18F-fluorodeoxyglucose (18F-FDG) positron emission tomography/computed tomography (PET/CT) imaging of pediatric neuroblastoma: a multi-omics parameters method to predict MYCN copy number category.

Background: The MYCN copy number category is closely related to the prognosis of neuroblastoma (NB). Therefore, this study aimed to assess the predictive ability of 18F-fluorodeoxyglucose (18F-FDG) positron emission tomography/computed tomography (PET/CT) radiomic features for MYCN copy number in NB. Methods: A retrospective analysis was performed on 104 pediatric patients with NB that had been confirmed by pathology. To develop the Bio-omics model (B-model), which incorporated clinical and biological aspects, PET/CT radiographic features, PET quantitative parameters, and significant features with multivariable stepwise logistic regression were preserved. Important radiomics features were identified through least absolute shrinkage and selection operator (LASSO) and univariable analysis. On the basis of radiomics features obtained from PET and CT scans, the radiomics model (R-model) was developed. The significant bio-omics and radiomics features were combined to establish a Multi-omics model (M-model). The above 3 models were established to differentiate MYCN wild from MYCN gain and MYCN amplification (MNA). The calibration curve and receiver operating characteristic (ROC) curve analyses were performed to verify the prediction performance. Post hoc analysis was conducted to compare whether the constructed M-model can distinguish MYCN gain from MNA. Results: The M-model showed excellent predictive performance in differentiating MYCN wild from MYCN gain and MNA, which was better than that of the B-model and R-model [area under the curve (AUC) 0.83, 95% confidence interval (CI): 0.74-0.92 vs. 0.81, 95% CI: 0.72-0.90 and 0.79, 95% CI: 0.69-0.89]. The calibration curve showed that the M-model had the highest reliability. Post hoc analysis revealed the great potential of the M-model in differentiating MYCN gain from MNA (AUC 0.95, 95% CI: 0.89-1). Conclusions: The M-model model based on bio-omics and radiomics features is an effective tool to distinguish MYCN copy number category in pediatric patients with NB.

Hypoxia-activated selectivity-improved anti-PKM2 antibody combined with prodrug TH-302 for potentiated targeting therapy in hepatocellular carcinoma.

Background: Hypoxia induces hepatocellular carcinoma (HCC) malignancies; yet it also offers treatment opportunities, exemplified by developing hypoxia-activated prodrugs (HAPs). Although HAP TH-302 combined with therapeutic antibody (Ab) has synergistic effects, the clinical benefits are limited by the on-target off-tumor toxicity of Ab. Here, we sought to develop a hypoxia-activated anti-M2 splice isoform of pyruvate kinase (PKM2) Ab combined with TH-302 for potentiated targeting therapy. Methods: Codon-optimized and hypoxia-activation strategies were used to develop H103 Ab-azo-PEG5k (HAP103) Ab. Hypoxia-activated HAP103 Ab was characterized, and hypoxia-dependent antitumor and immune activities were evaluated. Selective imaging and targeting therapy with HAP103 Ab were assessed in HCC-xenografted mouse models. Targeting selectivity, systemic toxicity, and synergistic therapeutic efficacy of HAP103 Ab with TH-302 were evaluated. Results: Human full-length H103 Ab was produced in a large-scale bioreactor. Azobenzene (azo)-linked PEG5k conjugation endowed HAP103 Ab with hypoxia-activated targeting features. Conditional HAP103 Ab effectively inhibited HCC cell growth, enhanced apoptosis, and induced antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) functions. Analysis of HCC-xenografted mouse models showed that HAP103 Ab selectively targeted hypoxic HCC tissues and induced potent tumor-inhibitory activity either alone or in combination with TH-302. Besides the synergistic effects, HAP103 Ab had negligible side effects when compared to parent H103 Ab. Conclusion: The hypoxia-activated anti-PKM2 Ab safely confers a strong inhibitory effect on HCC with improved selectivity. This provides a promising strategy to overcome the on-target off-tumor toxicity of Ab therapeutics; and highlights an advanced approach to precisely kill HCC in combination with HAP TH-302.

Hypoxia-activated ADCC-enhanced humanized anti-CD147 antibody for liver cancer imaging and targeted therapy with improved selectivity.

Therapeutic antibodies (Abs) improve the clinical outcome of cancer patients. However, on-target off-tumor toxicity limits Ab-based therapeutics. Cluster of differentiation 147 (CD147) is a tumor-associated membrane antigen overexpressed in cancer cells. Ab-based drugs targeting CD147 have achieved inadequate clinical benefits for liver cancer due to side effects. Here, by using glycoengineering and hypoxia-activation strategies, we developed a conditional Ab-dependent cellular cytotoxicity (ADCC)-enhanced humanized anti-CD147 Ab, HcHAb18-azo-PEG5000 (HAP18). Afucosylated ADCC-enhanced HcHAb18 Ab was produced by a fed-batch cell culture system. Azobenzene (Azo)-linked PEG5000 conjugation endowed HAP18 Ab with features of hypoxia-responsive delivery and selective targeting. HAP18 Ab potently inhibits the migration, invasion, and matrix metalloproteinase secretion, triggers the cytotoxicity and apoptosis of cancer cells, and induces ADCC, complement-dependent cytotoxicity, and Ab-dependent cellular phagocytosis under hypoxia. In xenograft mouse models, HAP18 Ab selectively targets hypoxic liver cancer tissues but not normal organs or tissues, and has potent tumor-inhibiting effects. HAP18 Ab caused negligible side effects and exhibited superior pharmacokinetics compared to those of parent HcHAb18 Ab. The hypoxia-activated ADCC-enhanced humanized HAP18 Ab safely confers therapeutic efficacy against liver cancer with improved selectivity. This study highlights that hypoxia activation is a promising strategy for improving the tumor targeting potential of anti-CD147 Ab drugs.

Predicting MYCN amplification in paediatric neuroblastoma: development and validation of a 18F-FDG PET/CT-based radiomics signature.

OBJECTIVES: To develop and validate an 18F-FDG PET/CT-based clinical-radiological-radiomics nomogram and evaluate its value in the diagnosis of MYCN amplification (MNA) in paediatric neuroblastoma (NB) patients. METHODS: A total of 104 patients with NB were retrospectively included. We constructed a nomogram to predict MNA based on radiomics signatures, clinical and radiological features. The multivariable logistic regression and the least absolute shrinkage and selection operator (LASSO) were used for feature selection. Radiomics models are constructed using decision trees (DT), logistic regression (LR) and support vector machine (SVM) classifiers. A clinical-radiological (C-R) model was developed using clinical and radiological features. A clinical-radiological-radiomics (C-R-R) model was developed using the C-R model of the best radiomics model. The prediction performance was verified by receiver operating characteristic (ROC) curve analysis, calibration curve analysis and decision curve analysis (DCA) in the training and validation cohorts. RESULTS: The present study showed that four radiomics signatures were significantly correlated with MNA. The SVM classifier was the best model of radiomics signature. The C-R-R model has the best discriminant ability to predict MNA, with AUCs of 0.860 (95% CI, 0.757-0.963) and 0.824 (95% CI, 0.657-0.992) in the training and validation cohorts, respectively. The calibration curve indicated that the C-R-R model has the goodness of fit and DCA confirms its clinical utility. CONCLUSION: Our research provides a non-invasive C-R-R model, which combines the radiomics signatures and clinical and radiological features based on 18F-FDGPET/CT images, shows excellent diagnostic performance in predicting MNA, and can provide useful biological information with stratified therapy. CRITICAL RELEVANCE STATEMENT: Radiomic signatures of 18F-FDG-based PET/CT can predict MYCN amplification in neuroblastoma. KEY POINTS: • Radiomic signatures of 18F-FDG-based PET/CT can predict MYCN amplification in neuroblastoma. • SF, LDH, necrosis and TLG are the independent risk factors of MYCN amplification. • Clinical-radiological-radiomics model improved the predictive performance of MYCN amplification.

18F-FDG PET/CT imaging of pediatric peripheral neuroblastic tumor: a combined model to predict the International Neuroblastoma Pathology Classification.

Background: The aim of this study was to evaluate the effect of a model combining a 18F-fluorodeoxyglucose positron emission tomography/computed tomography (18F-FDG PET/CT)-based radiomics signature with clinical factors in the preoperative prediction of the International Neuroblastoma Pathology Classification (INPC) type of pediatric peripheral neuroblastic tumor (pNT). Methods: A total of 106 consecutive pediatric pNT patients confirmed by pathology were retrospectively analyzed. Significant features determined by multivariate logistic regression were retained to establish a clinical model (C-model), which included clinical parameters and PET/CT radiographic features. A radiomics model (R-model) was constructed on the basis of PET and CT images. A semiautomatic method was used for segmenting regions of interest. A total of 1,016 radiomics features were extracted. Univariate analysis and the least absolute shrinkage selection operator were then used to select significant features. The C-model was combined with the R-model to establish a combination model (RC-model). The predictive performance was validated by receiver operating characteristic (ROC) curve analysis, calibration curves, and decision curve analysis (DCA) in both the training cohort and validation cohort. Results: The radiomics signature was constructed using 5 selected radiomics features. The RC-model, which was based on the 5 radiomics features and 3 clinical factors, showed better predictive performance compared with the C-model alone [area under the curve in the validation cohort: 0.908 vs. 0.803; accuracy: 0.903 vs. 0.710; sensitivity: 0.895 vs. 0.789; specificity: 0.917 vs. 0.583; net reclassification improvement (NRI) 0.439, 95% confidence interval (CI): 0.1047-0.773; P=0.01]. The calibration curve showed that the RC-model had goodness of fit, and DCA confirmed its clinical utility. Conclusions: In this preliminary single-center retrospective study, an R-model based on 18F-FDG PET/CT was shown to be promising in predicting INPC type in pediatric pNT, allowing for the noninvasive prediction of INPC and assisting in therapeutic strategies.

An injectable bioactive dressing based on platelet-rich plasma and nanoclay: Sustained release of deferoxamine to accelerate chronic wound healing

Delayed diabetic wound healing has placed an enormous burden on society. The key factors limiting wound healing include unresolved inflammation and impaired angiogenesis. Platelet-rich plasma (PRP) gel, a popular biomaterial in the field of regeneration, has limited applications due to its non-injectable properties and rapid release and degradation of growth factors. Here, we prepared an injectable hydrogel (DPLG) based on PRP and laponite by a simple one-step mixing method. Taking advantages of the non-covalent interactions, DPLG could overcome the limitations of PRP gels, which is injectable to fill irregular injures and could serve as a local drug reservoir to achieve the sustained release of growth factors in PRP and deferoxamine (an angiogenesis promoter). DPLG has an excellent ability in accelerating wound healing by promoting macrophage polarization and angiogenesis in a full-thickness skin defect model in type I diabetic rats and normal rats. Taken together, this study may provide the ingenious and simple bioactive wound dressing with a superior ability to promote wound healing.

Correlation analysis of sleep behavior and overweight and obesity in junior high school students / 中国学校卫生

Objective@#To analyze the association between different sleep behaviors and overweight and obesity of junior high school students in Yangzhou City, and to provide a basis for policies and interventions related to adolescent health management.@*Methods@#A total of 1 589 students in grades 7-9 from two middle schools in Yangzhou City were selected using the cluster sampling method and were administered with sleep time, bedtime, social jetlag difference, and sleep habits.@*Results@#Totally 64.38% were sleep deprived during the school days, 86.78% went to bed too late, 46.51% had a social jetlag of ≥1 h, and 37.44% took a nap every day(Incluldes holidays and school days). Social jetlag length was statistically different between grades( F =6.97, P < 0.01 ). Girls[(0.95±0.65)h] shown significantly higher social jetlag than the boys[(0.76±0.59)h]( t=6.19, P <0.01). Later bedtime on weekends, later wake up time on weekends and poor sleep behavior were risk factors for overweight and obesity in junior high school students( OR=1.20, 1.14, 1.04, P <0.05).@*Conclusion@#Junior high school students had less sleep and later bedtimes with the increase of grade, and weekend bedtimes,wake up times and poor sleep behavior were independently associated with the risk of overweight and obesity in junior high school students. Parents and schools should be instructed to pay attention to their sleep health and carry out adolescent sleep health guidance.

Generation of two induced pluripotent stem cell lines, GZHMCi009-A and GZHMCi010-A, derived from peripheral blood mononuclear cells of two SCA3 patients with 14/74 CAG repeats of the ATXN3 mutation.

Spinal cerebellar ataxia type 3 (SCA3), also known as Machado-Joseph disease, is the result of abnormal repeat amplification of CAG of the ATXN3 gene. It is one of the main types of autosomal dominant ataxia, with motor symptoms of cerebellar ataxia, mainly accompanied by non-motor symptoms, such as ocular symptoms, psychiatric symptoms, and nutritional disorders. Currently, no effective treatment is available for patients with SCA3. The construction of induced pluripotent stem cells (iPSCs) from two SCA3 patients (14/74 CAG repeats) will be an excellent tool for studying SCA3 disease mechanisms and for drug screening.

Recent progress on dissolution and absorption of amorphous solid dispersions / 药学学报

The amorphous solid dispersion is one of the most effective formulation approaches to enhance the oral bioavailability of poorly water-soluble drugs. However, the amorphous drugs tend to crystallize during storage or dissolution due to inadequate formulations, preparation techniques, storage and dissolution conditions, thus negating their advantages. Meanwhile, it is often difficult to establish in vitro-in vivo correlation for amorphous solid dispersions owing to the difference between dissolution media and physiological environments and between the apparent concentration and membrane transport flux, the dynamic process of the in vivo absorption, which put great challenges to the development of amorphous solid dispersion products. This review covers the recent progress on the mechanistic study of the in vitro dissolution and in vivo absorption of amorphous solid dispersions, aiming to provide guidance for the formulation development of poorly soluble drugs.

Combining ventilation efficiency and peak systolic blood pressure in prognostic assessment of patients with chronic heart failure / 中华全科医师杂志

Objective:To analyze the value of minute ventilation to carbon dioxide production slope (VE/VCO 2 slope) combined with peak systolic blood pressure (SBP) in predicting prognosis for patients with chronic heart failure (CHF). Methods:A total of 170 patients with CHF who visited the Cardiac Rehabilitation Center of Tongji Hospital Affiliated to Tongji University and completed cardiopulmonary exercise test from March 2007 to December 2018 were enrolled in the study. The clinical data, cardiopulmonary exercise testing results and follow-up information of patients were collected to explore the predictors of all-cause mortality in patients with CHF.Results:The median follow-up time was 647 (182-1 764) days. All-cause death occurred in 34 patients. Compared with surviving patients, the proportion of diabetes and angiotensin-converting enzyme inhibitor/angiotensin Ⅱ receptor blocker (ACEI/ARB) use in fatal patients was significantly higher ( P<0.01). The VE/VCO 2 slope and peak SBP*VE/VCO 2 in the fatal patients were significantly higher, and the peak oxygen consumption (peak VO 2) was lower than those in the surviving patients ( P<0.01). The areas under the receiver operating characteristic curve (AUC) of VE/VCO 2 slope and peak SBP*VE/VCO 2 in predicting all-cause mortality in patients with CHF were 0.648 ( P=0.008) and 0.681 ( P=0.001), respectively; the optimal thresholds were >40.95 ( P=0.008) and > 5 423.50 mmHg (1 mmHg=0.133 kPa, P=0.006), the sensitivity was 0.559 and 0.588, and the specificity was 0.728 and 0.735, respectively. Multivariate Cox regression analysis showed that after adjusting for age, gender, diabetes and ACEI/ARB use, VE/VCO 2 slope ( HR=2.12, P=0.036) and peak SBP*VE/VCO 2 ( HR=2.42, P=0.016) were independent risk factors for all-cause mortality in patients with CHF. Conclusion:Compared to the traditional index VE/VCO 2 slope, a novel index peak SBP* VE/VCO 2 provides a relatively better predictive value for all-cause death of CHF patients.

Elimination of hepatitis B virus infection in children: experience and challenge in China / 中华医学杂志(英文版)

Chronic hepatitis B virus (HBV) infection is a serious health issue because of its severe sequelae. Prevention of mother-to-child transmission (MTCT) of HBV is critical to eliminate chronic HBV infection. Here, we reviewed the progress toward the elimination of HBV infection in children in China in the recent decade. A universal hepatitis B vaccination program started from 2002 has been intensified, with the coverage of timely birth dose >95% of all newborn infants from 2012. Since 2011, China has taken a nationwide program to administer hepatitis B immunoglobulin (HBIG) with free of charge in all neonates of HBV-infected mothers, leading to a significant increment of timely use of HBIG. The prevalence of hepatitis B surface antigen (HBsAg) was declined from around 10% among children in 1980s to 2 × 105 U/mL during the third trimester is increasing, which will further reduce MTCT of HBV. However, there are some challenges in the elimination of HBV infection in children, which need to overcome by the concerted efforts. Nevertheless, it is anticipated that China will achieve the goal set by the World Health Organization that the prevalence of HBsAg in children aged <5 years is ≤0.1% by 2030.

Effect of mild moxibustion on cancer-related fatigue, serum ghrelin and adiponectin in patients undergoing chemotherapy after colorectal cancer surgery / 针灸推拿医学（英文版）

Objective: To observe the effect of mild moxibustion on cancer-related fatigue, serum ghrelin and adiponectin in patients undergoing chemotherapy after colorectal cancer surgery. Methods: Seventy patients were divided into a control group and an observation group according to the random number table method, with 35 cases in each group. Patients in both groups were treated with conventional FOLFOX4 chemotherapy regimen. Patients in the control group also received routine treatments such as symptomatic treatment of complications, health education, nutritional support, and exercise intervention. Patients in the observation group received mild moxibustion on the basis of interventions used in the control group. The Piper fatigue scale-revision (PFS-R), Karnofsky performance status (KPS) and spleen and kidney yang deficiency syndrome were scored, and serum ghrelin and adiponectin levels were measured before and after treatment. Results: After treatment, total score of PFS-R in the control group did not change significantly (P>0.05), while in the observation group it was significantly reduced (P<0.05) and lower than that in the control group (P<0.05); KPS scores in both groups were increased (both P<0.05), and it was significantly higher in the observation group than in the control group (P<0.05); total score of spleen and kidney yang deficiency syndrome in the control group did not change significantly (P>0.05), while it was significantly reduced in the observation group (P<0.05) and lower than that in the control group (P<0.05); serum ghrelin and adiponectin levels in the control group had no significant changes (both P>0.05), but the levels were significantly increased in the observation group (both P<0.05) and higher than those in the control group (both P<0.05). Conclusion: Based on the routine treatments, mild moxibustion can relieve fatigue, improve quality of life, and improve the symptoms of spleen and kidney yang deficiency in patients undergoing chemotherapy after colorectal cancer surgery, which may be related to the regulation of serum ghrelin and adiponectin levels.

Improvement Effects of Weijing Decoction on AECOPD Model Rats and Its Mechanism / 中国药房

OBJECTIVE：To study the improvement effects of Weijing deco ction on AECOPD model rats and its possibile mechanism. METHODS ：Totally 55 male SD rats were randomly divided into normal group ，model group ，Weijing decoction low-dose and high-dose groups （8.37，16.74 g/kg，by crude drug ），dexamethasone group （positive control group ，0.09 mg/kg），with 11 rats in each group. Except for normal group ，AECOPD model was induced by cigarettes combined with lipopolysaccharide in other groups. After modeling ，normal group and model group were given constant volume of water intragastrically ，and other groups were given relevant medicine intragastrocally ，twice a day ，for 14 days. After last intragastric administration ，the serum level of IL- 1 β was determined，and pathological changes of lung tissue and bronchus were observed in each group ；mRNA expression of MMP-9 and TIMP- 1 genes in lung tissue were detected ；protein expression of Ras homologous gene family member （RhoA）， dishevelled associated activator of morphogenesis- 1（DAAM1）and hyperplasic suppress gene （HSG）in lung tissue were also determined. RESULTS ：Compared with normal group ，the levels of IL- 1β in serum，mRNA expression of MMP- 9 and TIMP-1 as well as protein expression of RhoA and DAAM 1 in lung tissue were increased significantly in the model group（P＜0.05），while protein expression of HSG in lung tissue was decreased significantly （P＜0.05）；there were many chronic inflammatory cells infiltrating around the bronchus ，some airway mucosa epithelium exfoliating ，alveolar compensatory dilation，pulmonary septal capillary dilation and hyperemia. Compared with model group ，the levels of IL- 1β in serum，mRNA expression of MMP- 9 and TIMP- 1 in lung tissue were decreased significantly in Weijing decoction high-dose group （P＜0.05）；the protein expression of RhoA and DAAM 1 in lung tissue were decreased significantly in Weijing decoction low-dose and high-dose groups（P＜0.05），while the protein expression of HSG in lung tissue was increased （P＜0.05）；the pathological changes of Weijing decoction high-dose group ，such as inflammatory cells infiltrating around the bronchus and shedding of airway mucosa ， were improved significantly ， and there was complete alveolar epithelium structure but no obvious pulmonary dilation. CONCLUSIONS：Weijing decoction can improve AECOPD model rats to certain extent ；its mechanism may be associated with down-regulating mRNA expression of MMP- 9 and TIMP -1 as well as protein expression of RhoA and DAAM 1 in lung tissue ， up-regulating protein expression of HSG in lung tissue so as to inhibit the airway remodeling.

Association of TLR4 polymorphisms with primary open angle glaucoma in Chinese population / 中华实验眼科杂志

Objective:To detect whether Toll-like receptor 4 ( TLR4) polymorphisms contributed to primary open angle glaucoma (POAG) in a Chinese population. Methods:A Chinese cohort, including 799 unrelated POAG patients and 799 unrelated controls, was enrolled in our case-control association study. The data was collected at Sichuan Provincial People's Hospital from May 2014 to March 2018. TLR4 functional single nucleotide polymorphisms (SNPs), including rs4986790 and rs4986791, were genotyped by SNaPshot method. Genotype and allele frequencies of the two SNPs were evaluated. This study was approved by the Institutional Review Boards of the Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital (No.2016-58), and complied with the guidelines of the Declaration of Helsinki. Written informed consents were obtained from all subjects prior to the study. Results:Allelic association analysis revealed that there were no significant association detected in the allelic distributions between the POAG cases and controls for SNPs rs4986790 ( P=0.317) and rs4986791 ( OR=1.000, 95% CI =0.062 5-16.002 2, P=1.000) in the TLR4 gene. Conditional analysis of the two SNPs did not show any significant difference in genotype and allele frequency between the case and the control groups. No association of the two SNPs with POAG was detected under four different genetic models, including homozygote, heterozygote, dominant and recessive models. Conclusions:Polymorphisms rs4986790 and rs4986791 in the TLR4 gene are not related to POAG in the Chinese cohort.

An investigation into the mechanisms underlying the regulatory effect of the E2F6 transcription factor on proliferation and metastasis of malignant melanoma cells through β-catenin signaling pathway / 中华皮肤科杂志

Objective:To determine the expression of the E2F6 transcription factor in human malignant melanoma tissues and cell lines, and to evaluate the effect of E2F6 on proliferation, migration and invasion of a malignant melanoma cell line A375.Methods:Frozen tissues and paraffin-embedded tissue sections were collected from 50 cases of cutaneous malignant melanoma and 30 cases of pigmented nevus in Department of Dermatology, Chongqing Traditional Chinese Medicine Hospital from January 2012 to December 2017. Quantitative reverse transcription-PCR (qRT-PCR) was performed to determine the mRNA expression of E2F6 in the malignant melanoma and pigmented nevus tissues, as well as in 7 malignant melanoma cell lines (HM, A375, WM451, WM35, SK-MEL-1, Hs-695T and MDA-MB-435s) and pigmented nevus cells, and immunohistochemical study and Western blot analysis were conducted to determine the protein expression of E2F6 and β-catenin in the malignant melanoma tissues. An E2F6-inhibiting plasmid and a control plasmid were separately transfected into A375 cells by using a liposome-mediated transfection method, and the E2F6 gene-knockdown efficiency was verified by qRT-PCR and Western blot analysis. Cell counting kit-8 (CCK8) assay, soft-agar plate cloning assay, Transwell migration and invasion assays and 3D cell culture assay were conducted to evaluate the effect of E2F6 gene knockdown on the proliferation, migration and invasion of A375 cells, flow cytometry was performed to detect the cell cycle and apoptosis rate, and Western blot analysis was conducted to determine the protein expression of total β-catenin, activated β-catenin, c-Myc and cyclin D1. The comparison between two groups was carried out by t test, the comparison among several groups by one-way analysis of variance, and multiple comparisons by least significant difference t test; Pearson correlation coefficient was used to analyze the correlation between E2F6 and β-catenin expression in cutaneous malignant melanoma. Results:The E2F6 mRNA expression was significantly higher in the 7 malignant melanoma cell lines than in the pigmented nevus cells (all P < 0.001). qRT-PCR showed that the relative mRNA expression of E2F6 was significantly higher in the cutaneous malignant melanoma tissues (0.000 55 ± 0.000 17) than in the pigmented nevus tissues (0.000 18 ± 0.000 09, t = 3.22, P < 0.001). Both the immunohistochemical study and Western blot analysis showed significantly increased E2F6 protein expression, but decreased β-catenin protein expression in the cutaneous malignant melanoma tissues compared with the pigmented nevus tissues (all P < 0.001). Correlation analysis showed that E2F6 protein expression was negatively correlated with β-catenin expression in the malignant melanoma tissues (immunohistochemical study: r = -0.56, Western blot analysis: r = -0.63, both P < 0.01). After knockdown of the E2F6 gene in A375 cells, the mRNA and protein expression of E2F6 was significantly lower in the E2F6 inhibition group than in the control group ( t = 3.38, 2.76 respectively, both P < 0.001). CCK8 assay showed that the cellular proliferative ability was significantly lower in the E2F6 inhibition group than in the control group ( t = 4.58, P < 0.01) 48 hours after transfection; soft-agar plate cloning assay showed that the colony-formation ratio was significantly lower in the E2F6 inhibition group than in the control group ( t = 2.26, P < 0.001) ; Transwell migration and invasion assays showed that the number of cells crossing the chamber was significantly lower in the E2F6 inhibition group (165 ± 23, 96 ± 11 respectively) than in the control group (376 ± 22, 315 ± 31, t = 3.14, 2.12, respectively, both P < 0.01) ; 3D cell culture assay showed that the cell morphology markedly changed, and the invasive pseudopodia disappeared in the E2F6 inhibition group. Flow cytometry revealed that the proportion of cells at G0-G1 phase and apoptosis rate were significantly higher in the E2F6 inhibition group than in the control group (both P < 0.001). Western blot analysis showed significantly decreased protein expression of β-catenin, activated β-catenin and its downstream target proteins c-Myc and cyclin D1, but significantly increased protein expression of P21 in the E2F6 inhibition group compared with the control group (all P < 0.001) ; additionally, the E2F6 inhibition group showed significantly decreased protein expression of epithelial-mesenchymal transition-related molecules vimentin and N-cadherin, but significantly increased expression of E-cadherin compared with the control group (all P < 0.001) . Conclusions:The E2F6 transcription factor is highly expressed in malignant melanoma. Knockdown of the E2F6 gene in A375 cells can inhibit cell proliferation, migration and invasion by antagonizing the β-catenin signaling pathway.

GABRP regulates chemokine signalling, macrophage recruitment and tumour progression in pancreatic cancer through tuning KCNN4-mediated Ca2+ signalling in a GABA-independent manner.

BACKGROUND AND AIMS: Pancreatic ductal adenocarcinoma (PDAC) is a leading cause of cancer-related death worldwide. Neurotransmitter-initiated signalling pathway is profoundly implicated in tumour initiation and progression. Here, we investigated whether dysregulated neurotransmitter receptors play a role during pancreatic tumourigenesis. METHODS: The Cancer Genome Atlas and Gene Expression Omnibus datasets were used to identify differentially expressed neurotransmitter receptors. The expression pattern of gamma-aminobutyric acid type A receptor pi subunit (GABRP) in human and mouse PDAC tissues and cells was studied by immunohistochemistry and western blot analysis. The in vivo implications of GABRP in PDAC were tested by subcutaneous xenograft model and lung metastasis model. Bioinformatics analysis, transwell experiment and orthotopic xenograft model were used to identify the in vitro and in vivo effects of GABRP on macrophages in PDAC. ELISA, co-immunoprecipitation, proximity ligation assay, electrophysiology, promoter luciferase activity and quantitative real-time PCR analyses were used to identify molecular mechanism. RESULTS: GABRP expression was remarkably increased in PDAC tissues and associated with poor prognosis, contributed to tumour growth and metastasis. GABRP was correlated with macrophage infiltration in PDAC and pharmacological deletion of macrophages largely abrogated the oncogenic functions of GABRP in PDAC. Mechanistically, GABRP interacted with KCNN4 to induce Ca2+ entry, which leads to activation of nuclear factor κB signalling and ultimately facilitates macrophage infiltration by inducing CXCL5 and CCL20 expression. CONCLUSIONS: Overexpressed GABRP exhibits an immunomodulatory role in PDAC in a neurotransmitter-independent manner. Targeting GABRP or its interaction partner KCNN4 may be an effective therapeutic strategy for PDAC.

Epidemiology of allergic rhinitis in children / 中国实用儿科杂志

Allergic rhinitis is one of the most common chronic disease in children. According to the global epidemiological investigation,the incidence of allergic rhinitis in children is different among different ages and different regions,and the prevalence of allergic rhinitis shows slow increase trends all over the world. In addition,the distribution of allergens in different areas is different. In Europe,America and Xinjiang,plants species are the main allergens,while mites are the main allergens in other areas of China.

Spectrometric analyses of larotaxel and larotaxel liposomes quantification by high performance liquid chromatography / 北京大学学报(医学版)

OBJECTIVE@#Larotaxel is a new chemical structure drug, which has not been marketed worldwide. Accordingly, the standard identification and quantification methods for larotaxel remain unclear. The spectrometric analyses were performed for verifying weight molecular formula, molecular weight and chemical structure of larotaxel. Besides, a quantification method was developed for measuring larotaxel in the liposomes.@*METHODS@#The molecular formula, molecular weight and chemical structure of larotaxel were studied by using mass spectrometry (MS), infra-red (IR), nuclear magnetic resonance (NMR) and ultraviolet-visible (UV-vis) spectrometric techniques. The absorption wavelength of larotaxel was investigated by UV-vis spectrophotometry full-wavelength scanning. Besides, a quantification method was developed by high performance liquid chromatography (HPLC), and then validated by measuring the encapsulation efficacy of larotaxel liposomes.@*RESULTS@#The four spectral characteristics of larotaxel were revealed and the corresponding standard spectra were defined. It was confirmed that larotaxel had the structure of tricyclic diterpenoids, with the molecular formula of C45H53NO14, the molecular weight of 831.900 1, and the maximum absorption wavelength of 230 nm. The quantitative method of larotaxel was established by using HPLC with a reversed phase C18 column (5 μm, 250 mm×4.6 mm), a mobile phase of acetonitrile-water (75:25, volume/volume), and a detection wavelength of 230 nm. The validation study exhibited that the established HPLC method was stable, and had a high recovery and precision in the quantitative measurement of larotaxel in liposomes. In addition, a new kind of larotaxel liposomes was also successfully prepared. The particle size of the liposomes was about 105 nm, with an even size distribution. And the encapsulation efficiency of larotaxel in the liposomes was above 80%.@*CONCLUSION@#The present study offers reference standard spectra of larotaxel, including MS, IR, NMR, and UV-vis, and confirms the molecular formula, molecular weight and chemical structure of larotaxel. Besides, the study develops a rapid HPLC method for quality control of larotaxel liposomes.

Neutrophil to lymphocyte ratio predicts the outcomes in patients with acute intracerebral hemorrhage / 国际脑血管病杂志

Objective To investigate the predictive value of neutrophil to lymphocyte ratio (NLR) in peripheral blood for the outcomes in patients with acute intracerebral hemorrhage.Methods Consecutive inpatients with intracerebral hemorrhage diagnosed with the head CT were entolled.The modified Rankin Scale (mRS) was used to evaluate the functional outcomes at 90 d,0-2 wvas defined as good outcome,3-6 were defined as poor outcome,and 6 was defined as death.Univariate analysis was used to compare the demographic characteristics,baseline data,imaging,and laboratory findings between the groups.Multivariate logistic regression analysis was used to determine the independent correlation between NLR and the outcomes,and receiver operating characteristics (ROC) analysis was performed to assess the predictive value of NLR for the outcomes.Results A total of 205 patients with acute intracerebral hemorrhage were enrolled in the study,107 (52.2％) had poor outcome and 57 (27.8％) died.There were significant differences in age (P=0.038),Glasgow Coma Scale (GCS) scores (P=0.001),National Institutes of Health Stroke Scale (NIHSS) scores (P =0.001),neutrophil count (P =0.005),lymphocyte count (P =0.002),NLR (P =0.001),fasting blood glucose (P =0.012),hypersensitivity C-reactive protein (P=0.002),hematoma volume (P =0.005),and proportion of bleeding into the ventricles (P =0.002) between the poor outcome group and the good outcome group.There were significant differences in age (P =0.002),previous stroke (P =0.018),GCS scores (P =0.001),NIHSS scores (P =0.001),neutrophil count (P=0.008),lymphocyte count (P=0.001),NLR (P=0.001),fasting blood glucose (P=0.016),hematoma volume (P=0.001),and proportion of bleeding into ventricle (P=0.002) between the death group and the survival group.Multivariate logistic regression analysis showed that NLR was an independent predictive factor for poor outcome (odds ratio [OR] 2.405,95％ confidence interval [CI] 1.613-3.587;P=0.001) and death (OR 2.268,95％ CI 1.532-3.358;P =0.001) after adjusting for confounders.The ROC curve analysis showed that NLR had a higher predictive value for poor outcome at 90 d (area under the ROC curve 0.703,95％ CI 0.632-0.774;P ＜ 0.001).When the cutoff value was 2.3,the sensitivity and specificity were 61.7％ and 72.4％,respectively.NLR also had a predictive value for death within 90 d (area under the ROC curve 0.706,95％ CI 0.629-0.786;P =0.003).When the cutoff value was 2.2,the sensitivity and specificity were 63.2％ and 72.6％,respectively.Conclusion NLR may have certain predict value for outcomes in patients with acute intracerebral hemorrhage.

Correlation between neutrophil to lymphocyte ratio and cerebral microbleeds in patients with ischemic stroke / 国际脑血管病杂志

ObjectiveTo investigate the correlation between neutrophil to lymphocyte ratio (NLR) and cerebral microbleeds (CMBs) in patients with acute ischemic stroke.MethodsThe consecutive inpatients with acute ischemic stroke were prospectively enrolled.Gradient echo-T2*-weighted imaging was used to evaluate CMBs and their quantity.Univariate analysis was used to compare the baseline data between the CMB group and the non-CMB group.Multivariable logistic regression analysis was used to identify the independent correlation between NLR and CMBs.ResultsA total of 218 patients with acute ischemic stroke were prospectively enrolled, including 66 (30.3%) with CMBs.The age (64.7±6.6 years vs.66.9±8.6 years;t=2.052, P=0.041), high sensitive C-reactive protein (7.0[2.3-13.9] mg/L vs.8.9[4.0-28.1] mg/L;Z=2.008, P=0.045) and NLR (1.9[1.4-2.9] vs.2.3[1.7-3.6];Z=2.071, P=0.038) in the non-CMB group were significantly lower than those of the CMB group.Multivariate logistic regression analysis showed that NLR (odds ratio 1.276, 95% confidence interval 1.008-1.670;P=0.045) and age (odds ratio 1.044, 95% confidence interval 1.002-1.087;P=0.040) were the independent risk factor for CMBs.Spearman correlation analysis showed that NLR was significantly positively correlated with the severity of CMBs (r=0.210, P=0.007).ConclusionsIn patients with acute ischemic stroke, NLR was associated with CMBs and their severity, suggesting that inflammatory reaction might be involved in the occurrence of CMBs.