Clinical outcomes of laser hemorrhoidoplasty with feeding vessels suture ligation: a retrospective study in a single center.

BACKGROUND: Laser hemorrhoidoplasty has demonstrated significant therapeutic effectiveness. To diminish postoperative bleeding and enhance overall outcomes, we have additionally adopted suture ligating the feeding vessels. This study aimed to understand the treatment outcomes and any associated complications. METHODS: This study comprised patients with symptomatic grade II-III hemorrhoids who underwent laser hemorrhoidoplasty with feeding vessel suture ligation and Milligan-Morgan hemorrhoidectomy between 1 September 2020, and 31 August 2022. Surgical-related details, postoperative pain, discomfort after discharge, hemorrhoid recurrence, and any complications were collected from inpatient records, outpatient follow-ups, and telephone interviews. Initially, we will analyze the distinctions between the laser group and the traditional group, followed by an investigation into complications and satisfaction within the laser surgery subgroup. RESULTS: The study included 323 patients, with 173 undergoing laser hemorrhoidoplasty (LHP) and 150 undergoing Milligan-Morgan hemorrhoidectomy. Regarding pain assessment, the LHP group exhibited superior performance compared to traditional surgery at postoperative 4 h, before discharge, and during the first and second outpatient visits, with statistically significant differences. Additionally, the LHP group had a lower rate of urinary retention and experienced significantly less pain, with statistically significant differences. CONCLUSIONS: Laser hemorrhoidoplasty with feeding vessels suture ligation has been shown to reduce postoperative pain and appears to be a promising minimally invasive treatment option for symptomatic grade II and III hemorrhoids.

The Physical Resilience Instrument for Older Adults (PRIFOR) in Surgical Inpatients: Further Evidence for Its Factor Structure and Validity.

BACKGROUND: The Physical Resilience Instrument for Older Adults (PRIFOR) is a questionnaire for assessing physical resilience in older adults suffering from acute health stressors. Prior psychometric evidence of the PRIFOR showed that it has good criterion-related validity, known-group validity, predictive validity, and internal consistency. However, it is unclear whether the PRIFOR can be replicated in older adults suffering after surgical treatment. OBJECTIVES: This study aimed at evaluating whether the three-factor structure of the PRIFOR can be replicated in older adults suffering after surgical treatment. Moreover, the concurrent validity of the PRIFOR was examined using the association between the PRIFOR and measures of depression, cognition, activities of daily living, and frailty. DESIGN AND SETTING: A longitudinal study was adopted in a tertiary-care medical center in Taiwan. PARTICIPANTS: A total of 207 patients aged 65 years old and older who underwent surgery and if they were able to communicate independently. MEASUREMENTS: The PRIFOR, the 5-item Geriatric Depression Scale, the Short Portable Mental Status Questionnaire, the Katz Index of Independence in Activities of Daily Living and Clinical Frailty Scale were all assessed after surgery. RESULTS: The three-factor structure (positive thinking, cope and adjust lifestyle, and belief and hopeful mindset) was supported by the CFA results in the present sample. In addition, the PRIFOR showed good concurrent validity with depression (r = -0.470 to -0.542), cognition (r = 0.358 to 0.409), activities of daily living (r = 0.209 to 0.310), and frailty (r =-0.161 to -0.237). CONCLUSION: The PRIFOR can be recommended to measure physical resilience in older adults suffering after surgical treatment. For the adequate estimation of older adults' level of physical resilience postoperatively and to guide the implementation of individualized interventions, it is important to provide appropriate care for older adults to recover after surgery.

Impact of chemotherapy on surgical outcomes in ileostomy reversal: a propensity score matching study from a single centre.

PURPOSE: In patients with chemotherapy, there is no consensus on the timing of ileostomy closure. Ileostomy reversal could improve the quality of life and minimise the long-term adverse events of delayed closure. In this study, we evaluated the impact of chemotherapy on ileostomy closure and searched for the predictive factors for complications. METHODS: We retrospectively analysed 212 patients with rectal cancer who underwent ileostomy closure surgery during and without chemotherapy and were consecutively enrolled between 2010 and 2016. As a result of the heterogeneity of the two groups, propensity score matching (PSM) was performed with a 1:1 PSM cohort. RESULTS: A total of 162 patients were included in the analysis. The overall stoma closure-related complications (12.4% vs. 11.1%, p = 1.00) and major complications (2.5% vs. 6.2%, p = 0.44) were not significantly different between the two groups. Multivariate analysis demonstrated that chronic kidney disease and bevacizumab use are risk factors for major complications. CONCLUSION: Patients with oral or intravenous chemotherapy can safely have ileostomy closure with an adequate time delay from chemotherapy. When patients use bevacizumab, major complications related to ileostomy closure should still be cautioned.

Determining tolerance levels for quality assurance of 3D printed bolus for modulated arc radiotherapy of the nose.

Given the existing literature on the subject, there is obviously a need for specific advice on quality assurance (QA) tolerances for departments using or implementing 3D printed bolus for radiotherapy treatments. With a view to providing initial suggested QA tolerances for 3D printed bolus, this study evaluated the dosimetric effects of changes in bolus geometry and density, for a particularly common and challenging clinical situation: specifically, volumetric modulated arc therapy (VMAT) treatment of the nose. Film-based dose verification measurements demonstrated that both the AAA and the AXB algorithms used by the Varian Eclipse treatment planning system (Varian Medical Systems, Palo Alto, USA) were capable of providing sufficiently accurate dose calculations to allow this planning system to be used to evaluate the effects of bolus errors on dose distributions from VMAT treatments of the nose. Thereafter, the AAA and AXB algorithms were used to calculate the dosimetric effects of applying a range of simulated errors to the design of a virtual bolus, to identify QA tolerances that could be used to avoid clinically significant effects from common printing errors. Results were generally consistent, whether the treatment target was superficial and treated with counter-rotating coplanar arcs or more-penetrating and treated with noncoplanar arcs, and whether the dose was calculated using the AAA algorithm or the AXB algorithm. The results of this study suggest the following QA tolerances are advisable, when 3D printed bolus is fabricated for use in photon VMAT treatments of the nose: bolus relative electron density variation within [Formula: see text] (although an action level at [Formula: see text] may be permissible); bolus thickness variation within [Formula: see text] mm (or 0.5 mm variation on opposite sides); and air gap between bolus and skin [Formula: see text] mm. These tolerances should be investigated for validity with respect to other treatment modalities and anatomical sites. This study provides a set of baselines for future comparisons and a useful method for identifying additional or alternative 3D printed bolus QA tolerances.

Simultaneous Ipsilateral Vascularized Lymph Node Transplantation and Contralateral Lymphovenous Anastomosis in Bilateral Extremity Lymphedema with Different Severities.

BACKGROUND: Extremity lymphedema can occur bilaterally with different severities on each side. The aim of this study is to investigate the treatment outcomes of such patients with bilateral extremity lymphedema of different severities. PATIENTS AND METHODS: Between 2013 and 2017, patients with bilateral extremity lymphedema of different severities according to the Taiwan Lymphoscintigraphy Staging (TLS) system were retrospectively reviewed. Ipsilateral vascularized lymph node transplantation (VLNT) was indicated in TLS total obstruction and contralateral lymphovenous anastomosis (LVA) in TLS partial obstruction with patent lymphatic vessels on indocyanine green lymphography. Outcomes were assessed using circumference improvement, frequency of cellulitis, and lymphedema-specific quality of life (LYMQoL) questionnaires. RESULTS: A total of 10 patients with bilateral extremity lymphedema with median age of 63 (range 12-75) years were included. The median symptom duration of the lymphedematous limb was 60 (range 36-168) months and 12 (range 1-60) months in the VLNT and LVA group, respectively (p < 0.05). At average follow-up of 37.5 (range 14-58) months, the average limb circumference improvement was 2.4 (range - 3.3 to 7.8) cm in the VLNT group and 2.3 (range 0.3-7) cm in the LVA group (p = 1). The median episodes of cellulitis decreased significantly from 4 to 0.5 and 1 to 0 times/year in the VLNT and LVA group, respectively (p = 0.02, p = 0.06). The overall LYMQoL score improved from 4.5 preoperatively to 7.5 postoperatively (p < 0.01). CONCLUSIONS: Limb-specific VLNT and LVA selected by TLS effectively treated bilateral extremity lymphedema with different severities.

Helicobacter pylori infection and the risk of colorectal cancer: a nationwide population-based cohort study.

BACKGROUND: The role of Helicobacter pylori (H. pylori) infection in the development of colorectal neoplasia has been a matter of scientific debate with controversial findings. AIMS: This study examined the association between H. pylori infection and colorectal cancer (CRC) in a nationwide population-based Chinese cohort study. METHODS: A total of approximately 3936 individuals with newly diagnosed H. pylori infection (the H. pylori-infected cohort) and 15 744 age- and sex-matched patients with diagnoses absence of H. pylori infection (the comparison cohort) from 2000 to 2005 were identified from Taiwan's National Health Insurance Research Database. The Kaplan-Meier method was used for measuring the cumulative incidence of CRC in each cohort. Cox proportional hazards models were used to compute hazard ratios (HRs) and accompanying 95% confidence intervals (CIs) for the estimation of the association between H. pylori infection and CRC. RESULTS: The cumulative incidence of CRC was higher in H. pylori-infected cohort than that in the comparison cohort (log-rank test, P < 0.001). After adjustment for potential confounders, H. pylori infection was associated with a significantly increased risk of CRC (adjusted HR 1.87; 95% CI 1.37-2.57). In addition, the HR of CRC appeared to increase with increasing frequency of clinical visits for H. pylori infection. CONCLUSIONS: Our study demonstrated that H. pylori infection was associated with an increased risk of CRC, which warrants confirmation and exploration of the underlying biologic mechanisms by future studies.

[Pulmonary extranodal natural killer/T-cell lymphoma: clinical analysis of three cases].

Objective: To improve the understanding of pulmonary involvement of extranodal natural killer/T-cell lymphoma (ENKTL) by analyzing the clinical manifestations, imaging and pathological features of this disease. Methods: Three cases of ENKTL, proven by pathological diagnosis in Fuzhou General Clinical Medical College of Fujian Medical University, were retrospectively analyzed. Results: All of the 3 cases were males, aged 74, 61 and 49 years, respectively. The main clinical symptoms included cold and fever. One patient had nasal congestion and runny nose. Chest CT showed multiple lung nodules (n=3), masses (n=2) and patchy shadows (n=2), with irregular lesions and clear boundaries. The 3 cases had been misdiagnosed as community acquired pneumonia, and treated with intravenous injection of moxifloxacin. ENKTL was confirmed by CT-guided percutaneous biopsy with immunohistochemical staining, which showed that 3 cases were positive for CD(56), CD(2), in situ hybridization for EBV encoded RNA (EBER), while negative for CD(20), and 2 cases were positive for CD(3), Granzyme B; and 1 case was positive for T-cell intracytoplasmic antigen-1. All 3 patients received chemotherapy, but 2 patients died, one of rejection 1 month later after bone marrow transplantation. One patient had improved after chemotherapy with follow-up. Conclusion: ENKTL should be considered when patients presented with fever, multiple lung nodules or consolidations which were non-responsive to antibiotics. Lung biopsy was the key to diagnosis.

The expression and clinical significance of miRNA-99a and miRNA-224 in non-small cell lung cancer.

OBJECTIVE: To investigate the expression and clinical significance of miRNA-99a and miRNA-224 in the serum of patients with non-small cell lung cancer (NSCLC). PATIENTS AND METHODS: 83 patients with NSCLC, who were diagnosed and treated in our hospital from January 2014 to September 2017, were included in the experiment group. 79 patients, who made health check up, were included in the control group. The quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) technique was used to test the expressions of miRNA-99a and miRNA-224 in the serum of the patients in the two groups, and the relationship between the expression levels of miRNA-99a and miRNA-224 and the clinicopathological features of the patients with NSCLC was analyzed; the correlation between the expression of miRNA-99a and the expression of miRNA-224 in NSCLC was also analyzed. RESULTS: The expression level of miRNA-99a in the patients with NSCLC was significantly lower than that in the patients in the normal control group; the differences were statistically significant (p<0.001). The expression level of miRNA-224 in the patients with NSCLC was markedly higher than that in the patients in the normal control group; the differences were statistically significant (p<0.001). The expression level of miRNA-99a in the patients with NSCLC was remarkably correlated with pathological stage, the presence or absence of lymph node metastasis and tissue differentiation (p<0.001). The expression level of miRNA-224 in the patients with NSCLC was significantly correlated with pathological stage, the presence or absence of lymph node metastasis and pathological grade (p<0.001). The results of partial correlation analysis showed that the expression levels of miRNA-99a and miRNA-224 were negatively correlated with each other in NSCLC (r=-0.985, p<0.001). CONCLUSIONS: MiRNA-99a and miRNA-224 may be involved in the development and progression of NSCLC. MiRNA-99a is associated with NSCLC pathological stage, lymph node metastasis and tissue differentiation, while miRNA-224 is associated with NSCLC pathological stage, lymph node metastasis and pathological grade. MiRNA-99a and miRNA-224 can be used as clinical monitoring indicators for NSCLC.

Time-to-progression following conventional compared with drug-eluting-bead transcatheter arterial chemoembolisation in patients with large hepatocellular carcinoma.

AIMS: To identify the optimal transarterial chemoembolisation (TACE) approach in patients with large hepatocellular carcinoma (HCC; >5 cm) by comparing conventional TACE (cTACE) and drug-eluting-bead (DEB)-TACE. MATERIALS AND METHODS: This retrospective study included 63 consecutive HCC patients who received TACE at a single medical centre from September 2009 to October 2015. Primary endpoints were 3-year overall survival (OS) rate and time-to-progression (TTP). Hazard ratios (HRs) from Kaplan-Meier curves were calculated to compare survival estimates. RESULTS: The median OS was shorter in the cTACE group, but was not significantly different from the DEB-TACE group (33.9 versus 35.6 months, respectively; p=0.52). The mean TTP was shorter in the cTACE group than in the DEB-TACE group (13.9 versus 17.5 months, respectively; p=0.01). There was no difference in 3-year survival (HR=0.95, 95% confidence interval [CI]: 0.51-1.78; p=0.880) and TTP (HR=0.70, 95% CI: 0.42-1.16; p=0.147) between the groups; however, patients treated with DEB-TACE were more likely to have longer TTP in the first 2 years following treatment (HR=0.51, 95% CI: 0.29-0.88; p=0.009). CONCLUSION: Although DEB-TACE is not superior in terms of TTP or OS in patients with large HCC, it may have greater efficacy in the first 24 months following therapy.

Correction: The atypical ubiquitin ligase RNF31 stabilizes estrogen receptor α and modulates estrogen-stimulated breast cancer cell proliferation.

Since the publication of the above article, the authors have noted that the input data in Fig. 6E is incorrect. The correct data are included in the below Fig. 6E. The mistake does not affect the conclusions of the paper as the levels of input proteins remain similar between samples. We apologise for any inconvenience caused by this error.

Visualising the boundary sharpness of uterine zonal structures using high-resolution T2-weighted images during the menstrual cycle.

AIM: To investigate whether there is an optimal time in the menstrual cycle to obtain the best image quality of uterine zonal structures with high-field magnetic resonance imaging (MRI). MATERIALS AND METHODS: Thirty-eight normal volunteers with regular menstrual cycles underwent pelvic 3 T high-resolution T2-weighted three-dimensional (3D) turbo spin echo (TSE) with variable flip angle MRI examinations during the menstrual phase (MP), follicular phase (FP), peri-ovulatory phase (OP), and luteal phase (LP). Two radiologists blinded evaluated the boundary sharpness of the three zonal structures of the uterine corpus and cervix on mid-sagittal images using a three-point Likert-scale. The signal intensity (SI) on T2-weighted sequences of each zonal structure was measured and the ratio between the SI of adjacent structures was calculated. Paired Wilcoxon's test and repeated measurement analysis of variance were used to investigate the differences among the four phases. RESULTS: No variation during the menstrual cycle was found in 10.5% (4/38) of volunteers and their boundaries were all well-defined. The OP exhibited the clearest boundaries of the corpus zonal structures. For the endometrium to junctional zone, mean scores of boundary sharpness from high to low were 3 (OP), 2.97 (FP), 2.76 (LP), 2.74 (MP); that for the junctional zone to myometrium were 2.76 (OP), 2.42 (FP), 2.32 (LP), 2.11 (MP); which were consistent with the SI ratio results. The results for the cervix showed no statistical difference during the menstrual cycle (p>0.05), and was well-defined throughout. CONCLUSIONS: The OP is recommended as the best phase to investigate zonal-related uterine corpus diseases due to the best contrast. For cervical diseases, imaging could be performed when necessary at any time point, due to the limited influence of menstrual phases on cervical zone delineation.

Sequential gene regulatory events leading to glucocorticoid-evoked apoptosis of CEM human leukemic cells:interactions of MAPK, MYC and glucocorticoid pathways.

Gene expression responses to glucocorticoid (GC) in the hours preceding onset of apoptosis were compared in three clones of human acute lymphoblastic leukemia CEM cells. Between 2 and 20h, all three clones showed increasing numbers of responding genes. Each clone had many unique responses, but the two responsive clones showed a group of responding genes in common, different from the resistant clone. MYC levels and the balance of activities between the three major groups of MAPKs are known important regulators of glucocorticoid-driven apoptosis in several lymphoid cell systems. Common to the two sensitive clones were changed transcript levels from genes that decrease amounts or activity of anti-apoptotic ERK/MAPK1 and JNK2/MAPK9, or of genes that increase activity of pro-apoptotic p38/MAPK14. Down-regulation of MYC and several MYC-regulated genes relevant to MAPKs also occurred in both sensitive clones. Transcriptomine comparisons revealed probable NOTCH-GC crosstalk in these cells.

The effectiveness of a saline mouth rinse regimen and education programme on radiation-induced oral mucositis and quality of life in oral cavity cancer patients: A randomised controlled trial.

Radiation therapy (RT) and concurrent chemotherapy RT (CCRT) generate radiation-induced oral mucositis (OM) and lower quality of life (QOL). This study assessed the impact of a saline mouth rinse regimen and education programme on radiation-induced OM symptoms, and QOL in oral cavity cancer (OCC) patients receiving RT or CCRT. Ninety-one OCC patients were randomly divided into a group that received saline mouth rinses and an education programme and a control group that received standard care. OM symptoms and QOL were assessed with the WHO Oral Toxicity Scale, MSS-moo and UW-QOL. Data were collected at the first postoperative visit to the radiation department (T0) and at 4 weeks and 8 weeks after beginning RT or CCRT. Patients in both groups had significantly higher levels of physical and social-emotional QOL at 8 weeks after beginning RT or CCRT compared to the first visit. Patients in the saline rinse group had significantly better physical and social-emotional QOL as compared to the standard care group at 8 weeks. Radiation-induced OM symptoms and overall QOL were not different between the groups. We thus conclude the saline rinse and education programme promote better physical and social-emotional QOL in OCC patients receiving RT/CCRT.

Factors associated with healthcare professional's rating of disfigurement and self-perceived body image in female patients with head and neck cancer.

The purpose of this study was to determine factors associated with self-perceived body image in female patients with head and neck cancer (HNC), and factors associated with healthcare professional's rating of disfigurement, as well as the correlation between patient and observer ratings. This cross-sectional study recruited 105 women with HNC at a large medical centre. Measures of facial disfigurement and body image, as well as demographic and clinical characteristics, were collected. Multivariate multiple linear regression modelling was used to identify factors associated with healthcare professional's rating of disfigurement and patient self-perceived body image. Disfigurement ratings by healthcare professionals were positively associated with patient self-perceived body image. Medical treatment, cancer stage, radiation dose and cancer site were significantly associated with disfigurement. Medical treatment was an important predictor of perceived body image. These findings indicate a moderate prevalence of disfigurement among women with HNCs. Patients with more disfigurement were more likely to have dissatisfaction with their body image. Nursing professionals need to carefully assess the appearance of women with HNC. Camouflage interventions can be used to help appropriately cope with the disfigurement, and to achieve improved satisfaction with their body image.

HLA-A*33:74N, a novel HLA-A*33 variant, identified by sequence-based typing in a Taiwanese individual.

A one nucleotide replacement in codon 803 of HLA-A*33:03:01 results in a novel allele, HLA-A*33:74N.

Eradication of hepatitis C virus profoundly prolongs survival in hepatocellular carcinoma patients receiving transarterial chemoembolization.

Adjuvant pegylated interferon plus ribavirin treatment (PegIFN/RBV) reduces recurrence and prolongs survival in early stage hepatocellular carcinoma (HCC) patients with chronic hepatitis C (CHC) infection receiving resection or ablation. However, the impact of antiviral therapy in intermediate and advanced stage of CHC-HCC patients is uncertain. This study aimed to investigate the impact PegIFN/RBV treatment on recurrence-free interval and survival in patients with HCC receiving transarterial chemoembolization (TACE). From 2010 to 2013, 274 CHC patients from a 1073 patient-based cohort composed of freshly diagnosed HCC and receiving TACE treatment the Chang Gung Memorial Hospital, Linkou Medical Center were recruited. Propensity score matching (PSM) (age, gender, AST to Platelet Ratio Index (APRI), tumour size, tumour number and Child-Turcotte-Pugh score) with the ratio 1:2 for patients with and without PegIFN/RBV treatment was performed. Statistics were performed with SPSS V.20 (IBM, USA). After matching, 153 patients were analysed and 27 patients (17.6%) achieved sustained virologic response (SVR). The 2-year cumulative overall survival rate and recurrence-free survival rate among patients with SVR, non-SVR, and untreated were 85.2% vs 58.3% vs 69.6% (P=.001) and 73.3% vs 53.8% vs 58.5% (P=.013). By Cox regression analysis, non-SVR, untreated, increase CTP score and nonresponder to TACE were independent factors related to mortality. The SVR achieved by PegIFN/RBV treatment markedly improves survival and reduces tumour recurrence in CHC-HCC patients receiving TACE treatment after complete response.

Negative association between acrylamide exposure and body composition in adults: NHANES, 2003-2004.

BACKGROUND/OBJECTIVES: Acrylamide is present in mainstream cigarette smoke and in some food prepared at high temperature. Animal studies have shown that acrylamide exposure reduces body weight. Prenatal exposure to acrylamide also has been linked to reduced birth weight in human. Whether acrylamide exposure is associated with altered body compositions in adults is not clear. SUBJECTS/METHODS: We selected 3623 subjects (aged ⩾20 years) from a National Health and Nutrition Examination Survey (NHANES) in 2003-2004 to determine the relationship among hemoglobin adducts of acrylamide (HbAA), hemoglobin adducts of glycidamide (HbGA) and body composition (body measures, bioelectrical impedance analysis (BIA), dual energy x-ray absorptiometry (DXA)). Data were adjusted for potential confounding variables. RESULTS: The geometric means and 95% CI concentrations of HbAA and HbGA were 60.48 (59.32-61.65) pmol/g Hb and 55.64 (54.40-56.92) pmol/g Hb, respectively. After weighting for sampling strategy, we identified that one-unit increase in natural log-HbAA, but not HbGA, was associated with reduction in body measures (body weight, body mass index (BMI), subscapular/triceps skinfold), parameters of BIA (fat-free mass, fat mass, percent body fat, total body water) and parameters of DXA (android fat mass, android percent fat, gynoid fat/lean mass, gynoid percent mass, android to gynoid ratio). Subgroup analysis showed that these associations were more evident in subjects at younger age, male gender, whites, lower education level, active smokers and those with lower BMI. CONCLUSIONS: Higher concentrations of HbAA are associated with a decrease in body composition in the US general population. Further studies are warranted to clarify this association.

Phagocytosis and immune response studies of Macrophage-Nanodiamond Interactions in vitro and in vivo.

The applications of nanodiamond as drug delivery and bio-imaging can require the relinquishing ND-drug conjugate via blood flow, where interaction with immune cells may occur. In this work, we investigated the ND penetration in macrophage and the immune response using the tissue-resident murine macrophages (RAW 264.7). Confocal fluorescence imaging, immunofluorescence analysis of nuclear translocation of interferon regulatory factor IRF-3 and transcriptional factor NF-κΒ, analysis of pro-inflammatory cytokines production IL-1ß, IL-6 IL-10 with a reverse transcription-polymerase chain reaction technique were applied. The TNF-α factor production has been studied both in vitro at ND interaction with the macrophage and in vivo after ND injection in the mice blood system using immunoassay. The macrophage antibacterial function was estimated through E. coli bacterial colony formation. ND didn't stimulate the immune response and functionality of the macrophage was not altered. Using MTT test, ND was found negligibly cytotoxic to macrophages. Thus, ND can serve as a biocompatible platform for bio-medical applications. Left: Graphic representation of Nanodiamond internalization in macrophage. Right: (a) Fluorescence images of lysosomes, (b) nanodiamond and (c) merged image of nanodiamond internalization in macrophage.

Simultaneous induction of apoptosis and necroptosis by Tanshinone IIA in human hepatocellular carcinoma HepG2 cells.

Tanshinone IIA (Tan IIA), a constituent of the traditional medicinal plant Salvia miltiorrhiza BUNGE, has been reported to possess anticancer activity through induction of apoptosis in many cancer cells. Surprisingly, the present study finds that Tan IIA simultaneously causes apoptosis and necroptosis in human hepatocellular carcinoma HepG2 cells. We further find that apoptosis can be converted to necroptosis by pan-caspase inhibitor Z-VAD-fmk, and the two death modes can be blocked by necroptotic inhibitor necrostatin-1. The underlying mechanisms are revealed by analysis of the signaling molecules using western blotting. In control cells, FLICE inhibitory protein in short form (FLIPS) is expressed in relatively high levels and binds to caspase 8 in ripoptosome, which supposedly sustains cell survival. However, in Tan IIA-treated cells, FLIPS is down-regulated and may thus cause homodimer formation of cleaved caspase 8, cleavage of receptor-interacting serine/threonine-protein kinases 1, 3 (RIP1, RIP3), and mixed-lineage kinase domain-like (MLKL), in turn leads to cell apoptosis. In parallel, Tan IIA causes necroptosis by forming a suggested necrosomal complex composed of RIP1/RIP3. Regarding the inhibitors, z-VAD-fmk diminishes the cleaved caspase 8, RIP1, RIP3, and MLKL induced by Tan IIA, and reconstructs the ripoptosome complex, which marks cells moving from apoptosis to necroptosis. Nec-1 recovers the Tan IIA down-regulated FLIPS, consequently causes FLIPS to form heterodimer with caspase 8 and thus block apoptosis. Meanwhile, cleaved forms of RIP1 and RIP3 were observed preventing necroptosis. Intriguingly, the cytotoxicity of tumor necrosis factor-related apoptosis-inducing ligand to HepG2 cells is enhanced by Tan IIA in a pilot study, which may be attributed to low FLIPS levels induced by Tan IIA. In short, Tan IIA simultaneously induces both Nec-1 inhibition and FLIPS regulation-mediated apoptosis/necroptosis, which has not been previously documented. Moreover, the involvement of the cleavage type of MLKL in executing necroptosis warrants further investigation.