressiveness of the two groups were compared,and the relationship was analyzed between impulsivity,aggressiveness and different brain structures in patients with HLD.Results The total impulsive score,unplanned factor score,the total aggressive score and anger factor score of patients with cerebral HLD (61.74±9.82,26.08±5.06,82.71 ± 15.92,20.06± 5.74,respectively) were higher than those in patients with hepatic HLD (56.73±7.11,23.02±4.20,72.84± 11.15,16.64±5.01,respectively),and health control group(52.19±7.53,21.50± 3.93,64.64±9.83,14.27 ±4.38,respectively),and the differences were significant (F=3.193,4.646,11.830,8.270,all P＜0.05).Total impulsive score was positively correlated with aggressive score and physical aggression in HLD group(r=0.299,0.290,both P＜0.05).Unplanned score was positively correlated with aggressive total score (r=0.324) and physical aggression (r=0.320) in HLD group (P＜0.05).Frontal lobe injury was the influencing factor of total impulsive score(B=10.263,95％CI=0.467-19.946,P=0.008),attention score(B=2.837,95％CI=0.382-5.600,P=0.010) and unplanned factor score (B=3.977,95％ CI=0.848-8.502,P=0.046).Thalamus injury was the influencing factor of aggressive total score and its factor score.Caudate nucleus injury was the influencing factor of aggressive total score (B=10.030,95％ CI=3.351-18.039,P=0.017) and physical aggression score (B =4.432,95％ CI=1.193-7.729,P=0.016).Conclusion Patients with HLD have higher impulsive and aggressive tendencies,which are mainly manifested in unplanned impulsive and anger tendencies.Brain injury may be an important factor affecting impulsiveness and aggression in patients with HLD.Impulsiveness is related with frontal lobe injury and aggression to thalamus and caudate nucleus injury.Impulsiveness and aggressiveness in patients with HLD are not caused by damage to isolated brain areas,but are related to damage to multiple brain areas.

Objective The purpose of the present study was to develop an animal model of type II diabetic coro-nary heart disease in Zuker diabetic fatty ( ZDF) rats.Methods The ZDF rat model of type II diabetic coronary heart disease was prepared by high-fat diet feeding combined with continuous injection of isoprenaline hydrochloride (ISO) in a dose of 1 mg· mL-1 for 10 consecutive days.Serum creatine kinase (CK), and creatine kinase izozyme (CK-MB), ST segment in electrocardiogram ( ECG) and myocardial pathological changes were detected to evaluate the rat model.Results CK of both the control and model groups was gradually increased with ISO injections, while CK-MB increased first and then decreased.The ST segment in ECG part II had significant changes.The pathological examination found that about half of the myocardial cross section in the model group was necrotic after injections of ISO for 5 days and more than 3/4 of the my-ocardial cross section was necrotic after injection of ISO for 10 days.The results indicated that ISO caused myocardial inju-ry in ZDF rats.Conclusions The variation of CK-MB, CK, ST segments in ECG and myocardial necrosis indicate that the model is successfully established.The use of high-fat diet combined with continuous injection of isoprenaline hydrochlo-ride in a dose of 1 mg· mL is a simple way to develop a Zuker diabetic fatty rat model of type II diabetic coronary heart dis-ease.

Objective To explore the computed tomography (CT)manifestations of adrenocortical oncocytoma (ACO)for better understanding the disease and improving its diagnosis accuracy.Methods The CT manifestations of 9 cases with adrenocortical onco-cytomas confirmed by surgery were retrospectively reviewed and compared with pathological results.Results Five of cases were lo-cated in left side while 4 cases were in right side.Well defined and round or oval border could be found in all the cases.Of all the ca-ses,3 cases were solid lesions with equal density,< 3 cm in diameter,and mild consistent intensification after enhancement.Cystic lesion was detectd in 6 cases with > 3 cm in diameter.The mixed cystic and solid components with polycystic changes could be found on CT image,and remarkable intensification on solid lesion while no intensification on cystic lesion after the enhancement. Moreover,patchy or cotton-like shape could be found on solid lesion in 4 cases and 1 case with island like intensification;reticular pattern could be found in the central of lesion in 3 cases and 1 case with stellate shape.Pathological observation showed that the on-cotytomas were comprised of cells with abundant eosinophilic cytoplasm and necrotic and fibrous capsule without pathological mitosis could be found in some of lesion.Conclusion The characteristic CT features of adrenocortical oncotytoma has a worse specificity,its diagnosis was dependent on pathologic examination.

OBJECTIVETo investigate the expression and significance of CCL5 in patients with esophageal carcinoma.

METHODSUsing reverse transcriptase polymerase chain reaction (RT-PCR), the expressions of CCL5/CD8/granzyme B/perforin in tumor and corresponding adjacent tissues from esophageal carcinoma patients were examined. Flow cytometry (FACS) was used to detect the percentages of CD8(+) T cells and CCR5(+)CD8(+) T cells in TIL and PBMC from the patients. Transwell assay was performed to study the effect of CCL5 on the migration of T cells in vitro. T test and Spearman correlation analysis were performed.

RESULTSThe mRNA expressions of CCL5 and perforin were 0.348 2 ± 0.300 1 and 0.181 9 ± 0.118 6, respectively, in the tumor samples, while their expressions in adjacent samples were 0.279 6 ± 0.138 0 and 0.118 0 ± 0.109 8, respectively, with no statistically significant differences between them (P > 0.05 for both). The mRNA expressions of CD8 and granzyme B were significantly higher in the tumor tissues than in adjacent tissues (0.464 9 ± 0.300 8 vs. 0.279 0 ± 0.173 4, 0.648 7 ± 0.516 0 vs. 0.469 7 ± 0.259 1; P < 0.05 for both). The relative expression of CCL5 was positively correlated with that of CD8, perforin and granzyme B (r(CD8) = 0.272, P = 0.034; r(perforin) = 0.305, P = 0.026; r(granzymeB) = 0.108, P = 0.012) in the tumor sites. FACS data revealed that the proportions of CD8(+) T cells in TIL and PBMC were (45.86 ± 16.09)% and (34.05 ± 15.07)%, respectively, showing a significant difference (P = 0.022). Similarly, CCR5(+)CD8(+) T cells fraction in TIL (48.12 ± 26.75)% was much higher than that in PBMC (19.53 ± 13.67) % (P < 0.001). Transwell assay showed that CCL5 protein enhanced the migration of T cells, supporting that CCL5 is crucial for CD8(+) T cells recruitment in vivo. Intriguingly, CCL5 expression was down-regulated in advanced patients (stage IIb-IV). The accumulation of CD8(+) T cells and CCR5(+)CD8(+) T cells was strongly reduced in advanced patients, suggesting that CCL5 expression may be involved in the local control of the disease and its reduction may be involved in disease progression.

CONCLUSIONSThe current data indicate the involvement of CCL5 in the regulation of CD8(+) T cell entry into tumor lesions in esophageal carcinoma patients. This process may affect the disease status and potentially as a prognostic factor for cancer patients. Enhancing local CCL5 expression in tumor lesions may represent a novel strategy in esophageal cancer therapy.

CD8-Positive T-Lymphocytes ; Chemokine CCL5 ; metabolism ; Disease Progression ; Esophageal Neoplasms ; metabolism ; Flow Cytometry ; Humans ; Leukocytes, Mononuclear ; Lymphocytes, Tumor-Infiltrating

Objective To investigate the incidence,distribution patterns,and influencing factors of cerebral microbleed (CMB) in Chinese adult patients with moyamoya disease.Methods Thirty consecutive patients with moyamoya disease confirmed by digital subtraction angiography from the Nanking Stroke Registry Program were included.All patients performed conventional MRI sequences (3.0 T) and susceptibility-weighted imaging.The clinical data,such as medical history,systolic blood pressure,diastolic blood pressure,mean arterial pressure,and white matter lesions were collected.Their fasting blood glucose levels and fibrinogen levels were detected.The numbers of CMB lesion,distribution information,and their relationship with various clinical parameters in patients with moyamoya disease were analyzed.Results Among the 30 subjects included,14 CMBs were detected in 10 patients from 11 hemispheres.The distribution of the lesions was mainly in deep brains (71.4%),especially in the periventricular white matter (50.0%).There was no statistical difference in age,gender,hemorrhage symptoms,blood pressure,white mater lesions,and plasma fibrinogen levels between the CMB positive group and CMB negative group.However,the fasting blood glucose levels in the former was significantly higher than those in the latter (8.0 ± 4.1 mmol/Lvs.4.8 ± 0.4 mmol/L;P =0.035).Multivariate regression analysis showed that the increased fasting blood glucose level was an independent risk factor for the occurrence of CMBs in adult patients with moyamoya disease (OR = 10.992,95% CI 1.325-91.218;P=0.026).Conclusions The CMB lesions are susceptible to Chinese adult patients with moyamoya disease in deep brains,especially in the periventricular white matter.The fasting blood glucose level may influence the incidence of CMBs in patients with moyamoya disease.

Stroke is one of the diseases that seriously threats the health of the elderly, and atherosclerosis is the main cause of ischemic stroke. Became it is closely associated with stroke, carotid carotid atherosclerosis has been one of the research hot spots in neurology. In recent years, the development of MRI techniques has provided a new approach for the qualitative and quantitative analysis of atherosclerotic plaque, and has reflected its unique superiority, This article reviews the applications of MRI in the evaluation of carotid atherosclerotic plaques.

Objective To evaluate the clinical value and effectiveness of ultrasound screening for fetal chromosomal abnormalitie in the middle and late pregnancy. Methods Fetuses who were detected with abnormal ultrasound findings during the middle and late pregnancy, and high risk of maternal serum screening underwent amnioeentesis or eordocentesis for fetal chromosome karyotypes. Results (1) A total of 31 cases with fetal malformation diagnosed by ultrasound were analysed for fetal chromosome karyotypes, and 8 (25.8%) cases were proved with fetal abnormal chromosome karyotypes. There were 3 cases of cervical springwater cyst accompany with edema,and all were fetal abnormal chromosome karyotypes. There were 3 cases of cervical pachyderma,and 2 were fetal abnormal chromosome karyotypes. There was one case with multiple malformations, one with Dandy-Walker malformation and one with holoprosencephaly malformation,all were revealed fetal abnormal chromosome karyotypes. (2) A total of 516 cases with high risk of Down's syndrome and trisomy 18 by maternal serum screening were analysed for fetal chromosome karyotypes,and 14(2.710%) cases were proved with fetal abnormal chromosome karyotypes, which include 7 cases of Down's syndrome and 7 cases of other fetal abnormal chromosome karyotypes. (3) A total of 544 (516 + 28)cases with high risk by the combination of ultrasound and maternal serum screening were analysed for fetal chromosome karyotypes, and 21 (3.86%) cases were proved with fetal abnormal chromosome karyotypes, the rate of detection higher than only maternal serum screening 42.43%.Conclusions Fetal structure abnormalities were the effective ultrasound signs for fetus chromosomal abnormalities screening in the middle and late pregnancy. The combination of ultrasound and maternal serum screening can improve the rate of fetus chromosomal abnormalities screening and be an effective way to retrieve false-positive and lower risk of maternal serum screening.