Role of interferon alpha-inducible protein 6 in modulating the proliferation, apoptosis and senescence of oesophageal squamous cell carcinoma cells.

Oesophageal cancer is one of the most malignant tumors worldwide. Dysfunction of interferon alpha-inducible protein 6 (IFI6) has been implicated in numerous human diseases, including cancer. We performed the study to investigate the function and potential molecular pathways of IFI6 in oesophageal squamous cell carcinoma (ESCC) cells. IFI6 expression was analysed using databases-derived data and paraffin-embedded tissue samples. CCK-8-based analyses and EdU staining, colony formation, ß-galactosidase staining and Annexin V/PI double-staining assays were used to determine the influence of IFI6 on cell growth, senescence and apoptosis. Tumor growth in vivo was investigated in mouse xenograft models. RNA sequencing (RNA-seq) was performed to identify the transcripts and pathways affected by IFI6. The results showed that IFI6 expression was elevated in ESCC and correlated with poor clinical prognosis (P<0.05). IFI6 was overexpressed and silenced in TE-1 and TE-10 cells using lentiviruses. Upregulation of IFI6 promoted cell growth both in vitro and in vivo, whereas downregulation induced opposite effects. IFI6 overexpression inhibited cell senescence and apoptosis but did not influence cell cycle progression, while IFI6 downregulation increased cell senescence and apoptosis. RNA-seq revealed that 3 mRNAs (EPHA5, CLIP1 and GTF2F2) were consistently associated with both IFI6 overexpression and silencing. IFI6 appeared to modulate TE-1 cells via complex mechanisms. In conclusion, IFI6 plays a positive role in the proliferation of ESCC cells both in vitro and in vivo, which could be a novel therapeutic target for treating ESCC.

[Application of next-generation sequencing in detection of BRCA1/2 and homologous recombination repair pathway multi-genes germline mutation and correlation analysis].

Objective: To investigate the germline mutation status of related genes in breast cancer patients and high-risk individuals by next-generation sequencing. To analyze the correlations between homologous recombination repair (HR) pathway gene mutation status and clinicopathological characteristics of breast cancer patients. To supplement the database of breast cancer related gene mutations in Chinese population. Methods: This study is a cross-sectional study. From October 2020 to September 2021, whole blood samples were collected from 350 breast cancer patients and 49 high-risk individuals, admitted to Peking University People's Hospital and accepted genetic testing voluntarily. Germline mutations in 32 breast cancer related genes were detected by NGS. The clinicopathological characteristics, including age at the onset, family history, unilateral/bilateral tumor, Luminal typing (Luminal A subtype, Luminal B subtype, HER2-enriched subtype and triple negative breast cancer), tumor size and metastasis, were analyzed, and the correlations between HR pathway gene mutation status and clinicopathological characteristics were analyzed by Chi-squared test and Fisher's exact probability test. Results: Among 350 breast cancer patients, 64 (18.3%) cases carried gene pathogenic mutations (including pathogenic and likely pathogenic mutations), including 47 (13.4%) in BRCA1/2, 16 (4.6%) in non-BRCA1/2 genes, 1 (0.3%) in BRCA2 and FANCL. Among 49 high-risk individuals, 7 (14.3%) cases carried gene pathogenic mutations, including 6 (12.3%) in BRCA1/2 and 1 (2%) in ATM genes. BRCA1/2 pathogenic mutations were associated with age at the onset (18%, 8.7%, χ²=6.346, P=0.012), and the BRCA1/2 pathogenic mutation frequency was higher in patients diagnosed at age ≤45 years. HR pathway gene mutations (including pathogenic, likely pathogenic and uncertain significance mutations) were correlated with unilateral/bilateral tumor (49.5%, 68.4%, χ²=4.841, P=0.028) and Luminal typing (45.7%, 62.2%, 32%, 60%, χ²=12.004, P=0.007), and the HR mutation frequencies were higher in patients with bilateral tumor, Luminal B breast cancer and triple negative breast cancer (TNBC). Conclusion: The BRCA1/2 pathogenic mutation frequency in high-risk individuals is similar to that in breast cancer patients, and BRCA1/2 testing is helpful to guide breast cancer screening and prevention in high-risk individuals. Patients with early onset breast cancer, bilateral breast cancer, Luminal B breast cancer and TNBC have higher mutation frequencies of HR pathway genes, and HR pathway genes testing should be conducted as soon as possible to provide laboratory evidence for diagnosis, treatment, prognosis and risk evaluation of breast cancer.

[Lengths of the fixed loop and the adjustable loop in the coarse bone tunnel were compared to influence the widening of the femoral bone tunnel and the function of the knee joint].

OBJECTIVE: To evaluate the effects of two different femoral cortical suspension devices (fixation loop and adjustable loop) on tunnel widening and knee function in patients following anterior cruciate ligament reconstruction for 12 months. METHODS: A total of 60 patients who had undergone anterior cruciate ligament reconstruction were included in this study. According to the length of the loop(n)[n= total length of loop-(total length of femoral tunnel-total length of coarse tunnel)] in the rough bone tunnel, the patients were divided into A (adjustable loop was 0 mm in the coarse bone tunnel), B (fixation loop was greater than 0 mm and less than or equal to 10 mm in the coarse bone tunnel) and C (fixation loop was greater than 10 mm in the coarse bone tunnel) groups, of which 11 cases were in group A, 27 cases in group B and 22 cases in group C. In the three-dimensional reconstruction of the knee joint with multi-slice spiral CT, the widening of the bone tunnel in the three groups was compared. At the same time, IKDC, Lysholm and Tegner scores of the patients in the three groups were compared. RESULTS: There were differences in the widening degree of the femoral canal among groups A, B and C, and the median difference of the widening degree of the femoral tunnel 12 months and immediately after the surgery was A < B < C. The difference of femoral canal widening in group A was significantly different from that in groups B and C (P < 0.05).According to the linear regression the relationship between the difference of the width of the femoral canal and the change of the length (n) of the loop in the coarse canal, it was found that there was a linear relationship between the value of n and the difference of the width of the bone canal. With the increase of the value of n, the difference of the width of the bone canal gradually became larger. The median difference of the width of the middle and superior tunnel was negative, while the median difference of the width of the middle and inferior tunnel was positive. During the follow-up, we found that there were no statistical differences in IKDC, Lysholm and Tegner scores among the three groups one year after surgery (P > 0.05). CONCLUSION: Twelve months after surgery, compared with group B (fixed loop group) and group C (fixed loop group), group A (adjustable loop group) had less bone tunnel widening.In groups A, B and C, as the length of the loop in coarse bone tunnel gradually increased, the width of bone tunnel became more significant. At the end of 12 months follow-up after anterior cruciate ligament reconstruction, the medial and inferior femoral tunnel was significantly wider than immediately after surgery, and the medial and superior femoral tunnel had gradually begun to undergo tendon-bone healing. There was no significant difference in knee function scores among groups A, B, and C in the follow-up 12 months after surgery.

Heterogeneity of enhancement kinetics in dynamic contrast-enhanced MRI and implication of distant metastasis in invasive breast cancer.

AIM: To investigate the heterogeneity of enhancement kinetics for breast tumour in order to demonstrate the predictive power of dynamic contrast-enhanced (DCE) magnetic resonance imaging (MRI) features for distant metastasis (DM) in invasive breast cancer. MATERIALS AND METHODS: Time-signal intensity curve (TIC) patterns from 128 patients with invasive breast cancer were analysed by a pixel-based DCE-MRI analysis. This MRI technique enabled pixels with varying TIC patterns (persistent, plateau, washout and non-enhancement) to be categorised semi-automatically and the percentage of different TIC patterns in each breast tumour to be calculated. The percentage of TIC patterns was compared between the DM and non-DM groups. DM-free survival was estimated using Kaplan-Meier survival analysis. RESULTS: This study demonstrated a larger percentage of persistent TIC and non-enhancement TIC was associated with DM in invasive breast cancer. The cut-off values of persistent TIC and non-enhancement TIC were 22.5% and 2.5%. Combining TIC patterns and traditional predictors (tumour size and axillary lymph node status) can improve the prediction efficiency. The multivariable model yielded an area under the receiver operating characteristic curve (AUC) of 0.87 with 0.70 sensitivity and 0.87 specificity in leave-one-out cross-validation (LOOCV). These predictors showed significant differences in DM-free survival by Kaplan-Meier analysis. CONCLUSION: This study shows that breast tumours with higher heterogeneity are more likely to metastasise, and pixel-based TIC analysis has utility in predicting distant metastasis of invasive breast cancer.

[Correlation between breast density and age, breast cancer molecular subtype].

Objective: To explore the correlation between breast density and breast cancer molecular subtype. Methods: The data of 1 407 cases of breast cancer immunohistochemistry and preoperative mammography in Tianjin Medical University Cancer Institute and Hospital from August 2015 to August 2016 were retrospectively analyzed. All cases were divided into four groups: <45 years old, 45-54 years old, 55-64 years old and ≥ 65 years old, and all groups were divided into four molecular subtypes: Luminal A, Luminal B, HER2-enriched and triple-negative according to immunohistochemical results. Person correlation test was used to analyze the correlation between breast density and age; Kruskal-Wallis test was used to analyze the difference in breast density between different molecular subtypes. Results: There was a negative correlation between breast density and age (r=-0.55, P<0.01). Descriptive analysis results showed that the mean breast density of<45 years old, 45-54 years old, 55-64 years old and ≥65 years old was 20.0%±7.3%, 16.2%±8.4%, 10.5%±5.2%, and 7.9%±3.2%, respectively. The mean breast density of Luminal A, Luminal B, HER2-enriched and triple-negative was 14.0%±8.1%, 14.5%±7.6%, 14.8%±7.7% and 13.2%±7.3%, respectively. Kruskal-Wallis test showed significant statistical differences in breast density among molecular subtypes in the group of<45 years old (P<0.05), while no statistical significance in other groups (all P>0.05). Conclusion: There is correlation between breast density and breast cancer molecular subtype in young patients, but whether breast density has a potential evaluation effect on breast cancer molecular subtype still needs to be further studied.

[Preparation and comparative study of decellularized rat cricoarytenoid dorsalis muscle and gastrocnemius muscle scaffold].

Objective:To prepare and to compare the decellularized rat cricoarytenoid dorsalis muscle and gastrocnemius muscle scaffolds for providing a basis for tissue engineered cricoarytenoid dorsalis muscle.Method:Sprague Dawley rats were randomly divided into 6 groups with 10 rats in each group.The experimental groups were divided as 2, 4, 6, 8, and 10day groups after decellularization. Muscles were decellularized with 1% sodium dodecyl sulfate and observed by general observation, histological section with hematoxylin eosin staining and collagenous fiber staining, scanning electron microscope images with measurement of pore diameter and area, and DNA extraction and quantification. The control group was not decellularized.Result:The decellularized muscles in experimental groups showed a semitransparent appearance while in control group the fresh muscle showed a ruddy color.Muscle fibers were completely disappeared in cricoarytenoid dorsalis muscle after 4 days decellularization and in gastrocnemius muscle after 8 days decellularzation, the extracellular matrix reserved well and no DNA was detected. Scanning electron microscope revealed a different pore size between decellularized cricoarytenoid dorsalis muscle and gastrocnemius muscle,and the difference was statistically significant. The pore diameter in 4 days decellularized cricoarytenoid dorsalis muscle was larger than the one in 8 days decellularzed gastrocnemius muscle.Conclusion:Compare to gastrocnemius muscle,decellularized cricoarytenoid dorsalis muscle has advantages in pore form and size and more suitable as a scaffold for tissue engineered cricoarytenoid dorsalis muscle.

[Discussion of breast cancer screening model in China].

Cancer screening has been considered as double-edged sword with both advantages and disadvantages. For decades, there have been strong interests in screening strategies for the early detection of cancers to reduce the mortality, especially breast X-ray(mammography)screening. However, several evidences also suggested that the benefit of reduction of breast-cancer mortality with mammography might become a problem due to the repeat mammography, subsequent biopsies, and overdiagnosis. And different screening strategies with different models, different intervals, and different target populations also incurred debates. After systematical analysis and discussion, we suggested to focus on high-risk population, improve the accuracy of screening technique, conduct the informed consent of participants, and explore individual screening mode in the screening of breast cancer.

Integrated microRNA-mRNA analysis of pancreatic ductal adenocarcinoma.

The main aim of this study was to explore the underlying molecular mechanisms and potential target molecules of pancreatic adenocarcinoma. The miRNA (GSE32678) and mRNA (GSE32676) expression profiles of patients with pancreatic ductal adenocarcinoma and healthy controls were downloaded from the Gene Expression Omnibus database. Differentially expressed miRNA and differentially expressed genes were identified by analyzing the microarray algorithm after data preprocessing. Functional analysis was conducted by the Database for Annotation, Visualization and Integrated Analysis. miRNA-mRNA regulation pairs were obtained in TarMir database. The node degree of hsa-miR-200c, hsa-miR-429, and hsa-miR-200b (miRNA), and EFNB2, MYRIP, and PHF17 (mRNA) were extremely high in the miRNA-mRNA network, indicating that these miRNA and mRNA may play a key role in the development of pancreatic cancer. Our study screened out some target miRNAs and mRNAs for pancreatic ductal adenocarcinoma, which may be helpful in its diagnosis and treatment.

Study of human acellular amniotic membrane loading bone marrow mesenchymal stem cells in repair of articular cartilage defect in rabbits.

The aim of this study was to investigate the repair effect of human acellular amniotic membrane (HAAM) loading bone marrow mesenchymal stem cells (BMSCs) on articular cartilage defect in rabbits. Rabbit BMSCs were isolated and cultured, and they were then inoculated on HAAM to prepare the complex of HAAM and BMSCs. Twenty-four rabbits were randomly divided into groups A and B, with 12 animals in each group. The left and right sides were used as the experimental and control sides, respectively. The models of bilateral articular cartilage defect were established. The defect areas on the experimental side in groups A and B were implanted with the complex of HAAM and BMSCs and HAAM alone, respectively. The control sides of the two groups were not implanted with any material. In the 8th and 12th week after surgery, gross observation, histological examination and cartilage defect scoring were performed. In the 8th and 12th postoperative week, gross observation and histological observation showed that dense cartilage-like cells appeared in group A but not in group B, indicating preferable cartilage repair. The cartilage defect score on the experimental side in group A was 5.31 ± 0.68 in the 8th week and 3.23 ± 0.52 in the 12th week, and that in group A was significantly lower than in group B (P < 0.05). HAAM loading BMSCs has a good repair effect on articular cartilage defect under an in vitro environment.

MRI of the uterus, uterine cervix, and vagina: diagnostic performance of dynamic contrast-enhanced fast multiplanar gradient-echo imaging in comparison with fast spin-echo T2-weighted pulse imaging.

The aim of this study was to compare the diagnostic performance of contrast-enhanced fast multiplanar gradient-echo (GRE) and T2-weighted fast spin-echo (FSE) image sets in the assessment of uterus, cervix, and vagina. Fast (up to 20 contiguous sections in 23 s) multiplanar GRE and FSE images of 45 patients referred for imaging of the female pelvis were evaluated retrospectively with regard to overall image quality and the ability to detect normal anatomic structures, as well as lesion conspicuity. Results were compared with histologic findings (n = 29) or clinical follow-up. Furthermore, a quantitative assessment of contrast-to-noise ratios among normal uterine and cervical structures as well as uterine lesions was performed for both sequences. On GRE images, uterine and cervical differentiation was best seen on the image sets acquired 15 and 60 s following contrast enhancement and results were significantly better compared with delayed images (p < 0.05). Delineation of the junctional zone was significantly (p < 0.05) better on FSE compared with GRE images; no significant difference was seen for the other anatomic structures. Overall image quality of GRE and FSE images was similar. Sensitivity for lesion detection based on both GRE and FSE images was 96% with a sensitivity of 93% for GRE, and 81% for FSE images alone, respectively. Using the extended McNemar chi 2 test, the difference in diagnostic performance between FSE and GRE revealed no significant difference, whereas the combination of both techniques performed better than FSE imaging alone (p < 0.05). The presented data suggest that dynamic contrast-enhanced GRE imaging should be part of an MR examination of the female pelvis. Combined GRE and FSE imaging provide an excellent sensitivity in the assessment of uterine and vaginal pathologies.

Improved diagnostic accuracy in dynamic contrast enhanced MRI of the breast by combined quantitative and qualitative analysis.

To develop an optimized interpretation strategy of dynamic contrast enhanced breast MRI based on a combination of quantitative and qualitative criteria, the features of 120 histopathologically verified lesions were retrospectively analysed on contrast enhanced dynamically acquired (before and 1, 2, 4 and 7 min following injection of 0.2 mmol kg-1 gadolinium-DOTA) three-dimensional gradient echo images. Quantitative criteria, based on relative signal intensity measurements and qualitative morphological features, including lesion shape, margins and enhancement patterns were analysed in regard to differentiating malignant (n = 70) from benign (n = 50) lesions. Quantitative assessment of enhancement profiles was most accurate when analysis was based on early 1 min measurements. Using a 90% threshold, sensitivity and specificity in detecting breast cancer were 83% and 66%, respectively. When based on qualitative morphological analysis alone, sensitivity and specificity were 83% and 54%. Combined quantitative and qualitative assessment yielded a considerably higher sensitivity, specificity and accuracy of 93%, 74% and 85%, respectively. In conclusion, results from dynamic contrast enhanced breast MRI can be improved by basing the interpretation on both quantitative and qualitative criteria.

Clinical significance of CD44v mRNA detection by PCR in peripheral blood of patients with hepatocellular carcinoma.

AIM: To study the clinical significance of detecting the expression of CD44v mRNA in the blood of patients with hepatocellular carcinoma (HCC). METHODS: The expression of CD44v mRNA was detected in blood with RT and diploid PCR, and the clinical significance was discussed based on the result of pathological examination and follow-up. RESULTS: CD44v mRNA was detected in the blood of 10/15 patients, giving a positive rate of 66.67%. In the 13 patients who showed response in the follow up period, the CD44v mRNA expression was positive in 9 and negative in 4. Recurrence rate was higher in the patients with positive CD44v mRNA expression than in those with negative CD44v mRNA expression, and the clinical pathological indices were also higher in the former than in the latter. CONCLUSION: Detection of the expression level of CD44v mRNA in blood of the patients with HCC can be used as an adjuvant means for differential diagnosis, prediction and monitoring of HCC recurrence.

Effect of timolol on intraocular pressure elevation following argon laser iridotomy.

Argon laser iridotomy was performed in 123 eyes of 83 patients with primary angle-closure glaucoma. Sixty-three eyes received argon laser iridotomies with timolol pretreatment and the other 60 eyes served as controls. There were 43 eyes (71.7%) in the control group and 32 eyes (50.7%) in the timolol-pretreated group, whose intraocular pressure increased significantly over baseline level 2 hours after laser treatment. There was statistical difference in the incidence of intraocular pressure elevation in these two groups. Seven eyes (11.7%) in the control group and 4 eyes (6.2%) in the timolol-pretreated group had intraocular pressure elevation at least 11 mmHg over baseline. Additional 28 patients, who received argon laser iridotomy bilaterally, one eye with timolol pretreatment and the other eye served as a matched control, revealed that eyes which were timolol pretreated had statistically significant less intraocular pressure elevation.