[Treatment of Fournier's gangrene with "firebreak" drainage based on skin preservation].

Objective: To observe the clinical efficacy of a new type of "firebreak" drainage with skin preservation in the treatment of Fournier's gangrene. Methods: This technique is suitable for patients with perianal necrotizing fasciitis who can tolerate surgery without large area of skin blackness and necrosis. Procedure and key points: (1) The dividing line between inflammatory tissue and normal tissue was determined according to imaging examination and intraoperative exploration; (2) The abscess cavity was cut along the most obvious part of the abscess fluctuation, with a long diameter of 3~4 cm and a short diameter of 1~2 cm; (3) Necrotic tissue was discreetly separated and removed from the main incision to the outer edge of the infection. A fusiform incision was made every 3 to 5 cm, with a long diameter of 2 to 3 cm and a short diameter of 1 cm, and discreetly separated until the normal tissue, and a hose was hung between the adjacent incisions for drainage. (4) Each adjacent edge cut between the stealth separation and hanging hose drainage, forming a "firebreak"; (5) Rinse the wound repeatedly; (6) If the infection invades the rectum, colostomy is performed as required. The case data of 11 patients with perianal necrotizing fasciitis admitted to the Second Affiliated Hospital of Nanjing University of Chinese Medicine from July 2019 to February 2023 were retrospectively analyzed. All patients were treated with emergency surgical debridement by "firebreak" drainage with skin preservation. Results: All 11 cases were cured with 100%. One case underwent multiple operations. The hospitalization time was 11-46 days, with an average of 22 days. The wound healing time was 28-75 days, with an average of 43 days. Except for 1 patient with trauma, all the other patients had no significant anal function injury after surgery. All the 11 patients recovered and were discharged from hospital with a median follow-up of 136 (115-413) days. Conclusions: The "firebreak" drainage based on skin preservation has the advantages of less trauma and faster recovery, and do not cause obvious anal function damage.

Integrative analyses of radiation-related genes and biomarkers associated with breast cancer.

OBJECTIVE: In addition to significantly reducing breast cancer recurrence risk, radiotherapy also prolongs patients' lives. However, radiotherapy-related genes and biomarkers still remain poorly understood. The present study aimed to identify radiation-associated genes in breast cancer. MATERIALS AND METHODS: Breast cancer data were downloaded from Gene Expression Omnibus (GEO) and UCSC Xena database. The gene ontology (GO) enrichment and gene set enrichment analysis (GSEA) were performed for annotation and integrated discovery. Protein-protein interaction (PPI) network was constructed by STRING database and hub genes were identified. Then, immunohistochemistry and tissue expression of key genes was analyzed by using the Human Protein Atlas (HPA) and GEPIA database. Genes associated with prognosis were identified by performing univariate cox analysis. RESULTS: We identified 341 differentially expressed genes related to radiotherapy in breast cancer patients. PPI analysis revealed a total of 129 nodes and 516 interactions and identified five hub genes (EGFR, FOS, ESR1, JUN, and IL6). In addition, 11 SDEGs THBS1, SERPINA11, NFIL3, METTL7A, KCTD12, HSPA6, EGR1, DDIT4, CCDC3, C11orf96, and BCL2A1 candidate genes can be used as potential diagnostic markers. The calibration curve and ROC indicate good probability consistencies of 3-years and 5-year survival rates of patients between estimation and observation. CONCLUSIONS: Our findings provide novel insight into the functional characteristics of breast cancer through integrative analysis of GEO data and suggest potential biomarkers and therapeutic targets for breast cancer.

[Expression of SOX10 and GATA3 in breast cancer and their significance].

Objective: To investigate the expression characteristics of SOX10 and GATA3 in breast cancer and the value of their combination. Methods: A total of 360 breast cancer specimens with SOX10 immunohistochemical staining were collected from the Department of Pathology in Shenzhen People's Hospital from 2018 to 2021, including 268 cases with simultaneous SOX10 and GATA3 staining. The expression of SOX10 and GATA3 in primary and metastatic breast cancer was detected, and the correlations between SOX10 and GATA3 and the molecular types and clinicopathological features of breast cancer were compared, and the distribution differences among each group were statistically analyzed. Results: The overall expression of SOX10 and GATA3 in breast cancer were 25.8%(93/360) and 81.7%(219/268), and that in triple negative breast cancer (TNBC) were 83.3%(80/96) and 42.7%(32/75), respectively. SOX10 was strongly associated with TNBC (P<0.001), whereas GATA3 was highly expressed in luminal A, luminal B and HER2 over expression breast cancers (P<0.001). The expression of SOX10 and GATA3 was negatively correlated in TNBC, and the combined expression rates of SOX10 and GATA3 in breast cancer and TNBC could reach 97.8% (262/268) and 94.7%(71/75), respectively. In addition, the expression of SOX10 was closely correlated with high histological grade, high Ki-67 proliferation index and lymph node metastasis, and negatively correlated with AR. The expression of GATA3 was correlated with low histological grade and lymph node metastasis, and positively correlated with AR, and the difference was statistically significant. Conclusions: SOX10 is a sensitive marker of TNBC, while GATA3 is highly expressed in non-triple negative breast cancer. The two complementary, combined application of SOX10-GATA3 can improve the detection rate of breast cancer, especially TNBC. SOX10 is associated with malignant characteristics of the tumor, suggesting that SOX10 can be used as a prognostic marker and potential therapeutic target for breast cancer.

Histopathology-Based Diagnosis of Oral Squamous Cell Carcinoma Using Deep Learning.

Oral squamous cell carcinoma (OSCC) is prevalent around the world and is associated with poor prognosis. OSCC is typically diagnosed from tissue biopsy sections by pathologists who rely on their empirical experience. Deep learning models may improve the accuracy and speed of image classification, thus reducing human error and workload. Here we developed a custom-made deep learning model to assist pathologists in detecting OSCC from histopathology images. We collected and analyzed a total of 2,025 images, among which 1,925 images were included in the training set and 100 images were included in the testing set. Our model was able to automatically evaluate these images and arrive at a diagnosis with a sensitivity of 0.98, specificity of 0.92, positive predictive value of 0.924, negative predictive value of 0.978, and F1 score of 0.951. Using a subset of 100 images, we examined whether our model could improve the diagnostic performance of junior and senior pathologists. We found that junior pathologists were able to delineate OSCC in these images 6.26 min faster when assisted by the model than when working alone. When the clinicians were assisted by the model, their average F1 score improved from 0.9221 to 0.9566 in the case of junior pathologists and from 0.9361 to 0.9463 in the case of senior pathologists. Our findings indicate that deep learning can improve the accuracy and speed of OSCC diagnosis from histopathology images.

[Molecular mechanism of miR-369-3p regulating hepatocellular carcinoma cell proliferation and apoptosis by targeting ACTN4].

Objective: To investigate the effect of miR-369-3p targeting ACTN4 expression on proliferation and apoptosis of hepatocellular carcinoma cells. Methods: Real-time quantitative polymerase chain reaction (RT-qPCR) and western blot were used to detect the expression levels of miR-369-3p and ACTN4 in hepatocarcinoma tissues and adjacent tissues. MiR-369-3p mimics, miR-negative control (NC), si-ACTN4, and si-NC were transfected into hepatocellular carcinoma MHCC97H cells by liposome method. Cell proliferation was detected by 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-dipheny-ltetrazolium bromide (MTT) assay. Flow cytometry was used to detect cell cycle and apoptotic rates. The dual luciferase reporter assay was used to verify the targeted regulation of ACTN4 by miR-369-3p. Western blot was used to detect the expressions of cyclin D1, p21, Bcl-2 and Bax. Results: The expression level of miR-369-3p in liver cancer tissue was lower than that in adjacent tissues [(0.46±0.04) vs (1.00±0.08), P<0.001)], while the expression level of ACTN4 was higher than that in adjacent tissues [mRNA (3.12±0.29) vs (1.01±0.09); protein (0.61±0.06) vs (0.25±0.03), P<0.001]. Overexpression of miR-369-3p significantly decreased the cell viability[(0.71±0.06) vs (1.26±0.11), P<0.001)], increased cell apoptosis rate [(20.16±2.11)% vs (6.25±0.64)%, P<0.001], increased the proportion of cells in G(1) phase [(31.14±3.36)% vs (51.56±5.23)%, P<0.001], decreased the proportion of cells in S phase [(32.44±3.56)% vs (14.33) ±1.45)%, P<0.001], increased the levels of p21 and Bax protein (P<0.001), and decreased the levels of cyclin D1 and Bcl-2 protein (P<0.001). Inhibition of the expression of ACTN4 significantly reduced the cell viability [(0.78±0.07) vs (1.24±0.12), P<0.001], increased the apoptosis rate [(6.58±0.66)% vs (18.32±1.82)%, P<0.001], increased the proportion of cells in G(1) phase [(48.69±4.21)% vs (30.33±3.01)%, P<0.001], decreased the proportion of cells in S phase [(36.21±3.42)% vs (18.54±1.61)%, P<0.001], increased the protein levels of p21 and Bax (P<0.001), and decreased the levels of cyclin D1 and Bcl-2 protein (P<0.001). Compared with the miR-369-3p+ pcDNA group, overexpression of ACTN4 increased the proliferation ability of hepatocellular carcinoma MHCC97H cells at 72 hours of culture[(1.12±0.11) vs (0.68±0.06), P<0.001], significantly reduced the proportion of cells in G(1) stage [(38.81±3.24)% vs (51.80±4.57)%, P<0.001], significantly increased the proportion of S-phase cells [(31.65±3.11)% vs (15.69±1.44)%, P<0.001], decreased cell apoptosis rate [(13.86±1.37)% vs (22.69±2.24)%, P<0.001], increased protein expressions of cyclin D1 and Bcl-2 (P<0.001), decreased the protein expressions of p21 and Bax (P<0.001). Conclusion: MiR-369-3p can induce cell cycle arrest in G(1) phase, inhibit the proliferation and promote apoptosis of liver cancer cells by regulating the expression of ACTN4.

[Clinical and prognosis analysis of children with kidney retransplantation].

Objective: To analyze the clinical and prognosis of children with kidney retransplantation. Methods: Clinical data of 11 children who underwent kidney retransplantation from January 2011 to December 2020 in Department of Nephrology, Children's Hospital of Fudan University were retrospectilely analyzed. The clinical data including demographic parameters, primary diagnosis, characteristics in the follow-up of renal allograft were analyzed. Results: Totally 11 cases received secondary renal transplantation (male 6, female 5). They were initially diagnosed with chronic kidney disease at the age of 11.9 (7.4, 13.3) years. The median duration of dialysis was 22.1 (3.5, 36.5) months. In the first transplantation, recipient age was 13.9 (11.1, 15.2) years. Ten cases received donation from cardiac death donor (DCD) (9 cases received donors aged less than one year, 5 of them received whole kidney transplantation and one case received donor aged one to three years) and 1 case with living-related donor. Ten graft failures occurred within 1 month after renal transplantation and the other one occurred at the fifth month after transplantation. The causes included vascular factors (9 cases), rejection (1 case) and primary non-function (1 case). In the second transplantation, recipient age was 14.7 (11.7, 16.2) years. All the 11 children received dialysis (7 with PD and 4 with HD) and successfully completed the second transplantation. The median time between the two transplants was 210 (16, 1 041) days. Donors were all DCD donors from 3 years of age or older. The mean follow-up duration was (42±15) months. The estimated glomerular filtration rate was (85±34)ml/(min·1.73 m2) when the last investigation after kidney retransplantation with the kidney and patient all survived. Conclusions: Kidney retransplantation may have better prognosis in children. Dialysis transition during waiting period and DCD donor from 3 years of age or older can effectively ensure the success of kidney retransplantation.

[Necessity of repeated renal arteriography in the treatment of severe hemorrhage after percutaneous nephrolithotomy].

In order to discuss the necessity of repeated renal arteriography in the treatment of severe bleeding after percutaneous nephrolithotomy, this study retrospectively analyzed the clinical data of patients with severe bleeding after percutaneous nephrolithotomy in the Department of Urology Surgery of the First Hospital of China Medical University from August 2010 to July 2020, summarily analyzing treatments, outcomes and follow-up results of 27 patients who were treated by renal arteriography more than twice. Of these 27 patients who underwent repeated renal arteriography, 23 of them were treated by two times, 4 by three times, all of whom were diagnosed as renal vascular injury. And 15 of them were diagnosed as pseudoaneurysm, 4 of them renal arteriovenous fistula, and 8 of them pseudoaneurysm combined with renal arteriovenous fistula. After clear diagnosis, all these patients were performed with renal artery embolization, after which the symptoms of hematuria and lumbar discomfort were relieved or disappeared immediately. These patients were followed up from 6 months to 5 years, without corresponding symptoms recurring and with the renal function equivalent to that before embolization. The results showed that repeated renal arteriography was of great significance in the treatment of patients with severe bleeding after percutaneous nephrolithotomy, helping to clarify the cause of bleeding and giving appropriate and timely treatment.

[Establishment of human colon cancer transplantation tumor model in normal immune mice].

Objective: Establishment of a new model of human primary colon cancer transplantation tumor in normal immune mice and to provide a reliable experimental animal model for studying the pathogenesis of colon cancer under normal immunity. Methods: Human colon cancer cells come from colon cancer patients who underwent surgery in the Affiliated Hospital of Jining Medical College in 2017. The mice in the cell control group were inoculated with phosphate buffered solution (PBS) containing colon cancer cells, the microcarrier control group was inoculated with PBS containing microcarrier 6, and the cell-microcarrier complex group was inoculated with the PBS containing colon cancer cell-microcarrier complex. The cells of each group were inoculated under the skin of the right axilla of mice by subcutaneous injection, and the time, size, tumor formation rate and pathological changes under microscope were recorded. The transplanted tumor tissue was immunohistochemically stained with the EnVisiion two-step method, and the tumor formation rate of the transplanted tumor was judged according to the proportion of positive cells in the visual field. The polymerase chain reaction (PCR) method was used to detect the expression of human-specific Alu sequence in mice tumor tissue. Results: After inoculation with tumor cells, the mice in the cell control group and the microcarrier control group did not die and did not form tumors; the mice in the cell-microcarrier complex group had palpable subcutaneous tumors in the right axillary subcutaneously on the 5th to 7th days after inoculation, and tumor formation rate is 67% (10/15), and the tumor volume can reach about 500 mm(3) 2 to 3 weeks after vaccination. The immunohistochemistry results showed that CK20, CDX-2 and carcinoembryonic antigen were all positively expressed. The PCR results showed that the expression of human-specific Alu sequence can be detected in the transplanted tumor tissue of tumor-bearing mice. Conclusion: Human primary colon cancer cells used microcarrier 6 as a carrier to form tumors in normal immunized mice, and successfully established a new model of human colon cancer transplantation tumor in normal immune mice.

[Research progress on anti-tumor mechanism of tumor treating fields].

The anti-tumor mechanism of tumor treating fields (TTFields) is mainly through interfering with the dynamics of microtubule subunits in mitosis,which blocks the normal process of cell division and eventually leads to cell death.In recent years,relevant studies have found that TTFields still have immunological,molecular biological and other related anti-tumor mechanisms,and can induce reversible increase of cell membrane and blood-brain barrier permeability,which plays a synergistic role in combination with anti-tumor drugs.With the development of multi-system research,the specific treatment frequency,time and field strength of TTFields in different tumor treatments will be revealed.These research progress will further expand the application field of TTFields and benefit more patients.

LncRNA PCAT1 enhances cell proliferation, migration and invasion by miR-508-3p/NFIB axis in diffuse large B-cell lymphoma.

OBJECTIVE: In previous studies, PCAT1 has been proved to be a key carcinogenic driver in hepatocellular carcinoma. However, the regulatory mechanism of PCAT1 remains poorly understood in diffuse large B-cell lymphoma (DLBCL). PATIENTS AND METHODS: The expression of PCAT1, miR-508-3p and NFIB in DLBCL was detected by RT-qPCR assay. CCK-8 assay and transwell assay were used to measure cell proliferation, migration and invasion of DLBCL cells. Western blot assay was used to explore the protein expression of NFIB. Dual-Luciferase reporter assay was applied to measure the correlation between PCAT1, miR-508-3p and NFIB. RESULTS: PCAT1 was demonstrated to be upregulated in DLBCL tissues and cell lines. Besides, PCAT1 expression was associated with clinical stage and IPI score of DLBCL patients. Moreover, overexpression of PCAT1 promoted DLBCL cell proliferation, migration and invasion in vitro. Mechanistic investigation displayed that PCAT1 interplayed with miR-508-3p, while NFIB was a target gene of miR-508-3p. Further, miR-508-3p was in a downtrend while NFIB was increased in DLBCL tissues and cell lines. MiR-508-3p overexpression repressed DLBCL cell growth and metastasis, while PCAT1 overexpression reversed the inhibitory effect of miR-508-3p on the progression of DLBCL. Moreover, NFIB silencing suppressed DLBCL cell proliferation, migration and invasion, whereas PCAT1 vector or miR-508-3p knockdown destroyed the inhibitory of si-NFIB on the progression of DLBCL. CONCLUSIONS: Taken together, our findings validated that PCAT1 acted as completive endogenous RNA by sponging miR-508-3p and upregulating NFIB to facilitate DLBCL cell proliferation, migration and invasion.

Automated Radiographic Evaluation of Adenoid Hypertrophy Based on VGG-Lite.

Adenoid hypertrophy is a pathological hyperplasia of the adenoids, which may cause snoring and apnea, as well as impede breathing during sleep. The lateral cephalogram is commonly used by dentists to screen for adenoid hypertrophy, but it is tedious and time-consuming to measure the ratio of adenoid width to nasopharyngeal width for adenoid assessment. The purpose of this study was to develop a screening tool to automatically evaluate adenoid hypertrophy from lateral cephalograms using deep learning. We proposed the deep learning model VGG-Lite, using the largest data set (1,023 X-ray images) yet described to support the automatic detection of adenoid hypertrophy. We demonstrated that our model was able to automatically evaluate adenoid hypertrophy with a sensitivity of 0.898, a specificity of 0.882, positive predictive value of 0.880, negative predictive value of 0.900, and F1 score of 0.889. The comparison of model-only and expert-only detection performance showed that the fully automatic method (0.07 min) was about 522 times faster than the human expert (36.6 min). Comparison of human experts with or without deep learning assistance showed that model-assisted human experts spent an average of 23.3 min to evaluate adenoid hypertrophy using 100 radiographs, compared to an average of 36.6 min using an entirely manual procedure. We therefore concluded that deep learning could improve the accuracy, speed, and efficiency of evaluating adenoid hypertrophy from lateral cephalograms.

Supramolecular Solvent Extraction-Gas Chromatography-Tandem Mass Spectrometry for Detection of Benzodiazepines in Urines.

ABSTRACT: Objective To establish a method using supramolecular solvent and gas chromatography-tandem mass spectrometry （GC-MS/MS） to analyze 9 benzodiazepines in urines. Methods Urine samples containing 9 benzodiazepines reference substance were subjected to liquid-liquid extractions with supramolecular solvent, which consisted of tetrahydrofuran and 1-hexanol. The solvent layer was evaporated to dryness by stream of nitrogen. The residue was reconstituted with methanol, and GC-MS/MS analysis was performed on it. The way of data collection was multiple reaction monitoring （MRM） mode; internal standard method was employed for quantification. Results In urine samples, when the range of mass concentration was 1-100 ng/mL for diazepam, midazolam, flunitrazepam and clozapine, 5-100 ng/mL for lorazepam and alprazolam, 2-100 ng/mL for nitrazepam and clonazepam, and 0.2-100 ng/mL for estazolam, respectively, good linearities were obtained, correlation coefficients were 0.999 1-0.999 9, the lower limits of the quantifications ranged from 0.2 to 5 ng/mL, the extraction recovery rates were 81.12%-99.52%. The intra-day precision ［relative standard deviation （RSD）］ and accuracy （bias） were lower than 9.86% and 9.51%, respectively; the inter-day precision （RSD） and accuracy （bias） were lower than 8.74% and 9.98%, respectively. Nine drugs in urine samples showed good stability at ambient temperature and -20 ℃ within 15 days. The mass concentrations of alprazolam in urine samples obtained from 8 volunteers who took alprazolam tablets orally within 8-72 h after ingestions ranged from 6.54 to 88.28 ng/mL. Conclusion The supramolecular solvent extraction GC-MS/MS method for analysis of 9 benzodiazepines in urines provided by this study is simple, fast, accurate and sensitive, which can provide technical support for monitoring of poisoning by benzodiazepines for clinical treatment and judicial identification.

[Bibliometric and visual analysis of current status and trends of international research on keloids].

Objective: To conduct a bibliometric and visual analysis of the current status and trends of international research on keloids. Methods: The articles on keloid research from 2000 to 2020 in the core collection of Web of Science database were retrieved. The bibliometrics method was applied to analyze the number of articles published per year, journals and the first authors, countries and institutions, research fields, the annual citation frequency of cumulative published articles, highly cited articles, keywords. CiteSpace5.6.R2 software was applied to visually cluster keywords of the included articles, while VOSviewer1.6.13 software was applied to visually cluster keywords in titles and abstracts of the included articles in order to analyze research directions and development trends. Results: A total of 2 693 keloid-related articles were retrieved. From 2000 to 2020, the number of keloid-related articles published every year showed a significant upward trend. Totally 777 journals published keloid-related articles, of which Dermatologic Surgery published the most. Rei Ogawa published the largest number of keloid-related articles as the first author of 52 related articles. Keloid-related researches were conducted in a total of 98 countries, of which the United States published the largest number of related articles (613 articles), followed by China (524 articles) and Japan (107 articles). A total of 2 656 institutions conducted keloid-related research, and the institution with the largest number of related articles published was Shanghai Jiao Tong University of China (67 articles). According to the subject classification of the Web of Science database, the included articles involved 110 research fields, and the top three were dermatology, surgery, and medicine, research and experimental. The included articles were cited 47 746 times in total, and the citation frequency of cumulative published articles increased by year. The most frequently cited article (152 times) was published in 2011. There were a total of 45 571 keywords in the included literature. The top 5 keywords ranked according to the number of articles involved from high to low were keloid (588 articles), hypertrophic scar (385 articles), expression (198 articles), fibroblast (155 articles), and scar (133 articles). The keyword map visualized by CiteSpace5.6.R2 software further displayed that the research focused on the cause, performance, and composition of keloids. VOSviewer1.6.13 software analysis showed that the research direction of keloids was divided into two categories of clinical keloid management and keloid mechanism research, the initial research hotspots were mainly to explore the diagnosis and treatment of keloids from individual cases, with a preference for apparent research, while in the later stage, the focus was on the overall management of keloids, in which the mechanism research went to the molecular level. Conclusions: At present, international research interest on keloids is showing an upward trend. Both foreign (the United States, etc.) and domestic research institutions are conducting in-depth explorations of keloids. With dermatology as the leader, the research trend is gradually shifting from observational research to molecular research.

CLIC1 knockout inhibits invasion and migration of gastric cancer by upregulating AMOT-p130 expression.

PURPOSE: To explore the regulatory relationship between Chloride intracellular channel 1 (CLIC1) and Angiomotin (AMOT)-p130, and reveal the role of AMOT-p130 in gastric cancer (GC). METHODS: Immunohistochemistry was performed to analyze the expression of CLIC1 and AMOT-p130 in GC tissues and adjacent tissues. The expression of AMOT-p130 upon CLIC1 silencing was analyzed using RT-PCR, western blot, and immunofluorescence in GC cells. Transwell and wound-healing assays were performed to detect migration and invasion in GC cells. The changes in EMT-related proteins were detected using western blot. RESULTS: Our study found that high CLIC1 expression was significantly associated with low AMOT-p130 expression in GC tissues. Silencing CLIC1 expression in MGC-803 cells (MGC-803 CLIC1 KO) and AGS cells (AGS CLIC1 KO) decreased the invasive and migratory abilities of tumor cells, which were induced by the upregulation of AMOT-p130. Subsequently, we demonstrated that AMOT-p130 inhibits the invasive and migratory abilities of GC cells by inhibiting epithelial-mesenchymal transition. CONCLUSIONS: Our study suggests that AMOT-p130 could inhibit epithelial-mesenchymal transition in GC cells. CLIC1 may participate in the metastatic progression of GC by downregulating the expression of AMOT-p130.

Long non-coding RNA NEAT1 regulates Hodgkin's lymphoma cell proliferation and invasion via miR-448 mediated regulation of DCLK1.

OBJECTIVE: To explore whether long non-coding RNA nuclear enriched abundant transcript 1 (lncRNA NEAT1) could regulate Hodgkin's lymphoma (HL) cell proliferation and invasion through miR-448, which could target doublecortin like kinase 1 (DCLK1) and mediate DCLK1 expression. PATIENTS AND METHODS: Expressions of NEAT1, miR-448 and DCLK1 were evaluated by qRT-PCR or Western blot assay. Cell Counting Kit-8 (CCK-8) and transwell assay were utilized to detect cell proliferation and invasion capability in L428 cells respectively. The target relationship between NEAT1, miR-448 and DCLK1 was confirmed by Luciferase reporter assay. RESULTS: QRT-PCR results showed that NEAT1 expressed higher in HL tissues and cell lines than that in controls. In vitro experiments, NEAT1 downregulation could decrease cell proliferation and invasion capability in L428 cells. NEAT1 directly interacted with miR-448 and negatively regulated it. Moreover, DCLK1 was confirmed as a target of miR-448. DCLK1 expression was increased in L428 cells and positively regulated by NEAT1. NEAT1 overexpression upregulated the protein level of DCLK1 in L428 cells according to Western blot analysis. Additionally, DCLK1 overexpression could reverse the suppression on cell proliferation and invasion capability induced by NEAT1 knockdown or miR-448 overexpression. CONCLUSIONS: NEAT1 might be contributed to HL progression by promoting cell proliferation and invasion capability via miR-448 mediated DCLK1 expression.

SAPCD2 promotes invasiveness and migration ability of breast cancer cells via YAP/TAZ.

OBJECTIVE: The aim of this study was to explore whether SAPCD2 can affect the proliferation of breast cancer cells through YAP/TAZ, thus promoting the development of breast cancer (BCa). PATIENTS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) was performed to examine SAPCD2 expression level in BCa tissues collected from patients in different tumor TNM-stage. The correlation between SAPCD2 expression and clinicopathological features of patients were analyzed, and the Kaplan-Meier test was used for survival analysis. In addition, after knocking down SAPCD2 in cells, qRT-PCR and Western blot were applied to analyze the expression of the related genes and proteins, respectively. Moreover, Cell Counting Kit-8 (CCK-8) and transwell experiments were conducted to detect cell viability, migration, and invasion abilities. Furthermore, the changes in cell viability and migration and invasion abilities were examined after the simultaneous overexpression of YAP. RESULTS: It was found that SAPCD2 expression levels in BCa tissues were remarkably higher than those in the normal control samples; meanwhile, patients with tumor size >3 cm or in the T3+T4 stage had a relatively higher expression of SAPCD2 than those with tumor size <3 cm or in T1+T2 stage. At the same time, the overall survival rate of BCa patients with highly expressed SAPCD2 was remarkably lower than that of patients in the low expression group. Moreover, it was found that the SAPCD2 level was correlated to the tumor size, TNM stage, and lymph node metastasis. After knocking down SAPCD2 in cells, cell viability, migration, and invasion abilities, as well as the YAP/TAZ protein expression levels were all found to be remarkably attenuated, which, however, were reversed after simultaneous overexpression of YAP in cells. CONCLUSIONS: SAPCD2 may be able to enhance the proliferation ability of BCa cells via modulating the expression of YAP/TAZ, thereby prompting the progression of BCa.

Effects of miR-150-5p on cerebral infarction rats by regulating the Wnt signaling pathway via p53.

OBJECTIVE: The aim of this study was to screen differentially expressed micro ribonucleic acids (miRNAs) in the plasma of patients with cerebral infarction (CI). In addition, the role of miR-150-5p in the incidence of CI is mainly explored via animal models and molecular biology experiments. PATIENTS AND METHODS: Blood samples were collected from hospitalized patients diagnosed with CI, including 15 CI patients and 15 non-CI patients as negative controls. Differentially expressed miRNAs in the plasma of these subjects were screened by microarray analysis. TargetScan was applied to predict the target genes of miR-150-5p, which were subjected to GO and pathway enrichment analyses using WebGestalt. Sprague-Dawley rats were randomly divided into Sham group (n=20), Control group (n=20), and Experimental group (n=20). CI model in rats was established in the latter two groups. Rats in Experimental group and Control group were intravenously injected with miR-150-5p mimics or miR-negative control (NC), respectively. The expressions of vital genes in the Wnt signaling pathway, including p53, Cyclin D1 (CCND1), c-Myc, ß-catenin (CTNNB1) and Survivin were detected by Western blot in rats at 3 d after injection. RESULTS: A total of 3,568 differentially expressed miRNAs were detected in the peripheral blood between CI patients and controls, whose 2,100 were upregulated, including miR-150-5p (p<0.05). The target genes of miR-150-5p were involved in molecular pathways, such as the Wnt signaling pathway, carcinogenesis, endocrine regulation, and infection. Compared with rats in Control group, the protein expression of p53 was downregulated (p<0.05), while CCND1, c-Myc, CTNNB1 and Survivin were upregulated (p<0.05) in Experimental group. CONCLUSIONS: MiR-150-5p regulates the Wnt signaling pathway and participates in cell proliferation and apoptosis by downregulating p53, which may be a potential mechanism of CI induction.

KRAS gene silencing inhibits the activation of PI3K-Akt-mTOR signaling pathway to regulate breast cancer cell epithelial-mesenchymal transition, proliferation and apoptosis.

OBJECTIVE: Our study was performed to investigate the effect of KRAS gene silencing on epithelial-mesenchymal transition (EMT), proliferation, and apoptosis of breast cancer cells by mediating PI3K-Akt-mTOR signaling pathway. MATERIALS AND METHODS: The positive rate of KRAS protein expression was detected in tissues collected from breast cancer patients, associated with the analysis of the relationship between KRAS protein expression and clinicopathological features of patients. The expression of KRAS in breast cancer cell lines was tested to screen the suitable cell line. After cell transfection and grouping, qRT-PCR and Western blot were then used to detect the mRNA and protein expression in each group. MTT assay and flow cytometry detected cell proliferation, cell cycle, and apoptosis, respectively. RESULTS: The expression of KRAS in cancer tissue was much higher than that in paracancerous normal tissue, and its high expression was correlated statistically with lymph node metastasis, distant metastasis, and tumor infiltration level of patients (all p<0.05). KRAS was highly expressed in T47D and thus selected as suitable cell lines for subsequent analysis (all p<0.05). Compared with Blank group and NC group, there were significantly reduced mRNA and protein expression of KRAS, PI3K, Akt, mTOR, N-cadherin and Vimentin, increased PTEN and E-cadherin, decreased cell proliferation activity, and increased apoptosis in si-KRAS and Wortmannin groups (all p<0.05); while opposite trends were found in HA-KRAS group and Recilisib group (all p<0.05). However, there was no significant difference when compared si-KRAS+Recilisib with that in Blank and NC groups (all p>0.05). CONCLUSIONS: Silencing of KRAS gene expression may inhibit the activation of PI3K-Akt-mTOR signaling pathway, and thus inhibit EMT, proliferation and apoptosis of breast cancer cells. By contrast, activation of the studied signaling pathway can reverse the positive effect of KRAS gene silencing.