Diagnostic prediction of gastrointestinal graft-versus-host disease based on a clinical- CT- signs nomogram model.

OBJECTIVE: Gastrointestinal graft-versus-host disease (GI-GVHD) is one of the complications that can easily occur after hematopoietic stem cell transplantation (HSCT). Timely diagnosis and treatment are pivotal factors that greatly influence the prognosis of patients. However, the current diagnostic method lacks adequate non-invasive diagnostic tools. METHODS: A total of 190 patients who suspected GI-GVHD were retrospectively included and divided into training set (n = 114) and testing set (n = 76) according to their discharge time. Least absolute shrinkage and selection operator (LASSO) regression was used to screen for clinically independent predictors. Based on the logistic regression results, both computed tomography (CT) signs and clinically independent predictors were integrated in order to build the nomogram, while the testing set was verified independently. The receiver operating characteristic (ROC), area under the curve (AUC), decision curve, and clinical impact curve were used to measure the accuracy of prediction, clinical net benefit, and consistency of diagnostic factors. RESULTS: Four key factors, including II-IV acute graft-versus-host disease (aGVHD), the circular target sign, multifocal intestinal inflammation, and an increased in total bilirubin, were identified. The combined model, which was constructed from CT signs and clinical factors, showed higher predictive performances. The AUC, sensitivity, and specificity of the training set were 0.867, 0.787, and 0.811, respectively. Decision curve analysis (DCA), net reclassification improvement (NRI), and integrated discrimination improvement (IDI) showed that the developed model exhibited a better prediction accuracy than the others. CONCLUSIONS: This combined model facilitates timely diagnosis and treatment and subsequently improves survival and overall outcomes in patients with GI-GVHD. CRITICAL RELEVANCE STATEMENT: GI-GVHD is one of the complications that can easily occur after HSCT. However, the current diagnostic approach lacks adequate non-invasive diagnostic methods. This non-invasive combined model facilitates timely treatment and subsequently improves patients with GI-GVHD survival and overall outcomes. KEY POINTS: • There is currently lacking of non-invasive diagnostic methods for GI-GVHD. • Four clinical CT signs are the independent predictors for GI-GVHD. • Association between the CT signs with clinical factors may improve the diagnostic performance of GI-GVHD.

Causal relationship between psychological factors and hepatocellular carcinoma as revealed by Mendelian randomization.

PURPOSE: The impact of psychological factors on the incidence of hepatocellular carcinoma (HCC) in humans remains unclear. Mendelian randomization (MR) study is a novel approach aimed at unbiased detection of causal effects. Therefore, we conducted a two-sample MR to determine if there is a causal relationship between psychological distress (PD), participation in leisure/social activities of religious groups (LARG), and HCC. METHODS: The genetic summary data of exposures and outcome were retrieved from genome-wide association studies (GWAS). We used PD and LARG as exposures and HCC as outcome. Five MR methods were used to investigate the causal relationship between PD, LARG, and HCC. The result of inverse variance weighted (IVW) method was deemed as principal result. Besides, we performed a comprehensive sensitivity analysis to verify the robustness of the results. RESULTS: The IVW results showed that PD [odds ratio (OR) 1.006, 95% confidence interval (CI) 1.000-1.011, P = 0.033] and LARG (OR 0.994, 95% CI 0.988-1.000, P = 0.035) were causally associated with the incidence of HCC. Sensitivity analysis did not identify any bias in the results. CONCLUSION: PD turned out to be a mild risk factor for HCC. In contrast, LARG is a protective factor for HCC. Therefore, it is highly recommended that people with PD are seeking positive leisure activities such as participation in formal religious social activities, which may help them reduce the risk of HCC.

Narrative review of magnetic resonance imaging in quantifying liver iron load.

Objective: To summarize the research progress of magnetic resonance imaging (MRI) in quantifying liver iron load. Methods: To summarize the current status and progress of MRI technology in the quantitative study of liver iron load through reviewing the relevant literature at home and abroad. Results: Different MRI sequence examination techniques have formed a series of non-invasive methods for the examination of liver iron load. These techniques have important clinical significance in the imaging diagnosis of liver iron load. So far, the main MRI methods used to assess liver iron load are: signal intensity measurement method (signal intensity, SI) [signal intensity ratio (SIR) and difference in in-phase and out-of-phase signal intensity], T2/R2 measurement (such as FerriScan technique), ultra-short echo time (UTE) imaging technique, and susceptibility weighted imaging (including conventional susceptibility weighted imaging) (SWI), quantitative susceptibility mapping (QSM), T2*/R2* measurement, Dixon and its derivative techniques. Conclusion: MRI has become the first choice for the non-invasive examination of liver iron overload, and it is helpful to improve the early detection of liver injury, liver fibrosis, liver cirrhosis and liver cancer caused by liver iron overload.

Magnetic resonance imaging assessment of the changes of cardiac and hepatic iron load in thalassemia patients before and after hematopoietic stem cell transplantation.

To investigate the value of T2* technique on 3.0 T magnetic resonance imaging (MRI) in evaluating the changes of cardiac and hepatic iron load before and after hematopoietic stem cell transplantation (HSCT) in patients with thalassemia (TM), the 141 TM patients were divided into 6 group for subgroup analysis: 6, 12, 18, 24 and > 24 months group, according to the postoperative interval. The T2* values of heart and liver (H-T2*, L-T2*) were quantified in TM patients before and after HSCT using 3.0 T MRI T2* technology, and the corresponding serum ferritin (SF) was collected at the same time, and the changes of the three before and after HSCT were compared. The overall H-T2* (P = 0.001) and L-T2* (P = 0.041) of patients after HSCT were higher than those before HSCT (mean relative changes = 19.63%, 7.19%). The H-T2* (P < 0.001) and L-T2* (P < 0.001) > 24 months after HSCT were significantly higher than those before HSCT (mean relative changes = 69.19%, 93.73%). The SF of 6 months (P < 0.001), 12 months (P = 0.008), 18 months (P = 0.002) and > 24 months (P = 0.001) were significantly higher than those before HSCT (mean relative changes = 57.93%, 73.84%, 128.51%, 85.47%). There was no significant improvement in cardiac and liver iron content in TM patients within 24 months after HSCT, while the reduction of cardiac and liver iron content in patients is obvious when > 24 months after HSCT.

A multicenter study on the quantification of liver iron concentration in thalassemia patients by means of the MRI T2* technique.

Objective: To investigate the feasibility and accuracy of quantifying liver iron concentration (LIC) in patients with thalassemia (TM) using 1.5T and 3T T2* MRI. Methods: 1.5T MRI T2* values were measured in 391 TM patients from three medical centers: the T2* values of the test group were combined with the LIC (LICF) provided by FerriScan to construct the curve equation. In addition, the liver 3T MRI liver T2* data of 55 TM patients were measured as the 3T group: the curve equation of 3T T2* value and LICF was constructed. Results: Based on the test group LICF (0.6-43 mg/g dw) and the corresponding 1.5T T2* value, the equation was LICF = 37.393T2*∧(-1.22) (R2 = 0.971; P < 0.001). There was no significant difference between LICe - 1.5T and LICF in each validation group (Z = -1.269, -0.977, -1.197; P = 0.204, 0.328, 0.231). There was significant consistency (Kendall's W = 0.991, 0.985, 0.980; all P < 0.001) and high correlation (rs = 0.983, 0.971, 0.960; all P < 0.001) between the two methods. There was no significant difference between the clinical grading results of LICe - 1.5T and LICF in each validation group (χ2 = 3.0, 4.0, 2.0; P = 0.083, 0.135, 0.157), and there was significant consistency between the clinical grading results (Kappa's K = 0.943, 0.891, 0.953; P < 0.001). There was no statistical correlation between the LICF (≥14 mg/g dw) and the 3T T2* value of severe iron overload (P = 0.085). The LICF (2-14 mg/g dw) in mild and moderate iron overload was significantly correlated with the corresponding T2* value (rs = -0.940; P < 0.001). The curve equation constructed from LICF and corresponding 3T T2* values in this range is LICF = 18.463T2*∧(-1.142) (R2 = 0.889; P < 0.001). There was no significant difference between LICF and LICe - 3T in the mild to moderate range (Z = -0.523; P = 0.601), and there was a significant correlation (rs = 0.940; P < 0.001) and significant consistency (Kendall's W = 0.970; P = 0.008) between them. LICe - 3T had high diagnostic efficiency in the diagnosis of severe, moderate, and mild liver iron overload (specificity = 1.000, 0.909; sensitivity = 0.972, 1.000). Conclusion: The liver iron concentration can be accurately quantified based on the 1.5T T2* value of the liver and the specific LIC-T2* curve equation. 3T T2* technology can accurately quantify mild-to-moderate LIC, but it is not recommended to use 3T T2* technology to quantify higher iron concentrations.

Quantification of cardiac iron in patients with thalassemia with 3-T MRI calibrated by 1.5-T MRI.

BACKGROUND: Due to the small sample size of many studies, it remained unclear what standardized reference range the T2* cutoff at 3 T would be used to assess the severity of cardiac iron load. In addition, the number of patients with moderate to severe cardiac iron load was small in some studies, especially the sample of patients with severe cardiac iron load. PURPOSE: To explore the feasibility, reproducibility, and reliability of using T2* values in quantifying cardiac iron load in patients with thalassemia at 3 T. MATERIAL AND METHODS: A total of 122 patients with thalassemia underwent cardiac T2* imaging at both 1.5 T and 3 T. Cardiac R2* (1000/T2*) values of the 100 patients at 3 T were fitted against the values at 1.5 T using linear regression and the prediction equation was derived. The remaining 22 cases were used to test the prediction accuracy of the equation. RESULTS: The combined R2* values exhibited a strong linear relationship between 1.5 T and 3 T (r = 0.830，P<0.001). At the center, it had a slope of 1.348 and an intercept of 37.279. According to the equation, the truncated T2* values of cardiac iron overload and cardiac heavy iron overload at 3 T were <10 ms and <6 ms, respectively. The two truncated T2* values were used to diagnose different levels of cardiac iron overloaded of 22 patients at 3 T; the accuracy rates were 95.5% and 100.0%, respectively. CONCLUSION: T2* quantification of cardiac iron load at 3 T MRI resulted to be feasible, reproducible, and reliable.

Quantitative Analysis of Liver Iron Deposition Based on Dual-Energy CT in Thalassemia Patients.

Background: To explore the feasibility and accuracy of liver iron deposition based on dual-energy CT in thalassemia patients. Materials and methods: 105 thalassemia patients were examined with dual-energy CT and MR liver scanning. Dual-energy CT was performed to measure CT values on 80kVp, 140kVp, and virtual iron content (VIC) imaging; ΔH was figured out by the difference in CT values between 80kVp and 140kVp. Using the liver iron concentration (LIC) obtained by FerriScan as a gold standard, the correlation between CT measurements and LIC was evaluated. Receiver operating characteristic (ROC) analysis was used to evaluate the diagnostic performance for dual-energy CT in liver iron quantification and stratification. Results: The correlation analysis between CT measurements and LIC showed that 80kVp, 140kVp, VIC, and ΔH all had a high positive correlation with LIC (P<0.001). The correlation analysis among different degree groups of VIC, ΔH, and LIC showed that the normal, moderate, and severe groups of VIC and ΔH had moderate or high positive correlations with that of LIC (P<0.01), but the mild group had no correlation (P>0.05). ROC analysis revealed that the corresponding optimal cutoff value of VIC was -2.8, 6.3,11.9 HU (corresponds to 3.2,7.0,15.0 mg/g dry weight) respectively, while the ΔH were 5.1, 8.4, 17.8HU, respectively. The area under the receiver operating characteristic curves (AUCs) for both VIC and ΔH increased with LIC thresholds. Conclusion: Dual-energy CT can accurately quantify and stratify liver iron deposition, contributing to predicting the status of liver iron deposition in thalassemia patients.

Incidence and Risk Factors of Surgical Complications and Anastomotic Leakage After Transanal Total Mesorectal Excision for Middle and Low Rectal Cancer.

PURPOSE: Transanal total mesorectal excision (taTME) is a promising surgical procedure for middle and low rectal cancer; however, it is linked to significant morbidity. This study aimed to determine the incidence of postoperative surgical complications and anastomotic leakage following taTME and to identify their associated risk factors. METHODS: The prospective clinical data of 114 patients, who underwent taTME and primary anastomosis for mid-low rectal cancer between November 2016 and June 2022, were retrospectively analyzed. Univariate and multivariate analyses were performed to identify clinical characteristics and risk factors for predicting surgical complications and anastomotic leakage. RESULTS: Surgical complications occurred in 40 (35.1%) patients within the first 30 days following surgery. Based on the Clavien-Dindo classification, minor complications (Clavien-Dindo grades I + II) accounted for 30.7%, while major complications (Clavien-Dindo grades III + IV) accounted for only 4.4%. None of the patients died within 30 days. The incidence of anastomotic leakage was 15.8%: 4.4% as grade A (5 cases), 9.6% as grade B (11 cases), and 1.8% as grade C (2 cases). Preoperative T3-4 was identified as an independent risk factor for surgical complications (p = 0.031) by multivariate analysis. American Society of Anesthesiology score ≥ 3 (P = 0.021) and incomplete total mesorectal excision specimens (P = 0.030) were significantly associated with the risk of anastomotic leakage. CONCLUSIONS: In this study, the incidence of surgical complications and anastomotic leakage in taTME aligned with previously reported rates. Preoperative T3-4 was significantly associated with surgical complications. American Society of Anesthesiology score ≥ 3 and incomplete TME specimens independently increased the risk of anastomotic leakage.

Prognostic nutrition index predicts short-term surgical complications in patients with rectal cancer after laparoscopic surgery.

Purpose: Surgical complications following laparoscopic rectal cancer surgery remain a major clinical problem. The prognostic nutritional index (PNI) is reportedly associated with postoperative outcomes. We aimed to evaluate the correlation between PNI and short-term surgical complications in patients with rectal cancer after laparoscopic surgery. Methods: The prospective clinical data of 225 patients with rectal cancer receiving laparoscopic surgery between January 2021 and April 2022 were retrospectively analyzed. The cut-off values and diagnostic accuracy of PNI preoperatively and on postoperative day (POD) 1 were determined using receiver operating characteristic (ROC) curves. Univariate and multivariate analyses were performed to identify clinical characteristics and risk factors for surgical complications. Results: In total, 81 (36.0%) patients developed surgical complications. The optimal cut-off value for preoperative PNI was 40.15, and that for PNI on POD 1 was 35.28. The DeLong test found no statistically between-group difference in the area under the ROC curve (P = 0.598). Multivariate analysis identified that a preoperative PNI ≤40.15 [odds ratio (OR): 2.856, 95% confidence interval (CI): 1.287-6.341, P = 0.010] and PNI on POD 1 ≤35.28 (OR: 2.773, 95% CI: 1.533-5.016, P = 0.001) were independent risk factors for surgical complications. Patients with a preoperative PNI ≤40.15 or PNI on POD 1 ≤35.28 were more likely to have surgical complications after laparoscopic surgery for rectal cancer (61.1% vs. 31.2%, P = 0.001; 53.0% vs. 28.9%, P = 0.001). Conclusion: Preoperative and POD 1 PNI were independent predictors of short-term surgical complications after laparoscopic surgery for rectal cancer.

The taTME learning curve for mid-low rectal cancer: a single-center experience in China.

PURPOSE: As transanal total mesorectal excision (taTME) is performed worldwide, the optimization of existing training and guidance programs to enhance new taTME learners' competence in performing this procedure is warranted. This study aimed to evaluate the taTME learning curve in patients with mid-low rectal cancer. METHODS: Patients who underwent taTME for mid-low rectal cancer between October 2015 and August 2021 at a single center were included. A cumulative sum (CUSUM) learning curve analysis was performed with the total operation time as the study outcome. The learning curve was analyzed using risk-adjusted CUSUM analysis, with postoperative complications and anastomotic leakage (AL) as outcomes. RESULTS: In total, 104 consecutive patients were included in this study. The CUSUM learning curve for total operative time started declining after 42 cases (309.1 ± 84.4 vs. 220.2 ± 46.4, P < 0.001). The risk-adjusted CUSUM (RA-CUSUM) learning curve for postoperative complications fluctuated in cases 44-75 and declined significantly after case 75. The RA-CUSUM learning curve for AL declined after 68 cases. CONCLUSIONS: taTME had learning curves of 42, 75, and 68 cases for total operative time, postoperative complications, and AL, respectively. A surgeon may require 42 and 75 cases to achieve "proficiency" and "mastery" in taTME procedures, respectively.

Blocking autophagy with chloroquine aggravates lipid accumulation and reduces intracellular energy synthesis in hepatocellular carcinoma cells, both contributing to its anti-proliferative effect.

PURPOSE: The autophagy inhibitor chloroquine enhances the effect of targeted therapy using tyrosine kinase inhibitor in liver cancer. We would like to further understand the specific mechanism by which chloroquine inhibits the proliferation of tumor cells. METHODS: We used a human hepatocarcinoma cell line (HepG2) as cell culture model. In contrast to the control groups (treated only with complete medium), cells in experimental groups were treated either with complete medium + 40 ng/ml Hepatocyte growth factor (HGF), or with complete medium + 60 µM chloroquine or with complete medium + 40 ng/ml HGF + 60 µM chloroquine for 24 h. Cell number and ATP content were investigated using spectrophotometric assays. Cell proliferation and apoptosis were detected by immunohistochemistry. Cell morphological alterations were examined by Giemsa and H&E staining. Cellular lipid content was determined by Oil Red O staining and Triglyceride quantification assay. Autophagy-related proteins (LC3B and p62) and hepatocyte proliferation-related protein (S6K1) were examined using western blot. The autophagic flux of cells was assessed by mRFP-EGFP-LC3 transfection assay. RESULTS: We found that chloroquine inhibited the proliferation of HepG2 cells, as evidenced by a decrease in cellular ATP content, Ki-67 and S6K1 protein expression and a reduction in cell number. This finding was associated with an increase in lipid content. As expected, chloroquine inhibited autophagy of HepG2 cells, as evidenced by the accumulation of LC3B-II and the significant upregulation of p62. mRFP-EGFP-LC3 transfection assay showed that indeed chloroquine blocked the autophagic flux in HepG2 cells. CONCLUSION: Chloroquine impaired proliferation of HepG2 cells might be due to intracellular accumulation of lipids and inhibition of energy synthesis.

Clinical benefits of PD-1/PD-L1 inhibitors in patients with metastatic colorectal cancer: a systematic review and meta-analysis.

BACKGROUND: Immunotherapy for colorectal cancer has developed rapidly in the past decade. Many high-quality clinical trials examining the application of PD-1/PD-L1 inhibitors in patients with metastatic colorectal cancer (mCRC) have been conducted in recent years. However, the clinical benefits, including the efficacy and safety of these treatments against mCRC, remain controversial. Hence, we conducted this meta-analysis on the clinical benefits of PD-1/PD-L1 inhibitors in patients with mCRC. METHODS: We searched online databases including MEDLINE, Embase, Cochrane Library, and Web of Science, from inception to January 4, 2021. The outcomes related to efficacy and safety were extracted and analyzed. Subgroup analyses were conducted according to the categories of dMMR-MSI-H (tumors with mismatch repair deficiency and high levels of microsatellite instability) ≥ 5% vs. dMMR-MSI-H < 5%, monotherapy vs. combination therapy, PD-1 inhibitors vs. PD-L1 inhibitors, and nivolumab vs. pembrolizumab. RESULTS: Fourteen studies including 1129 subjects were included in our systematic review. The overall complete response (CR), partial response (PR), stable disease (SD), and progression of disease (PD) rates were 0.01 (95% CI 0.00-0.04), 0.04 (95% CI 0.05-0.26), 0.27 (95% CI 0.22-0.32), and 0.44 (95% CI 0.30-0.58), respectively. The overall objective response rate (ORR) and disease control rate (DCR) were 0.16 (95%CI 0.06-0.31) and 0.50 (95%CI 0.35-0.65), respectively. The overall rate of adverse events (AEs) and severe adverse responses (SAEs) were 0.84 (95% CI 0.72-0.92) and 0.30 (95% CI 0.20-0.41), respectively. The ORRs of the dMMR-MSI-H ≥ 5% and dMMR-MSI-H < 5% subgroups were 0.40 (95% CI 0.30-0.51) and 0.04 (95% CI 0.00-0.09), respectively. CONCLUSIONS: PD-1/PD-L1 inhibitors produced encouraging clinical benefits including the response rate in the treatment of dMMR-MSI-H mCRC. They actually have been influenced by the present state of mCRC therapy including pMMR-MSI-L mCRC. Nevertheless, additional multi-center prospective studies are still expected. TRIAL REGISTRATION: We have registered this study in the International Prospective Register of Systematic Reviews (PROSPERO), and the registration number is CRD42021249601 .

Additional partial hepatectomy at the time of portal vein ligation accelerates the regeneration of the future liver remnant.

Portal vein ligation (PVL) has been adopted to induce hypertrophy of the future liver remnant (FLR) in patients with primarily irresectable liver tumor. However, regeneration of the FLR is not always sufficient to allow curative resection of the portally-deprived tumor-bearing liver lobe. We hypothesize that simultaneous hepatectomy (PHx) and PVL augments regeneration of the FLR and that the effect is related to the extent of the additional resection. Seventy-two Lewis rats were enrolled into 3 groups: 20%PVL + 70%PHx; 70%PVL + 20%PHx; 90%PVL. Animals were observed for 1, 2, 3 and 7 days postoperatively (n = 6/time point). Liver enzymes, caudate liver/body-weight-ratio, BrdU-proliferation-index (PI), proliferating-cell-nuclear-antigen (PCNA)-mRNA-expression level and autophagy-related-proteins were evaluated. Compared with 90% PVL, additional PHx induced significantly more hypertrophy during the observation time, which was confirmed by significantly higher PI and higher level of PCNA-mRNA expression. Similarly, the additional PHx induced more autophagy in the FLR compared with PVL alone. However, both effects were not clearly related to the extent of additional resection. Additional resection augmented liver regeneration and autophagy substantially compared with PVL alone. Therefore, we concluded that autophagy might play a critical role in regulating hepatocyte proliferation and the size of the FLR after simultaneous PVL + PHx.

Modulation of Autophagy: A Novel "Rejuvenation" Strategy for the Aging Liver.

Aging is a natural life process which leads to a gradual decline of essential physiological processes. For the liver, it leads to alterations in histomorphology (steatosis and fibrosis) and function (protein synthesis and energy generation) and affects central hepatocellular processes (autophagy, mitochondrial respiration, and hepatocyte proliferation). These alterations do not only impair the metabolic capacity of the liver but also represent important factors in the pathogenesis of malignant liver disease. Autophagy is a recycling process for eukaryotic cells to degrade dysfunctional intracellular components and to reuse the basic substances. It plays a crucial role in maintaining cell homeostasis and in resisting environmental stress. Emerging evidence shows that modulating autophagy seems to be effective in improving the age-related alterations of the liver. However, autophagy is a double-edged sword for the aged liver. Upregulating autophagy alleviates hepatic steatosis and ROS-induced cellular stress and promotes hepatocyte proliferation but may aggravate hepatic fibrosis. Therefore, a well-balanced autophagy modulation strategy might be suitable to alleviate age-related liver dysfunction. Conclusion. Modulation of autophagy is a promising strategy for "rejuvenation" of the aged liver. Detailed knowledge regarding the most devastating processes in the individual patient is needed to effectively counteract aging of the liver without causing obvious harm.

The Role of Autophagy for the Regeneration of the Aging Liver.

Age is one of the key risk factors to develop malignant diseases leading to a high incidence of hepatic tumors in the elderly population. The only curative treatment for hepatic tumors is surgical removal, which initiates liver regeneration. However, liver regeneration is impaired with aging, leading to an increased surgical risk for the elderly patient. Due to the increased risk, those patients are potentially excluded from curative surgery. Aging impairs autophagy via lipofuscin accumulation and inhibition of autophagosome formation. Autophagy is a recycling mechanism for eukaryotic cells to maintain homeostasis. Its principal function is to degrade endogenous bio-macromolecules for recycling cellular substances. A number of recent studies have shown that the reduced regenerative capacity of the aged remnant liver can be restored by promoting autophagy. Autophagy can be activated via multiple mTOR-dependent and mTOR-independent pathways. However, inducing autophagy through the mTOR-dependent pathway alone severely impairs liver regeneration. In contrast, recent observations suggest that inducing autophagy via mTOR-independent pathways might be promising in promoting liver regeneration. Conclusion: Activation of autophagy via an mTOR-independent autophagy inducer is a potential therapy for promoting liver regeneration, especially in the elderly patients at risk.

Diosmetin Induces Cell Apoptosis by Regulating CYP1A1/CYP1A2 Due to p53 Activation in HepG2 Cells.

It was explored that CYP1 family of cytochromes P450 were over-expressed in several types of cancer. Our study aimed to characterize anti-proliferative activity and metabolism of the natural flavonoid diosmetin in the human hepatoma cell HepG2, expressing CYP1 family. Diosinduced cell apoptosis could be reversed due to p53 blockade and the cellular P53 and CYP1A1/CYP1A2 proteins levels were examined. P53 and CYP1A1/CYP1A2 proteins were upregulated by Dios; when PFT-α was added into cells, the P53 levels were down-regulated accompanied with up-regulated CYP1A1/CYP1A2. Meanwhile, when cells were co-treated with Dios and PFT-α, P53 was down-regulated and CYP1A1/CYP1A2 up-regulated controlled with that of Dios treated cells. The data reveal the new evidence that cytochrome P450 CYP1A regulation by P53 enzyme plays an important role in Diosmetin anti-cancer activity of HepG2 cells.

Dihydromyricetin induces mouse hepatoma Hepal-6 cell apoptosis via the transforming growth factor-ß pathway.

Dihydromyricetin (DHM) is a flavonoid compound which possesses potent antitumor activity. In the present study, it was demonstrated that DHM significantly inhibited proliferation and induced apoptosis in mouse hepatocellular carcinoma Hepal­6 cells. Transforming growth factor ß (TGF­ß) is recognized as a major profibrogenic cytokine and is therefore a common target for drugs in the treatment of liver disease. The present study aimed to investigate whether TGF­ß was involved in DHM­triggered cell­viability inhibition and apoptosis induction. An MTT assay was used to evaluate the viability of Hepal­6 cells following DHM treatment. TGF­ß signalling is mediated by Smads and nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4) is a crucial regulator of reactive oxygen species ROS production. TGF­ß, Smad3, phosphorylated (p)­Smad2/3 and NOX4 protein expression levels were evaluated by western blot analysis. TGF­ß and NOX4 gene expression levels were determined by quantitative polymerase chain reaction. The results indicated that DHM downregulated TGF­ß, Smad3, p­Smad2/3 and NOX4 in a concentration­dependent manner. A cell counting assay indicated that DHM also inhibited Hepal­6 cell growth in a concentration­dependent manner. TGF­ß expression was significantly decreased following DHM treatment. In conclusion, the results of the present study defined and supported a novel function for DHM, indicating that it induced cell apoptosis by downregulating ROS production via the TGF­ß/Smad3 signaling pathway in mouse hepatocellular carcinoma Hepal­6 cells.

Structures of novel norstilbene dimer, longusone A, and three new stilbene dimers, longusols A, B, and C, with antiallergic and radical scavenging activities from Egyptian natural medicine Cyperus longus.

The methanolic extract of the whole plant of Cyperus longus originating in Egypt was found to show antiallergic effect on ear passive cutaneous anaphylaxis reactions in mice. By bioassay-guided separation, 11 stilbenes and stilbene dimers including a novel norstilbene dimer, longusone A, and three new stilbene dimers, longusols A, B, and C, were isolated. Their structures were elucidated on the basis of chemical and physicochemical evidence. Among the isolates, longusol B (IC(50)=96 µM), luteolin (3.0 µM), resveratrol (17 µM), piceatannol (24 µM), and cassigarols E (84 µM) and G (84 µM) were found to inhibit the release of ß-hexosaminidase, as a marker of antigen-induced degranulations, in rat basophilic leukemia (RBL-2H3) cells. In addition, the methanolic extract and the constituents showed 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity (SC(50)=22 µg/ml and 2.8-29 µM, respectively).

Melanogenesis inhibitors from the desert plant Anastatica hierochuntica in B16 melanoma cells.

The methanolic extract from the whole plants of Anastatica hierochuntica, an Egyptian herbal medicine, was found to inhibit melanogenesis in theophylline-stimulated murine B16 melanoma 4A5 cells. Among the constituents isolated, anastatin A, silybin A, isosilybins A and B, eriodictyol, luteolin, kaempferol, quercetin, hierochins A and B, (2R,3S)-2,3-dihydro-2-(3,4-dimethoxyphenyl)-3-hydroxymethyl-5-(2-formylvinyl)-7-hydroxybenzofuran, (+)-dehydrodiconiferyl alcohol, (+)-balanophonin, 1-(4-hydroxy-3-methoxyphenyl)-2-{4-[(E)-3-hydroxy-1-propenyl]-2-methoxyphenoxy}-1,3-propanediol, and 3,4-dihydroxybenzaldehyde substantially inhibited melanogenesis with IC(50) values of 6.1-32 microM. With regard to the mechanism of action of silybins and isosilybins, the inhibition of tyrosinase activity suggested to be important. In addition, isosilybins A and B inhibited the mRNA expression of TRP-2, but silybins A and B oppositely enhanced the mRNA expression of tyrosinase and TRP-1 and -2 at 10 and/or 30 microM, and the inhibition of phosphorylation of extracellular signal-regulated kinases (ERK1/2) is involved in the enhanced expression of mRNA, at least in part, similar to that of PD98059.

Structures of new flavonoids and benzofuran-type stilbene and degranulation inhibitors of rat basophilic leukemia cells from the Brazilian herbal medicine Cissus sicyoides.

Three new flavonoid glycosides (cissosides I, II, and III) and a new benzofuran-type stilbene (cissusin) were isolated from the methanolic extract of the aerial parts of Cissus sicyoides cultivated in Brazil. Their structures were elucidated on the basis of chemical and physicochemical evidence. The inhibitory effects of the isolated constituents on the release of beta-hexosaminidase as a marker of degranulation in rat basophilic leukemia (RBL-2H3) cells were examined. Cissusin, flavonols (kaempferol, quercetin), flavones (7,3',4'-trihydroxyflavone, lanceolatin B), pterocarpanes (homopterocarpin), chalcones (isoliquiritigenin, E-7-O-methylpongamol), and tryptanthrin markedly inhibited the release of beta-hexosaminidase.