Objective To isolate and identify stem-like cells (CSCs) from human ACHN renal cell carcinoma cell line.Methods ACHN cancer stem cells CSCs were enriched by serum-free culture condition.Methyl thiazolyl tetrazolium (MTT) method was performed to draw the growth curve.Flow cytometry was performed to evaluate the expressions of two CSCs markers,CD133 and CD44.Real-time polymerase chain reaction (RT-PCR) was performed to evaluate the expression of stem-related genes (Oct3/4,Bmi,Nanog,and 3-catenin).The tumorigenicity in vivo was also examined.Results ACHN CSCs were formed by sphere-forming culturing.The data demonstrated that there were stronger capacities of proliferation in ACHN tumor spheres than that of ACHN cells.And the expressions of CD133 and CD44 were also significandy enhanced in tumor spheres.The data showed that four genes (Oct3/4,Bmi,Nanog,and β-catenin)were upregulated in sphere-forming cells.Sphere-forming cells had a higher tumorigenic potential in vivo than incubated cells.Conclusions Human ACHN renal cell carcinoma cell line sphere-forming cells can be enriched by serum-free culture condition and exhibits several characteristic cancer stem cells properties.

Objective:To investigate the effects of tert-butyl hydroperoxide (TBH) on the expression of second mitochondria-derived activator of caspase (Smac) and X-linked inhibitor of apoptosis (XIAP) in mitochondrial pathway after cerebral ischemia/reperfusion injury in rats.Methods:From March to December in 2019, 45 healthy male Sprague-Dawley rats were randomly divided into sham-operation, model and TBH groups. Rat models of cerebral ischemia/reperfusion injury were established by ligation of the left carotid artery. Rat neurological function was evaluated to exclude the rats that failed in cerebral ischemia/reperfusion injury induction. Ten rats were left in each group. At 0.5 and 12 hours after cerebral ischemia/reperfusion injury, rats in the TBH group were treated by intragastric administration of 12.5 mg/kg TBH and those in the sham-operation and model groups were identically treated by intragastric administration of equal volume of 0.9% sodium chloride injection. After 24 hours of reperfusion, rat neurological function was assessed in each group. Then the rats were killed and the brains were harvested. Apoptosis of nerve cells was detected by terminal deoxynucleotidyl transferase dUTP nick end-labeling (TUNEL) assay. The levels of superoxide dismutase (SOD), malondialdehyde (MDA), tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in the brain tissue were detected by enzyme-linked immunosorbent assay. XIAP- and Smac-positive cell count and protein expression were determined by immunohistochemical staining and western blot assay, respectively.Results:Rat neurological function score in the TBH group was significantly lower than that in the model group [(1.36 ± 0.49) points vs. (3.73 ± 0.97) points, t = 6.896, P < 0.001]. In the TBH group, a large number of apoptotic nerve cells were found in the ischemic cerebral cortex, but the number of apoptotic nerve cells in the TBH group was significantly smaller than that in the model group. In the model group, SOD level was significantly lower, MDA, TNF-α and IL-1β levels were significantly higher compared with the sham-operation group [SOD: (51.94 ± 3.46) U/mg vs. (70.68 ± 2.67) U/mg, t = 13.560, P < 0.001; MDA: (5.69 ± 0.78) nmol/mg vs. (1.20 ± 0.96) nmol/mg, t = 11.479, P < 0.001; TNF-α: (89.36 ± 9.84) pg/mg vs. (40.53 ± 4.35) pg/mg, t = 14.353, P < 0.001; IL-1β: (41.35 ± 6.79) pg/mg vs. (17.22 ± 2.31) pg/mg, t = 10.639, P < 0.001]. In the TBH group, SOD level was significantly higher, MDA, TNF-α and IL-1β levels were significantly lower compared with the model group [SOD: (51.94 ± 3.46) U/mg vs. (68.84 ± 5.03) U/mg, t = 8.754, P < 0.001; MDA: (5.69 ± 0.78) nmol/mg vs. (2.46 ± 0.48) nmol/mg, t = 11.153, P < 0.001; TNF-α: (89.36 ± 9.84) pg/mg vs. (57.64 ± 6.22) pg/mg, t = 8.617, P < 0.001; IL-1β: (41.35 ± 6.79) pg/mg vs. (23.84 ± 5.48) pg/mg, t = 6.346, P < 0.001]. XIAP- and Smac-positive cell count and protein expression in the model group were significantly greater than those in the sham-operation group [XIAP-positive cell count: (22.63 ± 4.37) vs. (12.39 ± 3.18), t = 5.992, P < 0.001, Smac-positive cell count: (47.58 ± 6.94) vs. (5.64 ± 1.35), t = 18.759, P < 0.001; XIAP protein expression: (0.53 ± 0.08) vs. (0.24 ± 0.05), t = 9.721, P < 0.001; Smac protein expression: (0.92 ± 0.15) > ( 0.36 ± 0.05), t = 11.200, P < 0.001 ]. In the TBH group, XIAP-positive cell count and XIAP protein expression were significantly higher and Smac-positive cell count and Smac protein expression were significantly lower compared with the model group [XIAP-positive cell count: (36.78 ± 5.26) vs. (22.63 ± 4.37), t = 6.543, P < 0.001, Smac-positive cell count: (31.74 ± 4.26) vs. (47.58 ± 6.94), t = 6.151, P < 0.001; XIAP protein expression: (0.79 ± 0.10) vs. (0.53 ± 0.08), t = 6.420, P < 0.001, Smac protein expression: (0.70 ± 0.09) vs. (0.92 ± 0.15), t = 3.977, P < 0.001]. Conclusion:TBH can effectively reduce neuronal apoptosis, oxidative stress and inflammatory reaction after cerebral ischemia/reperfusion injury, which may be related to the regulation of XIAP and Smac signaling pathways.

Objective To evaluate the appearance of CT in patients with severe acute respiratory syndrome (SARS) in the recovery phase, and to study the correlation of CT findings with pulmonary function.Methods From June to August in 2003, 100 patient with confirmed SARS accepted examination in our hospital. Among them, 91 patients (39 men, 52 women, mean age 36.4 years, age range 19- 66 years) received CT examination and pulmonary function test on the same day. The interval between SARS onset and the examination ranged from 52 to 125 days (mean 87.4 days). CT appearances of pulmonary parenchymal abnormalities including distribution and extent of involvement were quantitatively analyzed, and four levels on CT scan including the aortic arch, the tracheal carina, the pulmonary venous confluence, and the dome of right diaphragm were selected to score the lesions. The correlation of CT scores with the results of pulmonary function tests was studied.Results Of the 91 cases, 47 patients had normal CT appearance in the recovery phase, whereas the other 44 patients still had parenchymal abnormalities, including residual ground-glass opacification and reticular shadow. CT visual score had correlation with DLco% ( r =-0.618, P

OBJECTIVETo explore the role of ompT gene in uropathogenic E. coli (UPEC) CFT073 strain in urinary tract infection (UTI).

METHODSAn ompT deletion mutant (COTD) was generated by λ Red recombineering in the UPEC CFT073 strain, which was characterized by PCR and sequencing. C57B/L6 mouse models of acute UTI with the mutant and wild-type strains were established to compare the colonization abilities of the two strains in the bladder. The adhesion of CFT073 mutant to human unthelial 5637 cells was also investigated in vitro.

RESULTSPCR and DNA sequencing confirmed the loss of ompT gene in the mutant COTD. The in vitro adhesion rate of the mutant strain COTD to 5637 cells was (6.7±2.2)%, significantly lower than that of (8.3±1.9)% of the wild-type strain (P<0.05). In the murine models of acute UTI, the mutant strain showed a mean colonization number of about (17±8)×10⁴ cfu, which was significantly lower than that of (7∓2)×10⁵ cfu of the wide-type CFT073 strain (P<0.05).

CONCLUSIONOmpT gene can be involved in the colonization of UPEC in the bladder tissue and plays an important role in the pathogenesis of UPEC-induced UTI.

Animals ; Bacterial Proteins ; genetics ; Cell Line ; Escherichia coli Infections ; microbiology ; Escherichia coli Proteins ; genetics ; Gene Knockout Techniques ; Humans ; Mice ; Mice, Inbred C57BL ; Porins ; genetics ; Urinary Tract Infections ; microbiology ; Uropathogenic Escherichia coli ; genetics

Objective:To observe the incidence, risk factors of malignant brain edema (MBE) and the influence of MBE on outcomes after early successful recanalization of acute large vascular occlusion stroke (ALVOS).Methods:A total of 149 patients (age (68±11) years, male 85 (57.0%)) with ALVOS who underwent early endovascular treatment and achieved successful recanalization at the First Affiliated Hospital of Wannan Medical College from July 2014 to February 2019 were retrospectively analyzed. Baseline data, perioperative data, and 90-day prognostic information were collected from patients enrolled in the study. Univariate and multivariate analyses were used to explore the relationship between MBE and outcomes, and the risk factors of MBE.Results:Among the 149 patients, baseline National Institutes of Health Stroke Scale score was 16 (13, 20), baseline Alberta Stroke Project early CT score was 9 (8, 10), the time of onset-to-puncture was (248.3±61.3) minutes, and the onset-to-recanalization time was (312.4±69.7) minutes. MBE occurred in 23 patients (15.4%, 23/149). The 90-day favorable outcome (90-day modified Rankin Scale score≤ 2) in patients with MBE was significantly lower than those without MBE (17.4% (4/23) vs 61.1% (77/126), χ 2=14.985, P<0.001), and the 90-day mortality in patients with MBE was significantly higher than those without MBE (43.5% (10/23) vs14.3% (18/126), χ 2=10.861, P=0.003). MBE was shown to be an independent predictor of 90-day poor outcome (adjusted OR=12.078, 95 %CI 1.934-75.443, P=0.008) and death (adjusted OR=4.146, 95 %CI 1.060-16.216, P=0.041). Multivariate Logistic regression analysis showed that the collateral circulation status was related to the incidence of MBE in patients with ALVOS after successful recanalization (level 2 vs level 0, adjusted OR=0.109, 95 %CI 0.021-0.563, P=0.008). Conclusions:MBE is an independent risk factor of ALVOS patients with poor outcome or death in 90 days. For patients with ALVOS, even if the occlusive vessels have been successfully recanalized after early endovascular treatment, MBE is still not uncommon. The collateral circulation state is an independent predictive factor of the development of MBE after recanalization by early endovascular treatment in patients with ALVOS.

OBJECTIVE: To develop a deep features-based model to classify benign and malignant breast lesions on full- filed digital mammography. METHODS: The data of full-filed digital mammography in both craniocaudal view and mediolateral oblique view from 106 patients with breast neoplasms were analyzed. Twenty-three handcrafted features (HCF) were extracted from the images of the breast tumors and a suitable feature set of HCF was selected using -test. The deep features (DF) were extracted from the 3 pre-trained deep learning models, namely AlexNet, VGG16 and GoogLeNet. With abundant breast tumor information from the craniocaudal view and mediolateral oblique view, we combined the two extracted features (DF and HCF) as the two-view features. A multi-classifier model was finally constructed based on the combined HCF and DF sets. The classification ability of different deep learning networks was evaluated. RESULTS: Quantitative evaluation results showed that the proposed HCF+DF model outperformed HCF model, and AlexNet produced the best performances among the 3 deep learning models. CONCLUSIONS The proposed model that combines DF and HCF sets of breast tumors can effectively distinguish benign and malignant breast lesions on full-filed digital mammography.
Breast Neoplasms ; classification ; diagnostic imaging ; Deep Learning ; Diagnosis, Computer-Assisted ; methods ; Female ; Humans ; Mammography ; methods

【Objective】 To evaluate which factors may affect magnetic resonance imaging (MRI) performance in the detection of pathologic complete response (pCR) after neoadjuvant chemotherapy (NAC). 【Methods】 This retrospective study involved 89 patients diagnosed with invasive breast carcinoma who received NAC at The Second Affiliated Hospital of Xi’an Jiaotong University from July 2019 to December 2021. Breast MRI was performed before and after NAC. Based on the pathological results obtained surgery after the completion NAC and using Miller-Payne classification as the evaluation standard, the patients were divided into two subgroups: pCR and non-pathological complete response (npCR). Chi-square test was used to compare the MRI characteristics of pre-NAC lesions between the two groups. ROC curve analysis was made to analyze the accuracy, sensitivity, specificity, positive predictive value, and negative predictive value of MRI after NAC; the diagnostic performance of MRI in predicting pCR in different tumor subtypes was analyzed. We made univariate and multivariate analyses of factors affecting radiographic complete response (rCR) and pCR concordance. 【Results】 MRI analysis after NAC showed rCR in 20 cases (22.5%) and pCR in 28 cases (31.5%). Considering rCR as a "positive" result of MRI analysis, MRI assessment was accurate in 79 cases, including 19 true positive cases and 60 true negative cases. MRI assessment was inaccurate in 10 cases, including 9 false negative cases and 1 false positive case. The sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of MRI assessment of pCR were 67.86%, 98.36%, 88.76%, 95.00%, and 86.96%. MRI had the lowest diagnostic efficiency in evaluating pCR of ERBB2+ breast cancer after NAC. Single factor analysis showed that estrogen receptor (ER), clinical stage, background parenchymal enhancement, and maximum tumor diameter all affected the consistency of rCR and pCR (P<0.05). Multivariate Logistic regression analysis showed that the independent influencing factor affecting the consistency of rCR and pCR was clinical stage. 【Conclusion】 MRI demonstrated good accuracy in predicting pCR after NAC in the breast cancer patients examined. Pre-treatment MRI characteristics and tumor subtypes may be related to the diagnostic accuracy of post-NAC MRI in breast cancer patients.