Circulating tumor DNA association with residual cancer burden after neoadjuvant chemotherapy in triple-negative breast cancer in TBCRC 030.

BACKGROUND: We aimed to examine circulating tumor DNA (ctDNA) and its association with residual cancer burden (RCB) using an ultrasensitive assay in patients with triple-negative breast cancer (TNBC) receiving neoadjuvant chemotherapy. PATIENTS AND METHODS: We identified responders (RCB 0/1) and matched non-responders (RCB 2/3) from the phase II TBCRC 030 prospective study of neoadjuvant paclitaxel versus cisplatin in TNBC. We collected plasma samples at baseline, 3 weeks and 12 weeks (end of therapy). We created personalized ctDNA assays utilizing MAESTRO mutation enrichment sequencing. We explored associations between ctDNA and RCB status and disease recurrence. RESULTS: Of 139 patients, 68 had complete samples and no additional neoadjuvant chemotherapy. Twenty-two were responders and 19 of those had sufficient tissue for whole-genome sequencing. We identified an additional 19 non-responders for a matched case-control analysis of 38 patients using a MAESTRO ctDNA assay tracking 319-1000 variants (median 1000 variants) to 114 plasma samples from 3 timepoints. Overall, ctDNA positivity was 100% at baseline, 79% at week 3 and 55% at week 12. Median tumor fraction (TFx) was 3.7 × 10-4 (range 7.9 × 10-7-4.9 × 10-1). TFx decreased 285-fold from baseline to week 3 in responders and 24-fold in non-responders. Week 12 ctDNA clearance correlated with RCB: clearance was observed in 10 of 11 patients with RCB 0, 3 of 8 with RCB 1, 4 of 15 with RCB 2 and 0 of 4 with RCB 3. Among six patients with known recurrence, five had persistent ctDNA at week 12. CONCLUSIONS: Neoadjuvant chemotherapy for TNBC reduced ctDNA TFx by 285-fold in responders and 24-fold in non-responders. In 58% (22/38) of patients, ctDNA TFx dropped below the detection level of a commercially available test, emphasizing the need for sensitive tests. Additional studies will determine whether ctDNA-guided approaches can improve outcomes.

Bifocal 532/1064 nm alternately illuminated photoacoustic microscopy for capturing deep vascular morphology in human skin.

BACKGROUND: As a promising technology, photoacoustic microscopy (PAM) plays a critical role in diagnosis and assessment of dermatological conditions by providing subtle vascular networks non-invasively. However, the established PAMs are insufficient for clinical dermatology when faced with complex structures of human skin instead of animal models owing to high melanin content and superimposed vasculature for Asians, which cannot balance the spatial resolution and the imaging depth. OBJECTIVES: To evaluate the ability of bifocal 532/1064-nm alternately illuminated photoacoustic microscopy (BF-PAM) to non-invasively reveal the morphological structure of human skin for improving the diagnosis and therapeutic efficacy of skin diseases. METHODS: A BF-PAM was developed to capture biopsy-like information of human skin from epidermis to hypodermis. The optical foci of the two excitation beams are staggered in the axial direction to form an extended depth-of-field, which can maintain the lateral resolution and the contrast of PA image. RESULTS: The imaging capability of the BF-PAM was demonstrated by depicting the vascular morphology of multilayered skin with imaging depth of ˜3 mm. Furtherly, vascular malformations in port-wine stains skin were quantitatively assessed without the need for any contrast agent, and the distribution, depth and diameter of the ectatic vessels can determine an optimal treatment protocol for port-wine stains lesions. CONCLUSIONS: The quantitative vascular morphology in the dermis can be used to accurately assess vascular characteristics, in which case it enables clinicians to determine optimum treatment parameters in individual patients. As a non-invasive imaging technique, BF-PAM holds great potential to provide objective assessment to enhance the therapeutic efficacy. ETHICAL STATEMENT: The study was conducted in accordance with the Declaration of Helsinki (as revised in 2013). The study was approved by the Chinese Ethics Committee of Registering Clinical Trials (ChiECRCT20200184) and registered with Chinese Clinical Trial Registry (ChiCTR2000034400). Before skin imaging, written informed consent was taken from all individual participants.

[Status of HVPG clinical application in China in 2021].

Objective: The investigation and research on the application status of Hepatic Venous Pressure Gradient (HVPG) is very important to understand the real situation and future development of this technology in China. Methods: This study comprehensively investigated the basic situation of HVPG technology in China, including hospital distribution, hospital level, annual number of cases, catheters used, average cost, indications and existing problems. Results: According to the survey, there were 70 hospitals in China carrying out HVPG technology in 2021, distributed in 28 provinces (autonomous regions and municipalities directly under the central Government). A total of 4 398 cases of HVPG were performed in all the surveyed hospitals in 2021, of which 2 291 cases (52.1%) were tested by HVPG alone. The average cost of HVPG detection was (5 617.2±2 079.4) yuan. 96.3% of the teams completed HVPG detection with balloon method, and most of the teams used thrombectomy balloon catheter (80.3%). Conclusion: Through this investigation, the status of domestic clinical application of HVPG has been clarified, and it has been confirmed that many domestic medical institutions have mastered this technology, but it still needs to continue to promote and popularize HVPG technology in the future.

[Relationship between emotional apathy and motor symptoms, sleep and cognitive function in patients with early Parkinson's disease].

Objective: To study the relationship between emotional apathy and motor symptoms, sleep, and cognitive function in patients with early Parkinson's disease (PD). Methods: One hundred and twenty-nine early PD patients who were treated in the Second Affiliated Hospital of Soochow University from June to October 2020 were included, including 82 male and 47 female patients. The emotional apathy was assessed by modified apathy rating scale (MAES). The above 129 patients were divided into 67 patients in the PD with emotional apathy group (MAES>14 points) and 62 patients in the PD without emotional apathy group (MAES≤ 14 points). Age, gender, course of disease and levodopa equivalent dose were also collected. Hoehn-Yahr stage and unified Parkinson's disease rating scale PartⅢ(UPDRS-Ⅲ), Pittsburgh Sleep Quality Index (PSQI), polysomnography, and Montreal Cognitive Assessment Scale (MoCA) were used to evaluate the motor symptoms, sleep and cognitive functions of patients with early PD, and the clinical characteristics of patients with early PD with apathywere determined. Results: Compared with PD patients without apathy, those with apathy had longer disease duration [M(Q1,Q3)][5.0 (3.0, 7.0) years vs 3.0 (2.0, 5.0) years, P=0.006] and severer motor symptoms [20.0 (10.0, 28.0) vs 14.0 (8.5, 23.0), P=0.047]. There was no significant difference in PSQI score between the two groups. Among the 33 patients who completed polysomnography, compared with PD patients without apathy (n=16), those with apathy (n=17) had a longer rapid eye movement (REM) sleep latency [150 (124, 184) min vs 87 (57, 133) min, P=0.035)] and more frequent periodic limb movements in the REM phase(P=0.042).The REM sleep ratio (r=0.373, P=0.042), apnea-hypopena index (AHI)(r=0.374, P=0.046) and oxygen deficit index (r=0.409, P=0.025) were positively correlated with the degree of apathy in PD patients. PD patients with apathy had relatively poorer performance in cognition assessment than those without apathy and total MoCA score was inversely correlated with the degree of apathy (r=-0.231, P=0.017). Conclusion: Early PD patients with apathy have objective sleep disorders dominated by REM sleep disorders, which can have a negative impact on cognitive function.

[Transcranial sonographic characteristics of Parkinson's disease with symptoms of restless legs syndrome].

Objective: To evaluate the transcranial sonographic characteristics in patients with Parkinson's disease (PD) with symptoms of restless legs syndrome (RLS). Methods: Patients with diagnosis of definite PD from the Second Affiliated Hospital of Soochow University and 3 other participating hospitals between September 2018 and December 2019 were consecutively enrolled. Concurrent RLS symptoms were determined using Non-motor Symptoms Questionnaire. Transcranial sonography (TCS) and clinical assessments were performed during the same time and the related variables were compared between the two groups using t-test, non-parametric test, Chi-square test and Spearman correlation analysis, respectively. Results: Among 349 patients with PD, the prevalence of RLS symptoms was 22.6%. Compared to patients without RLS symptoms, those with RLS had longer disease duration (43.0 (24.0, 91.0) months vs 37.0 (20.0, 60.0) months, P<0.05) and higher Hoehn-Yahr stage (2.5 (2.0, 3.0) vs 2.0 (1.5, 2.5), P<0.01).TCS revealed that patients with RLS symptoms were more likely to have abnormality in the raphe nucleus (21.50% vs 7.78%, χ²=15.9, P<0.001) and increased third ventricle width ((6.22±1.97) mm vs (5.16±1.90) mm, P<0.001). No significant differences were found regarding parameters of substantia nigra. Conclusions: Concurrent RLS symptoms are common in PD patients. Abnormal echogenicity of raphe nucleus and increased third ventricle width could be characteristics of TCS in PD patients with RLS symptoms.

[Association between high expression of intrahepatic programmed death-1 and liver inflammation in patients with autoimmune hepatitis].

Objective: To investigate the expression of programmed death-1 (PD-1) in liver tissue and its association with liver pathology in patients with autoimmune hepatitis (AIH). Methods: A total of 54 AIH patients (38 in the active stage and 16 in the remission stage) were enrolled, and 9 healthy volunteers were enrolled as control group. Immunohistochemistry combined with quantitative image analysis was used to measure the expression of PD-1 in liver tissue. The t-test, rank sum test, one-way analysis of variance, least significant difference t-test, Mann-Whitney U test, and Pearson relation analysis were used for statistical analysis of different types of data. Results: The AIH group had a significantly higher positive rate of PD-1 in liver tissue than the control group (13.57%±6.84% vs 2.22%±0.66%, P < 0.01), and the patients in the active stage of AIH had a significantly higher positive rate of PD-1 in liver tissue than those in the remission stage (16.53%±7.72% vs 6.56%±3.16%, P < 0.01). The positive rate of PD-1 in liver tissue was 6.56%±3.16% in G0 group, 14.33%±5.08% in G1-2 group, and 19.23%±5.41% in G3-4 group (P < 0.01), but there was no significant difference in the positive rate of PD-1 between S0, S1-2, and S3-4 groups (P > 0.05). In AIH patients, the positive rate of PD-1 in liver tissue was positively correlated with the levels of total bilirubin, alanine aminotransferase, aspartate aminotransferase, and IgG (r = 0.665, 0.721, 0.711, and 0.813, all P < 0.01). Conclusion: AIH patients have regulated PD-1 expression in liver tissue, which is closely associated with liver inflammation and is not associated with fibrosis degree, suggesting that PD-1 is involved in the development and progression of inflammation in AIH patients.

Brain functional connectivity and cognition in mild traumatic brain injury.

INTRODUCTION: The aim of this study was to analyze brain functional connectivity and its relationship to cognition in patients with mild traumatic brain injury (mTBI). METHODS: Twenty-five patients with mTBI and 25 healthy control subjects were studied using resting-state functional MRI (rs-fMRI). Amplitudes of low-frequency fluctuations (ALFFs) and functional connectivity (FC) were calculated and correlated with cognition. RESULTS: Compared with the normal control group, the mTBI patients showed a significant decrease in working memory index (WMI) and processing speed index (PSI), as well as significantly decreased ALFFs in the cingulate gyrus, the middle frontal gyrus and superior frontal gyrus. In contrast, the mTBI patients' ALFFs in the left middle occipital gyrus, the left precuneus, and lingual gyrus increased. Additionally, FC significantly decreased in the thalamus, caudate nucleus, and right hippocampus in the mTBI patients. Statistical analysis further showed a significant positive correlation between the ALFF in the cingulate gyrus and the WMI (R (2) = 0.423, P < 0.05) and a significant positive correlation between the FC in the left thalamus and left middle frontal gyrus and the WMI (R (2) = 0.381, P < 0.05). CONCLUSION: rs-fMRI can reveal the functional state of the brain in patients with mTBI. This finding differed from observations of the normal control group and was significantly associated with clinical cognitive dysfunction. Therefore, rs-fMRI offers an objective imaging modality for treatment planning and prognosis assessment in patients with mTBI.

Two completely linked polymorphisms in the PPARG transcriptional regulatory region significantly affect gene expression and intramuscular fat deposition in the longissimus dorsi muscle of Erhualian pigs.

This article is the first to report the significant effect of PPARG promoter polymorphisms on the expression and intramuscular fat (IMF) content in the longissimus dorsi (LD) muscle of Erhualian pigs. Polymorphic sites for the PPARG upstream transcriptional regulatory region were scanned by direct sequencing of two DNA pools consisting of individuals with significantly different IMF content (P < 0.05). Two SNPs (c.-1633C>T and c.-1572G>A) were identified, with different allele distribution between these two groups. A total of 66 Erhualian pigs and 24 Yorkshire × Large White (Y × L) pigs were genotyped for these two SNPs using PCR-SSCP (single-strand conformation polymorphism), which exhibited that only the T-A and C-G haplotypes existed. The T-A type promoter was found to have higher activity than G-C type promoter for PPARG gene transcription in the induced preadipocytes using luciferase assay. In addition, the frequency of the T-A type in Erhualian pigs was much higher than that in the Y × L pigs (35.6% vs. 8.3%), and in fact, no T-A/T-A samples were found in Y × L pigs. Therefore, we conclude that the T-A haplotype of PPARG is a desirable form that might contribute to the relatively higher IMF content in Erhualian pigs.

Global comparison of gene expression between subcutaneous and intramuscular adipose tissue of mature Erhualian pig.

Adipose tissue plays an important role in energy metabolism and related diseases. The content of intramuscular fat significantly influences the pork quality. In this study, the whole gene expression of dorsal subcutaneous (s.c.) adipose tissue and intramuscular (i.m.) adipose tissue isolated from longissimus dorsi muscle tissue were compared using Affymetrix Gene-Chip microarray technology. The result revealed that 1228 genes were more highly expressed in s.c. adipose tissue, whereas 965 genes were higher expressed in i.m. adipose tissue. We found that the s.c. adipose tissue had a stronger capacity of lipid metabolism and fatty acid metabolism compared with i.m. adipose tissue, and angiopoietin-like-4, neuronatin, neuron-derived orphan receptor-1 alfa, and chloride intracellular channel 5 may play important roles in the regulation of fat deposition between i.m. and s.c. adipose tissues.

Rescue from dominant follicle atresia by follicle-stimulating hormone in mice.

We investigated the effects of follicle-stimulating hormone (FSH) on atresia of the dominant follicle and changes in relevant apoptosis genes in granulosa cells of dominant follicles regulated by FSH in vivo. Four-week-old mice were administered FSH by intraperitoneal injection to induce follicular maturation. Granulosa cells of dominant follicles were collected at 48, 72, and 96 h after the first FSH injection. Phosphate-buffered saline was injected as a control. The mRNA levels of relevant granulosa cell apoptosis genes were examined by real-time quantitative polymerase chain reaction, and apoptosis of granulosa cells in dominant ovarian follicles was determined by the terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay. Apoptosis in granulosa cells of dominant follicles was almost TUNEL-negative at 48, 72-66, 72, and 96-90 h after the first FSH injection, but granulosa cell apoptosis in dominant follicles was clearly detected at 96, 102, and 102-96 h by TUNEL. The BIM, caspase-3, and caspase-9 mRNA expression levels were significantly lower after FSH treatment at 72-66 and 96-90 h, compared with that at 72 and 96 h (P < 0.05). Caspase-8 and FasL mRNA expressions did not respond to FSH. FSH rescued granulosa cells from apoptosis when the relevant apoptosis genes were upregulated in early atretic follicles. FSH did not rescue granulosa cells from apoptosis if the DNA was cut into fragments by endonucleases. Thus, the rescue by FSH of granulosa cells from apoptosis and dominant follicle atresia may be accomplished by inhibition of apoptosis in mitochondria.

Clinical feasibility of deep learning-based automatic head CBCT image segmentation and landmark detection in computer-aided surgical simulation for orthognathic surgery.

The purpose of this ambispective study was to investigate whether deep learning-based automatic segmentation and landmark detection, the SkullEngine, could be used for orthognathic surgical planning. Sixty-one sets of cone beam computed tomography (CBCT) images were automatically inferred for midface, mandible, upper and lower teeth, and 68 landmarks. The experimental group included automatic segmentation and landmarks, while the control group included manual ones that were previously used to plan orthognathic surgery. The qualitative analysis of segmentation showed that all of the automatic results could be used for computer-aided surgical simulation. Among these, 98.4% of midface, 70.5% of mandible, 98.4% of upper teeth, and 93.4% of lower teeth could be directly used without manual revision. The Dice similarity coefficient was 96% and the average symmetric surface distance was 0.1 mm for all four structures. With SkullEngine, it took 4 minutes to complete the automatic segmentation and an additional 10 minutes for a manual touchup. The results also showed the overall mean difference between the two groups was 2.3 mm for the midface and 2.4 mm for the mandible. In summary, the authors believe that automatic segmentation using SkullEngine is ready for daily practice. However, the accuracy of automatic landmark digitization needs to be improved.

Structural stability, elasticity, thermodynamics, and electronic structures of L12-type Ni3X (X = Al, Ti, V, Nb) phases under external pressure condition.

In this paper, the effect of pressure on the structural stability, elasticity, thermodynamics, and associated electronic structure of L12-type Ni3X (X = Al, Ti, V, Nb) phases is investigated using a first-principles approach. It is shown that pressure leads to volume compression of the Ni3X phase and reduction of the lattice parameters. The increase of pressure promotes the increase of elastic constants, bulk modulus, shear modulus, and Young's modulus. And there is an extremely strong linear correlation between the pressure and the elastic constants. The calculated elastic constants indicate that the pressure leads to strong mechanical stability and ductility of the Ni3X phase. Mechanical anisotropy of the Ni3X phase also increases with increasing pressure. The electronic analysis shows that the increase in pressure leads to enhanced Ni-d-orbitals and X-d-orbitals hybridization and increased electron transfer. The order in terms of electron accumulation intensity is Ni3Ti > Ni3Nb > Ni3V > Ni3Al. It is more directly reflected in the charge density difference diagram. This is in agreement with the results of the enthalpy of formation (ΔH) and Debye temperature (ΘD) analysis.

Expression of lncRNA AK058003 in esophageal carcinoma and analysis of its intervention effect.

OBJECTIVE: The aim of this study was to investigate the expression of long non-coding ribonucleic acid (lncRNA) AK058003 in esophageal carcinoma (EC) tissues, and to analyze its intervention effect. PATIENTS AND METHODS: The expression of lncRNA AK058003 in EC tissues and para-carcinoma tissues from 130 EC patients was detected via quantitative Polymerase Chain Reaction (qPCR). EC cell lines were selected for exogenous interference in lncRNA AK058003. Subsequently, the expression of lncRNA AK058003 in normal esophageal epithelial cell line (Het-1A) and EC cell lines (EC109, EC9706, KYSE-150, KYSE-30, and TE-1) was detected by qPCR. EC9706 cell lines with the highest expression of lncRNA AK058003 were selected and transfected with lncRNA AK058003 siRNA and lncRNA AK058003 control, respectively. After transfection, the expression of lncRNA AK058003 was determined using PCR. The changes in cell growth and proliferation were analyzed via cell growth curve and cell cycle assay. Meanwhile, the changes in cell migration and invasion were analyzed through wound healing assay. Protein expressions of matrix metalloproteinase-1 (MMP1) and MMP2 were determined by Western blot. Clinical data were collected from EC patients, and the association between lncRNA AK058003 expression and tumor-node-metastasis (TNM) stage was finally analyzed. RESULTS: LncRNA AK058003 was highly expressed EC tissues compared with para-carcinoma tissues (p<0.01). Compared with Het-1A cells, the expression of lncRNA AK058003 was significantly higher in EC109, EC9706, KYSE-150, KYSE-30, and TE-1 cells, with highest level in EC9706 cells (p<0.05). The expression of lncRNA AK058003 remarkably declined in lncRNA AK058003 siRNA group compared with lncRNA AK058003 control group (p<0.001). Compared with lncRNA AK058003 control group, the proliferation of EC cells was significantly weakened in lncRNA AK058003 siRNA group, with the greatest difference at 3 d. Flow cytometry results revealed that cell cycle was arrested in G0/G1 phase in lncRNA AK058003 siRNA group. Wound healing assay indicated that the intercellular distance became large, and cell migration ability was evidently enhanced in lncRNA AK058003 siRNA group with time (p<0.05). Besides, the protein expressions of MMP1 and MMP2 were remarkably lower in lncRNA AK058003 siRNA group than those in lncRNA AK058003 control group. This indicated remarkably declined invasion and metastasis ability. In addition, the postoperative prognosis was significantly worse in patients with higher expression of lncRNA AK058003 (p<0.05). All these findings suggested that lncRNA AK058003 could serve as a biomarker for EC prognosis. CONCLUSIONS: LncRNA AK058003 is highly expressed in EC patients, which promotes proliferation, migration, invasion, and metastasis of EC cells. In addition, the postoperative prognosis of EC patients with high expression of lncRNA AK058003 is relatively poor.

Vision-related quality of life and emotional impact in children with strabismus: a prospective study.

The potential impact of the surgical correction of strabismus on vision-related quality of life (VRQOL) and the symptoms of anxiety and depression in children with strabismus remain unclear. The present study included 60 children with strabismus: 30 with heterophoria and 30 with heterotropia. A healthy age- and gender-matched control group (n = 60) was also recruited. The psychological instruments that were used were the short-form 25-item National Eye Institute Visual Functioning Questionnaire (NEI-VFQ-25) and the Hospital Anxiety and Depression Scale (HADS). The results demonstrated that eight of the 12 NEI-VFQ-25 subscales were significantly impaired in children with strabismus compared with matched controls. Compared with pre-operative values, significant improvements were noted after surgery in the NEI-VFQ-25 summary score, and the anxiety and depression scores. This study demonstrated that the NEI-VFQ-25 instrument can be used in strabismus children and that surgical interventions can improve VRQOL, anxiety and depression in strabismus patients.

Chemokines stimulate human T lymphocyte transendothelial migration to utilize VLA-4 in addition to LFA-1.

Lymphocyte infiltration in inflammation is induced by the dual actions of chemokines and cell adhesion molecules. The role of LFA-1 and VLA-4 in chemokine-induced T cell transendothelial migration (TEM) across cytokine-activated endothelium has not been examined. LFA-1, but not VLA-4, mediated blood T cell TEM to RANTES, macrophage inflammatory protein-1alpha (MIP-1alpha), and stromal cell-derived factor-1 (SDF-1), and across tumor necrosis factor alpha (TNF-alpha) or interferon-gamma (IFN-gamma) -stimulated endothelial cells (EC). Chemokine stimulation in combination with TNF-alpha activation of EC induced TEM, which was partially mediated by VLA-4. SDF-1 increased a beta1-integrin activation epitope on T cells and enhanced VLA-4-mediated adhesion. Thus, LFA-1 mediates TEM under most conditions, but VLA-4 can also mediate TEM, although, in contrast to LFA-1, this requires exogenous chemokines and EC activation. In addition, an LFA-1- and VLA-4-independent pathway of lymphocyte TEM can also be induced by SDF-1.

Regulation of chemokine-induced transendothelial migration of T lymphocytes by endothelial activation: differential effects on naive and memory T cells.

Human T lymphocyte transendothelial migration (TEM) was examined in response to chemokines across cytokine-activated endothelium. Monocyte chemotactic protein-1 (MCP-1), RANTES, and macrophage inflammatory protein-1alpha (MIP-1alpha) induced TEM by memory T cells, while stromal cell-derived factor-1 (SDF-1) induced TEM by both naive and memory T cells. Tumor necrosis factor alpha (TNF-alpha) and interleukin-1 (IL-1) increased endothelial adhesion molecule (CAM) expression, whereas interferon-gamma (IFN-gamma) induced little up-regulation of CAM. However, both TNF-alpha and IFN-gamma strongly facilitated T cell migration, which was completely inhibited by pertussis toxin and both greatly increased TEM to RANTES, MIP-1alpha, and SDF-1 selectively of memory but not naive T cells. Thus, the dual selective effect on memory T cells of endothelial activation and these chemokines promotes the preferential recruitment of memory T cells to inflammatory sites. However, the enhanced chemokine-induced migration by memory T cells across activated endothelium appears to be independent of the increase in endothelial CAM expression. G-protein-linked stimuli may play an important part in T cell TEM across cytokine-activated endothelium.

Synthesized ultrathin MoS2 nanosheets perpendicular to graphene for catalysis of hydrogen evolution reaction.

We synthesize the ultrathin MoS2 nanosheets perpendicular to reduced graphene oxides (MoS2⊥RGO) as an electro-catalyst, which exhibits excellent catalytic activity and good stability for the hydrogen evolution reaction (HER) in acidic medium.