Granzyme B Promotes Proliferation, Migration and EMT Process in Gastric Cancer.

Granzyme B (GZMB), a critical member of the Gr gene family, is known to play an essential role in diverse physiological and pathological processes such as inflammation, acute and chronic inflammatory diseases, and cancer progression. In this study, we delve deeper into the role of GZMB within the context of gastric cancer (GC) to examine its expression patterns and functional implications. To accomplish this, we applied a combination of quantitative real-time polymerase chain reaction, western blotting, and immunohistochemistry techniques. These methodologies allowed us to accurately gauge GZMB expression levels in GC tissues and investigate their correlation with various clinical-pathological variables. Our secondary focus was to discern the regulatory influence of GZMB on GC cell biology. We used an array of assays including cell counting kit-8 (CCK-8), colony formation, 5-ethynyl-2'-deoxyuridine, and migration assays. The effect of GZMB on gastric cancer progression was further validated through a subcutaneous xenograft mouse model. Our findings underscored that GZMB mRNA and protein levels were upregulated in GC tissues, a feature that showed a significant correlation with GC staging. We also discovered that a decrease in GZMB expression via knockdown experiments suppressed the proliferation and migration capabilities of GC cells. This effect was manifested through diminished expression levels of epithelial-mesenchymal transition (EMT) markers. In stark contrast, the overexpression of GZMB through plasmid transfection appeared to enhance the proliferation and migration abilities of GC cells. This was coupled with an upregulation in EMT expression. Our study concludes by emphasizing that GZMB promotes the growth, migration, and EMT processes in gastric cancer. In vitro, cell-based experiments and in vivo xenograft mouse models confirm this. Our findings provide a more comprehensive understanding of GZMB's role in gastric cancer pathogenesis, potentially opening doors for novel therapeutic strategies targeting this molecular pathway.

miR-20a/Foxj2 Axis Mediates Growth and Metastasis of Colorectal Cancer Cells as Identified by Integrated Analysis.

BACKGROUND MicroRNAs (miRNAs) have a significant regulatory effect on the proliferation, migration, and invasion of cells, and have been widely reported to have oncogenic or tumor-suppressive impacts on various tumors. In the present study we assessed the regulation and function of miR-20a on colorectal cancer (CRC) cell lines. MATERIAL AND METHODS qPCR was used to quantify miR-20a expression. Luciferase reporter assay was conducted to confirm Foxj2 3'UTR associations. In addition, the function of miR-20a and Foxj2 in CRC was detected using MTT, colony formation, transwell assays, and cell cycle analysis. RESULTS Our data revealed that miR-20a expression was elevated in the CRC cell lines, and cell migration, proliferation, and invasion abilities were promoted by the overexpression of miR-20a. Moreover, Foxj2 was authenticated as a direct target gene of miR-20a in CRC cells. Furthermore, we found that the ectopic Foxj2 dramatically suppressed miR-20a-promoted proliferation, migration, invasion, and xenografts in vitro and in vivo, and induced cell cycle arrest at G1 stage. CONCLUSIONS Our results showing the roles of miR-20a/Foxj2 in carcinogenesis of CRC may help improve treatment of CRC.

[Optimization for preparation technology of shangke jiefu lotion by orthogonal test].

OBJECTIVE: To optimize the preparation technology of Shangke Jiefu lotion. METHODS: The extraction process was optimized by orthogonal test, with water addition, extraction times and time used for extraction as factors of investigation. In refined process test, alcohol precipitation concentration, time, and the relative density of extract were studied. Each factor had three levels. The content of sophorcarpidine and the yield of dry extract were used as the evaluation indexes. The content of sophorcarpidine was determined by HPLC, and dry extract rates were determined by drying method. RESULTS: The best extraction condition was as follows: the amount of water was 10 times of the medicinal materials, the decoction duration was 2 h and for 3 times. The optimum purification process was: alcohol precipitation concentration was 50%, time was 15 hours, relative density of extract was 1.05 g/mL. CONCLUSION: The optimized preparation technology of Shangke Jiefu lotion is stable, feasible and convenient. It provides a theoretical basis for standardized production.

The Number of Lymph Nodes Examined is Associated with Survival Outcomes of Neuroendocrine Tumors of the Jejunum and Ileum (siNET): Development and Validation of a Prognostic Model Based on SEER Database.

PURPOSE: The number of neuroendocrine tumors (NETs) is gradually increasing worldwide, and those located in the small intestine (siNETs) are the most common. As some biological and clinical characteristics of tumors of the jejunum and the ileum differ, there is a need to assess the prognosis of individuals with siNETs of the jejunum and ileum separately. We generated a predictive nomogram by assessing individuals with siNETs from the Surveillance, Epidemiology, and End Results (SEER) database. METHODS: We used univariate Cox regression analysis to determine both the overall survival (OS) and the cancer-specific survival (CSS) of 2501 patients with a pathological confirmation of siNETs of the jejunum and ileum. To predict 3-, 5-, and 10-year OS of siNETs, a nomogram was generated based on a training cohort and validated with an external cohort. Accuracy and clinical practicability were evaluated separately by Harrell's C-indices, calibration plots, and decision curves. The correlation was examined between dissected lymph nodes and positive lymph nodes. RESULTS: Dissection of 7 or more lymph nodes significantly improved patient OS and was found to be a protective factor for patients with siNETs. In Cox regression analyses, age, primary site, tumor size, N stage, M stage, and regional lymph node examination were significant predictors in the nomogram. A significant positive correlation was found between dissected lymph nodes and positive lymph nodes. CONCLUSIONS: Patients with 7 or more dissected lymph nodes showed an accurate tumor stage and a better prognosis. Our nomogram accurately predicted the OS of patients with siNETs.

Upregulated Linc01836 in Serum Promisingly Serving as a Diagnostic and Prognostic Biomarker for Colorectal Cancer.

Objectives: Colorectal cancer (CRC) is a common carcinoma of the gastrointestinal tract with high incidence and mortality worldwide. Studies have shown that long noncoding RNAs (lncRNAs) play important roles in CRC. Our purpose is to investigate the potential of serum Linc01836 as a diagnostic and prognostic marker in CRC. Methods: We evaluated the expression of Linc01836 via quantitative real-time polymerase chain reaction (qRT-PCR). The serum CEA, CA19-9, Cyfra21-1, and CA72-4 concentrations were measured by Architect I4000 SR. Receiver operating characteristic (ROC) curves were plotted to estimate the diagnostic value in CRC. Relationship between serum Linc01836 expression and clinicopathological characteristics of CRC cases was analyzed via chi-square test. The underlying mechanism of Linc01836 on the development and prognosis in CRC was predicted by bioinformatic analysis. Results: The method of qRT-PCR for Linc01836 detection was confirmed with high precision and specificity. Serum Linc01836 expression in CRC patients was significantly higher than that in healthy donors (p < 0.0001) and benign patients (p < 0.0001), and declined after resection (p < 0.01). High expression of Linc01836 was associated with histological stage (p = 0.002) and lymph node metastasis (p = 0.006). In addition, serum Linc01836 could effectively differentiate CRC patients from the healthy folks, with favorable area under the curve (AUC) of 0.809 (95% CI: 0.757-0.861, p < 0.001). What is more, the combination of serum Linc01836, CEA, and Cyfra21-1 could improve diagnostic sensitivity (92.0%). Linc01836 was averagely located in the nucleus and cytoplasm, suggesting that it might participate in CRC progression and prognosis through the crosstalk among lncRNAs, miRNAs, and mRNAs. Conclusion: Linc01836 may serve as a valuable noninvasive biomarker for population screening, early detection, and clinical surveillance of CRC.

The lncRNA B3GALT5-AS1 Functions as an HCC Suppressor by Regulating the miR-934/UFM1 Axis.

Accumulating evidence has demonstrated that long noncoding RNA (lncRNA) is importantly related to the occurrence and development of cancer. According to reports, the expression of B3GALT5-AS1 in hepatocellular carcinoma (HCC) is downregulated; however, the role of B3GALT5-AS1 in HCC is not yet clear. In this study, our purpose is to explore the biological function of B3GALT5-AS1 in HCC and its coupling mechanism with miR-934 and ubiquitin-fold modifier 1 (UFM1). We found that the B3GALT5-AS1 expression level was of significant reduction in both HCC tissues and cell lines; B3GALT5-AS1 overexpression (ov) may inhibit the malignant features of HCC. In addition, we demonstrated that miR-934 mimics could reverse the effect of B3GALT5-AS1 ov, which proved miR-934 was the downstream regulator of B3GALT5-AS1. Furthermore, si-UFM1 could reverse the effect of miR-934 inhibitor, which revealed the connection between them. Moreover, we found that B3GALT5-AS1 could keep down the PI3K/AKT pathway through UFM1. Our results demonstrated that B3GALT5-AS1 was an excellent HCC suppressant by regulating miR-934 and UFM1 to achieve negative regulation of HCC cell proliferation, invasion, and metastasis, indicating that B3GALT5-AS1 is a promising potential therapeutic target for HCC treatment.

Outcomes of revision neurolysis of the ulnar nerve and ulnar groove plasty for persistent and recurrent cubital tunnel syndrome-A retrospective study of 21 cases.

BACKGROUND: Patients undergoing revision surgical treatment of the ulnar nerve at the elbow for cubital tunnel syndrome (CuTS) will have worse results compared to patients successfully treated with primary surgery. OBJECTIVE: The purpose of this study is to evaluated clinical outcomes of revision neurolysis and ulnar groove plasty for recurrent and persistent cubital tunnel syndrome after failed surgical treatment. METHODS: This retrospective investigation included patients presented with recurrent and persistent CuTS who were treated surgically with combination of revision neurolysis and ulnar groove plasty at a single institution from May 2006 to Oct 2016 with postoperative follow-up more than 24 months. Demographic data of all patients including age, sex, months to revision surgery, presenting symptoms after index surgery, previous surgical procedure and intraoperative findings were all recorded and pre-operative and post-operative data were compared. McGowen grading was used to evaluated functional impairment before and after revision surgery. RESULTS: There were 28 patients were identified with recurrent and persistent CuTS after primary surgery and 21 patients (75%) were completed in this study with an average age was 56 years, mean duration of symptoms was 17.24 months, and mean postoperative follow-up was 35.38 months. 17 patients had McGowan stage III and 4 had stage II preoperatively. The most common cause of recurrent and persistent CuTS was perineural fibrosis with or without kink which accounts for 86.36% according to intraoperative findings. McGowan grading improved after revision neurolysis and ulnar groove plasty is 80.95%. Improvement of Visual Analogue Scale (VAS) and 2-point discrimination test were 81.25% and 85.71%, respectively. Patients satisfaction after revision neurolysis and ulnar groove plasty was 95.24%. CONCLUSION: The favorable results of this study demonstrated that revision neurolysis and ulnar groove plasty as the treatment of choice for recurrent or persistent cubital tunnel syndrome.

[Coumarins from Peucedanum harry-smithii var. subglabrum].

The root of Peucedanum harry-smithii var. subglabrum was extracted with methanol, then separated with solvents at different polarity into four fractions: aqueous (H2O), ethyl acetate (AcOEt), chloroform (CHCl3) and petroleum ether (DAB-6). From AcOEt psoralen, bargapten, xanthotoxin, marmesin, umbelliferone, scopoletin, (+/-) peuformosin, Pd-I b, (+/-) selinidin, praeruptorin D were isolated by column chromatography on silica gel, using petroleum ether/ethyl acetate as eluent. The structures of the coumarins were identified by 1H-NMR and 13C-NMR.

Overexpression of proteinase inhibitor 9 is associated with poor prognosis in human hepatocellular carcinoma and with proliferation and apoptosis in HepG2 cells in vitro.

Excessive proteinase inhibitor 9 (PI9) levels predict a poor outcome for patients with several tumor types. We compared the expression of PI9 in HepG2 cells and in 21 pairs of tumor and peritumor tissue samples using western blotting. Immunohistochemical staining was used to detect PI9 expression in 105 cases of hepatocellular carcinoma (HCC) and in 20 adjacent normal liver tissues. Changes in the degree of apoptosis and proliferation were determined before and after transfection with pcDNA3.1-PI9 and small interfering (si)RNA-PI9 using MTT analysis, colony formation assay, and flow cytometry. The correlation between PI9 expression and prognosis was determined in a large HCC patient cohort (n=105). Western blotting showed that PI9 expression was significantly higher in tumor tissues than in adjacent tissues. PI9 immunohistochemical staining was positive in 78.1% of HCC tissues, which was significantly higher than that seen in adjacent normal liver tissue (35%). PI9 expression in HCC correlated closely with the extent of tumor differentiation, tumor-node-metastasis staging, and tumor size. Cox regression analysis demonstrated that PI9 is an independent predictor of prognosis in patients with HCC, and is related to overall survival. The apoptosis of HepG2 cells was significantly increased following PI9 up-regulation and significantly decreased after siRNA interference against PI9 expression. Cell proliferation was significantly decreased following PI9 up-regulation and significantly increased after siRNA interference of PI9 expression. PI9 appears to contribute to the apoptosis of HCC, and could be an independent predictor of recurrence and a suitable pharmaceutical target in patients with HCC.