[Colorectal cancer with ß-catenin protein expression deficiency: a clinicopathological analysis].

Objective: To investigate the clinicopathological features and molecular characteristics of ß-catenin-deficient colorectal cancer. Methods: The clinical, pathological and molecular features of 11 colorectal cancers with ß-catenin protein loss diagnosed at the 960th Hospital of People's Liberation Army of China, from January 2012 to November 2022 were analyzed. Results: Among the 11 patients, 3 were males and 8 were females. Their age ranged from 43 to 74 years, with the median age of 59 years. Six were in the left colon and 5 were in the right colon. One of the 11 cases had lymph node metastasis, 10 cases were well and moderately differentiated adenocarcinoma, and 1 was mucinous adenocarcinoma. Eight cases were of TNM stage T4, 2 of T1 stage and 1 of Tis stage. ß-catenin protein was not detected using immunohistochemistry. Sanger sequencing revealed the presence of fragment-deletion mutation in exon 3 of CTNNB1 gene, resulting in loss of ß-catenin protein expression. Conclusion: ß-catenin deficiency is present in a small number of colorectal cancers and may be associated with exon 3 mutations of CTNNB1 gene.

Bi-paratopic and multivalent VH domains block ACE2 binding and neutralize SARS-CoV-2.

Neutralizing agents against SARS-CoV-2 are urgently needed for the treatment and prophylaxis of COVID-19. Here, we present a strategy to rapidly identify and assemble synthetic human variable heavy (VH) domains toward neutralizing epitopes. We constructed a VH-phage library and targeted the angiotensin-converting enzyme 2 (ACE2) binding interface of the SARS-CoV-2 Spike receptor-binding domain (Spike-RBD). Using a masked selection approach, we identified VH binders to two non-overlapping epitopes and further assembled these into multivalent and bi-paratopic formats. These VH constructs showed increased affinity to Spike (up to 600-fold) and neutralization potency (up to 1,400-fold) on pseudotyped SARS-CoV-2 virus when compared to standalone VH domains. The most potent binder, a trivalent VH, neutralized authentic SARS-CoV-2 with a half-maximal inhibitory concentration (IC50) of 4.0 nM (180 ng ml-1). A cryo-EM structure of the trivalent VH bound to Spike shows each VH domain engaging an RBD at the ACE2 binding site, confirming our original design strategy.

Engineered ACE2 receptor traps potently neutralize SARS-CoV-2.

An essential mechanism for severe acute respiratory syndrome coronavirus 1 (SARS-CoV-1) and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection begins with the viral spike protein binding to the human receptor protein angiotensin-converting enzyme II (ACE2). Here, we describe a stepwise engineering approach to generate a set of affinity optimized, enzymatically inactivated ACE2 variants that potently block SARS-CoV-2 infection of cells. These optimized receptor traps tightly bind the receptor binding domain (RBD) of the viral spike protein and prevent entry into host cells. We first computationally designed the ACE2-RBD interface using a two-stage flexible protein backbone design process that improved affinity for the RBD by up to 12-fold. These designed receptor variants were affinity matured an additional 14-fold by random mutagenesis and selection using yeast surface display. The highest-affinity variant contained seven amino acid changes and bound to the RBD 170-fold more tightly than wild-type ACE2. With the addition of the natural ACE2 collectrin domain and fusion to a human immunoglobulin crystallizable fragment (Fc) domain for increased stabilization and avidity, the most optimal ACE2 receptor traps neutralized SARS-CoV-2-pseudotyped lentivirus and authentic SARS-CoV-2 virus with half-maximal inhibitory concentrations (IC50s) in the 10- to 100-ng/mL range. Engineered ACE2 receptor traps offer a promising route to fighting infections by SARS-CoV-2 and other ACE2-using coronaviruses, with the key advantage that viral resistance would also likely impair viral entry. Moreover, such traps can be predesigned for viruses with known entry receptors for faster therapeutic response without the need for neutralizing antibodies isolated from convalescent patients.

Validation and use of the Second Victim Experience and Support Tool questionnaire: a scoping review.

OBJECTIVES: Patient safety incidents can impact not only patients and families but also healthcare providers, who may experience negative emotions and symptoms, such as anxiety, guilt, stress, and loss of confidence. To identify and support these "second victims," a screening tool called the Second Victim Experience and Support Tool (SVEST) has been developed. This scoping review aims to map our current knowledge of the SVEST in terms of its scope of use, validation and limitations. STUDY DESIGN: Scoping review. METHODS: In accordance with the framework outlined by Arksey and O'Malley and the Preferred Reporting Items for Systematic Reviews and Meta-Analysis extension for Scoping Reviews, we conducted a literature search in MEDLINE, CINAHL, Cochrane Library, SCOPUS, Embase and PsycINFO databases from database inception up till 1 March 2023. RESULTS: A total of 31 studies were reviewed. The SVEST has been cross-culturally adapted from English into other languages. The SVEST has been successfully used in different contexts and with various healthcare professionals, including doctors, nurses, allied health professionals, midwives and pharmacists. The tool has been used to assess the impact of second victim experiences and the effectiveness of support interventions in addressing the phenomenon. Validity assessment of translated versions of SVEST in the reviewed studies revealed good content validity in most cases, although some studies did not report clear values for scale-level Content Validity Index. On the whole, SVEST is generally a reliable and valid tool, although further refinements and modifications may improve its validity and reliability. CONCLUSIONS: The review highlights the significance of SVEST as a crucial resource for healthcare providers and organisations that prioritise well-being and safety in health care. It also underscores the importance of recognising the needs of second victims and offering them appropriate interventions to manage the aftermath of adverse events.

Enhanced photocatalytic activity of Fe-, S- and N-codoped TiO2 for sulfadiazine degradation.

The composite material based on N-, S-, and Fe-doped TiO2 (NSFe-TiO2) synthesized by wet impregnation was used as a photocatalyst to rapidly degrade sulfadiazine. The photocatalytic degradation behavior and mechanism of sulfadiazine on NSFe-TiO2 were investigated for revealing the role of degradation under ultraviolet light. The results showed that compared with TiO2, NSFe-TiO2 markedly improved the efficiency in photocatalytic degradation of sulfadiazine: more than 90% of sulfadiazine could be removed within 120 min by NSFe-TiO2 dosage of 20 mg L-1. The process conformed to first-order reaction kinetics model. The parameters such as loaded amount of NSFe-TiO2, solution pH value, humic acid concentration and recycle numbers on removal efficiency were also studied. Compared to neutral and alkaline conditions, acidic condition was not conducive to the photocatalysis. HA, Ca2+, Cu2+ and Zn2+ in the actual water body had mild inhibition on sulfadiazine degradation in UV/NSFe-TiO2 system. Fragments screened by high-resolution mass spectrometry were conducted to explore the oxidation mechanism and pathways of sulfadiazine degradation. On the whole, UV/NSFe-TiO2 photocatalysis has a good effect on sulfadiazine removal. Supplementary Information: The online version contains supplementary material available at 10.1007/s13762-023-04771-6.

Adaptor-Specific Antibody Fragment Inhibitors for the Intracellular Modulation of p97 (VCP) Protein-Protein Interactions.

Protein-protein interactions (PPIs) form complex networks to drive cellular signaling and cellular functions. Precise modulation of a target PPI helps explain the role of the PPI in cellular events and possesses therapeutic potential. For example, valosin-containing protein (VCP/p97) is a hub protein that interacts with more than 30 adaptor proteins involved in various cellular functions. However, the role of each p97 PPI during the relevant cellular event is underexplored. The development of small-molecule PPI modulators remains challenging due to a lack of grooves and pockets in the relatively large PPI interface and the fact that a common binding groove in p97 binds to multiple adaptors. Here, we report an antibody fragment-based modulator for the PPI between p97 and its adaptor protein NSFL1C (p47). We engineered these antibody modulators by phage display against the p97-interacting domain of p47 and minimizing binding to other p97 adaptors. The selected antibody fragment modulators specifically disrupt the intracellular p97/p47 interaction. The potential of this antibody platform to develop PPI inhibitors in therapeutic applications was demonstrated through the inhibition of Golgi reassembly, which requires the p97/p47 interaction. This study presents a unique approach to modulate specific intracellular PPIs using engineered antibody fragments, demonstrating a method to dissect the function of a PPI within a convoluted PPI network.

A musculoskeletal ultrasound program as an intervention to improve disease modifying anti-rheumatic drugs adherence in rheumatoid arthritis: a randomized controlled trial.

Objectives: To evaluate the effect of a musculoskeletal ultrasound programme (MUSP) applying real-time ultrasonography with reinforcement of findings by a rheumatologist on improving disease-modifying anti-rheumatic drugs (DMARDs) adherence in rheumatoid arthritis (RA).Method: Eligible RA patients with low adherence score (< 6) on the 8-item Morisky Medication Adherence Scale (MMAS-8) were randomized to either an intervention group (receiving MUSP at baseline) or a control group (no MUSP), and followed up for 6 months. Adherence measures (patient-reported and pharmacy dispensing records) and clinical efficacy data were collected. The MUSP's feasibility and acceptability were assessed.Results: Among 132 recruited RA patients, six without baseline visits were excluded; therefore, 126 patients were analysed (62 intervention and 64 control). The primary outcome (proportion of patients with 1 month MMAS-8 score < 6) was significantly smaller (p = 0.019) in the intervention (35.48%) than the control group (56.25%). However, 3 and 6 month adherence and clinical efficacy outcomes were not significantly different between the two groups (all p > 0.05). All 62 patients completed the MUSP (mean time taken, 9.2 min), with the majority reporting moderately/very much improved understanding of their joint condition (71%) and the importance of regularly taking their RA medication(s) (79%). Most patients (90.3%) would recommend the MUSP to another RA patient.Conclusions: The MUSP improved RA patients' DMARDs adherence in the short term and was feasible and well accepted by patients. Future studies could evaluate whether repeated feedback using MUSP could help to sustain the improvement in DMARD adherence in RA patients, and whether this may be clinically impactful and cost-effective.

[The application value of plasma heterogeneous nuclear ribonucleoprotein A2/B1, Aß42 and P-tau in the preoperative diagnosis of mild cognitive dysfunction].

Objective: To explore the application value of plasma heterogeneous nuclear ribonucleoprotein A2/B1(hnRNP A2B1), ß-amyloid 42(Aß42) and phosphorylated tau protein(P-tau) levels in elderly patients in the preoperative diagnosis of mild cognitive impairment(MCI). Methods: A total of 200 patients who underwent elective surgery at Tianjin Third Central Hospital from June 2020 to March 2021were Enrolled, regardless of gender, age 65-80 years old. According to the international MCI working group standards and the European Alzheimer's Disease Federation working group standards, patients were divided into MCI group and control group. There were 58 males and 42 females in each group. The patient's plasma hnRNP A2/B1, Aß42 and P-tau levels were detected before operation. The sensitivity, specificity and accuracy of the diagnosis of MCI were calculated. The receiver operating characteristic curve were drew to evaluate the diagnostic value of each index. Results: The plasma levels of hnRNP A2/B1, Aß42 and P-tau in the MCI group were 310.0 (275.1, 344.2), 34.5 (24.9, 42.5), 190.4 (150.4, 301.7) ng/L, respectively, which were significantly higher than those of the control group [272.7 (239.6, 291.5), 18.7 (14.7, 26.6), 140.0 (101.8, 217.5) ng/L]. The differences were statistically significant (all P<0.05). Taking the international MCI working group standard as the gold standard, the sensitivity, specificity and area under the ROC curve (AUC) of plasma hnRNP A2/B1 for predicting MCI were 80%, 61%, and 0.781, respectively. The sensitivity, specificity and AUC of plasma Aß42 for predicting MCI were 78%, 73%, and 0.744. The sensitivity, specificity, and AUC of P-tau for predicting MCI were 51%, 79%, and 0.675, respectively. The sensitivity, specificity and AUC of hnRNP A2/B1 and Aß42 in predicting MCI were not statistically significant (all P>0.05), but the sensitivity of both were higher than P-tau (all P<0.001). Compared with P-tau, the AUC of plasma hnRNP A2/B1 was higher when predicting MCI (P<0.05). When the three indicators were combined, the sensitivity was 82%, and the AUC was 0.842, both of which were the highest, but the specificity reduced (71%) (all P<0.05). Conclusions: Plasma hnRNP A2/B1 combined with Aß42 and P-tau levels can improve the sensitivity and accuracy of MCI diagnosis in elderly MCI patients before surgery, and have the greatest diagnostic efficiency. It has certain application value.

[Application of superb microvascular imaging in focal liver lesions].

Objective: To investigate the ability of superb-microvascular imaging (SMI) to detect microvascular characteristics of focal liver lesions (FLLs) and analyze the relationship between vascular index (VI) and microvascular density (MVD) and Ki-67 levels. Methods: The imaging data of patients diagnosed as FLLs at Tianjin Medical University Cancer Hospital in 2018 were collected. A total of 166 FLLs patients were divided into non-hepatocellular liver cancer (non-HCC group, 96) and HCC group (70), respectively. The whole group of patients were subjected to color Doppler blood flow imaging (CDFI) and SMI examination. The patient's Adler's semi-quantitative grading (0 to 3 levels) and vascular morphological characteristics (a-f type) were analyzed. The receiver operating characteristic (ROC) curve was used to evaluate the detection ability of HCC with SMI and CDFI blood flow characteristics, The Pearson correlation analysis was used to evaluate the correlation between HCC patients VI and MVD and the Spearman correlation analysis was used to evaluate the correlation between VI and Ki-67. Results: In HCC group, SMI detected 50 cases of high-level blood flow (Adler's semi-quantitative grade 2 to 3) patients, higher than 22 cases of CDFI (P=0.033). In HCC group, SMI detected 52 cases of blood-rich mode (e, f type), higher than 18 cases of CDFI (P<0.001). In non-HCC group, the difference of blood flow characteristics detection between CDFI and SMI was not statistically significant. In HCC group, SMI detected 52 cases of rich blood supply patterns, which was higher than 14 cases of non-HCC group (P<0.001). The area under the ROC curve of SMI was 0.760 (sensitivity was 74.3%, specificity was 85.4%), and the SMI rich blood supply mode had the best diagnostic effect on HCC based on the blood-rich mode as the HCC diagnostic standard. In HCC group, VI was positively correlated with MVD and Ki-67 (r=0.698 and r=0.669, respectively, P<0.05). Conclusions: SMI has better detection ability than CDFI for HCC microvascular characteristics, HCC has more blood-rich mode than non-HCC. In HCC, VI is positively correlated with MVD and Ki-67 expression levels.

[Application value of superb microvascular imaging for diagnosis of different size renal solid tumors].

Objective: To discuss the diagnostic value of superb microvacular imaging (SMI) in renal solid tumors of different sizes. Methods: A total of 142 patients with 146 renal tumors detected by ultrasound in Tianjin Medical University Cancer Institute and Hospital from September 2017 to March 2019 were retrospectively analyzed. The maximum diameter of lesions was 0.8-7.3 cm, and patients were divided into the maximum diameter ≤3.0 cm group (61 patients, 64 lesions) and the maximum diameter >3.0 cm group (61 patients, 82 lesions). All of the tumors were separately evaluated by SMI and color Doppler flow imaging (CDFI) with Adler grade, vascular morphology and peripheral blood flow. Results: In the group with maximum diameter ≤3.0 cm, 50 lesions were malignant and 14 were benign. In the group with maximum diameter >3.0 cm, 62 lesions were malignant and 20 were benign. In the group with maximum diameter ≤3.0 cm, there were no significant differences in location, boundary, echo, homogeneity and small cystic area between malignant tumors and benign tumors (P>0.05). In the group with maximum diameter >3.0 cm, there were significant differences in echo, homogeneity and small cystic area between malignant tumors and benign tumors (P<0.05). For all of the benign tumors, there were no significant difference between CDFI and SMI in evaluating Adler grade, vascular morphology and peripheral blood flow (P>0.05). For malignant tumors with maximum diameter ≤3.0 cm, 43 lesions with Adler grade 2-3 were detected by SMI, which was higher than CDFI (32, P<0.05). There were 38 lesions with dendritic and irregular vascular morphology detected by SMI, which was higher than CDFI (7, P<0.05). The detected rate of peripheral annular or semi-annular blood flow in lesions was 80.0% (40/50), higher than CDFI (18/50, P<0.05). While in malignant tumors with maximum diameter >3 cm, the lesion with peripheral annular or semi-annular blood flow detected by SMI was 38, higher than 22 of CDFI (P<0.05). The area under the receiver operating characteristic (ROC) curve for CDFI and SMI diagnosis of renal solid tumors with the maximum diameter ≤3.0 cm was 0.627 (sensitivity: 54.0%, specificity: 71.4%) and 0.791 (sensitivity: 94.0%, specificity: 64.3%), respectively, with statistically significant difference (P=0.039). The area under the ROC curve for CDFI and SMI diagnosis of renal solid tumors with the maximum diameter>3.0 cm was 0.852 (sensitivity: 85.5%, specificity: 85.0%) and 0.860 (sensitivity: 91.9%, specificity: 80.0%), respectively, without statistically significance (P=0.858). Conclusions: SMI is superior to CDFI in detecting low-velocity blood flow and microvessels in both malignant and benign renal tumors, and can effectively improve the display rate of peripheral blood flow in malignant tumors. Otherwise, SMI can provide better diagnostic efficiency for renal tumors with the maximum diameter ≤3.0 cm.

Targeting Phosphotyrosine in Native Proteins with Conditional, Bispecific Antibody Traps.

Engineering sequence-specific antibodies (Abs) against phosphotyrosine (pY) motifs embedded in folded polypeptides remains highly challenging because of the stringent requirement for simultaneous recognition of the pY motif and the surrounding folded protein epitope. Here, we present a method named phosphotyrosine Targeting by Recombinant Ab Pair, or pY-TRAP, for in vitro engineering of binders for native pY proteins. Specifically, we create the pY protein by unnatural amino acid misincorporation, mutagenize a universal pY-binding Ab to create a first binder B1 for the pY motif on the pY protein, and then select against the B1-pY protein complex for a second binder B2 that recognizes the composite epitope of B1 and the pY-containing protein complex. We applied pY-TRAP to create highly specific binders to folded Ub-pY59, a rarely studied Ub phosphoform exclusively observed in cancerous tissues, and ZAP70-pY248, a kinase phosphoform regulated in feedback signaling pathways in T cells. The pY-TRAPs do not have detectable binding to wild-type proteins or to other pY peptides or proteins tested. This pY-TRAP approach serves as a generalizable method for engineering sequence-specific Ab binders to native pY proteins.

Minimal-access retroperitoneal pancreatic necrosectomy for infected necrotizing pancreatitis: a multicentre study of a step-up approach.

BACKGROUND: Various minimally invasive approaches have been described for infected necrotizing pancreatitis. This article describes a modified minimal-access retroperitoneal pancreatic necrosectomy (MARPN) procedure assisted by gas insufflation. METHODS: This retrospective, observational study documented patients who had undergone a step-up MARPN between 1 January 2010 and 31 December 2016. A minimum follow-up of 1 year was required for inclusion. The step-up approach involved percutaneous catheter drainage followed by the modified MARPN and necrosectomy. If more than one access site was needed it was categorized as complex MARPN. RESULTS: Of 212 patients with infected necrotizing pancreatitis, 164 (77·4 per cent) underwent a step-up approach. The median number of percutaneous catheter drains and MARPN procedures was 3 (range 1-7) and 1 (1-6) respectively. Ninety patients (54·9 per cent) underwent complex MARPN. For residual necrosis after MARPN, three patients (1·8 per cent) underwent sinus tract gastroscopy, and 11 (6·7 per cent) had sinography combined with a tube change. However, operations in 13 patients (7·9 per cent) required conversion to open surgery. Postoperative complications developed in 103 patients (62·8 per cent). The mortality rate was 6·1 per cent (10 deaths). CONCLUSION: A step-up approach using a modified MARPN for infected necrotizing pancreatitis is a reasonable option.

ANTECEDENTES: Los procedimientos mínimamente invasivos se han convertido en los más frecuentes para el tratamiento de necrosis pancreáticas infectadas. El objetivo de este estudio fue presentar un procedimiento de necrosectomía pancreática retroperitoneal de acceso mínimo (minimal-access retroperitoneal pancreatic necrosectomy, MARPN) modificado y asistido mediante insuflación de gases, así como evaluar su seguridad y eficacia. MÉTODOS: Se realizó un análisis retrospectivo y observacional de los datos de un hospital desde el 1 de enero de 2010 hasta el 31 de diciembre de 2016. Se incluyeron en el análisis todos los pacientes en los que realizó un abordaje por etapas, que consistía en el drenaje percutáneo mediante la colocación de un catéter seguido de un procedimiento MARPN modificado, en los que se dispusiese de un seguimiento postoperatorio mínimo de 1 año. El MARPN en el lado derecho y la necrosectomía realizada a través de más de un acceso se clasificaron como MARPN complejo. Se evaluaron los resultados radiológicos y quirúrgicos. RESULTADOS: De 212 pacientes con necrosis pancreática infectada, en 164 (77,4%) se realizó un abordaje por etapas. La mediana del número de drenajes percutáneos y procedimientos MARPN fue 3 (rango, 1-7) y 1 (rango, 1-6), respectivamente. En 90 pacientes (54,9%) se realizó un MARPN complejo. Para la exéresis de necrosis residual después de un MARPN, en 3 pacientes (1,8%) se realizó mediante gastroscopia y en 11 pacientes (6,7%) con un recambio de drenaje bajo control radiológico. En 13 pacientes (7,9%) fue necesaria la reconversión a cirugía abierta. Hubo complicaciones postoperatorias en 103 pacientes (62,8%). La tasa de mortalidad fue del 6,1% (n = 10). CONCLUSIÓN: El abordaje por etapas con un MARPN modificado es seguro y efectivo en el tratamiento de la necrosis pancreática infectada.

[UPLC quantitative fingerprint research on Gardeniae Fructus based on chemical pattern recognition technology].

Twenty-six batches of Gardeniae Fructus from different producing area were collected for the development of the fingerprint, and the main components of Gardeniae Fructus were identified by liquid chromatography-mass spectrometry. The producing areas of Gardeniae Fructus were distinguished by chemical pattern recognition technology, and the index components of Gardeniae Fructus were quantitated. An UPLC wavelength switching method was adopted, and the separation was carried out on a Waters Acquity UPLC HASS C_(18)(2.1 mm×100 mm, 1.7 µm) column using the mobile phase of acetonitrile-0.5% formic acid water for gradient elution. Principal component analysis(PCA) and orthogonal partial least square discriminant analysis(OPLS-DA) were used for the data ana-lysis. The results showed that the similarity of 26 batches of Gardeniae Fructus was more than 0.89, and ten common peaks were defined. Sixteen compounds including monoterpenes, iridoids and diterpenoids were identified by reference identification, literature comparison and high-resolution mass spectrometry data analysis. The distinguishment of origin of Gardeniae Fructus was realized by PCA and OPLS-DA analysis, and two quality differential markers were screened as geniposide and crocin Ⅰ. The contents of crocin Ⅰ, crocin Ⅱ and geniposide in Gardeniae Fructus from different places were different. These results will provide reference for the geographical origin traceability of Gardeniae Fructus.

Fluorescent indicators for simultaneous reporting of all four cell cycle phases.

A robust method for simultaneous visualization of all four cell cycle phases in living cells is highly desirable. We developed an intensiometric reporter of the transition from S to G2 phase and engineered a far-red fluorescent protein, mMaroon1, to visualize chromatin condensation in mitosis. We combined these new reporters with the previously described Fucci system to create Fucci4, a set of four orthogonal fluorescent indicators that together resolve all cell cycle phases.

Skeletal development of the chondrocranium in the tongue sole Cynoglossus semilaevis (Pleuronectiformes: Cynoglossidae).

This study provides a comprehensive description of chondrocranial development before, during and after larval metamorphosis in the tongue sole Cynoglossus semilaevis, a commercially valuable flatfish in China. Samples were collected at regular intervals ranging from 1 to 23 days post hatching (dph). Based on observations of cleared and double-stained specimens and images from sections stained with safranin O-fast green, major morphological events during early development were described. No cartilaginous structure was visible at hatching. From 2 dph onwards, cartilaginous structures such as the trabecular bar and some elements of the mandibular, hyoid and branchial arches appeared. At this time also, cartilaginous structures of the neurocranium started to form. Hypertrophic chondrocytes could be observed in many splanchnocranium elements at 5 dph. The start of ossification was indicated by alizarin red stain visible at 14 dph. At 17 dph, most of the cartilaginous skeleton was ossified. Soon after, the right eye started to migrate and pass through a slit beneath the dorsal-fin base and above the skull. Metamorphosis was complete at 20 dph, at which time the dorsal-fin base cartilage extended onto the anterior region of the head. Meanwhile, extremities of the hyoid and branchial arch elements remained cartilaginous. At 23 dph, endochondral ossification of the splanchnocranium was nearly complete. Unlike previous observations of other Pleuronectiformes, our study indicates that endochondral ossification of C. semilaevis skull cartilage occurs before metamorphosis.

[Analysis of the relationship between ultrasonographic features and cervical lymph node skip metastasis of papillary thyroid micro-carcinoma].

Objective: To investigate the correlation between cervical lymph node skip metastasis with ultrasonographic characteristics of papillary thyroid micro- carcinoma (PTMC). Methods: We reviewed ultrasonographic features of 385 primary PTMC and cervical lymph node metastasis, confirmed by pathology in Tianjin Medical University Cancer Institute and Hospital, to evaluate the efficacy of ultrasonography in the diagnosis of cervical lymph node metastasis of PTMC patients. The relationship between ultrasonographic features of primary lesions and skip metastasis of cervical lymph nodes was analyzed by χ(2) test and multiple factor Cox regression. Results: Among the 385 cases of PTMC patients with cervical lymph node metastasis, 231 cases were central lymph node metastasis alone, 31 cases were lateral cervical lymph node metastasis alone, 123 cases were both central and lateral cervical lymph node metastasis. Among the 354 cases without skip metastasis of cervical lymph nodes, 48 cases were level Ⅱ, 92 cases were level Ⅲ, 83 cases were level Ⅳ, 9 cases were level Ⅴ, 354 cases were level Ⅵ. Among the 31 cases with skipping metastasis of cervical lymph nodes, 12 cases were level Ⅱ, 14 cases were level Ⅲ, 14 cases were level Ⅳ, 1 case was level Ⅴ. The sensitivity and specificity of preoperative ultrasonography in the diagnosis of central cervical lymph node metastasis were 46.3% and 66.7%, respectively, and those of lateral cervical lymph node were 91.0% and 87.8%, respectively. Univariate analysis showed that the abutment/perimeter, diameter and location of PTMC were significantly associated with skip metastasis (P<0.05), multivariate analysis showed that abutment/perimeter and location of PTMC were significantly associated with skip metastasis (P<0.05). Conclusions: The sensitivity and specificity of preoperative ultrasound diagnosis for lateral cervical lymph node metastasis of PTMC is higher than that of central metastasis. PTMC with abutment/perimeter ≥1/4 and upper portion location are prone to skip metastasis.

Etanercept reduces anxiety and depression in psoriasis patients, and sustained depression correlates with reduced therapeutic response to etanercept.

BACKGROUND: The purpose of this study was to explore the correlation of anxiety and depression with therapeutic response to etanercept in psoriasis patients. PATIENTS AND METHODS: One hundred and thirty-three patients with moderate-to-severe plaque psoriasis undergoing etanercept treatment were consecutively enrolled in this prospective cohort study, with all patients receiving etanercept treatment for 6 months. Psoriasis Area and Severity Index (PASI) score was evaluated at baseline (M0) and at month 1 (M1), M3 and M6 after treatment, and PASI 75/90 responses were calculated. The Hospital Anxiety and Depression Scale-Anxiety (HADS-A) score and the HADS-Depression (HADS-D) score were used to evaluate patients' anxiety and depression at M0, M1, M3 and M6. Sustained anxiety/depression were defined as HADS-A/D score≥8points both at M0 and M1. RESULTS: Female gender and higher PASI score were associated with high risk of anxiety, while female gender, higher PASI score and longer disease duration were correlated with increased depression risk. After 6 months of etanercept treatment, 65.4% and 36.1% patients achieved PASI 75 and PASI 90 responses respectively, and both HADS-A and HADS-D scores were decreased. Most importantly, no correlation of baseline anxiety and depression with PASI 75 or PASI 90 response after 6 months of treatment was noted, while sustained depression, though not sustained anxiety, was observed to be correlated with decreased PASI 75 and PASI 90 responses. CONCLUSIONS: Etanercept reduces anxiety and depression in psoriasis patients, and sustained depression correlates with reduced therapeutic response to etanercept.

[Comparative analysis of superb microvascular imaging and contrast-enhanced ultrasound in diagnosis of small renal masses].

Objective: To explore the value of superb microvascular imaging (SMI) and contrast- enhanced ultrasound (CEUS) in differential diagnosis of small renal masses. Methods: A total of 84 patients with 86 lesions of renal masses were collected from October 2016 to March 2018 in Tianjin Medical University Cancer Institute and Hospital. There were 56 males and 28 females,aged from 29 to 76 years old,with an average age of (54±10) years old. These tumors were all examined by using SMI and CEUS. And the imaging diagnostic results were compared based on the pathological results. Adler semi quantitative analysis of blood flow grading of the tumors were evaluated in SMI,and the evaluation standard was as follows:0-1 grade was benign, 2-3 grade was malignant. Besides,the patterns and enhancement of the small renal masses were observed in CEUS. The main diagnostic criteria of malignant tumors were "fast forward" , "rapid regression" and "high perfusion" . Some special types of renal cell carcinoma were diagnosed by "slow in" , "rapid regression" and "hypo-perfusion" . Benign tumors were diagnosed by "slow in" , "slow down" and "hypo-perfusion" . Results: Among 86 cases, 76 cases were confirmed as malignant masses and 10 cases were confirmed as benign masses by pathological diagnosis. By SMI, Adler grade of 89.5% (68/76) malignant tumors were 2-3, Adler grade of 6/10 benign tumors were 0-1. By CEUS, 77.6% (59/76) of malignant tumors were "fast forward" , 82.9% (63/76) were "rapid regression" , 76.3% (58/76) were "high perfusion" ; 7/10 of benign tumors were "slow in" , 5/10 were "slow down" and 6/10 were "low perfusion" . The accuracy, sensitivity, specificity, positive predictive value, negative predictive value and AUC of SMI were 86.0%,89.5%, 60.0%, 94.4%,42.9% and 0.747 respectively. The values for CEUS were 89.5%, 92.1%, 70.0%,95.9%, 53.8% and 0.811 respectively.There was no statistical difference in diagnostic efficacy between CEUS and SMI (P=0.288). Conclusions: Although there is no statistical significance between SMI and CEUS, the accuracy, sensitivity and positive predictive value of SMI in the diagnosis of small renal masses are higher,which can clearly show micro-vessels and exquisitely detect the low-velocity blood flow in small renal masses. In brief,SMI provides a new method in the differential diagnosis of small renal masses.

[Non-surgical treatment of acute pancreatitis with colonic fistula].

Objective: To examine the effectiveness of non-operative of colonic fistula following acute pancreatitis. Methods: Retrospective analysis of 354 patients with acute pancreatitis who were admitted to Department of Hepatobiliary Surgery of Chinese People's Liberation Army General Hospital from January 2013 to December 2018. Age of the patients was (46±14) years (range: 14-85 years); 249 cases (70.3%) were males. There were 41 cases of acute edematous pancreatitis and 313 cases of acute necrotising pancreatitis. Two hundred and fifteen cases were diagnosed as moderate severe acute pancreatitis and 139 were diagnosed as severe acute pancreatitis. Among 313 cases of acute necrotising pancreatitis, 62 cases underwent non-surgical treatment, 251 cases underwent surgical treatment in which 218 of minimal access retroperitoneal pancreatic necrosectomy underwent percutaneous nephroliguectomy with peripancreatic necrotic tissue removal technique. Results: There were 15 cases of colon fistula following acute necrotising pancreatitis, and the incidence rate was 4.2%(15/354). There were 7 males and 8 females, with age of (39±8) years (range: 27 to 50 years). The median interval between acute pancreatitis onset and diagnosis of colonic fistula was 71 days(27-134) days. Two cases occurred at the hepatic flexure of the colon, 4 cases at transverse colon, and 9 cases at splenic flexure of colon. Of the 354 patients, 39 cases died and the mortality was 11.0%. Two patients underwent laparotomy, and one of them died. The remaining 13 patients underwent non-surgical treatment and were discharged. Conclusion: Acute pancreatitis with colonic fistula can be treated with non-surgical treatment and can achieve good prognosis.

Engineering Improved Antiphosphotyrosine Antibodies Based on an Immunoconvergent Binding Motif.

Phosphotyrosine (pY) is one of the most highly studied posttranslational modifications that is responsible for tightly regulating many signaling pathways in eukaryotes. Pan-specific pY antibodies have emerged as powerful tools for understanding the role of these modifications. Nevertheless, structures have not been reported for pan-specific pY antibodies, greatly impeding the further development of tools for integrating this ubiquitous posttranslational modification using structure-guided designs. Here, we present the first crystal structures of two widely utilized pan-specific pY antibodies, PY20 and 4G10. The two antibodies, although developed independently from animal immunizations, have surprisingly similar modes of recognition of the phosphate group, implicating a generic binding structure among pan-specific pY antibodies. Sequence alignments revealed that many pY binding residues are predominant in the mouse V germline genes, which consequently led to the convergent antibodies. On the basis of the convergent structure, we designed a phage display library by lengthening the CDR-L3 loop with the aid of computational modeling. Panning with this library resulted in a series of 4G10 variants with 4 to 11-fold improvements in pY binding affinities. The crystal structure of one improved variant showed remarkable superposition to the computational model, where the lengthened CDR-L3 loop creates an additional hydrogen bond indirectly bound to the phosphate group via a water molecule. The engineered variants exhibited superior performance in Western blot and immunofluorescence.