Virus-derived circular RNAs populate hepatitis C virus-infected cells.

It is known that pre-mRNAs in eukaryotic cells can be processed to circular RNAs by a backsplicing mechanism. Circular RNAs have great stability and can sequester proteins or small RNAs to exert functions on cellular pathways. Because viruses often exploit host pathways, we explored whether the RNA genome of the cytoplasmic hepatitis C virus is processed to yield virus-derived circRNAs (vcircRNAs). Computational analyses of RNA-seq experiments predicted that the viral RNA genome is fragmented to generate hundreds of vcircRNAs. More than a dozen of them were experimentally verified by rolling-circle amplification. VcircRNAs that contained the viral internal ribosome entry site were found to be translated into proteins that displayed proviral functions. Furthermore, two highly abundant, nontranslated vcircRNAs were shown to enhance viral RNA abundance. These findings argue that novel vcircRNA molecules modulate viral amplification in cells infected by a cytoplasmic RNA virus.

Particulate matter 2.5 exposure induces epithelial-mesenchymal transition via PI3K/AKT/mTOR pathway in human retinal pigment epithelial ARPE-19 cells.

Exposure to particulate matter 2.5 (PM2.5) has been linked to ocular surface diseases, yet knowledge of the molecular mechanism impacted on retina pathogenesis is limited. Therefore, the purpose of this study was to explore the effects and involved factors of PM2.5 exposure in human retinal pigment epithelial APRE-19 cells. Our data revealed a decreased cell viability and an increased migratory ability in APRE-19 cells after PM2.5 stimulation. The MMP-2 and MMP-9 protein levels were markedly increased while the MMPs regulators TIMP-1 and TIMP-2 were significantly reduced in PM2.5-exposed APRE-19 cells. PM2.5 also increased pro-MMP-2 expression in the cell culture supernatants. Additionally, PM2.5 promoted the EMT markers through the activation of PI3K/AKT/mTOR pathway. Moreover, the ICAM-1 production was also remarkably increased by PM2.5 but reduced by PI3K/AKT inhibitor LY294002 in APRE-19 cells. Taken together, these results suggest that PM2.5 promotes EMT in a PI3K/AKT/mTOR-dependent manner in the retinal pigment epithelium.

Host-derived circular RNAs display proviral activities in Hepatitis C virus-infected cells.

Viruses subvert macromolecular pathways in infected host cells to aid in viral gene amplification or to counteract innate immune responses. Roles for host-encoded, noncoding RNAs, including microRNAs, have been found to provide pro- and anti-viral functions. Recently, circular RNAs (circRNAs), that are generated by a nuclear back-splicing mechanism of pre-mRNAs, have been implicated to have roles in DNA virus-infected cells. This study examines the circular RNA landscape in uninfected and hepatitis C virus (HCV)-infected liver cells. Results showed that the abundances of distinct classes of circRNAs were up-regulated or down-regulated in infected cells. Identified circRNAs displayed pro-viral effects. One particular up-regulated circRNA, circPSD3, displayed a very pronounced effect on viral RNA abundances in both hepatitis C virus- and Dengue virus-infected cells. Though circPSD3 has been shown to bind factor eIF4A3 that modulates the cellular nonsense-mediated decay (NMD) pathway, circPSD3 regulates RNA amplification in a pro-viral manner at a post-translational step, while eIF4A3 exhibits the anti-viral property of the NMD pathway. Findings from the global analyses of the circular RNA landscape argue that pro-, and likely, anti-viral functions are executed by circRNAs that modulate viral gene expression as well as host pathways. Because of their long half-lives, circRNAs likely play hitherto unknown, important roles in viral pathogenesis.

Robotic versus laparoscopic surgery for rectal cancer after neoadjuvant chemoradiotherapy: A propensity-score matching analysis.

BACKGROUND: We aimed to investigate the advantages of robotic versus laparoscopic surgery for rectal cancer after neoadjuvant chemoradiotherapy as these remains unclear. METHODS: We retrospectively recruited eligible patients with rectal cancer undergoing robotic or laparoscopic surgery following neoadjuvant chemoradiotherapy. We compared the surgical outcomes between patients undergoing either robotic surgery or laparoscopic surgery was based on the propensity-score matching analysis. RESULTS: A total of 171 patients were recruited, including 76 who underwent robotic surgery and 95 who underwent laparoscopic surgery. There were no significant differences in clinical and pathological characteristics between the groups after propensity-score matching (56 matched pairs). Longer operation times (324.964 ± 83.435 vs. 246.232 ± 111.324 min, p < 0.001) and more blood loss (187.679 ± 176.615 vs. 98.214 ± 107.011, p < 0.001) were observed in the robotic group. The major complication rates were similar between the treatment groups after propensity matching (p = 0.086). There were no significant differences in disease-free survival rates (p = 0.205) and overall survival rates (p = 0.837) between the groups. CONCLUSION: Robotic surgery is associated with similar technical safety and oncologic efficacy compared to laparoscopic surgery for the treatment of rectal cancer after neoadjuvant chemoradiotherapy; it is an acceptable option for patients requiring minimally invasive surgery. Nevertheless, the longer operation times and greater blood loss seen in the present study are a stark reminder that the convenience and surgical precision, on which the marketing of robotic surgery is rooted, are yet to be proven and require further investigation.

Ramucirumab plus triplet chemotherapy as an alternative salvage treatment for patients with metastatic colorectal cancer.

BACKGROUND: Ramucirumab is indicated for salvage treatment after failure of first-line treatment for metastatic colorectal cancer (mCRC). However, the application of ramucirumab at later-line treatment in real-world practice has not received much discussion. METHODS: In this retrospective study, we enrolled 70 patients with mCRC who received ramucirumab plus chemotherapy at National Taiwan University Hospital between 2018 and 2019. RESULTS: Compared with those who received third- or later-line ramucirumab treatment, patients who received second-line ramucirumab treatment had significantly longer median time to treatment discontinuation (mTTD; 6.7 vs 3.6 months, P = .004) and median overall survival (mOS; not reached vs 7.6 months, P = .009). Multivariate analyses revealed that second-line ramucirumab and triplet chemotherapy backbone were the only independent predictive factors for long mTTD and mOS. Patients who received ramucirumab with triplet chemotherapy had a significantly longer mOS than did patients who received ramucirumab with doublet chemotherapy (not reached vs 5.6 months, P = .002). Among those receiving second-line ramucirumab treatment, combination with triplet chemotherapy led to a longer mTTD than did combination with doublet chemotherapy, but the difference was non-significant (not reached vs 4.4 months, P = .108). By contrast, in patients receiving fourth- or later-line ramucirumab, combination with triplet chemotherapy led to significantly longer mTTD than did combination with doublet chemotherapy (8.0 vs 2.9 months, P = .032). CONCLUSION: Ramucirumab plus triplet chemotherapy may be an alternative regimen in patients with mCRC, particularly as a later-line treatment modality.

Technical feasibility of robotic vs. laparoscopic surgery for locally advanced colorectal cancer invading the urinary bladder.

BACKGROUND: The aim of this study was to determine if robotic surgery can reproduce the technical advantages and oncologic outcomes of laparoscopic surgery for the treatment of locally advanced colorectal cancer invading the urinary bladder. METHODS: We retrospectively reviewed the prospectively maintained data of patients with locally advanced colorectal cancer invading the urinary bladder undergoing robotic or laparoscopic surgery between June 2006 and November 2020. Clinicopathologic features, surgical outcomes, and oncologic efficacy were compared between patient groups of robotic or laparoscopic surgery. All patients underwent surgery with the intent of R0 resection for the primary tumor. Major surgical complications were defined as Clavien-Dindo grade ≥ III. Multivariate regression analysis was performed to identify risk factors. RESULTS: A total of 41 patients (M:F = 32:9; median age: 63 [42-88] years) were analysed; 32 underwent laparoscopic surgery and 9 underwent robotic surgery. There was no statistically significant difference between the two groups in baseline demographic and clinicopathologic features. There were no significant differences in terms of mean operative time (353.24 vs. 387.33 min), mean blood loss (315.00 vs. 171.11 mL), mean number of lymph nodes harvested (27.16 vs. 23.50), R0 resection (89.7 vs. 66.7%), conversion (12.5 vs. 11.1%), major complication rate (9.4 vs. 22.2%), mean time to flatus passage (4.8 vs. 4.1 days), mean postoperative length of hospital stay (18.9 vs. 19.8 days), 5-year disease-free survival rate (64.6 vs. 62.5%) and overall survival rate (75.3 vs. 83.3%). Multivariate analysis showed that R1 resection was the only independent prognostic factor for reduced disease-free survival (hazard ratio 21.386; 95% confidence interval 1.991-229.723; p = 0.0115). CONCLUSIONS: Robotic surgery can reproduce the technical advantages and oncologic outcomes of laparoscopic surgery for the treatment of locally advanced colorectal cancer invading the urinary bladder. However, larger studies are mandatory to clarify the role of robotic surgery in such a scenario.

[Developing and Evaluating the Effectiveness of a Leadership Integrated Educational Program for Master's Nursing Students in Taiwan].

BACKGROUND: Nursing students with master degrees have the strong potential to serve as future leaders in medical teams. Implementing a well-developed and integrated educational program for nursing leadership at the master's level can strengthen the leadership of advanced practice nurses and promote a positive nursing practice environment. PURPOSE: To develop a leadership integrated educational program for master's nursing students and conduct a preliminary evaluation of the effectiveness of this program in cultivating leadership competencies in these students. METHODS: Phase 1: A modified Delphi survey conducted on 14 experts with clinical or academic backgrounds was used to identify the teaching objectives and strategies of the leadership integrated educational program. Phase 2: These teaching objectives and strategies were embedded into nine compulsory courses within a current training program for master's nursing students at a national university. The core elements of the leadership integrated educational program were incorporated into each compulsory course. The objectives of each compulsory course directly reflected the objectives of the integrated program. The leadership integrated educational program was implemented for one academic year, and its effectiveness was evaluated using a quasi-experimental test with a single group pre- and post-test design. A self-developed, 10-item "Master Nursing Student's Leadership Competence Scale" covering four core elements was applied to measure the self-reported leadership competencies of the participants. A paired sample t-test was applied to analyze the differences in leadership competencies between pre- and post-intervention. RESULTS: A consensus on the teaching objectives and strategies of the leadership integrated educational program was achieved in the first round of the Delphi survey. The overarching teaching objective of the leadership integrated educational program was to "lead the healthcare team with the leadership and competencies, and demonstrate the advanced nursing practice skills for improving quality of care." In addition, the four core elements under the overarching goal, i.e., personal characteristics, leading people, business management, and vision building, were proposed. Forty-eight master's nursing students participated in this study. The results showed the average total score of leadership competency was 42.33 ± 12.16 (potential range: 10 - 70), indicating that the participants had a middle level of leadership competency prior to program participation. After participating in the Leadership Integrated Educational Program for one academic year, the average total score for leadership competency increased to 51.27 ± 9.74, indicating that the participants still had a middle level of leadership competency. Nevertheless, the 8.94 increase in the post-intervention score was statistically significant (p < .01). Moreover, the scores for each subscale (personal characteristics, leading people, business management, and vision building) had all increased significantly increased from 13.52 to 15.71, 12.65 to 15.35, 8.15 to 10.31, and 8.02 to 9.90, respectively (p < .01). CONCLUSIONS / IMPLICATIONS FOR PRACTICE: This study offers proactive recommendations for reforming master's degree programs in nursing. The proposed multidisciplinary-expert-informed leadership integrated educational program may be used to strengthen leadership competencies in this student population. Furthermore, the findings provide a benchmark for developing an effective nursing leadership integrated educational program that may be incorporated into domestic master's degree programs.

Metronomic chemotherapy with tegafur-uracil following radical resection in stage II colorectal cancer.

BACKGROUND: Stage II colorectal cancer has a relatively good prognosis. Adjuvant chemotherapy following surgery is the standard treatment for stage III colorectal cancer but is not routinely recommended for all stage II colorectal cancer patients. We aimed to evaluate the clinical outcomes, treatment results, and prognostic factors in stage II colorectal cancer patients who underwent curative surgery with/without oral tegafur-uracil (UFT). METHODS: We included stage II colorectal cancer patients who underwent curative surgery and were followed up for at least 5 years after surgery at the National Taiwan University Hospital between January 2008 and December 2012. Excluding patients receiving neoadjuvant therapy, adjuvant therapy other than UFT, and those lost follow-up, patients treated with UFT (UFT group) and those without adjuvant therapy (surgery alone group) were analyzed for their clinical outcomes and prognostic factors. RESULTS: A total of 233 patients were recruited. Of these, 104 (44.64%) underwent only surgery while 129 (55.36%) received adjuvant chemotherapy with oral UFT following surgery. Recurrence or death occurred within 5 years in 60 patients (25.75%), with a significant difference between the surgery alone (36/104, 34.62%) and UFT groups (24/129, 18.61%) (p = 0.007). The UFT group demonstrated significantly superior 5-year disease-free (p = 0.003) and overall survival rates (p = 0.001), respectively. Patient age of ≤35.3 or ˃72.7 years, UFT duration of <486.8 days, 7.1 cm < tumor size ≤13.2 cm, number of harvested lymph nodes ≤13.5, and mucinous adenocarcinoma were associated with poorer 5-year overall survival. CONCLUSION: The present data suggest that UFT following curative surgery may be associated with lower recurrence and improved survival in patients with stage II colorectal cancer.

Precursor microRNA-122 inhibits synthesis of Insig1 isoform mRNA by modulating polyadenylation site usage.

The insulin-induced gene 1 protein (Insig1) inhibits the cholesterol biosynthesis pathway by retaining transcription factor SREBP in the endoplasmic reticulum, and by causing the degradation of HMGCR, the rate-limiting enzyme in cholesterol biosynthesis. Liver-specific microRNA miR-122, on the other hand, enhances cholesterol biosynthesis by an unknown mechanism. We have found that Insig1 mRNAs are generated by alternative cleavage and polyadenylation, resulting in specific isoform mRNA species. During high cholesterol abundance, the short 1.4-kb Insig1 mRNA was found to be preferentially translated to yield Insig1 protein. Precursor molecules of miR-122 down-regulated the translation of the 1.4-kb Insig1 isoform mRNA by interfering with the usage of the promoter-proximal cleavage-polyadenylation site that gives rise to the 1.4-kb Insig1 mRNA. These findings argue that precursor miR-122 molecules modulate polyadenylation site usage in Insig1 mRNAs, resulting in down-regulation of Insig1 protein abundance. Thus, precursor microRNAs may have hitherto undetected novel functions in nuclear gene expression.

Standardize the Surgical Technique and Clarify the Relevant Anatomic Concept for Complete Mobilization of Colonic Splenic Flexure Using da Vinci Xi® Robotic System.

BACKGROUND: The present study is to set up a standardized approach for complete mobilization of colonic splenic flexure using da Vinci Xi® robotic system, based on clarification of the mesenteric structures of distal transverse colon. METHODS: The surgical outcomes and relevant anatomic structures of 104 consecutive patients undergoing robotic resection of primary colorectal cancer with the intent of complete mobilization of colonic splenic flexure using da Vinci Xi® robotic system were retrospectively reviewed. RESULTS: Complete mobilization of colonic splenic flexure can be efficiently performed by the Xi® robotic system, as demonstrated by short operation time, minimal intra-operative blood loss, and few surgical complications. Xi® robotic system has overcome the drawbacks of Si® robotic system for the mobilization of colonic splenic flexure. The present study defined the following anatomic hallmarks for the colonic splenic flexure: (1) The transverse mesocolon distal to the inferior mesenteric vein adheres to the low border of pancreas by the avascular fibrous connective tissues, which have been inappropriately named as "mesenteric root"; (2) The colonic splenic flexure abuts closely to spleen with an acute angle in 78.85% (n = 82/104); (3) Only a minority of patients presented with the Riolan branch (15.38%, n = 16/104) or the Moskowitz artery (8.65%, n = 9/104). CONCLUSION: With increased maneuverability of Xi® robotic arms and the clarification of relevant anatomic concept, the surgical technique for the complete mobilization of colonic splenic flexure can be standardized; and the standardization of surgical technique is the first step toward the enhanced automation in the rapidly evolving robotic systems.

Supporting Role for GTPase Rab27a in Hepatitis C Virus RNA Replication through a Novel miR-122-Mediated Effect.

The small GTPase Rab27a has been shown to control membrane trafficking and microvesicle transport pathways, in particular the secretion of exosomes. In the liver, high expression of Rab27a correlates with the development of hepatocellular carcinoma. We discovered that low abundance of Rab27a resulted in decreased hepatitis C virus (HCV) RNA and protein abundances in virus-infected cells. Curiously, both cell-associated and extracellular virus yield decreased in Rab27a depleted cells, suggesting that reduced exosome secretion did not cause the observed effect. Instead, Rab27a enhanced viral RNA replication by a mechanism that involves the liver-specific microRNA miR-122. Rab27a surrounded lipid droplets and was enriched in membrane fractions that harbor viral replication proteins, suggesting a supporting role for Rab27a in viral gene expression. Curiously, Rab27a depletion decreased the abundance of miR-122, whereas overexpression of miR-122 in Rab27a-depleted cells rescued HCV RNA abundance. Because intracellular HCV RNA abundance is enhanced by the binding of two miR-122 molecules to the extreme 5' end of the HCV RNA genome, the diminished amounts of miR-122 in Rab27a-depleted cells could have caused destabilization of HCV RNA. However, the abundance of HCV RNA carrying mutations on both miR-122-binding sites and whose stability was supported by ectopically expressed miR-122 mimetics with compensatory mutations also decreased in Rab27a-depleted cells. This result indicates that the effect of Rab27a depletion on HCV RNA abundance does not depend on the formation of 5' terminal HCV/miR-122 RNA complexes, but that miR-122 has a Rab27a-dependent function in the HCV lifecycle, likely the downregulation of a cellular inhibitor of HCV gene expression. These findings suggest that the absence of miR-122 results in a vulnerability not only to exoribonucleases that attack the viral genome, but also to upregulation of one more cellular factor that inhibit viral gene expression.

Revisiting Rectosacral and Waldeyer's Fascia by Laparoscopic or Robotic Approach - Video Vignette.

During the clinical practice of total mesorectal excision (TME) for the treatment of middle and low rectal cancer, posterior mobilisation of the rectum is along the holy plane, which consists of loose areolar connective tissues [1]. With further posterior downward mobilisation, a thick tough fascia will be encountered, generally known as rectosacral fascia, and failure to recognise and divide the rectosacral fascia can perforate the mesorectum or lead to severe presacral haemorrhage. This article is protected by copyright. All rights reserved.

Revisiting Toldt Fascia Through Robotic Top-down and Medial-to-lateral Apporach - Video Vignette.

In this video (Toldt fascia.wmv), we demonstrated anatomic features of Toldt fascia, based on the meticulous dissection and high-resolution images inherent in robotic surgery. Toldt fascia is sandwiched by the overlying mesothelial layer of the mesocolon and underlying mesothelial layer of the retroperitoneum, comprised loose fibrous tissues with minute vessels inside, and is contiguous from the ileocecal mesentery radix to the upper rectum. The medial-to-lateral surgical dissection plane is readily developed within the Toldt fascia; however, any attempt to dissect along the interface between Toldt fascia and the overlying mesocolon or underlying retroperitoneum failed. This article is protected by copyright. All rights reserved.

Impact of marital coping on the relationship between body image and sexuality among breast cancer survivors.

PURPOSE: The aims of the study were (1) to understand the relationship between women's marital coping efforts and body image as well as sexual relationships and (2) to test a hypothesized model suggesting that marital coping efforts have a mediating effect on the relationship between body image and sexual relationships among breast cancer survivors. METHODS: A total of 135 breast cancer survivors who had finished cancer treatment completed a self-reported questionnaire concerning body image, marital coping efforts, and sexual relationship. RESULTS: Body image, marital coping, and sexual relationship were found to be significantly correlated with each other. The final path model showed that negative marital coping efforts, including avoidance and self-blame, significantly mediated the effect of women's body image on their sexual relationships. Although a positive approach did not correlate with body image, it did significantly correlate with women's sexual relationships. CONCLUSIONS: Our study demonstrated that negative marital coping using self-blame and avoidance mediated the association between body image and sexual relationship. Future interventions to address the body image and sexual life of breast cancer survivors should be considered using positive approaches that prevent disengaged avoidance or self-blame coping efforts intended to deal with marital stress.

Impact of positron-emission tomography on the surgical treatment of locoregionally recurrent colorectal cancer.

BACKGROUND: The effect of positron emission tomography (PET) on the surgical treatment of locoregionally recurrent colorectal cancer (LRRCRC) remains unclear and warrants further investigation. MATERIAL AND METHODS: A total of 193 patients with LRRCRC were identified from a prospectively maintained institutional database, of whom 134 LRRCRCs were deemed resectable and underwent resection with curative intent, whereas the remaining 59 LRRCRCs were unresectable. Patients with resectable LRRCRC were further classified according to whether recurrence was detected solely by PET (n = 35, PET-only group) or by a combination of computed tomography (CT)/magnetic resonance imaging (MRI) and PET (n = 99, CT/MRI/PET group). Clinicopathologic features, operative morbidity/mortality, and overall survival were compared between the patient groups based on long-term follow-up for at least 5 years. RESULTS: Patients in the PET-only group tended to have less extensive organ resection (p = 0.0074), less blood loss (p < 0.0001), and shorter operation time and hospitalization (p < 0.0001), but surgical complication and readmission rates were not significantly different (p > 0.05) compared with the CT/MRI/PET group. Although the PET-only group had significantly higher R0 resection rate (80 % vs. 54.55 %, p = 0.0079), they also had a higher risk (17.14 % vs. 2.02 %, p = 0.0011) of sham operation. The estimated 5-year and 10-year survival rates significantly decreased in order (p < 0.0001) from PET-only (85.71 % and 57.98 %) and CT/MRI/PET (41.41 % and 15.93 %) to unresectable group of patients (16.95 % and 1.88 %). Subset analysis of the CT/MRI/PET group indicated that PET improved surgical decision-making because 24 (24.2 %) LRRCRCs that manifested on CT/MRI as equivocal lesions were later confirmed by PET as resectable recurrences, while 18 (19.4 %) LRRCRCs that manifested on CT/MRI as resectable lesions were later diagnosed by PET as more disseminated unresectable recurrences and precluded futile surgery. CONCLUSION: PET alone can identify a subset (20.9 %) of LRRCRCs with less tumor burden for timely surgery; PET in combination with CT/MRI can better define the resectability of LRRCRCs. The positive impacts of PET can translate into better surgical outcomes, with enhanced safety and patient survival.