Magneto-Acoustic Theranostic Approach: Integration of Magnetomotive Ultrasound Shear Wave Elastography and Magnetic Hyperthermia.

OBJECTIVES: Although magnetically induced hyperthermia has shown great efficiency in the treatment of solid tumors, it is still a challenge to avoid incomplete ablation or overtreatment. In this study, we applied magnetomotive ultrasound shear wave elastography (MMUS-SWE) as a tool for real-time image guidance and feedback in the magnetic hyperthermia (MH) process. We called this new method as magneto-acoustic theranostic approach (MATA). METHODS: In MATA, a ferromagnetic particle (fMP) was simultaneously used as a thermoseed for MH and a shear wave source for MMUS-SWE. The fMP was excited by a high-frequency magnetic field to induce the heating effect for MH. Meanwhile, the fMP was stimulated by a pulsed magnetic field to generate shear wave propagation for MMUS-SWE. Thus, the changes in elastic modulus surrounding fMP can be used to estimate the therapy effect of MH. RESULTS: The phantom and in vitro experiments were conducted to verify the feasibility of MATA, which has good performance in magnetothermal conversion and treatment efficacy feedback. The shear wave speed of the isolated pork liver changed significantly after the MH process, which varied from about 1.36 to 4.85 m/s. CONCLUSIONS: Preliminary results proved that changes in elastic modulus could be useful to estimate the therapy effect of MH. We expect that MATA, which is the integration of MMUS-SWE and MH, will be a novel theranostic method for clinical translation.

Real-world experience of postoperative adjuvant chemoimmunotherapy in patients with perihilar cholangiocarcinoma at high risk of recurrence.

BACKGROUND: This study aimed to assess whether postoperative adjuvant chemoimmunotherapy could lead to better clinical outcomes for high-risk patients with perihilar cholangiocarcinoma (pCCA). METHODS: In the cohort study, we retrospectively reviewed patients who received surgical resection for pCCA with curative intent from January 2018 to December 2021 at the Sun Yat-sen Memorial Hospital. The patients at high risk for relapse were further analyzed. Among them, 20 patients received adjuvant chemoimmunotherapy, 28 patients received adjuvant chemotherapy, and 33 patients received surgery alone. The oncological outcomes and drug-associated adverse events were evaluated. RESULTS: The 2-year overall survival (OS) rates in patients treated with adjuvant chemoimmunotherapy, adjuvant chemotherapy, and surgery alone were 80.0%, 49.4% and 22.6%, respectively. Univariable and multivariable Cox analyses showed that the treatment regimen and TNM stage were associated with adverse OS. Adjuvant chemoimmunotherapy led to an increase in OS compared with adjuvant chemotherapy [hazard ratio (HR) = 3.253; 95% confidence interval (CI) 1.072-9.870; P = 0.037] or surgery alone (HR = 7.560; 95% CI 2.508-22.785; P < 0.001). The median recurrence-free survival was 22.0 months for the adjuvant chemoimmunotherapy group, 17.0 months for the adjuvant chemotherapy group, and 13.2 months for the surgery alone group (P = 0.177); these differences were not significant. The chemoimmunotherapy group was associated with more frequent hematological side effects than the chemotherapy group, but the difference was not statistically significant. CONCLUSION: Postoperative adjuvant chemoimmunotherapy for resected pCCA patients showed improved OS compared with adjuvant chemotherapy or surgery alone, and further prospectively randomized controlled trials are necessary to validate these results.

CCL3 secreted by hepatocytes promotes the metastasis of intrahepatic cholangiocarcinoma by VIRMA-mediated N6-methyladenosine (m6A) modification.

BACKGROUND: Intrahepatic cholangiocarcinoma (ICC) is a malignant disease characterized by onset occult, rapid progression, high relapse rate, and high mortality. However, data on how the tumor microenvironment (TME) regulates ICC metastasis at the transcriptomic level remains unclear. This study aimed to explore the mechanisms and interactions between hepatocytes and ICC cells. METHODS: We analyzed the interplay between ICC and liver microenvironment through cytokine antibody array analysis. Then we investigated the role of N6-methyladenosine (m6A) modification and the downstream target in vitro, in vivo experiments, and in clinical specimens. RESULTS: Our study demonstrated that cytokine CCL3, which is secreted by hepatocytes, promotes tumor metastasis by regulating m6A modification via vir-like m6A methyltransferase associated (VIRMA) in ICC cells. Moreover, immunohistochemical analyses showed that VIRMA correlated with poor outcomes in ICC patients. Finally, we confirmed both in vitro and in vivo that CCL3 could activate VIRMA and its critical downstream target SIRT1, which fuels tumor metastasis in ICC. CONCLUSIONS: In conclusion, our results enhanced our understanding of the interaction between hepatocytes and ICC cells, and revealed the molecular mechanism of the CCL3/VIRMA/SIRT1 pathway via m6A-mediated regulation in ICC metastasis. These studies highlight potential targets for the diagnosis, treatment, and prognosis of ICC.

Cancer-specific survival in patients with cholangiocarcinoma after radical surgery: a Novel, dynamic nomogram based on clinicopathological features and serum markers.

BACKGROUND: This study aims to (1) identify preoperative testing-based characteristics associated with enhanced prognosis and survival for cholangiocarcinoma patients, and (2)create a distinctive nomogram to anticipate each patient's cancer-specific survival (CSS). METHODS: Retrospective analysis was performed on 197 CCA patients who underwent radical surgery at Sun Yat-sen Memorial Hospital; they were divided into a 131-person "training cohort" and a 66-person "internal validation cohort." The prognostic nomogram was created following a preliminary Cox proportional hazard regression search for independent factors influencing the patients' CSS. Its applicable domain was examined via an external validation cohort, which included 235 patients from the Sun Yat-sen University Cancer Center. RESULTS: The median follow-up period for the 131 patients in the training group was 49.3 months (range, 9.3 to 133.9 months). One-, three-, and five-year CSS rates were 68.7%, 24.5%, and 9.2%, respectively, with the median CSS length being 27.4 months (range: 1.4 to 125.2 months). PLT, CEA, AFP, tumor location, differentiation, lymph node metastasis, chemotherapy, and TNM stage were determined to be independent risk factors for CCA patients by univariate and multivariate Cox proportional hazard regression analysis. We were able to accurately predict postoperative CSS after incorporating all of these characteristics into a nomogram. The AJCC's 8th edition staging method's C-indices were statistically substantially (P < 0.001) lower than the nomogram's C-indices (0.84, 0.77, and 0.74 in the training, internal and external validation cohorts respectively). CONCLUSIONS: A realistic and useful model for clinical decision-making and the optimization of therapy is presented as a nomogram that includes serum markers and clinicopathologic features for predicting postoperative survival in cholangiocarcinoma.

Visualization of photothermal therapy by semiconducting polymer dots mediated photoacoustic detection in NIR II.

Visualization of photothermal therapy mediated by photothermal transduction agents (PTAs) is important to promote individual treatment of patients with low side effects. Photoacoustic detection has emerged as a promising noninvasive method for the visualization of PTAs distribution but still has limitations in temperature measurement, including poor measurement accuracy and low tissue penetration depth. In this study, we developed biocompatible semiconducting polymer dots (SPD) for in situ coupling of photothermal and photoacoustic detection in the near-infrared II window. SPD has dual photostability under pulsed laser and continuous-wave laser irradiation with a photothermal conversion efficiency of 42.77%. Meanwhile, a strong correlation between the photoacoustic signal and the actual temperature of SPD can be observed. The standard deviation of SPD-mediated photoacoustic thermometry can reach 0.13 °C when the penetration depth of gelatin phantom is 9.49 mm. Preliminary experimental results in vivo show that SPD-mediated photoacoustic signal has a high signal-to-noise ratio, as well as good performance in temperature response and tumor enrichment. Such a study not only offers a new nanomaterial for the visualization of photothermal therapy but will also promote the theranostic platform for clinical applications.

Dynamic Speed of Sound Adaptive Transmission-Reflection Ultrasound Computed Tomography.

Ultrasound computed tomography (USCT) can visualize a target with multiple imaging contrasts, which were demonstrated individually previously. Here, to improve the imaging quality, the dynamic speed of sound (SoS) map derived from the transmission USCT will be adapted for the correction of the acoustic speed variation in the reflection USCT. The variable SoS map was firstly restored via the optimized simultaneous algebraic reconstruction technique with the time of flights selected from the transmitted ultrasonic signals. Then, the multi-stencils fast marching method was used to calculate the delay time from each element to the grids in the imaging field of view. Finally, the delay time in conventional constant-speed-assumed delay and sum (DAS) beamforming would be replaced by the practical computed delay time to achieve higher delay accuracy in the reflection USCT. The results from the numerical, phantom, and in vivo experiments show that our approach enables multi-modality imaging, accurate target localization, and precise boundary detection with the full-view fast imaging performance. The proposed method and its implementation are of great value for accurate, fast, and multi-modality USCT imaging, particularly suitable for highly acoustic heterogeneous medium.

Synthetic Conjugated Oligoelectrolytes Are Effective siRNA Transfection Carriers: Relevance to Pancreatic Cancer Gene Therapy.

Conjugated oligoelectrolyte COE-S6 contains an elongated conjugated core with three cationic charges at each termini of the internal core. As an analogue of bolaamphiphiles, these structural attributes lead to the formation of spherical nanoplexes with Dh = 205 ± 5.0 nm upon mixing with small interfering RNA (siRNA). COE-S6/siRNA nanocomplexes were shown to be protective toward RNase, stimulate endosome escape, and achieve transfection efficiencies comparable to those achieved with commercially available LIP3000. Moreover, COE-S6/siRNA nanocomplexes enabled efficient silencing of the K-ras gene in pancreatic cancer cells and significant inhibition of cancer tumor growth with negligible in vitro toxicities. More importantly, cell invasion and colony formation of the Panc-1 cells were significantly inhibited, and apoptosis of the pancreatic cancer cells was also promoted. We also note that COE-S6 is much less toxic relative to commercial lipid formulations, and it provides optical signatures that can enable subsequent mechanistic work without the need to label nucleotides. COE-S6-based nanoplexes are thus a promising candidate as nonviral vectors for gene delivery.

Viscoelasticity measured by shear wave elastography in a rat model of nonalcoholic fatty liver disease: comparison with dynamic mechanical analysis.

BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) is rapidly becoming one of the most common liver diseases. Ultrasound elastography has been used for the diagnosis of NAFLD. However, clinical research on steatosis by elastography technology has mainly focused on steatosis with fibrosis or non-alcoholic steatohepatitis (NASH), while steatosis without fibrosis has been poorly studied. Moreover, the relationship between liver viscoelasticity and steatosis grade is not clear. In this study, we evaluated the degree of liver steatosis in a simple steatosis rat model using shear wave elastography (SWE). RESULTS: The viscoelasticity values of 69 rats with hepatic steatosis were measured quantitatively by SWE in vivo and validated by a dynamic mechanical analysis (DMA) test. Pathological sections were used to determine the steatosis grade for each rat. The results showed that the elasticity values µ obtained by the two methods followed the same trend, and µ is significantly correlated with liver steatosis. The Pearson's correlation coefficients indicate that [Formula: see text] obtained by SWE is positively linear correlated with DMA (r = 0.628, p = 7.85 × 10-9). However, the viscosity values [Formula: see text] obtained by SWE were relatively independent of those obtained by DMA with a correlation coefficient of - 0.01. The combined Voigt elasticity measurements have high validity in the prediction of steatosis (S0 vs. S1-S4), with an AUROC of 0.755 (95% CI 0.6175-0.8925, p < 0.01) and the optimal cutoff value was 2.08 kPa with a sensitivity of 78% and specificity of 63%. CONCLUSION: SWE might have the feasibility to be introduced as an auxiliary technique for NAFLD patients in clinical settings. However, the viscosity results measured by SWE and DMA are significantly different, because the two methods work in different frequency bands.

In vivo assessment of the mechanical properties of crystalline lenses in a rabbit model using ultrasound elastography: Effects of ultrasound frequency and age.

PURPOSE: The occurrence of presbyopia and cataract is closely related to changes in the mechanical properties of the crystalline lens. There are no established methods so far for in vivo assessment. By introducing ultrasound elastography, we proposed group velocity (Vg) of an induced shear wave as a new biomarker to characterize the mechanical properties of the lens in our previous study. Here, we investigated the effect of the ultrasound frequency on measurement accuracy and validated the results with a conventional ex vivo compression testing. We also demonstrated a change trend in Vg and its correlation with age in a rabbit model. METHODS: Eight New Zealand white rabbits were fed normally from the fourth to seventh month. An ultrasound elastography system was developed to measure Vgin vivo on every eye once per month. The performances when using a high-frequency (L22-11v) and low-frequency (L11-4v) probe were compared. Rabbits were sacrificed after in vivo measurements by the end of the seventh month and this was followed by ex vivo ultrasound measurements and conventional compression tests on the extracted lenses. RESULTS: The results demonstrated that there were no significance differences in Vg between measurements with high-frequency (USE-HF) and low-frequency (USE-LF) probes in the same month-age group. The mean Vg and the standard deviation of four rabbits that were 7 months old were 2.37 ±â€¯0.24  m/s, 2.36 ±â€¯0.25 m/s, 2.43 ±â€¯0.26 m/s and 2.44 ±â€¯0.38 m/s, with USE-HF for ex vivo and in vivo measurements and USE-LF for ex vivo and in vivo measurements, respectively. There were no significant differences (p > 0.05) and they were all in agreement with the results of compression tests, which was 16.16 ±â€¯1.84 kPa in Young's modulus. The results also showed that Vg increased with age. In combination with the results of our previous study, Vg showed a relatively sharp increase from 2 to 5 months, while it had a slight increase from 5 to 7 months. CONCLUSIONS: The USE-HF and USE-LF has comparable accuracy in Vg measurements while USE-HF had an advantage regarding better spatial resolution. The change trend of Vg was in accord with the growth phase of New Zealand white rabbits, which usually results in sexual maturity at 5 months old. This implies that Vg can be used as a biomarker parameter for evaluating the mechanical properties of the lens undergoing physiological changes.

KRAS promotes tumor metastasis and chemoresistance by repressing RKIP via the MAPK-ERK pathway in pancreatic cancer.

Oncogenic KRAS plays a crucial role in pancreatic ductal adenocarcinoma (PDAC) development and progression. However, the mechanism has not been clearly elucidated. RKIP is a tumor repressor, and loss of RKIP has been shown in PDAC. Here, we found that KRAS expression was inversely correlated with RKIP expression in PDAC fresh tissue regardless of the KRAS mutant status. The negative correlation between KRAS and RKIP was further confirmed in our PDAC tissue microarray. KRAS overexpression and RKIP downregulation were associated with poor clinical outcomes. Knockdown or overexpression of KRAS in PDAC cell lines robustly increased or decreased, respectively, RKIP protein and mRNA levels. Furthermore, the MAPK-ERK pathway was involved in the regulation of RKIP. KRAS-regulated RKIP expression, which in turn affected the expression of pivotal epithelial-mesenchymal transition (EMT) and apoptosis factors. The biological function of the KRAS-RKIP axis was demonstrated in human pancreatic cancer cells in vitro and in vivo. KRAS knockdown increased RKIP expression and inhibited metastasis and chemoresistance. Moreover, the feature of metastasis and chemoresistance was rescued in the KRAS-knockdown cells through the inhibition of RKIP by RNA interference. In conclusion, our studies demonstrate how KRAS inhibits the tumor suppressor RKIP, thus offering novel justification for targeting RKIP as a strategy to overcome KRAS-induced tumor metastasis and chemoresistance in PDAC.

Noninvasive assessment of age-related stiffness of crystalline lenses in a rabbit model using ultrasound elastography.

BACKGROUND: The pathological or physiological changes of a crystalline lens directly affect the eye accommodation and transmittance, and then they increase the risk of presbyopia and cataracts for people in the middle and old age groups. There is no universally accepted quantitative method to measure the lens' mechanical properties in vivo so far. This study aims to investigate the possibility of assessing the age-related stiffness change of crystalline lens by acoustic-radiation-force-based ultrasound elastography (ARF-USE) in a rabbit model in vivo. METHODS: There were 13 New Zealand white rabbits that were divided into four groups and fed normally until they were 60 (n = 4), 90 (n = 2), 120 (n = 4), and 150 (n = 3) days old, respectively. An ARF-USE platform was built based on the Verasonics™ Vantage 256 system. The shear waves were excited and traced in the lens by a linear ultrasound probe after a rabbit was anaesthetized. RESULTS: The average group velocities were 1.38 ± 0.2 m/s, 2.06 ± 0.3 m/s, 2.07 ± 0.29 m/s, and 2.30 ± 0.28 m/s, respectively, for the four groups of rabbits. The results shows that the group velocity has a strong correlation with the day age (r = 0.84, p < 1 × 10-7) and the weight (r = 0.83, p < 1×10-7) of the rabbits while the maximum displacement has no correlations with the day age (r = 0.27, p > 0.1) and the weight (r = 0.32, p > 0.1). CONCLUSION: This study demonstrated that the group velocity measured by ARF-USE had a strong correlation with age-related stiffness in a rabbit model, suggesting that group velocity is a good biomarker to characterize the stiffness of a crystalline lens. This study also demonstrated the feasibility of using this USE technique to assess the mechanical properties of the lens in vivo for clinical or research purposes.

A 2D Magneto-Acousto-Electrical Tomography Method to Detect Conductivity Variation Using Multifocus Image Method.

As magneto-acoustic-electrical tomography (MAET) combines the merits of high contrast and high imaging resolution, and is extremely useful for electrical conductivity measurement, so it is expected to be a promising medical imaging modalities for diagnosis of early-stage cancer. Based on the Verasonics system and the MC600 displacement platform, we designed and implemented a MAET system with a chirp pulse stimulation (MAET-CPS) method and a focal probe was utilized for stepscan focus excitation to enhance the imaging resolution. The relevant experiments were conducted to explore the influence of excitation positions of the single-focus point, and the effect of the excitation position on the amplitudes of the conductivity variation was clearly demonstrated. In order to take advantage of the merits of multifocus imaging, we firstly proposed a single focus MAET system with a chirp pulse stimulation (sfMAET-CPS) method and a multifocus MAET system with a chirp pulse stimulation (mfMAET-CPS) method for high-resolution conductivity imaging, and a homogenous gelatin phantom with a cuboid-shaped hole was used to investigate the accuracy of mfMAET-CPS. Comparative experiments were carried out on the same uniform phantom by the sfMAET-CPS and the mfMAET-CPS, respectively. The results showed that: (1) the electrical conductivity distributions of the homogenous phantom with a cuboid-shaped hole were detected by the sfMAET-CPS but were easily affected by the focal point, which demonstrated that the sfMAET-CPS had a low imaging resolution. (2) Compared with the sfMAET-CPS, the imaging effect of the mfMAET-CPS was much better than that of the sfMAET-CPS. (3) A linear interpolation algorithm was used to process the 2D conductivity distribution; it increased the smoothness of the conductivity distribution and improved the imaging effect. The stepscan focus excitation and the linearly frequency-modulated theory provide an alternative scheme for the clinical application of MAET.

[Design of magneto-acoustic-electrical detection system and verification of its linear sweep theory].

Clinical studies had demonstrated that early diagnosis of lesion could significantly reduce the risk of cancer. Magneto-acoustic-electrical tomography (MAET) is expected to become a new detection method due to its advantages of high resolution and high contrast. Based on thinking of modular design, a low-cost, digital magneto-acoustic conductivity detection system was designed and implemented in this study. The theory of MAET using chirp continuous wave excitation was introduced. The results of homogeneous phantom experiment with 0.5% NaCl clearly showed that the conductivity curve of homogeneous phantom was highly consistent with the actual physical size, which indicated that the chirp excitation theory in our proposed system was correct and feasible. Besides, the resolution obtained by 1 000 µs sweep time was better than that obtained by 500 µs and 1 500 µs, which means that sweep time is an important factor affecting the detection resolution of the conductivity. The same result was obtained in the experiments carried out on homogeneous phantoms with different concentrations of NaCl, which demonstrated the repeatability of our proposed MAET system.

Vps4A functions as a tumor suppressor by regulating the secretion and uptake of exosomal microRNAs in human hepatoma cells.

UNLABELLED: The deregulation of microRNAs (miRNAs) plays an important role in human hepatocarcinogenesis. In this study, we highlight exosomes as mediators involved in modulating miRNA profiles in hepatocellular carcinoma (HCC) cells. First, we examined the different miRNA expression profiles in HCC cells and HCC cell-derived exosomes. Next, coculture experiments indicated that HCC cell-derived exosomes promoted the cell growth, migration, and invasion of HCC cells and had the ability to shuttle miRNAs to recipient cells. Further, our data showed that Vps4A, a key regulator of exosome biogenesis, was frequently down-regulated in HCC tissues. The reduction of Vps4A in HCC tissues was associated with tumor progression and metastasis. In vitro studies revealed that Vps4A repressed the growth, colony formation, migration, and invasion of HCC cells. We further investigated the role and involvement of Vps4A in suppressing the bioactivity of exosomes and characterized its ability to weaken the cell response to exosomes. By small RNA sequencing, we demonstrated that Vps4A facilitated the secretion of oncogenic miRNAs in exosomes as well as accumulation and uptake of tumor suppressor miRNAs in cells. A subset of Vps4A-associated miRNAs was identified. Kyoto Encyclopedia of Genes and Genomes pathway analysis indicated that the phosphatidylinositol-3-kinase/Akt signaling pathway was the most likely candidate pathway for modulation by these miRNAs. Indeed, we proved that the phosphatidylinositol-3-kinase/Akt pathway was inactivated by Vps4A overexpression. CONCLUSION: Exosome-mediated miRNA transfer is an important mechanism of self-modulation of the miRNA expression profiles in HCC cells, and Vps4A may function as a tumor suppressor, which utilizes exosomes as mediators to regulate the secretion and uptake of miRNAs in hepatoma cells; these observations provide new insights into the development of HCC.

Optimal linear combination of ARFI, transient elastography and APRI for the assessment of fibrosis in chronic hepatitis B.

BACKGROUND & AIMS: Accurate assessment of liver fibrosis in patients with chronic hepatitis B (CHB) is necessary not only to predict the long-term clinical course but also to determine an appropriate antiviral therapy scheme. Several noninvasive approaches - serum markers and elastography - have been proposed as alternatives for the histopathological analysis of liver biopsies. The aim of this study was to evaluate two ultrasound elastography methods (ARFI and TE) and one biochemical test (APRI), as well as their optimal linear combination, in the assessment of liver fibrosis in CHB. METHODS: Ninety five patients with CHB and 16 volunteers underwent ARFI, TE and APRI; and liver fibrosis was staged in the patients by a liver biopsy. An optimal linear combination of the three methods was developed, and its diagnostic performance was evaluated by a 10-fold cross-validation. RESULTS: The accuracy of the linear combination was 83.86% and 91.88% for significant fibrosis (≥F2) and cirrhosis (F4), respectively, higher than those obtained for ARFI (83.50%, 88.76%), TE (75.27%, 87.61%) and APRI (73.29% and 81.67%). The combination also increased the sensitivity and the negative predictive values for the diagnosis of significant fibrosis and cirrhosis. CONCLUSIONS: The optimal linear combination algorithm is effective for noninvasive staging of liver fibrosis in CHB. However, linear combination has its own limitations; nonlinear methods may eventually reveal even clearer diagnostic results.

Iterative Reconstruction Algorithms in Magneto-Acousto-Electrical Computed Tomography (MAE-CT) for Image Quality Improvement.

Magneto-acousto-electrical computed tomography (MAE-CT) is a recently developed rotational magneto-acousto-electrical tomography (MAET) method, which can map the conductivity parameter of tissues with high spatial resolution. Since the imaging mode of MAE-CT is similar to that of CT, the reconstruction algorithms for CT are possible to be adopted for MAE-CT. Previous studies have demonstrated that the filtered back-projection (FBP) algorithm, which is one of the most common CT reconstruction algorithms, can be used for MAE-CT reconstruction. However, FBP has some inherent shortcomings of being sensitive to noise and non-uniform distribution of views. In this study, we introduced iterative reconstruction (IR) method in MAE-CT reconstruction and compared its performance with that of the FBP. The numerical simulation, the phantom, and in vitro experiments were performed, and several IR algorithms (ART, SART, SIRT) were used for reconstruction. The results show that the images reconstructed by the FBP and IR are similar when the data is noise-free in the simulation. As the noise level increases, the images reconstructed by SART and SIRT are more robust to the noise than FBP. In the phantom experiment, noise and some stripe artifacts caused by the FBP are removed by SART and SIRT algorithms. In conclusion, the IR method used in CT is applicable in MAE-CT, and it performs better than FBP, which indicates that the state-of-the-art achievements in the CT algorithm can also be adopted for the MAE-CT reconstruction in the future.

Cancer-Associated Fibroblasts Promote Lymphatic Metastasis in Cholangiocarcinoma via the PDGF-BB/PDGFR-ß Mediated Paracrine Signaling Network.

Patients with cholangiocarcinoma (CCA) with lymph node metastasis (LNM) have the worst prognosis, even after complete resection; however, the underlying mechanism remains unclear. Here, we established CAF-derived PDGF-BB as a regulator of LMN in CCA. Proteomics analysis revealed upregulation of PDGF-BB in CAFs derived from patients with CCA with LMN (LN+CAFs). Clinically, the expression of CAF-PDGF-BB correlated with poor prognosis and increased LMN in patients with CCA, while CAF-secreted PDGF-BB enhanced lymphatic endothelial cell (LEC)-mediated lymphangiogenesis and promoted the trans-LEC migration ability of tumor cells. Co-injection of LN+CAFs and cancer cells increased tumor growth and LMN in vivo. Mechanistically, CAF-derived PDGF-BB activated its receptor PDGFR-ß and its downstream ERK1/2-JNK signaling pathways in LECs to promote lymphoangiogenesis, while it also upregulated the PDGFR-ß-GSK-P65-mediated tumor cell migration. Finally, targeting PDGF-BB/PDGFR-ß or the GSK-P65 signaling axis prohibited CAF-mediated popliteal lymphatic metastasis (PLM) in vivo. Overall, our findings revealed that CAFs promote tumor growth and LMN via a paracrine network, identifying a promising therapeutic target for patients with advanced CCA.

Anticancer drugs cause release of exosomes with heat shock proteins from human hepatocellular carcinoma cells that elicit effective natural killer cell antitumor responses in vitro.

Failure of immune surveillance related to inadequate host antitumor immune responses has been suggested as a possible cause of the high incidence of recurrence and poor overall survival outcome of hepatocellular carcinoma. The stress-induced heat shock proteins (HSPs) are known to act as endogenous "danger signals" that can improve tumor immunogenicity and induce natural killer (NK) cell responses. Exosome is a novel secretory pathway for HSPs. In our experiments, the immune regulatory effect of the HSP-bearing exosomes secreted by human hepatocellular carcinoma cells under stress conditions on NK cells was studied. ELISA results showed that the production of HSP60, HSP70, and HSP90 was up-regulated in both cell lines in a stress-specific manner. After exposure to hepatocellular carcinoma cell-resistant or sensitive anticancer drugs (hereafter referred to as "resistant" or "sensitive" anticancer drug), the membrane microvesicles were actively released by hepatocellular carcinoma cells, differing in their ability to present HSPs on the cell surface, which were characterized as exosomes. Acting as a decoy, the HSP-bearing exosomes efficiently stimulated NK cell cytotoxicity and granzyme B production, up-regulated the expression of inhibitory receptor CD94, and down-regulated the expression of activating receptors CD69, NKG2D, and NKp44. Notably, resistant anticancer drugs enhanced exosome release and generated more exosome-carried HSPs, which augmented the activation of the cytotoxic response. In summary, our findings demonstrated that exosomes derived from resistant anticancer drug-treated HepG2 cells conferred superior immunogenicity in inducing HSP-specific NK cell responses, which provided a clue for finding an efficient vaccine for hepatocellular carcinoma immunotherapy.

The clinical effect of primary duct closure and T-tube drainage: A propensity score matched study.

BACKGROUND: Although laparoscopic common bile duct exploration (LCBDE) is considered a safe and effective method for the removal of bile duct stones, selecting primary duct closure (PDC) or T-tube drainage (TTD) following choledochotomy remains controversial. This study aims to explore the clinical effects of PDC and TTD after LCBDE. METHODS: We retrospectively analyzed clinical data of 348 patients with choledocholithiasis treated with LCBDE from January 2016 to October 2020. All patients were divided into PDC (225 cases) and TTD (123 cases) groups. Propensity score matching (PSM) was performed. We compared operative parameters and outcomes. RESULTS: After matching (n = 116/group), no significant difference was observed between the two groups (P > 0.05) regarding intra-abdominal infection, incision infection, bile leakage, and retained stones. In terms of operation time, intraoperative blood loss, postoperative hospital stay, postoperative exhaust time, postoperative antibiotic use time, and postoperative abdominal drainage time, PDC group was obviously superior to TTD group (P < 0.05). CONCLUSION: Primary closure following LCBDE is considered a safe and effective alternative to T-tube drainage.

Automatic Diagnosis of Significant Liver Fibrosis From Ultrasound B-Mode Images Using a Handcrafted-Feature-Assisted Deep Convolutional Neural Network.

The accurate diagnosis of significant liver fibrosis ( ≥ F2) in patients with chronic liver disease (CLD) is critical, as ≥ F2 is a crucial factor that should be considered in selecting an antiviral therapy for these patients. This article proposes a handcrafted-feature-assisted deep convolutional neural network (HFA-DCNN) that helps radiologists automatically and accurately diagnose significant liver fibrosis from ultrasound (US) brightness (B)-mode images. The HFA-DCNN model has three main branches: one for automatic region of interest (ROI) segmentation in the US images, another for attention deep feature learning from the segmented ROI, and the third for handcrafted feature extraction. The attention deep learning features and handcrafted features are fused in the back end of the model to enable more accurate diagnosis of significant liver fibrosis. The usefulness and effectiveness of the proposed model were validated on a dataset built upon 321 CLD patients with liver fibrosis stages confirmed by pathological evaluations. In a fivefold cross validation (FFCV), the proposed model achieves accuracy, sensitivity, specificity, and area under the receiver-operating-characteristic (ROC) curve (AUC) values of 0.863 (95% confidence interval (CI) 0.820-0.899), 0.879 (95% CI 0.823-0.920), 0.872 (95% CI 0.800-0.925), and 0.925 (95% CI 0.891-0.952), which are significantly better than those obtained by the comparative methods. Given its excellent performance, the proposed HFA-DCNN model can serve as a promising tool for the noninvasive and accurate diagnosis of significant liver fibrosis in CLD patients.