Trastuzumab resistance in HER2-positive breast cancer: Mechanisms, emerging biomarkers and targeting agents.

Trastuzumab is a standard molecular targeted therapy for human epidermal growth factor receptor 2(HER2) -positive breast cancer, which can significantly improve the survival of patients with this molecular subtype of breast cancer. However, the clinical problem of onset or secondary resistance to trastuzumab has limited its efficacy. Therefore, it is very important to explore the mechanism of trastuzumab resistance and formulate countermeasures. Our study described the underlying molecular mechanism of trastuzumab resistance including ERBB2 mutations and nuclear localization, transcriptional and post-translational alterations of ERBB2, over-activation of bypass signaling pathways activation and so on. Then summarize the potential emerging predicting biomarkers and therapeutic strategies for trastuzumab resistance, in order to provide research direction for reversing trastuzumab resistance.

Netting Gut Disease: Neutrophil Extracellular Trap in Intestinal Pathology.

Many gut disease etiologies are attributed to the presence of robust inflammatory cell recruitment. The recruitment of neutrophils plays a vital role in inflammatory infiltration. Neutrophils have various antimicrobial effector mechanisms, including phagocytosis, oxidative burst, and degranulation. It is suggested that neutrophils could release neutrophil extracellular traps (NETs) to kill pathogens. However, recent evidence indicates that neutrophil infiltration within the gut is associated with disrupted local immunological microenvironment and impaired epithelial barrier. Growing evidence implies that NETs are involved in the progression of many diseases, including cancer, diabetes, thrombosis, and autoimmune disease. Increased NET formation was found in acute or chronic conditions, including infection, sterile inflammation, cancer, and ischemia/reperfusion injury (IRI). Here, we present a comprehensive review of recent advances in the understanding of NETs, focusing on their effects in gut disease. We also discuss NETs as a potential therapeutic target in gut disease.

Tyrosinase@HKUST-1: a super stable biocatalyst efficient for catecholic product synthesis.

Although metal-organic frameworks (MOFs) have been considered as promising matrices for enzyme immobilization, HKUST-1, constructed from copper acetate (CuAc2) and benzene 1,3,5-tricarboxylate (BTC), has rarely been explored for this application. In this study, mushroom tyrosinase (EC 1.14.18.1) was immobilized in the form of tyrosinase@HKUST-1 following a simple reaction procedure by mixing BTC with the enzyme prior to addition of CuAc2. The resultant biocatalyst was characterized in both structural features and catalytic properties. Upon incorporation into the HKUST-1 frameworks, the enzyme gained a prominent enhancement in stability against pH, temperature and storage: When incubated at 50 °C and pH 6.0, tyrosinase@HKUST-1 presented a half-life of 32.6 h, which is 77-fold and over tenfold higher than that of the free enzyme and its other immobilization forms, respectively; and the catalyst fully maintained its activity for at least 2 months when stored at 30 °C. The applicability of this new biocatalyst was demonstrated by employing it as catalyst for regioselective ortho-hydroxylation reactions to produce catecholic products with huge pharmacological effects, i.e., hydroxytyrosol and L-DOPA, with excellent yields and productivities. This study has thus offered a facile immobilization method to prepare a novel biocatalyst with super stability, and tyrosinase@HKUST-1 so formed from crude mushroom extract provides an efficient catalyst which can be applied to the production of catecholic products with health benefits.

Pseudomonas putida Represses JA- and SA-Mediated Defense Pathways in Rice and Promotes an Alternative Defense Mechanism Possibly through ABA Signaling.

The signaling pathways induced by Pseudomonas putida in rice plants at the early plant-rhizobacteria interaction stages, with and without inoculation of Xanthomonas oryzae pv. oryzae, were studied. In the absence of pathogen, P. putida reduced ethylene (ET) production, and promoted root and stem elongation. Interestingly, gene OsHDA702, which plays an important role in root formation, was found significantly up-regulated in the presence of the rhizobacterium. Although X. oryzae pv. oryzae inoculation enhanced ET production in rice plants, P. putida treatment repressed ET-, jasmonic acid (JA)- and salicylic acid (SA)-mediated defense pathways, and induced the biosynthesis of abscisic acid (ABA), and the overexpression of OsHDA705 and some pathogenesis-related proteins (PRs), which in turn increased the susceptibility of the rice plants against the pathogen. Collectively, this is the first work on the defense signaling induced by plant growth-promoting rhizobacteria in plants at the early interaction stages, and suggests that rhizobacteria stimulate an alternative defense mechanism in plants based on ABA accumulation and OsHDA705 signaling.

LCZ696 Ameliorates Oxidative Stress and Pressure Overload-Induced Pathological Cardiac Remodeling by Regulating the Sirt3/MnSOD Pathway.

AIMS: We aimed to investigate whether LCZ696 protects against pathological cardiac hypertrophy by regulating the Sirt3/MnSOD pathway. METHODS: In vivo, we established a transverse aortic constriction animal model to establish pressure overload-induced heart failure. Subsequently, the mice were given LCZ696 by oral gavage for 4 weeks. After that, the mice underwent transthoracic echocardiography before they were sacrificed. In vitro, we introduced phenylephrine to prime neonatal rat cardiomyocytes and small-interfering RNA to knock down Sirt3 expression. RESULTS: Pathological hypertrophic stimuli caused cardiac hypertrophy and fibrosis and reduced the expression levels of Sirt3 and MnSOD. LCZ696 alleviated the accumulation of oxidative reactive oxygen species (ROS) and cardiomyocyte apoptosis. Furthermore, Sirt3 deficiency abolished the protective effect of LCZ696 on cardiomyocyte hypertrophy, indicating that LCZ696 induced the upregulation of MnSOD and phosphorylation of AMPK through a Sirt3-dependent pathway. CONCLUSIONS: LCZ696 may mitigate myocardium oxidative stress and apoptosis in pressure overload-induced heart failure by regulating the Sirt3/MnSOD pathway.

[Limonoids from seeds of Azadirachta indica and their antibacterial activity].

Fifteen limonoids were isolated from 95% ethanol extracts of the dry seeds of neem( Azadirachta indica) by various column chromatography techniques including silica gel,Pharmadex LH-20 gel and ODS resin. Based on spectroscopic analysis,their structures were determined as nimbocinol( 1),17ß-hydroxynimbocinol( 2),1α,3α,7α-triacetylvilasinin( 3),7α-benzoyltrichilinin( 4),1,3-diacetyl-7-tigloyl-12-hydroxyvilasinin( 5),3-deacetylsalannin( 6),1-O-acetyl-1-detigloylsalannin( 7),2'( R),3'-dihydrosalannin( 8),2'( S),3'-dihydrosalannin( 9),2,3-dihydronimbolide( 10),6-homodesacetylnimbin( 11),gedunin( 12),7-deacetyl-7-epi-dihydrogedunin( 13),7-deacetoxy-7α-hydroxygedunin( 14) and nimbinene( 15). Compound 7 is a new natural product. 4,8,9,13 and 14 are isolated from the genus Azadirachta for the first time. Compound 2 showed inhibitory activity against Escherichia coli and Staphylococcus epidermidis,with MIC values of 32 and 128 mg·L~(-1),respectively. Compound 10 showed moderate inhibitory activity against S. epidermidis with a MIC value of 64 mg·L~(-1). Compound 11 inhibited the growth of E. coli and Pseudomonas aeruginosa,both with MIC values of 128 mg·L~(-1). Compound 15 exhibited inhibitory activity against P. aeruginosa,with a MIC value of128 mg·L~(-1).

Long noncoding RNA H19 is a critical oncogenic driver and contributes to epithelial-mesenchymal transition in papillary thyroid carcinoma.

BACKGROUND: Growing evidence has indicated that the long noncoding RNA H19 (lncRNA H19), frequently deregulated in almost all tumor types tested, acted as a pivotal contributor to both cancer initiation and progression. However, the role of lncRNA H19 in human papillary thyroid carcinoma (PTC) remains controversial. The aim of the study was to investigate the expression and potential function of lncRNA H19 in human PTC. PATIENTS AND METHODS: The lncRNA H19 level was determined by quantitative real-time (RT)-PCR analyses in 58 PTC tissue samples and their paired paracancerous tissue samples. RNA interference, RT-PCR analysis, and Western blot assay were used to determine the impact of lncRNA H19 on epithelial-mesenchymal transition (EMT) markers in human PTC cells. The migratory and invasive capacities of PTC cells were determined by wound-healing and transwell migration and invasion assays. RESULTS: lncRNA H19 expression was 2.417-fold higher in PTC tissues than their paired paracancerous tissue (95% CI: 1.898-2.935, P<0.0001). Higher level of lncRNA H19 was correlated to elevated expression of Vimentin, ZEB2, Twist, and Snail2. Inhibition of lncRNA H19 resulted in upregulation of E-cadherin and downregulation of Vimentin both at mRNA and protein levels. Conversely, enforced expression of the exogenous lncRNA H19 led to E-cadherin mRNA and protein downregulation and relative upregulation of Vimentin. Moreover, wound-healing and transwell migration and invasion assays showed that lncRNA H19 could promote the migratory and invasive abilities of PTC cells. CONCLUSION: The level of lncRNA H19 was significantly higher in PTC tissues than paired paracancerous tissue or normal tissues. Overexpression of lncRNA H19 was correlated with higher tumor burden of PTC. It also contributes to EMT process, as well as promotes migration and invasion of PTC cells.

Prognostic values of the inhibitor of DNA­binding family members in breast cancer.

The inhibitor of DNA­binding (ID) proteins are dominant­negative modulators of transcription factors with basic helix­loop­helix (bHLH) structures, which control a variety of genes in cell cycle regulation. An increasing volume of evidence has demonstrated that the deregulated expression of IDs in several types of malignancy, including breast carcinoma, has been proven to serve crucial regulatory functions in tumorigenesis and the development of breast cancer (BC). The present study evaluated the prognostic values of the ID family members by investigating a set of publicly accessible databases, including Oncomine, bc­GenExMiner, Kaplan­Meier plotter and the Human Protein Atlas. The results demonstrated that mRNA levels of distinct IDs exhibited diverse profiles between BC and normal counterparts. The mRNA expression level of ID2 was significantly higher in breast cancer than normal tissues, while the mRNA expression levels of ID1, ID3 and ID4 were significantly lower in breast cancer tissues than in normal tissues. Furthermore, higher mRNA expression levels of ID1 and ID4 were associated with subgroups with lower pathological grades and fewer lymph node metastases. Survival analysis revealed that elevated mRNA levels of ID1 and ID4 predicted an improved survival in all patients with BC. Increased ID1 mRNA levels were associated with higher relapse­free survival rates in all patients with BC, particularly in those with ER positive and Luminal A subtype tumors. Increased ID4 mRNA expression predicted longer survival times in all patients with BC, particularly in those with hormone receptor­positive tumors or those treated with endocrine therapy. These results indicated that IDs are essential prognostic indicators in BC. Future studies on the effect of IDs on the pathogenesis and development of BC are warranted.

[Limonoids from seeds of Azadirachta indica and their cytotoxic activity].

Eight limonoids were isolated from 95% ethanol extracts of neem(Azadirachta indica) seeds by various chromatographic methods. By comparison of their spectroscopic data with those reported in the literatures, these limonoids were determined as salannin(1), 1-detigloyl-1-isobutylsalannin(2), salannol-3-acetate(3), salannol(4), spirosendan(5), 1-detigloyloxy-3-deacetylsalannin-1-en-3-one(6), nimbin(7) and 6-deacetylnimbin(8). Compounds 2 and 5 were firstly isolated from this genus and 5 represented the only example of its type. And 6 is a new natural product. 6 showed inhibitory activity against HeLa and HL-60 cells, with IC50 of(21.61±4.37) and(27.33±5.74) µmol·L⁻¹, respectively. Both 7 and 8 mildly inhibited the growth of HeLa cells, with IC50 of (33.15±5.24) and (38.56±6.41) µmol·L⁻¹, respectively.

FoxM1 is a promising candidate target in the treatment of breast cancer.

Forkhead box protein M1(FoxM1) is a member of forkhead superfamily transcription factors. Emerging evidences have progressively contributed to our understanding on a central role of FoxM1 in human cancers. However, perspectives on the function of FoxM1 in breast cancer (BC) remain conflicting, and mostly were from basic research. Here, we explored the expression profile and prognostic values of FoxM1 based on analysis of pooled clinical datasets derived from online accessible databases, including ONCOMINE, Breast Cancer Gene-Expression Miner v4.0, and Kaplan-Meier plotter. It was found that, FoxM1 mRNA expression was significantly higher in breast tumor versus normal control. FoxM1expression profile presented a distinct pattern in different molecular subtypes of BC patients. Higher expression of FoxM1 was correlated to low mRNA expression of estrogen receptor 1 (ESR1), erb-B2 receptor tyrosine kinase 2 (ERBB2), and was inversely associated with the expression of classical luminal regulators forkhead box protein A1 (FoxA1) and GATA binding protein 3 (GATA3). Elevated FoxM1 expression predicted shorter distance metastasis free survival (DMFS) in BC patients, particularly with estrogen receptor (ER) positive and Luminal A, Luminal B subtypes of BC. More interestingly, elevated FoxM1 expression predicted poor survival in breast cancer patients, especially in the ER (+), progesterone receptor (PR) (+) subgroups and BC patients received adjuvant chemotherapy only or treated with tamoxifen only. These results implied that FoxM1 is an essential prognostic factor and promising candidate target in the treatment of breast cancer.

Predictive Value of Preoperative Multidetector-Row Computed Tomography for Axillary Lymph Nodes Metastasis in Patients With Breast Cancer.

Introduction: Axillary lymph nodes (ALN) status is an essential component in tumor staging and treatment planning for patients with breast cancer. The aim of present study was to evaluate the predictive value of preoperative multidetector-row computed tomography (MDCT) for ALN metastasis in breast cancer patients. Methods: A total of 148 cases underwent preoperative MDCT examination and ALN surgery were eligible for the study. Logistic regression analysis of MDCT variates was used to estimate independent predictive factors for ALN metastasis. The prediction of ALN metastasis was determined with MDCT variates through receiver operating characteristic (ROC) analysis. Results: Among the 148 cases, 61 (41.2%) cases had ALN metastasis. The cortical thickness in metastatic ALN was significantly thicker than that in non-metastatic ALN (7.5 ± 5.0 mm vs. 2.6 ± 2.8 mm, P < 0.001). Multi-logistic regression analysis indicated that cortical thickness of >3 mm (OR: 12.32, 95% CI: 4.50-33.75, P < 0.001) and non-fatty hilum (OR: 5.38, 95% CI: 1.51-19.19, P = 0.009) were independent predictors for ALN metastasis. The sensitivity, specificity and AUC of MDCT for ALN metastasis prediction based on combined-variated analysis were 85.3%, 87.4%, and 0.893 (95% CI: 0.832-0.938, P < 0.001), respectively. Conclusions: Cortical thickness (>3 mm) and non-fatty hilum of MDCT were independent predictors for ALN metastasis. MDCT is a potent imaging tool for predicting ALN metastasis in breast cancer. Future prospective study on the value of contrast enhanced MDCT in preoperative ALN evaluation is warranted.

CCR10 activation stimulates the invasion and migration of breast cancer cells through the ERK1/2/MMP-7 signaling pathway.

CCR10, a member of the chemokine receptor subfamily, is overexpressed in several tumors and play a crucial role in cancer development and progression. However, the functions of CCR10 in breast cancer are unknown. Here, we detected the protein levels of CCR10 in breast cancer cells by western blotting, and examined CCR10 expression in breast cancer tissues via immunohistochemical assay. The results showed that CCR10 expression was elevated in breast cancer MCF7, BT-474 and MDA-MB-231 cells. Further, 63 of 89 cases (70.8%) had positive CCR10 staining, and the CCR10 level was closely related to capsular invasion, lymph node metastasis and tumor stage. Moreover, CCL27, the ligand of CCR10, dose-dependently stimulated the invasion and migration of breast cancer cells, and promoted MMP-7 expression and ERK1/2 activation. CCR10 knockdown in breast cancer cells through siRNA transfection attenuated CCL27-induced cell invasiveness, and suppressed MMP-7 expression and ERK1/2 activation. Additionally, blocking the ERK1/2 pathway inhibited the CCL27/CCR10-promoted cell invasion of breast cancer cells. Together, these data suggest that CCL27/CCR10 interaction induces the ERK1/2 pathway, which then increases MMP-7 expresion and subsequently promotes breast cancer cell invasion and migration. Thus, CCR10 may be a key regulator in breast cancer cell invasion and migration.

[A retrospective analysis of 621 cases with craniomaxillofacial fractures].

PURPOSE: To investigate the epidemiological characteristics of 621 hospitalized patients with craniomaxillofacial fractures by a retrospective study. METHODS: From July 2015 to August 2016, the medical records of 621 patients with craniomaxillofacial fractures in Shanghai Ninth People's Hospital were analyzed statistically according to age, gender, etiology and site of fracture. Chi-square test was used to analyze the causes of single and multiple fractures with SPSS 20.0 software package. RESULTS: The male to female ratio was 2.18: 1. The average age was 34.7±15.5 years with 19-29 years old group accounted for the most (28.3%). Traffic accident was the top cause of injury (49.0% ); There were 319 cases of orbital fractures (51.4%), followed by 292 cases of zygomatic complex (47.0%) and 247 cases of mandibular fractures (39.8%). CONCLUSION: The quantity and location of fractures are related to the cause of injury, the nature of stress and anatomical structure of corresponding areas. Establishing a sound system of traffic accident management and related laws is an urgent problem to be solved.

Impact of Intraoperative Blood Pressure Control and Temporary Parent Artery Blocking on Prognosis in Cerebral Aneurysms Surgery.

Objective In cerebral aneurysm clipping and embolization, blood pressure control and temporary parent artery blocking are common methods to prevent aneurysm rupture. Their influence on the prognosis is uncertain. In this study, we try to find out the association between methods above and prognostic indicators.Methods We held a retrospective analysis on patients' medical records of cerebral aneurysms surgical clipping and endovascular coiling , and recorded gender, age, diagnosis, Hunt-Hess grade, Glasgow coma scale score, treatment methods, a history of hypertension, preoperative systolic blood pressure, with or without controlled hypotension, systolic blood pressure difference before and after controlled hypotension, with or without temporary artery blocking, with or without hypertension after treated aneurysm, prognostic indicators including mortality after 1 month, intensive care unit (ICU) stay time of survivors, discharged Glasgow outcome scale (GOS) score. Prognostic indicators were regarded as dependent variable, all the factors were regarded as independent variable, and the strength analysis of influence factors on prognostic indicators was made by binary logistic regression.Results Total cases were 165, including 68 males and 97 females, with an average age of 56 (12-85) years. The mortality after 1 month was 10.9% (18 cases). The ICU stay time of survivors was 7.35 (0-67) days. GOS score at discharge was 1-3 in 40 (24.2%) patients and 4-5 in 125 (75.8%) patients. Systolic blood pressure difference before and after controlled hypotension was an independent factor influencing mortality (t=2.273, P=0.024), and the greater the difference was, the higher the mortality would be. Timely hypertension after aneurysm treated was an independent factor affecting ICU stay time of survivors and patients with hypertension had shorter ICU stay time (χ2=10.017, P=0.001). Blood pressure control (χ2=0.088, P=0.767) and temporary blocking (χ2=1.307, P=0.253) did not show significant influence on GOS score at discharge.Conclusions Timely controlled hypertension after aneurysm clipping and embolization can significantly shorten the stay time in ICU. The degree of controlled hypotension associates with postoperative mortality, the greater systolic blood pressure difference before and after antihypertensive treatment is, the higher the mortality will be.

Noninvasive Blood Glucose Analysis Based on Near-Infrared Reflectance Spectroscopy.

Noninvasive glucose detection is highly required for more convenient and less pain glycaemic monitoring. Most of currently used methods are invasive. In this paper, a near-infrared reflectance spectroscopy (NIRS) is proposed to detect blood glucose to protect patient absent of pain. NIRS is a safe, simple and efficient technology applied in many fields. Experiments, based on Oral Glucose Tolerance Test (OGTT), were conducted to collect data modeling with partial least squares (PLS) regression. 42 samples of fingertip blood and palm were measured by commercially available blood glucose meter and NIRS separately at the same time. The glucose concentration range is between 5 and 12 mmol·L-1. With leave-one-out cross-validation, we obtained a result of root mean square error of cross-validation (RMSECV) of 1.16 mmol·L-1 for all the data. With the pre-processing methods of normalization and un-informative variables elimination reducing noise and eliminating some additional effects, we get a better result of 0.79 mmol·L-1. A RMSECV of 0.41 mmol·L-1 for individual modeling is much less than the total modeling. It has a broad application prospect in individual customization.

Synthesis and cytotoxic activities of 2-substituted (25R)-spirostan-1,4,6-triene-3-ones via ring-opening/elimination and 'click' strategy.

To develop more effective antitumor steroidal drugs, we synthesized a library including twenty-two novel cytotoxic 2-alkyloxyl substituted (25R)-spirostan-1,4,6-triene-3-ones and corresponding 1,2,3-triazoles through an abnormal monoepoxide ring-opening/elimination and 'click' reactions. After the cytotoxic evaluations against HepG2, Caski and HeLa cell lines, three steroidal triazoles 5b, 5f and 5m in this library were found to possess potent anti-proliferative effects against Caski cells with the half-inhibitory concentrations (IC50) of 9.4-11.8 µM. The high-efficient and straightforward process was attractive feature for facile preparation of anti-tumor steroidal triazoles.

A facile strategy to decorate Cu9S5 nanocrystals on polyaniline nanowires and their synergetic catalytic properties.

Here, we demonstrated a novel method to decorate Cu9S5 nanocrystals on polyaniline (PANI) nanowires using the dopant of mercaptoacetic acid (MAA) in the PANI matrix as the sulfur source under a hydrothermal reaction. TEM images showed that Cu9S5 nanocrystals with a size in the range of 5-20 nm were uniformly formed on the surface of PANI nanowires. Significantly, the as-prepared PANI/Cu9S5 composite nanowires have been proven to be novel peroxidase mimics toward the oxidation of the peroxidase substrate 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of H2O2. Due to the synergetic effects between polyaniline nanowires and Cu9S5 nanocrystals, the obtained PANI/Cu9S5 composite nanowires exhibit superior catalytic activity over the independent components. This work not only presents a simple and versatile method to decorate semiconductor nanocrystals on the surface of conducting polymer nanostructures, but also provides fundamental guidelines for further investigations into the synergetic effect between conducting polymers and other materials.

A novel protocol for three-dimensional reconstruction of pharyngeal airway on computed tomography.

Three-dimensional measurement of the pharyngeal airway has been widely used, but the three-dimensional reconstruction of pharyngeal airway has been performed in various ways, especially during the anterior boundary demarcation of the nasopharyngeal airway and oropharyngeal airway. This would inevitably affect the measurement and lead to noncomparison between different studies. Our study provided a novel method for anterior boundary demarcation of pharyngeal airway that defined the anterior boundary of nasopharyngeal airway as the "choana" according to the anatomical definition and defined the anterior boundary of oropharyngeal airway as a plane perpendicular to the long axis of soft palate and through the intersections of the lateral space and inferior space to soft palate according to the physiologic characteristics of soft palate. By 2-step segmentation, a three-dimensional image of pharyngeal airway was eventually reconstructed.Ten computed tomographic scans of pharyngeal airway were included for the anterior boundary demarcation and three-dimensional reconstruction by a medical imaging software (Surgicase 5.0; Materialise Interactive Medical Image Control System, Leuven, Belgium), with the volume and surface area being calculated. By using intraclass correlation coefficient, the reliability between intra- and interobservers of this method was well tested.The method established in this study for anterior boundary demarcation and three-dimensional reconstruction of pharyngeal airway is highly reliable and could more veritably reflect the intrinsic anatomical characteristics of the pharyngeal airway.