A near-infrared fluorescent probe for the detection of Cu2+ in Chinese herbal medicine and imaging in living cells.

Copper is one of the common among the heavy metal pollution in Chinese herbal medicine (CHM). So, it is essential to develop rapid and accurate testing method to quantify the Cu2+ content in CHM. Herein, we prepared a coordination-based near-infrared fluorescent probe (NRh6G-FA) by introducing a hemicyanine dye in rhodamine 6G scaffold. NRh6G-FA had a high sensitivity, anti-interference performance, fast response (within 60 s), visualization (from light yellow to green) for Cu2+ and excellent sensing performance for the detection of Cu2+ at low concentrations (LOD = 0.225 µM). The most likely mechanism was verified on the basis of Job's plot, ESI-HRMS and DFT calculations. NRh6G-FA could be successfully applied for the detection and "naked eye" recognition of Cu2+ in CHM samples. Moreover, NRh6G-FA was used to visualize Cu2+ in living MCF-7 cells by confocal fluorescence imaging.

[Distribution Characteristics of Near Surface Ozone Volume Fraction in Shanxi Province Based on Atmospheric Composition Observation Network].

Based on the continuous data of O3， NO， NO2， and NOx and the meteorological data from March 2019 to February 2020 at six atmospheric composition observation stations in Shanxi Province， the characteristics and influence factors of O3 volume fractions were studied using statistical analysis and backward trajectory analysis. The results showed that O3 volume fractions were generally higher from April to September and lower from October to the following March. During the study period， O3 pollution represented by φ（MDA8O3）， i.e.， the maximum daily 8-h average of O3 volume fractions， was the most serious at the Jincheng and Linfen stations in the south of Shanxi， followed by that in the Wutaishan， Shuozhou， and Datong stations in the north， with the least pollution occurring at the Taiyuan station in the middle. There were differences between the urban and alpine stations， although their seasonal O3 volume fractions were both summer > spring > autumn > winter. O3 volume fractions at the urban station were usually lower than those at the alpine station； O3 at the urban station might have been influenced by photochemical reactions with precursor NOx； however， this was not the main source of high O3 at the alpine station. The peak and valley values appeared at 15：00 and 06：00， respectively， at the urban station， whereas they appeared at 20：00 and 10：00， respectively， at the alpine station， representing diametrically opposite diurnal variation patterns. Further， the daily amplitude of O3 at the urban station was much larger than that at the alpine station. For urban stations specifically， temperature was the most important meteorological factor affecting O3 volume fraction， compared with sunlight hours， precipitation， and total cloud cover. The NO2 volume fraction in the daytime affected the daily amplitude of O3； although the photochemical generation potential of O3 at the Taiyuan station was good， the O3 volume fractions were the lowest among urban stations due to strong NO titration. The higher O3 corresponded to lower NOx in which NO2 was dominant， and the higher NOx was largely composed of NO， under which conditions O3 would be depleted completely. The surface wind that affected O3 volume fractions of all stations primarily came from the southeast， south， and southwest， and specific wind speed led to the increase in O3 volume fraction. The geographical situation of the station would cause the difference in the transport of atmospheric pollutants， whereas the horizontal transmissions of high O3 from the North China Plain and Fenwei Plain were likely to be the common reason for the increase in O3 volume fraction in Shanxi.

Influencing factors on the psychosomatic health of medical staff since the normalized stage of COVID-19 in mainland China.

BACKGROUND: During the coronavirus disease 2019 (COVID-19) pandemic, there were under-recognized and unaddressed psychosomatic health problems among medical staff. The purpose of this study was to investigate their psychosomatic status. METHODS: An online questionnaire was performed to the medical staff of major hospitals in Jinan in January 2023. In total, 1244 valid questionnaires were collected, and their psychosomatic status was assessed by the Psychosomatic Symptom Scale (PSSS) and Perceived Stress Scale-10 (PSS-10). Finally, we analyzed the influencing factors for their psychosomatic status. RESULTS: The psychological health of the medical staff was poor, and the level of stress perception was intense, accompanied by obvious psychosomatic symptoms. Regression analysis indicated that age, gender, frontline involvement, work experience, marriage, presence of disease history and COVID-19 infection history were risk factors for psychosomatic symptoms, while education, frontline involvement and presence of disease history were risk factors for stress feeling. CONCLUSION: Medical staff often showed obvious psychosomatic symptoms and intense stress. Psychological health education and intervention should be given in order to improve their working quality.

Novel tetranuclear grid-like Zn(II) complexes derived from dihydrazone pyrimidine derivatives as antitumor agents.

Due to the antitumor properties, Zn(II) complexes have attracted more and more attention. Herein, three novel tetranuclear Zn(II) complexes 1-3 based on dihydrazone pyrimidine derivatives H2L1-H2L3 were synthesized and characterized using IR spectroscopy, 1H NMR spectroscopy, single crystal X-ray diffraction analysis, XRD, TG and elemental analysis. Single crystal X-ray diffraction analysis revealed that 1-3 all displayed a [2 × 2] grid-like topology. The stability in solution, lipophilicity, confocal imaging and antitumor activities were investigated. Complexes 1-3 displayed high structural stability, membrane permeability and different lipophilicities. They can target mitochondria due to the cation charge. The MTT assay indicated that all of them exhibited stronger antiproliferative activity than the corresponding derivatives H2L1-H2L3 and the well-known cisplatin against all the selected tumor cells (BGC-823, BEL-7402, MCF-7 and A549), with IC50 values ranging from 2.83 µM to 7.97 µM. AO/EB double staining, flow cytometry and ROS detection suggested that complexes 1 and 2 could induce BGC-823 apoptosis in a dose-dependent manner. UV-Vis spectra, CD spectra, viscosity analysis and molecular docking revealed that complexes 1 and 2 interact with DNA mainly via partial intercalation and groove binding. Tetranuclear [2 × 2] grid-like Zn(II) complexes have the potential to be promising antitumor agents in the future.

Babesia gibsoni Whole-Genome Sequencing, Assembling, Annotation, and Comparative Analysis.

The intracellular protozoan parasite Babesia gibsoni infects canine erythrocytes and causes babesiosis. The hazards to animal health have increased due to the rise of B. gibsoni infections and medication resistance. However, the lack of high-quality full-genome sequencing sets has expanded the obstacles to the development of pathogeneses, drugs, and vaccines. In this study, the whole genome of B. gibsoni was sequenced, assembled, and annotated. The genomic size of B. gibsoni was 7.94 Mbp in total. Four chromosomes with the size of 0.69 Mb, 2.10 Mb, 2.77 Mb, and 2.38 Mb, respectively, 1 apicoplast (28.4 Kb), and 1 mitochondrion (5.9 Kb) were confirmed. KEGG analysis revealed 2,641 putative proteins enriched on 316 pathways, and GO analysis showed 7,571 annotations of the nuclear genome in total. Synteny analysis showed a high correlation between B. gibsoni and B. bovis. A new divergent point of B. gibsoni occurred around 297.7 million years ago, which was earlier than that of B. bovis, B. ovata, and B. bigemina. Orthology analysis revealed 22 and 32 unique genes compared to several Babesia spp. and apicomplexan species. The metabolic pathways of B.gibsoni were characterized, pointing to a minimal size of the genome. A species-specific secretory protein SA1 and 19 homologous genes were identified. Selected specific proteins, including apetala 2 (AP2) factor, invasion-related proteins BgAMA-1 and BgRON2, and rhoptry function proteins BgWH_04g00700 were predicted, visualized, and modeled. Overall, whole-genome sequencing provided molecular-level support for the diagnosis, prevention, clinical treatment, and further research of B. gibsoni. IMPORTANCE The whole genome of B. gibsoni was first sequenced, annotated, and disclosed. The key part of genome composition, four chromosomes, was comparatively analyzed for the first time. A full-scale phylogeny evolution analysis based on the whole-genome-wide data of B. gibsoni was performed, and a new divergent point on the evolutionary path was revealed. In previous reports, molecular studies were often limited by incomplete genomic data, especially in key areas like life cycle regulation, metabolism, and host-pathogen interaction. With the whole-genome sequencing of B. gibsoni, we provide useful genetic data to encourage the exploration of new terrain and make it feasible to resolve the theoretical and practical problems of babesiosis.

Rare Recurrent EWSR1-PLAGL1 Rearranged Intracranial Tumor With Biphasic Epithelioid Differentiation: One Case Report With Literature Review.

EWSR1-rearranged tumors encompass a rare and heterogeneous group of entities with features of the central nervous system (CNS) mesenchymal and primary glial/neuronal tumors. EWSR1-PLAGL1 gene fusion is a particularly rare form of rearrangement. We presented a recurrent intracranial EWSR1-PLAGL1 rearranged tumor and reviewed the relevant literature. In this case, histopathology and immunohistochemistry (IHC) were evaluated for both the primary and relapsed tumors. Fluorescence in situ hybridization (FISH) and next-generation sequencing (NGS) were performed for the relapsed tumor. We compared the morphology, IHC results and molecular features with the previously reported EWSR1-PLAGL1 rearranged CNS tumors. Our case exhibited a unique feature with a variable biphasic pattern of epithelioid differentiation, which differed from the two reported groups. The primary and relapsed tumors both expressed cytokeratin of the focal area with epithelioid differentiation. The recurrent tumor showed an increased proliferation index (average Ki-67 index of 15%) compared with the primary tumor (average Ki-67 index of 5%). NGS showed that TERT promoter mutation was the only molecular change besides EWSR1-PLAGL1 fusion. Our study provides further insight into intracranial tumors with EWSR1-PLAGL1 fusion, representing a distinct CNS tumor with no-reported histological and immunohistochemical features. Future studies, particularly for the biphasic differentiation and the role of TERT promoter mutation were needed to clarify this unusual chromosomal rearrangement in the CNS tumor.

p53 Missense Mutation is Associated with Immune Cell PD-L1 Expression in Triple-Negative Breast Cancer.

The programmed death ligand 1 (PD-L1) is a pivotal biomarker of immunotherapy in triple negative breast cancer (TNBC). TP53 is reported as a positive regulatory predictor of immune efficacy. The correlation of p53 expression or mutation and PD-L1 expression is explored. By immunohistochemistry, PD-L1 expression between p53 mutation (missense and nonsense) and wild type; p53 no-expression/loss vs. expression were compared. There was a significant association between p53 mutation, especially missense mutation with higher histological grade, and PD-L1 expression in immune cells (ICs). Both p53 missense mutation and PD-L1 expression may be potential targets for improving immunotherapy response in TNBC.

miR-638 Serves as a Biomarker of 5-Fluorouracil Sensitivity to Neoadjuvant Chemotherapy in Breast Cancer.

PURPOSE: Neoadjuvant chemotherapy (NAC) is widely used to treat breast cancer (BC). The prediction and evaluation of chemotherapy responses remains a significant challenge. METHODS: MicroRNAs (miRNAs) play a crucial role in cancer drug resistance. We used a miRNA microarray and identified that miR-638 is downregulated in chemoresistant cases. However, the exact role of miR-638 and the underlying mechanisms of chemoresistance remain unclear. Using real-time quantitative reverse transcription polymerase chain reaction, we found significant downregulation of miR-638 in chemoresistant patients compared with chemosensitive patients. To explore the function of miR-638, we overexpressed and inhibited miR-638 expression in MDA-MB-231 and MCF-7 cells by transfecting them with miR-638 mimics and miR-638 inhibitor, respectively. Cell proliferation and apoptosis were measured using MTS and flow cytometry, respectively. A minimal patient-derived xenograft (MiniPDX™) model was established to evaluate the chemosensitivity to different drugs. RESULTS: The results showed that cell proliferation decreased and cell apoptosis increased in cells transfected with the miR-638 mimic, and cell proliferation and apoptosis were reversed with transfection of miR-638 inhibitor compared with the control group. Among patients who received 5-fluorouracil (5-FU), miR-638 expression levels were lower in the chemoresistant group than in the chemosensitive group. The MiniPDX™ model showed that MDA-MB-231 cells overexpressing miR-638 were more susceptible to 5-FU treatment in vivo. CONCLUSION: We provided evidence of acquired resistance to 5-FU caused by miR-638 deficiency. Alterations in miR-638 may be used with 5-FU chemotherapy during NAC for BC.

Clinicopathological Characteristics and Molecular Phenotypes of Primary Hepatic Lymphoma.

Primary hepatic lymphoma (PHL) is a rare malignant tumor, occurring in 0.016% of non-Hodgkin's lymphoma (NHL). The common histological subtype is diffuse large B-cell lymphoma (DLBCL). Due to the rarity of tumor, clinicopathological characteristics and molecular phenotypes of PHL are limited. Seven patients with PHL (primary liver DLBCL) and 13 cases of liver involvement by DLBCL diagnosed between 2014 and 2021 in our hospital were included. The genetic features were also compared between the two groups by next-generation sequencing (NGS). Differential gene expression and pathway enrichment analysis were also performed. There were some discrepancies on presenting symptoms, pathological characteristics, laboratory data, and prognosis between PHL and DLBCL-liver groups. No same mutation was found between PHL and DLBCL-liver groups by NGS. Differential gene expression analysis discovered some up- and downregulated genes in PHL compared with the DLBCL-liver group. Upregulated genes were enriched in metabolic pathways, and downregulated genes were enriched in the HTLV-1 infection pathway. PHL is a distinct entity, with unique molecular features compared to liver involvement of systemic lymphoma. Kaplan-Meier analysis showed that the prognosis of the PHL group was better than that of the DLBCL-liver group. Understanding the clinicopathological and molecular features of PHL would help to direct clinical treatment.

Appraisal Concerns for the Clinical Research and Development of Drugs for Rare Diseases / 罕见病研究

The incidence of each of the rare disease is very low. The complexity and diagnosis difficulty of the rare disease lead to the difficulties in the clinical research and development (R&D) of drugs for rare diseases. There is an urgent clinical need for the drug development of rare diseases in China. Encouraging R&D of new drugs, particularly the innovative drugs with China's own independent intellectural property is the basis for solving the predicament in drug shortage in China.. In order to further improve the efficiency of clinical R&D of drugs for rare diseases, the National Medical Products Administration (NMPA), Center for Drug Evaluation (CDE) issued Technical Guidance for Clinical Research and Development of Drugs for Rare Diseases. This is the first guidance for rare diseases in China that is drafted from the standpoint of the clinical technology research and development.The guidance is the scientifitc thinking and framework for the drug developing enterprises to research and develop drugs for rare disease efficiently and appropriately by following drug developing protocols and relating to the special features of rare disease.This paper presents the concepts and rationale in the guidance for the appraisal of rare disease drug research and development.

The Risk Stratification for Cervical Cancer and Precursors of Domestic HPV Testing With HPV 16/18 Genotyping in Women With NILM Cytology in CentralChina: A Cohort Study.

OBJECTIVE: To evaluate the clinical performance and utility for risk stratification of DH3 HPV assay in women (≥30 years) with NILM cytology. METHODS: A prospective cohort was established in Central China between November 8 to December 14, 2016 which consisted of 2180 women aging 30-64 years with NILM cytology. At baseline, all women were screened using DH3 HPV assay. HPV 16/18 positive women would be assigned to colposcopy and biopsied if necessary. Then, hr-HPV positive women without CIN2+ lesions would be followed up by cytology every 12 months for two years. In the 3rd year of follow up, all women that were not biopsy proven CIN2+ would be called back and screened by cytology again. In follow-up period, women with ASC-US and above were referred to colposcopy and biopsied if clinically indicated. CIN2+ was the primary endpoint in analysis. The clinical performance and utility for risk stratification of DH3 HPV assay were assessed by SPSS 22.0 and SAS 9.4. RESULTS: Of 2180 qualified women, the prevalence of hr-HPV was 8.5% (185/2180), 45(2.1%) were HPV 16/18 positive. The clinical performance for HPV16/18 was 91.7% for sensitivity, 98.4% for specificity, respectively against CIN2+ detection at baseline. In four years of study, the corresponding rates of HPV 16/18 were 51.5% and 98.7%, respectively. The cumulative absolute risk for the development of CIN2+ was as high as 37.8% for HPV 16/18 positive women, followed by hr-HPV positive (14.6%), other hr-HPV positive (11.0%) and HPV negative (0.3%) in three years. The relative risk was 125.6 and 3.4 for HPV 16/18 positive group when compared with HPV negative and other hr-HPV positive group, respectively. CONCLUSIONS: DH3 HPV assay demonstrated excellent clinical performance against CIN2+ detection in cervical cancer screening and utility of risk stratification by genotyping to promote scientific management of women with NILM cytology.

Identification of miRNA Signature in Breast Cancer to Predict Neoadjuvant Chemotherapy Response.

Background: Chemotherapy failure causes high breast cancer recurrence and poor patient prognosis. Thus, we studied a cohort of novel biomarkers to predict chemotherapeutic response in breast cancer. In this study, miRNA expression profiling was performed on 10 breast cancer punctured specimens sensitive to chemotherapy (MP grade 4, 5) and 10 chemotherapy resistant (MP grade 1). Differentially expressed miRNAs were verified by qRT-PCR in 60 initial samples, 59 validated samples and 71 independent samples. A miRNA signature was generated using a Logistic regression model. A receiver operating characteristic (ROC) test was used to assess specificity and sensitivity of single miRNA and miRNA signature. Target genes regulated by miRNAs and their involved signaling pathways were analyzed using GO enrichment and KEGG software. MiRNAs expression were separately compared with ER, PR, HER2 immunohistochemical staining and different drugs. qRT-PCR showed that the high expression of miR-23a-3p, miR-200c-3p, miR-214-3p and the low expression of miR-451a and miR-638 were closely related to chemoresistance. According to the formula for calculating the drug resistance risk, patients in the high-risk group were more likely to develop chemotherapy resistance than the low-risk group. Bioinformatics analysis showed that 5 miRNAs and target genes are mainly involved in p53, ubiquitin-mediated proteolysis, mTOR, Wnt, cells skeletal protein regulation, cell adhesion and ErbB signaling pathways. miR-451a expression was associated with ER, HER-2 status and anthracyclines. A miRNA signature of chemotherapeutic response may be clinically valuable for improving current chemotherapy regimens of individual treatment for patients with breast cancer.

Hypertension, Arterial Stiffness, and Clinical Outcomes: A Cohort Study of Chinese Community-Based Population.

[Figure: see text].

Elevated ZBTB7A expression in the tumor invasive front correlates with more tumor budding formation in gastric adenocarcinoma.

PURPOSE: Tumor budding (TB) is reported to predict nodal involvement and recurrence in multiple human malignancies. However, it is not clear how TB forms. The purpose of this study is to find markers related to TB formation in gastric cancer and to investigate the underlying mechanisms. METHODS: TB was scored on hematoxylin-eosin staining slides in 122 gastric cancer cases. Immunostaining score of CREB1, GAGE12I, CTNND1, KIF26B and ZBTB7A both at the invasive front and in the center of the tumor were assigned to each case. Spearman's correlation with the TB score was performed to find the TB-related markers. In vitro study and RNA-seq using gastric cancer cell lines were done to unveil the mechanisms. RESULTS: TB could predict lymph node metastasis and is negatively associated with overall survival of the patients. The expression of ZBTB7A in the invasive front, rather than the other four markers, was much higher than that in the tumor center and was positively correlated with TB score. ZBTB7A could enhance migration and invasion of gastric cancer cells in vitro. RNA-seq data followed by RT-qPCR and western blot verification demonstrated the activation of EGFR-MAPK-ERK and PI3K-AKT-mTOR pathways and increased expression of EMT related markers upon ZBTB7A over-expression. CONCLUSION: Higher ZBTB7A expression in the tumor margin may contribute to the dissociation of tumor cells from the tumor mass to form TB by initiating EMT via EGFR-MEK-ERK and PI3K-AKT-mTOR pathway.

CXADR-like membrane protein protects against heart injury by preventing excessive pyroptosis after myocardial infarction.

Myocardial infarction (MI) results in cardiomyocyte death and ultimately leads to heart failure. Pyroptosis is a type of the inflammatory programmed cell death that has been found in various diseased tissues. However, the role of pyroptosis in MI heart remains unknown. Here, we showed that CXADR-like membrane protein (CLMP) was involved in pyroptosis in the mouse MI heart. Our data showed that CLMP was strongly expressed in fibroblasts of the infarcted mouse hearts. The Clmp+/- mice showed more serious myocardial fibrosis and ventricular dysfunction post-MI than wild-type (Clmp+/+ ) mice, indicating a protective effect of the fibroblast-expressed CLMP against MI-induced heart damage. Transcriptome analyses by RNA sequencing indicated that Il-1ß mRNA was significantly increased in the MI heart of Clmp+/- mouse, which indicated a more serious inflammatory response. Meanwhile, cleaved caspase-1 and Gasdermin D were significantly increased in the Clmp+/- MI heart, which demonstrated enhanced pyroptosis in the Clmp knockdown heart. Further analysis revealed that the pyroptosis mainly occurred in cardiac fibroblasts (CFs). Compared to wild-type fibroblasts, Clmp+/- CFs showed more serious pyroptosis and inflammatory after LPS plus nigericin treatment. Collectively, our results indicate that CLMP participates in the pyroptotic and inflammatory response of CFs in MI heart. We have provided a novel pyroptotic insight into the ischaemic heart, which might hold substantial potential for the treatment of MI.

LncRNA-Safe contributes to cardiac fibrosis through Safe-Sfrp2-HuR complex in mouse myocardial infarction.

Rationale: As a hallmark of various heart diseases, cardiac fibrosis ultimately leads to end-stage heart failure. Anti-fibrosis is a potential therapeutic strategy for heart failure. Long noncoding RNAs (lncRNAs) have emerged as critical regulators of heart diseases that promise to serve as therapeutic targets. However, few lncRNAs have been directly implicated in cardiac fibrosis. Methods: The lncRNA expression profiles were assessed by microarray in cardiac fibrotic and remote ventricular tissues in mice with myocardial infarction. The mechanisms and functional significance of lncRNA-AK137033 in cardiac fibrosis were further investigated with both in vitro and in vivo models. Results: We identified 389 differentially expressed lncRNAs in cardiac fibrotic and remote ventricular tissues in mice with myocardial infarction. Among them, a lncRNA (AK137033) we named Safe was enriched in the nuclei of fibroblasts, and elevated in both myocardial infarction and TGF-ß-induced cardiac fibrosis. Knockdown of Safe prevented TGF-ß-induced fibroblast-myofibroblast transition, aberrant cell proliferation and secretion of extracellular matrix proteins in vitro, and mended the impaired cardiac function in mice suffering myocardial infarction. In vitro studies indicated that knockdown of Safe significantly inhibited the expression of its neighboring gene Sfrp2, and vice versa. The Sfrp2 overexpression obviously disturbed the regulatory effects of Safe shRNAs in both the in vitro cultured cardiac fibroblasts and myocardial infarction-induced fibrosis. Dual-Luciferase assay demonstrated that Safe and Sfrp2 mRNA stabilized each other via their complementary binding at the 3'-end. RNA electrophoretic mobility shift assay and RNA immunoprecipitation assay indicated that RNA binding protein HuR could bind to Safe-Sfrp2 RNA duplex, whereas the knockdown of HuR dramatically reduced the stabilization of Safe and Sfrp2 mRNAs, down-regulated their expression in cardiac fibroblasts, and thus inhibited TGF-ß-induced fibrosis. The Safe overexpression partially restrained the phenotype change of cardiac fibroblasts induced by Sfrp2 shRNAs, but not that induced by HuR shRNAs. Conclusions: Our study identifies Safe as a critical regulator of cardiac fibrosis, and demonstrates Safe-Sfrp2-HuR complex-mediated Sfrp2 mRNA stability is the underlying mechanism of Safe-regulated cardiac fibrosis. Fibroblast-enriched Safe could represent a novel target for anti-fibrotic therapy in heart diseases.

[miRNA-148b-3p Influences Glucose Metabolism of Offspring with Maternal Cholestasis by Regulating GLUT1 Expression in Placental Trophoblast Cells].

OBJECTIVE: To investigate the expression of miRNA-148b-3p and its target gene in the placenta between normal pregnant women and pregnant women with intrahepatic cholestasis of pregnancy (ICP) and to explore the possible mechanism of glucose metabolism of offspring with maternal cholestasis. METHODS: There were 30 cases of normal pregnant women and 30 cases of pregnant women with ICP recruited in the study, all of whom underwent cesarean delivery from Mar. 2017 to Jan. 2018. Placenta tissues, maternal blood and cord blood were collected in each case. Maternal blood and cord blood were sent for biochemical detection. miRNA of placenta tissues was extracted and qRT-PCR was used to measure the expression of miR-148b-3p in the placenta. Normal HTR-8 cells were transfected with miR-148b-3p inhibitor/mimics wrapped with lipofectaine3000. qRT-PCR was used to measure the expression of miR-148b-3p, and Western blot was used to measure the expression of glucose transporter 1 (GLUT1) after transfection. RESULTS: Maternal fasting blood glucose (FPG) and its fetal cord blood insulin levels in the ICP group were significantly higher than those of control. The expression of miR-148b-3p in the placenta of ICP group was lower than that of control group ( P<0.05). Compared with inhibitor control group, the expression of miR-148b-3p was decreased in HTR-8 cells transfected with miR-148b-3p inhibitor ( P<0.05), while the expression of GLUT1 was increased ( P<0.05). Compared with mimics control group, the expression of miR-148b-3p was increased in HTR-8 cells transfected with miR-148b-3p mimics ( P<0.05), while the expression of GLUT1 was decreased ( P<0.05). CONCLUSION: miR-148b-3p might participate in glucose metabolism of offspring with maternal cholestasis through the negative regulation of GLUT1 expression in placental trophoblast cells.

[Effect of otopoint pellet-pressing combined with medication on clinical symptoms of migraine patients and changes of plasma 5-HT and CGRP contents].

OBJECTIVE: To investigate the short-term and long-term clinical effects of otopoint pellet-pressing combined with medication in the treatment of patients with migraine without aura and its impact on plasma 5-hydroxytryptamine(5-HT) and calcitonin gene-related peptide(CGRP) contents. METHODS: Patients with migraine without aura were randomly divided into medication(control) group(n=48) and otopoint pellet-pressing plus medication (treatment) group(n=49). Patients of the control group were given oral Flunarizine capsules(10 mg/time) twice a day, and those of the treatment group received same dosage of Flunarizine and pellet-pressing of otopoints Nao(Brain), Nie (Temporal), Shenmen(Shenmen), Jiaogan(Sympathy) and Pizhixia(Subcortex), 2 min/point, 3 times a day, simultaneously. The treatment was conducted for 1 month. The short-term and long-term clinical effects were evaluated according to Yang and colleagues' methods, and "Guiding principles for clinical research of new TCM drugs (trial)". The contents of plasma 5-HT and CGRP were detected by ELISA. RESULTS: After one month's treatment, of the 48 and 49 patients in the control and treatment groups, 10(20.83%)and 17(34.69%) were under control, 19(39.59%)and 23(46.94%) experienced marked improvement, 10(20.83%)and 7(14.29%)were effective, 9(18.75%) and 2(4.08%) failed, with the total effective rates being 81.25% and 95.92%, respectively. Six months' follow-up survey showed that of the 48 and 49 patients in the control and treatment groups, 4(8.33%)and 11(22.45%) were under control, 20(41.67%)and 24(48.98%)experienced marked improvement, 11(22.92%)and 9(18.37%)were effective, and 13(27.08%) and 5(10.20%)failed, with the total effective rates being 72.92% and 89.80%, respectively. The number of headache attacks, duration of each attack and the degree of headache were significantly decreased after 1 and 6 months' treatment in both groups in comparison with their own pre-treatment (P<0.05). The contents of plasma 5-HT at the time-points of 1 and 6 months were markedly increased (P<0.05), and those of plasma CGRP at the two time points markedly decreased in both groups in comparison with their own pre-treatment (P<0.05). The therapeutic effects of the treatment group were obviously superior to those of the control group in lowering the number of headache attacks, duration of each attack and the degree of headache and plasma CGRP content, as well as in increasing plasma 5-HT levels after 1 and 6 months' treatment (P<0.05). CONCLUSION: Otopoint pellet-pressing combined with oral administration of Flunarizine can significantly improve the clinical symptoms in patients with migraine without aura, and possess a stable long-term clinical effect, which may be associated with its effect in increasing plasma 5-HT and decreasing CGRP levels.

Human embryonic stem cell-derived cardiomyocyte therapy in mouse permanent ischemia and ischemia-reperfusion models.

BACKGROUND: Ischemic heart diseases are still a threat to human health. Human pluripotent stem cell-based transplantation exhibits great promise in cardiovascular disease therapy, including heart ischemia. The purpose of this study was to compare the efficacy of human embryonic stem cell-derived cardiomyocyte (ESC-CM) therapy in two heart ischemia models, namely, permanent ischemia (PI) and myocardial ischemia reperfusion (IR). METHODS: Human embryonic stem cell-derived cardiomyocytes were differentiated from engineered human embryonic stem cells (ESC-Rep) carrying green fluorescent protein (GFP), herpes simplex virus-1 thymidine kinase (HSVtk), and firefly luciferase (Fluc). Two different heart ischemia models were generated by the ligation of the left anterior descending artery (LAD), and ESC-Rep-derived cardiomyocytes (ESC-Rep-CMs) were transplanted into the mouse hearts. Cardiac function was analyzed to evaluate the outcomes of ESC-Rep-CM transplantation. Bioluminescence signal analysis was performed to assess the cell engraftment. Finally, the inflammation response was analyzed by real-time PCR and ELISA. RESULTS: Cardiac function was significantly improved in the PI group with ESC-Rep-CM injection compared to the PBS-injected control, as indicated by increased left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS), as well as reduced fibrotic area. However, minimal improvement by ESC-Rep-CM injection was detected in the IR mouse model. We observed similar engraftment efficiency between PI and IR groups after ESC-Rep-CM injection. However, the restricted inflammation was observed after the injection of ESC-Rep-CMs in the PI group, but not in the IR group. Transplantation of ESC-Rep-CMs can partially preserve the heart function via regulating the inflammation response in the PI model, while little improvement of cardiac function in the IR model may be due to the less dynamic inflammation response by the mild heart damage. CONCLUSIONS: Our findings identified the anti-inflammatory effect of ESC-CMs as a possible therapeutic mechanism to improve cardiac function in the ischemic heart.