Contemporary trends and outcomes after liver transplantation and resection for intrahepatic cholangiocarcinoma.

BACKGROUND: Liver transplantation (LT) has been shown to be superior to resection in highly selected patients with perihilar cholangiocarcinoma (CCA), yet has traditionally been contraindicated for intrahepatic CCA (iCCA). Herein, we aimed to examine contemporary trends and outcomes for surgical resection and LT for iCCA. METHODS: The National Cancer Database was queried for patients presenting with stage I-III iCCA between 2010 and 2018 who underwent resection or LT. Overall survival (OS) was compared with Kaplan-Meier and multivariable Cox proportional hazards methods stratified by management. Secondary analysis of patients undergoing transplant for CCA was performed with the United Network for Organ Sharing database. RESULTS: Of 2565 patients, 2412 (94.0%) underwent resection and 153 (5.96%) LT of whom 84 (54.9%) received neoadjuvant therapy. Utilization of LT remained between 3.9% and 7.8% annually. Unadjusted 5-year OS was higher for LT than resection (59.8% vs 39.9%, P = .0067), yet adjusted analysis revealed no significant difference in mortality (hazard ratio, 0.91; 95% CI, 0.66-1.27; P = .58). On secondary analysis including 437 patients with all subtypes of CCA, unadjusted 5-year OS was higher for non-CCA indications (79% vs 52%-54%, P < .001). CONCLUSION: Utilization of LT for iCCA remains low and many cases are likely incidental. Although partial hepatectomy remains the standard of care for patients with resectable disease, our findings suggest that highly selected patients with unresectable iCCA may achieve favorable outcomes after LT. Granular, prospective data are needed to identify patients most likely to benefit from transplant and allocate scarce liver grafts.

Tumor size and survival in intrahepatic cholangiocarcinoma treated with surgical resection or ablation.

OBJECTIVES: We performed a retrospective analysis within a national cancer registry on outcomes following resection or ablation for intrahepatic cholangiocarcinoma (iCCA). METHODS: The National Cancer Database was queried for patients with clinical stage I-III iCCA diagnosed during 2010-2018, who underwent resection or ablation. Overall survival (OS) was compared with Kaplan-Meier and multivariable Cox proportional hazards methods. RESULTS: Of 2140 patients, 1877 (87.7%) underwent resection and 263 (12.3%) underwent ablation, with median tumor sizes of 5.5 and 3 cm, respectively. Overall, resection was associated with greater median OS (41.2 months (95% confidence interval [95% CI]: 37.6-46.2) vs. 28 months (95% CI: 15.9-28.6) on univariable analysis (p < 0.0001). There was no significant difference on multivariable analysis (p = 0.42); however, there was a significant interaction between tumor size and management. On subgroup analysis of patients with tumors <3 cm, there was no difference in OS between resection versus ablation. However, ablation was associated with increased mortality for tumors ≥3 cm. CONCLUSION: Although resection is associated with improved OS for tumors ≥3 cm, we observed no difference in survival between management strategies for tumors < 3 cm. Ablation may be an alternative therapeutic strategy for small iCCA, particularly in patients at risk for high surgical morbidity.

Comparing Survival After Resection, Ablation, and Radiation in Small Intrahepatic Cholangiocarcinoma.

BACKGROUND: Hepatectomy is the cornerstone of curative-intent treatment for intrahepatic cholangiocarcinoma (ICC). However, in patients unable to be resected, data comparing efficacy of alternatives including thermal ablation and radiation therapy (RT) remain limited. Herein, we compared survival between resection and other liver-directed therapies for small ICC within a national cancer registry. PATIENTS AND METHODS: Patients with clinical stage I-III ICC < 3 cm diagnosed 2010-2018 who underwent resection, ablation, or RT were identified in the National Cancer Database. Overall survival (OS) was compared using Kaplan-Meier and multivariable Cox proportional hazards methods. RESULTS: Of 545 patients, 297 (54.5%) underwent resection, 114 (20.9%) ablation, and 134 (24.6%) RT. Median OS was similar between resection and ablation [50.5 months, 95% confidence interval (CI) 37.5-73.9; 39.5 months, 95% CI 28.7-58.4, p = 0.14], both exceeding that of RT (20.9 months, 95% CI 14.1-28.3). RT patients had high rates of stage III disease (10.4% RT vs. 1.8% ablation vs. 11.8% resection, p < 0.001), but the lowest rates of chemotherapy utilization (9.0% RT vs. 15.8% ablation vs. 38.7% resection, p < 0.001). In multivariable analysis, resection and ablation were associated with reduced mortality compared with RT [hazard ratio (HR) 0.44, 95% CI 0.33-0.58 and HR 0.53, 95% CI 0.38-0.75, p < 0.001, respectively]. CONCLUSION: Resection and ablation were associated with improved survival in patients with ICC < 3 cm compared with RT. Acknowledging confounders, anatomic constraints of ablation, limitations of available data, and need for prospective study, these results favor ablation in small ICC where resection is not feasible.

Risk factors of postoperative acute kidney injury in patients with complex congenital heart disease and significance of early detection of serum transcription factor Nkx2.5.

OBJECTIVE: This study was designed to investigate the risk factors of postoperative acute kidney injury (AKI) in patients with complex congenital heart disease (CHD) and the significance of early detection of serum transcription factor Nkx2.5. METHODS: A total of 121 CHD patients admitted to the Shengli Clinical Medical College of Fujian Medical University were selected as study participants, among whom 69 patients with AKI after cardiac surgery were set as the research group (RG), and the rest of the 52 patients without AKI were set as the control group (CG). Cardiopulmonary bypass (CPB) duration, aortic occlusion time, postoperative creatinine (Cr) level and mechanical ventilation (MV) time were compared between the two groups. The expression and clinical significance of Nkx2.5 in the two groups were detected. Intensive Care Unit (ICU) residence time and total hospital stay were compared, and the risk factors were analyzed. RESULTS: The RG presented remarkably longer CPB duration and aortic occlusion time, evidently higher postoperative Cr level and longer MV time, and observably lower Nkx2.5 level in comparison to the CG (all P<0.05). According to the analysis of receiver operating characteristic (ROC) curves, Nkx2.5 displayed a favorable diagnostic value in predicting the occurrence of CHD complicated with AKI. ICU residence time and total hospital stay were longer in the RG than in the CG (P<0.05). CPB time and aortic occlusion time were independent risk factors for AKI in CHD patients, while surgical methods and Nkx2.5 detection were independent protective factors (P<0.05). CONCLUSIONS: CPB time, aortic occlusion time and surgical methods, as well as Nkx2.5 detection are independent factors affecting AKI in patients with CHD. Early detection of serum transcription factor Nkx2.5 is of particular importance for clinical diagnosis of CHD patients complicated with AKI.

Uni-Port Total Thoracoscopic Surgery Versus Median Sternotomy for Redo Tricuspid Valve Replacement: A Retrospective Study.

BACKGROUND: This study compared the perioperative and follow-up period data of patients who underwent redo tricuspid valve replacements performed via thoracoscopic surgery or median sternotomy. The purpose was to evaluate the feasibility, safety, and surgical outcomes of redo tricuspid valve replacement via uni-port thoracoscopic surgery. METHODS: Forty-nine patients with severe tricuspid valve regurgitation after left-side valve replacement underwent redo tricuspid valve replacements in our hospital from April 2012 to September 2019. Twenty-six patients underwent uni-port total thoracoscopy surgery, whereas 23 patients had the surgery performed via median sternotomy. We collected perioperative and 3- to 36-month postoperative data. RESULTS: No deaths occurred in the intraoperative period. Time of cardiopulmonary bypass in the study group significantly was longer than that in the control group (P < .05), but the operative times in the study and control groups were not significantly different. Thoracic drainage, length of ICU stay, postoperative hospital stay, and complication rates in the study group were significantly different from those in the control group (P < .05). Throughout the follow-up period, uni-port total thoracoscopic TVR was not inferior to traditional surgery with respect to cardiac function and recurrence of tricuspid valve regurgitation. CONCLUSIONS: Uni-port total thoracoscopic tricuspid valve replacement is safe,  feasible and effective, and that can be considered as a primary treatment strategy for patients with severe TR after previous left-sided valve procedure.

Transient Incomplete Separation Facilitates Finding Accurate Equilibrium Dissociation Constant of Protein-Small Molecule Complex.

Current practical methods for finding the equilibrium dissociation constant, Kd , of protein-small molecule complexes have inherent sources of inaccuracy. Introduced here is "accurate constant via transient incomplete separation" (ACTIS), which appears to be free of inherent sources of inaccuracy. Conceptually, a short plug of the pre-equilibrated protein-small molecule mixture is pressure-propagated in a capillary, causing fast transient incomplete separation of the complex from the unbound small molecule. A superposition of signals from these two components is measured near the capillary exit and used to calculate a fraction of unbound small molecule, which, in turn, is used to calculate Kd . Herein the validity of ACTIS is proven theoretically, its accuracy is verified by computer simulation, and its practical use is demonstrated. ACTIS has the potential to become a reference-standard method for determining Kd  values of protein-small molecule complexes.

Preparation, optimisation, and in vitro-in vivo evaluation of febuxostat ternary solid dispersion.

The research aimed to prepare febuxostat (FEB) solid dispersion through solvent evaporation. Optimised solid dispersion composed of FEB, polyvinylpyrrolidone (PVP K30) and poloxamer at a ratio of 1:3:3 was characterised. Powder X-ray diffraction (XRD) and differential scanning calorimetry (DSC) indicated FEB was transformed from crystalline into the amorphous state in solid dispersion and scanning electron microscopy (SEM) revealed the morphology. Fourier transform infrared spectroscopy (FT-IR) suggested the interactions formed between FEB and polymers. A remarkable increase was observed of the optimised formulation in saturation solubility, dissolution studies (96.17 ± 0.79% in pH 6.0), and bioavailability (Cmax 18.25 ± 2.44 vs. 7.72 ± 0.48 µg/mL and AUC0-∞ 53.62 ± 7.63 vs. 34.76 ± 2.45 µg·h/mL). Besides, the FEB solid dispersion showed great stability after 90 days storage. Thus, the present study supports the rationality of PVP K30 and poloxamer188 as co-carriers for the preparation of FEB solid dispersion.

Aptamer facilitated purification of functional proteins.

DNA aptamers are attractive capture probes for affinity chromatography since, in contrast to antibodies, they can be chemically synthesized and, in contrast to tag-specific capture probes (such as Nickel-NTA or Glutathione), they can be used for purification of proteins free of genetic modifications (such as His or GST tags). Despite these attractive features of aptamers as capture probes, there are only a few reports on aptamer-based protein purification and none of them includes a test of the purified protein's activity, thus, leaving discouraging doubts about method's ability to purify proteins in their active state. The goal of this work was to prove that aptamers could facilitate isolation of active proteins. We refined a complete aptamer-based affinity purification procedure, which takes 4 h to complete. We further applied this procedure to purify two recombinant proteins, MutS and AlkB, from bacterial cell culture: 0.21 mg of 85%-pure AlkB from 4 mL of culture and 0.24 mg of 82%-pure MutS from 0.5 mL of culture. Finally, we proved protein activity by two capillary electrophoresis based assays: an enzymatic assay for AlkB and a DNA-binding assay for MutS. We suggest that in combination with aptamer selection for non-purified protein targets in crude cell lysate, aptamer-based purification provides a means of fast isolation of tag-free recombinant proteins in their native state without the use of antibodies.

Video-assisted thoracic surgical technique versus conventional surgical technique for mitral valve replacement: A case control study / 中国胸心血管外科临床杂志

@#Objective    To compare video-assisted thoracic surgical technique (VATS) and conventional surgical technique (CSM) in mitral valve replacement (MVR). Methods    We retrospectively analyzed clinical data of 93 patients in our hospital with mitral valve replacement between January 2010 and January 2015. The patients were divided into two groups including a VATS group and a CSM group.There were 43 patients with 25 males and 18 females at age of 57.43±5.65 years in the VATS group, and 50 patients with 27 males and 23 females at age of 56.40±6.32 years in the CSM group.The clinical outcomes of the two groups were compared. Results    There was no mortality. Echocardiography was normal in both groups during 1-year follow-up. There was no significant difference between the two groups in the operative time, aortic clamping time, cardiopulmonary bypass (CPB) time, or ventilation time. As compared with the CSM group, the patients in the VATS group had a significantly lower complication rate, shorter chest incision length (5.23±1.36 cm vs. 18.21±3.89 cm), less blood transfusion (1.75±0.25 U vs. 3.15±1.50 U), less chest drainage (202.34±12.12 ml vs. 412.32±21.56 ml) and lower pain score (1.26±0.86 vs. 3.01±1.13), shorter time of postoperative hospital stay (8.20±2.36 d vs. 12.10±3.26 d). Conclusion    MVR under VATS is not only technically feasible, but also with excellent clinical results.

Original Research: Establishment of an early embolus-related cerebral injury model after cardiopulmonary bypass in miniature pigs.

Embolus-related cerebral injury is still a serious adverse event after cardiopulmonary bypass (CPB). But there is no stable animal model for basic and clinical research purposes. We chose miniature pig to establish a stable animal model of embolus-related cerebral injury after CPB and verified the validity of results by correlating the histopathological findings with those of diffusion-weighted magnetic resonance imaging (DW-MRI). Based on different treatment regimens, 24 male miniature pigs were randomly assigned into four groups: Control, CPB, embolus, and CPB-embolus groups. DW-MRI was performed before and after surgery to diagnose and locate the brain lesions. Histopathological changes in brain tissues were examined using H&E and Nissl staining. All surgical procedures were uneventful with 100% postoperative survival of pigs. Two animals in the Embolus group and six animals in the CPB-embolus group showed signs of ischemic penumbra on DW-MRI performed 6 h after surgery. Consistent with the results of DW-MRI, histopathological examination showed necrosis and ischemic lesions. In this paper, we demonstrate the feasibility and validity of a pig model of embolus-related cerebral injury associated with CPB. This model may be used in the future for basic and translational research.

Predicting Electrophoretic Mobility of Protein-Ligand Complexes for Ligands from DNA-Encoded Libraries of Small Molecules.

Selection of target-binding ligands from DNA-encoded libraries of small molecules (DELSMs) is a rapidly developing approach in drug-lead discovery. Methods of kinetic capillary electrophoresis (KCE) may facilitate highly efficient homogeneous selection of ligands from DELSMs. However, KCE methods require accurate prediction of electrophoretic mobilities of protein-ligand complexes. Such prediction, in turn, requires a theory that would be applicable to DNA tags of different structures used in different DELSMs. Here we present such a theory. It utilizes a model of a globular protein connected, through a single point (small molecule), to a linear DNA tag containing a combination of alternating double-stranded and single-stranded DNA (dsDNA and ssDNA) regions of varying lengths. The theory links the unknown electrophoretic mobility of protein-DNA complex with experimentally determined electrophoretic mobilities of the protein and DNA. Mobility prediction was initially tested by using a protein interacting with 18 ligands of various combinations of dsDNA and ssDNA regions, which mimicked different DELSMs. For all studied ligands, deviation of the predicted mobility from the experimentally determined value was within 11%. Finally, the prediction was tested for two proteins and two ligands with a DNA tag identical to those of DELSM manufactured by GlaxoSmithKline. Deviation between the predicted and experimentally determined mobilities did not exceed 5%. These results confirm the accuracy and robustness of our model, which makes KCE methods one step closer to their practical use in selection of drug leads, and diagnostic probes from DELSMs.

Prediction of protein-DNA complex mobility in gel-free capillary electrophoresis.

Selection of protein binders from highly diverse combinatorial libraries of DNA-encoded small molecules is a highly promising approach for discovery of small-molecule drug leads. Methods of kinetic capillary electrophoresis provide the high efficiency of partitioning required for such selection but require the knowledge of electrophoretic mobility of the protein-ligand complex. Here we present a theoretical approach for an accurate estimate of the electrophoretic mobility of such complexes. The model is based on a theory of the thin double layer and corresponding expressions used for the mobilities of a rod-like short oligonucleotide and a sphere-like globular protein. The model uses empirical values of mobilities of free protein, free ligand, and electroosmotic flow. The model was tested with a streptavidin-dsDNA complex linked through biotin (small molecule). The deviation of the prediction from the experimental mobility did not exceed 4%, thus confirming that not only is the model adequate but it is also accurate. This model will facilitate reliable use of KCE methods for selection of drug leads from libraries of DNA-encoded small molecules.

Pre-equilibration kinetic size-exclusion chromatography with mass spectrometry detection (peKSEC-MS) for label-free solution-based kinetic analysis of protein-small molecule interactions.

Here we introduce pre-equilibration kinetic size-exclusion chromatography with mass-spectrometry detection (peKSEC-MS), which is a label-free solution-based kinetic approach for characterizing non-covalent protein-small molecule interactions. In this method, a protein and a small molecule are mixed outside the column and incubated to approach equilibrium. The equilibrium mixture is then introduced into the SEC column to initiate the dissociation process by separating small molecules from the complex inside the column. A numerical model of a 1-dimensional separation was constructed to simulate mass chromatograms of the small molecule for varying rate constants of binding.

Kinetic size-exclusion chromatography with mass spectrometry detection: an approach for solution-based label-free kinetic analysis of protein-small molecule interactions.

Studying the kinetics of reversible protein-small molecule binding is a major challenge. The available approaches require that either the small molecule or the protein be modified by labeling or immobilization on a surface. Not only can such modifications be difficult to do but also they can drastically affect the kinetic parameters of the interaction. To solve this problem, we present kinetic size-exclusion chromatography with mass spectrometry detection (KSEC-MS), a solution-based label-free approach. KSEC-MS utilizes the ability of size-exclusion chromatography (SEC) to separate any small molecule from any protein-small molecule complex without immobilization and the ability of mass spectrometry (MS) to detect a small molecule without a label. The rate constants of complex formation and dissociation are deconvoluted from the temporal pattern of small molecule elution measured with MS at the exit from the SEC column. This work describes the concept of KSEC-MS and proves it in principle by measuring the rate constants of interaction between carbonic anhydrase and acetazolamide.

Volatile kinetic capillary electrophoresis for studies of protein-small molecule interactions.

Kinetic capillary electrophoresis (KCE) is a toolset of homogeneous affinity methods for studying kinetics of noncovalent binding. Sensitive KCE measurements are typically done with fluorescence detection and require a fluorescent label on a smaller-sized binding partner. KCE with fluorescence detection is difficult to use for study of protein-small molecule interactions since labeling small molecules is cumbersome and can affect binding. A combination of KCE with mass-spectrometry (KCE-MS) has been recently suggested for label-free studies of protein-small molecule interactions. The major obstacle for studies by KCE-MS is a buffer mismatch between KCE and MS; MS requires volatile buffers while KCE of protein-ligand interactions is always run in near-physiological buffers. Here we asked a simple question: can protein-ligand interactions be studied with KCE in a volatile buffer? We compared three volatile buffers (ammonium acetate, ammonium bicarbonate, and ammonium formate) with a near-physiological buffer (Tris-acetate) for three protein-ligand pairs. The volatile buffers were found not to significantly affect protein-ligand complex stability; moreover, when used as CE run buffers, they facilitated good-quality separation of free ligands from the protein-ligand complexes. The use of volatile buffers instead of Tris-acetate in detection of small molecules by MS improved the detection limit by approximately 2 orders of magnitude. These findings prove the principle of "volatile" KCE, which can be easily coupled with MS to facilitate label-free kinetic studies of protein-small molecule interactions.

Comparison of long-term clinical outcomes and costs between video-assisted thoracoscopic surgery and transcatheter amplatzer occlusion of the patent ductus arteriosus.

The current study was designed to compare long-term clinical outcomes and costs between video-assisted thoracoscopic surgery (VATS) and transcatheter Amplatzer occlusion (TAO). This study enrolled 294 patients with isolated patent ductus arteriosus (PDA) from April 2002 to April 2007, and 290 of these patients were followed up until April 2010. Of the 294 patients, 196 underwent VATS and 98 accepted TAO for PDA closure. The two groups were similar in terms of demographics and preoperative clinical characteristics. No cardiac deaths occurred in either group. All the patients in the VATS group had successful PDA closure, and 94 patients (94/98, 95.9%) in the TAO group had successful PDA occlusion. The incidence of acute procedure-related complications recorded was 1.5% in the VATS group compared with 10.2% in TAO group (P < 0.05). The cost per patient was $1,309.40 ± $312.20 in the VATS group and $3,415.80 ± $637.30 in the TAO group (P < 0.05). There were no cardiac deaths or newly occurring arrhythmias in either group during the fellow-up period. Up to the latest follow-up, no late recanalization or residual shunting was documented, and heart structure returned to normal level in the VATS group. However, residual shunting was detected in four more TAO patients. This study confirmed the long-term safety and efficacy of VATS clipping of PDA. Compared with TAO, PDA interrupted with VATS can achieve both excellent clinical results and satisfying cost effectiveness. The cost for VATS is only a little more than one third the cost for TAO.

Label-free solution-based kinetic study of aptamer-small molecule interactions by kinetic capillary electrophoresis with UV detection revealing how kinetics control equilibrium.

Here we demonstrate a label-free solution-based approach for studying the kinetics of biopolymer-small molecule interactions. The approach utilizes kinetic capillary electrophoresis (KCE) separation and UV light absorption detection of the unlabeled small molecule. In this proof-of-concept work, we applied KCE-UV to study kinetics of interaction between a small molecule and a DNA aptamer. From the kinetic analysis of a series of aptamers, we found that dissociation rather than binding controls the stability of the complex. Because of its label-free features and generic nature, KCE-UV promises to become a practical tool for challenging kinetic studies of biopolymer-small molecule interactions.

Kinetic capillary electrophoresis with mass-spectrometry detection (KCE-MS) facilitates label-free solution-based kinetic analysis of protein-small molecule binding.

Tandem tracker: Here we introduce a method for studying the kinetics of protein-small-molecule interactions based on kinetic capillary electrophoresis (KCE) separation and MS detection. Due to the variety of KCE methods and MS modes available, the KCE-MS tandem is a highly versatile platform for label-free, solution-based kinetic studies of affinity interactions.

Comparison of posterolateral thoracotomy and video-assisted thoracoscopic clipping for the treatment of patent ductus arteriosus in neonates and infants.

This study was designed to compare the long-term clinical outcomes and costs between video-assisted thoracic surgery (VATS) and posterolateral thoracotomy (PT) in neonates and infants. This study enrolled 302 patients with isolated patent ductus arteriosus (PDA) from January 2002 to 2007 and followed them up until April 2010. A total of 134 patients underwent total VATS (VATS group), and 168 underwent PDA closure through PT (PT group). The two groups were compared according to clinical outcomes and costs. The demographics and preoperative clinical characteristics of the patients were similar in the two groups. No cardiac deaths occurred, and the closure rate was 100% successful in both groups. The operating, recovery, and pleural fluid drainage times were significantly shorter in the VATS group than in the PT group. Statistically significant differences in length of incision, postoperative temperature, and acute procedure-related complications were observed between the two groups. The cost was $1,150.3 ± $221.2 for the VATS group and $2415.8 ± $345.2 for the PT group (P < 0.05). No cardiac deaths or newly occurring arrhythmias were detected in either group during the follow-up period. Statistically significant differences in the rate of residual shunt and scoliosis were observed between the two groups. The left ventricular end-diastolic diameter and the pulmonary artery diameter could be restored to normal in the VATS group but not in the PT group. The study confirmed that VATS offers a minimally traumatic, safe, and effective technique for PDA interruption in neonates and infants.