Perioperative Changes in Serum Transaminase Levels: Impact on Postoperative Morbidity Following Liver Resection of Hepatocellular Carcinoma.

OBJECTIVES: To define how dynamic changes in pre- versus post-operative serum aspartate aminotransferase (AST) and alanine aminotransaminase (ALT) levels may impact postoperative morbidity after curative-intent resection of hepatocellular carcinoma (HCC). BACKGROUND: Hepatic ischemia/reperfusion can occur at the time of liver resection and may be associated with adverse outcomes following liver resection. METHODS: Patients who underwent curative resection for HCC between 2010-2020 were identified from an international multi-institutional database. Changes in AST and ALT (CAA) on postoperative day (POD) 3 versus preoperative values () were calculated using the formula: based on a fusion index via Euclidean norm, which was examined relative to the comprehensive complication index (CCI). The impact of CAA on CCI was assessed by the restricted cubic spline regression and Random Forest analyses. RESULTS: A total of 759 patients were included in the analytic cohort. Median CAA was 1.7 (range, 0.9 to 3.25); 431 (56.8%) patients had a CAA<2, 215 (28.3%) patients with CAA 2-5, and 113 (14.9%) patients had CAA ≥5. The incidence of post-operative complications was 65.0% (n=493) with a median CCI of 20.9 (IQR, 20.9-33.5). Spline regression analysis demonstrated a non-linear incremental association between CAA and CCI. The optimal cutoff value of CAA=5 was identified by the recursive partitioning technique. After adjusting for other competing risk factors, CAA≥5 remained strongly associated with risk of post-operative complications (Ref. CAA<5, OR 1.63, 95%CI 1.05-2.55, P=0.03). In fact, the use of CAA to predict post-operative complications was very good in both the derivative (AUC 0.88) and external (ACU 0.86) cohorts (n=1137). CONCLUSIONS: CAA was an independent predictor of CCI after liver resection for HCC. Use of routine labs such as AST and ALT can help identify patients at highest risk of post-operative complications following HCC resection.

Donor-Derived Engineered Microvessels for Cardiovascular Risk Stratification of Patients with Kidney Failure.

Cardiovascular disease is the cause of death in ≈50% of hemodialysis patients. Accumulation of uremic solutes in systemic circulation is thought to be a key driver of the endothelial dysfunction that underlies elevated cardiovascular events. A challenge in understanding the mechanisms relating chronic kidney disease to cardiovascular disease is the lack of in vitro models that allow screening of the effects of the uremic environment on the endothelium. Here, a method is described for microfabrication of human blood vessels from donor cells and perfused with donor serum. The resulting donor-derived microvessels are used to quantify vascular permeability, a hallmark of endothelial dysfunction, in response to serum spiked with pathophysiological levels of indoxyl sulfate, and in response to serum from patients with chronic kidney disease and from uremic pigs. The uremic environment has pronounced effects on microvascular integrity as demonstrated by irregular cell-cell junctions and increased permeability in comparison to cell culture media and healthy serum. Moreover, the engineered microvessels demonstrate an increase in sensitivity compared to traditional 2D assays. Thus, the devices and the methods presented here have the potential to be utilized to risk stratify and to direct personalized treatments for patients with chronic kidney disease.

A facile fluid pressure system reveals differential cellular response to interstitial pressure gradients and flow.

Interstitial fluid pressure gradients and interstitial flow have been shown to drive morphogenic processes that shape tissues and influence progression of diseases including cancer. The advent of porous media microfluidic approaches has enabled investigation of the cellular response to interstitial flow, but questions remain as to the critical biophysical and biochemical signals imparted by interstitial fluid pressure gradients and resulting flow on resident cells and extracellular matrix (ECM). Here, we introduce a low-cost method to maintain physiological interstitial fluid pressures that is built from commonly accessible laboratory equipment, including a laser pointer, camera, Arduino board, and a commercially available linear actuator. We demonstrate that when the system is connected to a microfluidic device containing a 3D porous hydrogel, physiologic pressure is maintained with sub-Pascal resolution and when basic feedback control is directed using an Arduino, constant pressure and pressure gradient can be maintained even as cells remodel and degrade the ECM hydrogel over time. Using this model, we characterized breast cancer cell growth and ECM changes to ECM fibril structure and porosity in response to constant interstitial fluid pressure or constant interstitial flow. We observe increased collagen fibril bundling and the formation of porous structures in the vicinity of cancer cells in response to constant interstitial fluid pressure as compared to constant interstitial flow. Collectively, these results further define interstitial fluid pressure as a driver of key pathogenic responses in cells, and the systems and methods developed here will allow for future mechanistic work investigating mechanotransduction of interstitial fluid pressures and flows.

Advances in nucleic acid amplification techniques (NAATs): COVID-19 point-of-care diagnostics as an example.

Achieving superhigh sensitivity is the ultimate goal for bio-detection in modern analytical science and life science. Among variable signal amplification strategies, nucleic acid amplification technologies are revolutionizing the field of bio-detection, providing greater possibilities in novel diagnosis achieving high efficiency, specificity, and cost-effectiveness. Nucleic acid amplification techniques (NAATs), such as Polymerase Chain Reaction (PCR), Rolling Circle Amplification (RCA), Loop-Mediated Isothermal Amplification (LAMP), Recombinase Polymerase Amplification (RPA), CRISPR-related amplification, and others are dominating methods employed in research and clinical settings. They each provide distinctively unique features that can offer desirable performance in terms of sensitivity, specificity, simplicity, stability, and cost. NAATs are in unmet demand in molecular diagnosis, especially in point-of-care scenario. This review will discuss the principles and recent advancements of each NAAT, respectively, revealing their strengths and challenges in achieving rapid and accurate bio-detection with a focus on point-of-care diagnosis. Furthermore, this review will explore the application of each of the technologies through the contemporary COVID-19 pandemic, analyzing their ability in point-of-care diagnosis of the COVID-19 with high sensitivity to emphasize significance of developing NAATs based methods in battling COVID-19. Finally, advantages and potentials of each NAAT in enhancements of sensitivity and specificity in bio-detection from bench side to the bedside will be discussed, aiming for full exploitation of capability of each NAAT. This review will provide novel aspects in the selection and combination of usages of various NAATs based on their distinctive characteristics and limitations. A possible advancing direction of future accurate POCT is also proposed.

[Expressions of aromatase protein and sex hormone receptor in endometrial lesions].

OBJECTIVE: To investigate the expression of aromatase protein, estrogen receptor (ER), progesterone receptor (PR) and nuclear antigen associated with cell proliferation Ki67 in endometrial diseases and their clinical significance in diagnosis and endocrine therapy of endometrial diseases. METHOD: Expressions of aromatase, ER, PR and Ki-67 were detected with immunohistochemistry technic (streptavidin-peroxidase-biotin, SP) in 148 cases including 30 of endometrial hyperplasia, 30 of atypical proliferation and 88 of endometrial adenocarcinoma as observational group and 15 cases of proliferative endometrium and 15 cases of secretory endometrium as control group. RESULTS: Expression of aromatase protein and ER, PR, Ki67 in endometrial hyperplasia, atypical proliferation had no significant difference comparing with the proliferative endometrium group (P > 0.05). In endometrial adenocarcinoma, the expression of aromatase protein increased obviously (64%, 56/88), which was higher than in benign diseases [atypical proliferation group was 23% (7/30), endometrial hyperplasia group was 13% (4/30)] and control group significantly (P < 0.01). The positive expression of ER, PR in endometrial adenocarcinoma decreased [22% (19/88), 19% (17/88)], and Ki67 increased (41%, 36/88) and there was a significant difference compared with control group (P < 0.01). The positive rate of aromatase protein did not increase with the progress of clinical stage or grade of cellular differentiation. Aromatase was not consistent with ER, PR and Ki67 in endometrial adenocarcinoma. CONCLUSION: Aromatase protein is related to the incidence of endometrial adenocarcinoma, and the expression of proteins (aromatase, ER, PR and Ki67) provides theoretical basis for understanding biological behavior of endometrial adenocarcinoma.

[Study of tissue morphology and vascular endothelial growth factor and matrix metalloproteinase-9 gene expressions in nude mouse endometriosis model].

OBJECTIVE: To establish an experimental endometriosis model using nude mouse as a xenographic host for relative biological behavior study of endometriosis. METHODS: Nude mice of experimental group were implanted with the late secretory endometrium of patients with endometriosis (n = 24) or without endometriosis (n = 24) into pelvic and abdominal cavities. Nude mice of control group (n = 3) were implanted with the greater omentum. Nude mice were killed at 5, 15 and 30 d randomly to observe the growth of endometriotic lesions. The morphological changes of endometriotic lesions from different sources and at different time points of growth were examined by light microscopy and electron microscopy; the expressions of vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) of eutopic endometrium and endometriotic lesions were detected by RT-PCR method, and the expressions of steroid hormone receptors by immunohistochemistry method. RESULTS: Endometriotic lesions were found from 5 d with rich blood supply, endometrial glands and stroma under light microscope. And the adhesion to mouse tissues became more severe with time. Under electron microscope, proliferation and infiltration of inflammatory cells were most striking at 15 d. At 30 d the organellae of gland cells were disappeared and there was nuclear chromatin aggregation with apoptosis tendency, but secretory granules still could be seen. The expressions of VEGF and MMP-9 mRNA were increased in endometriotic lesions compared with eutopic endometrium (P < 0.05), and were strongest at 15 d (EM group: 1.11 +/- 0.13/1.00 +/- 0.11; NEM group: 1.10 +/- 0.11/0.99 +/- 0.08) and decreased at 30 d (EM group: 0.85 +/- 0.11/0.77 +/- 0.13; NEM group: 0.86 +/- 0.14/0.76 +/- 0.11). Immunochemistry revealed the positive rate of estrogen and progestogen receptors at 30 d (EM group: 4/8 and 4/8; NEM group: 5/8 and 4/8) was lower than that at 5 d (EM group: 7/8 and 7/8; NEM group: 8/8 and 7/8) and 15 d (EM group: 7/8 and 7/8; NEM group: 7/8 and 8/8), and the expressions of steroid hormone receptors were mostly weak. There was no differences in expression of VEGF and MMP-9 mRNA, estrogen and progestogen between endometriotic lesions implanted with normal endometrium and endometrium from patients with endometriosis. CONCLUSIONS: The nude mouse is an appropriate model for the study of the early phase of endometriosis, and the genesis and development of endometriotic lesions are similar to that of human endometriosis. Endometriosis is associated with multiple factors.