The influence of maternal prepregnancy weight and gestational weight gain on the umbilical cord blood metabolome: a case-control study.

BACKGROUND: Maternal overweight/obesity and excessive gestational weight gain (GWG) are frequently reported to be risk factors for obesity and other metabolic disorders in offspring. Cord blood metabolites provide information on fetal nutritional and metabolic health and could provide an early window of detection of potential health issues among newborns. The aim of the study was to explore the impact of maternal prepregnancy overweight/obesity and excessive GWG on cord blood metabolic profiles. METHODS: A case control study including 33 pairs of mothers with prepregnancy overweight/obesity and their neonates, 30 pairs of mothers with excessive GWG and their neonates, and 32 control mother-neonate pairs. Untargeted metabolomic profiling of umbilical cord blood samples were performed using UHPLC‒MS/MS. RESULTS: Forty-six metabolites exhibited a significant increase and 60 metabolites exhibited a significant reduction in umbilical cord blood from overweight and obese mothers compared with mothers with normal body weight. Steroid hormone biosynthesis and neuroactive ligand‒receptor interactions were the two top-ranking pathways enriched with these metabolites (P = 0.01 and 0.03, respectively). Compared with mothers with normal GWG, in mothers with excessive GWG, the levels of 63 metabolites were increased and those of 46 metabolites were decreased in umbilical cord blood. Biosynthesis of unsaturated fatty acids was the most altered pathway enriched with these metabolites (P < 0.01). CONCLUSIONS: Prepregnancy overweight and obesity affected the fetal steroid hormone biosynthesis pathway, while excessive GWG affected fetal fatty acid metabolism. This emphasizes the importance of preconception weight loss and maintaining an appropriate GWG, which are beneficial for the long-term metabolic health of offspring.

FvMYB108, a MYB Gene from Fragaria vesca, Positively Regulates Cold and Salt Tolerance of Arabidopsis.

MYB (myoblast) protein comes in large quantities and a wide variety of types and plays a role in most eukaryotes in the form of transcription factors (TFs). One of its important functions is to regulate plant responses to various stresses. However, the role of MYB TFs in regulating stress tolerance in strawberries is not yet well understood. Therefore, in order to investigate the response of MYB family members to abiotic stress in strawberries, a new MYB TF gene was cloned from Fragaria vesca (a diploid strawberry) and named FvMYB108 based on its structural characteristics and evolutionary relationships. After a bioinformatics analysis, it was determined that the gene belongs to the R2R3-MYB subfamily, and its conserved domain, phylogenetic relationships, predicted protein structure and physicochemical properties, subcellular localization, etc. were analyzed. After qPCR analysis of the expression level of FvMYB108 in organs, such as the roots, stems, and leaves of strawberries, it was found that this gene is more easily expressed in young leaves and roots. After multiple stress treatments, it was found that the target gene in young leaves and roots is more sensitive to low temperatures and salt stimulation. After these two stress treatments, various physiological and biochemical indicators related to stress in transgenic Arabidopsis showed corresponding changes, indicating that FvMYB108 may be involved in regulating the plant's ability to cope with cold and high-salt stress. Further research has found that the overexpression of this gene can upregulate the expression of AtCBF1, AtCOR47, AtERD10, and AtDREB1A related to low-temperature stress, as well as AtCCA1, AtRD29a, AtP5CS1, and AtSnRK2.4 related to salt stress, enhancing the ability of overexpressed plants to cope with stress.

The association between gestational weight trajectories in women with gestational diabetes and their offspring's weight from birth to 40 months.

AIMS: To identify the gestational weight gain (GWG) patterns in women with gestational diabetes mellitus (GDM) and evaluate their association with offspring weight status from birth to 40 months. MATERIALS AND METHODS: This study included 2,723 GDM-mother-child pairs from the Beijing Birth Cohort Study. The association between GWG trajectories identified by the latent class model and offspring weight outcomes from birth to 40 months were evaluated, after adjustment for maternal age, parity, pre-pregnancy body mass index, maternal height, and blood glucose levels. RESULTS: Three GWG rate groups, including the non-excessive GWG group (1,994/2,732), excessive GWG group (598 /2,732), and excessive early GWG group (140/2,732), were identified in women with GDM, respectively. Compared to the non-excessive GWG group, the adjusted OR (aOR) and 95% CI were 1.83 (1.35-2.47) and 1.79 (1.06-3.01) for macrosomia, 1.33 (1.07-1.66) and 1.48 (1.01-2.17) for large for gestational age (LGA) in the excessive GWG group and excessive early GWG group. Excessive GWG was also associated with an increased risk of BMI-for-age at 40 months (aOR = 1.66, 95% CI 1.14-2.42). CONCLUSIONS: Both excessive GWG and excessive early GWG increased the risk of macrosomia and LGA in women with GDM, but only the excessive GWG was associated with childhood overweight/obesity. The results suggest the long-term impact of GWG on offspring weight status in women with GDM and the potential benefits of GWG restriction after GDM diagnosis.

Associations of gestational weight gain with birth weight outcomes in early pregnancy weight loss women: Findings from the Beijing Birth Cohort Study.

OBJECTIVE: To explore the relationship between gestational weight gain (GWG) and birth weight outcomes and establish suggested GWG patterns in early pregnancy weight loss women. METHODS: This retrospective study was conducted based on the Beijing Birth Cohort Study from 2014 to 2021 and included 20 688 women. Weight change in early pregnancy was calculated using weight measurements within 16 weeks of gestation. Multivariable logistic regression was used to analyze the relationships of different GWG categories, based on the Chinese standard, and birth weight outcomes. The statistical-based approach was used to determine the optimal GWG ranges and weekly weight gain. RESULTS: Compared to 3313 women who gained appropriate weight in early pregnancy, 2614 women who lost weight in the same period increased the risk of small for gestational age (SGA) (OR = 1.43, 95% CI: 1.14-1.80, P = 0.002). However, the relationship disappeared after adjusting for total GWG. Among the early pregnancy weight loss women, both excessive GWG and inadequate GWG were associated with adverse birth weight outcomes after adjusting for confounders. The suggested GWG range and rate were 11.0 ~ 16.0 kg and 0.46 to 0.67 kg/week from 16 weeks to delivery for women with normal body mass index (BMI) and weight loss in early pregnancy. CONCLUSION: Weight loss in early pregnancy was not the independent risk factor of birth weight outcomes. GWG may offset the expected effects. To achieve optimal outcomes, women with normal BMI and weight loss in early pregnancy need to have a higher weight gain rate in mid-late pregnancy but similar total GWG ranges with the Chinese standard for general women.

PTX3 promotes IVIG resistance-induced endothelial injury in Kawasaki disease by regulating the NF-κB pathway.

Intravenous immunoglobulin (IVIG) resistance leads to serious complications in Kawasaki disease (KD) with no effective treatment. This study aimed to investigate the effects of pentraxin 3 (PTX3) on human coronary artery endothelial cells (HCAECs). PTX3 levels were measured using quantitative real-time PCR (qRT-PCR), enzyme-linked immunosorbent assay, and western blotting. Cell viability was detected using the MTT assay. Biological functions were analyzed using CCK-8, EdU, flow cytometry, TUNEL, and qRT-PCR. The levels of factors of the NF-κB pathway were examined using western blotting. The results demonstrated that PTX3 expression was highest in patients and HCAECs with IVIG-resistance. Knockdown of PTX3 promoted proliferation and suppressed apoptosis and inflammation of IVIG-resistant HCAECs, whereas PTX3 overexpression produced the opposite results. Moreover, PTX3 activated the NF-κB pathway in IVIG-resistant HCAECs. A rescue study showed that PTX3 modulated biological behaviors by regulating the NF-κB pathway. Overall, our findings demonstrate that PTX3 promotes IVIG resistance-induced endothelial injury by activating the NF-κB pathway, suggesting that PTX3 may become a novel therapeutic target for patients with IVIG-resistant KD.

Restoration of saturated outputs from microchannel plate photomultiplier tubes in sub-microsecond single-pulse-current mode.

Microchannel plate (MCP) photomultiplier tubes (PMTs) are frequently used in experimental diagnostics, where they are operated in single-pulse current measurement mode. However, considering the significant amplitude fluctuations in the measured signal, the resulting output signal from the MCP-PMT is inevitably distorted by gain saturation. Therefore, understanding the correlation between the MCP-PMT output signal and gain saturation is critical in assessing the extent of output signal distortion and determining the MCP-PMT saturation level. This knowledge allows for a more precise assessment of the input signal's features. In this paper, we present an experimental method for restoring the initial waveform from the saturated MCP-PMT signal. To correct the amplitude-drop caused by gain saturation, our technique involves calibrating the MCP-PMT's relative gain as a function of the accumulated output charge using a square-wave light source. We then applied this approach to restore a ∼500 ns saturated pulse from a double-layer 10 mm diameter MCP-PMT. The restored signal showed a deviation of less than 6% from the reference waveform, which validates the effectiveness of the technique.

Dynamic changes in blood lipid levels and their associations with hypertensive disorders of pregnancy in twin pregnancy: A retrospective study.

BACKGROUND: Little knowledge on the association of blood lipid levels with hypertensive disorders of pregnancy (HDP) in twin pregnancy. OBJECTIVE: To investigate the association of blood lipid levels with HDP in twin pregnancy. METHODS: This is a retrospective study in the Beijing Birth Cohort on patients followed between January 2014 and November 2021. A total of 2628 women pregnant with twins were included and divided into HDP (n = 565) and normal blood pressure (NBP, n = 2063) groups. HDP subtypes included gestational hypertension (GH, n = 190) and preeclampsia (PE, n = 375). Dynamic changes in blood lipid profiles and their associations with HDP were assessed. RESULTS: Compared to NBP group, higher triglyceride (TG) and low-density lipoprotein  cholesterol (LDL-c) in the first (T1) and second trimesters (T2) existed in women with PE. In addition, TG increased significantly from T1 to T2, and high-density lipoprotein cholesterol (HDL-c) decreased significantly since T2 in women with PE, especially in women with early-onset PE and severe PE. Elevated TG and LDL-c were associated with HDP, mainly PE. In a subgroup analysis, higher TG or LDL-c increased the risk of HDP for underweight, overweight and primipara women. CONCLUSIONS: In twin pregnancy, women with PE had higher TG and LDL-c, and elevated TG and LDL-c were associated with PE. A significant increase in TG or decrease in HDL-c were more prone to PE, especially early-onset PE and severe PE. It is helpful to monitor blood lipid levels in women pregnant with twins, especially in underweight, overweight, and primipara women.

Association between serum lipid profile during the first and second trimester of pregnancy as well as their dynamic changes and gestational diabetes mellitus in twin pregnancies: a retrospective cohort study.

BACKGROUND: Abnormal lipid metabolism is associated with gestational diabetes mellitus (GDM) in singleton pregnancies. Data were lacking on twin pregnancies with GDM. We explored the association between serum lipid profiles in the first and second trimesters as well as their dynamic changes and GDM in twin pregnancies. METHODS: This was a retrospective cohort study of 2739 twin pregnancies that underwent a 75-g oral glucose tolerance test (OGTT) and were selected from the Beijing Birth Cohort Study from June 2013 to May 2021. Cholesterol (CHO), triglyceride (TG), high-density lipoprotein (HDL) and low-density lipoprotein (LDL) levels were measured at mean 9 and 25 weeks of gestation. We described maternal lipid levels in different tertiles that were associated with the risk of GDM stratified for age, pre-BMI, and fertilization type. GDM patients were divided into two groups according to OGTT: elevated fasting plasma glucose only (FPG group) and the rest of the GDM (non-FPG group). We estimated the relative risk of GDM with multivariable logistic regression models. RESULTS: In this study, we found that 599 (21.9%, 599/2739) twin pregnancies developed GDM. They had increased CHO, TG, LDL, and LDL/HDL, decreased HDL levels in the first trimester, and increased TG as well as decreased HDL in the second trimester in univariate analyses, each P < 0.05. In multivariate analysis, when TG > 1.67 mmol/l (upper tertile) in elderly individuals, nonoverweight and ART groups increased the risk of GDM by 2.7-fold, 2.3-fold and 2.2-fold, respectively, compared with TG < 0.96 mmol/l (lower tertile). This effect remained in the abovementioned groups in the second trimester. Moreover, high TGs increased the risk of GDM in the FPG group (OR = 2.076, 95% CI 1.130-3.815) and non-FPG group (OR = 2.526, 95% CI 1.739-3.67) in the first trimester when TG > 1.67 mmol/l, and the rising risk in the non-FPG group as the TG tertile increased remained in the second trimester. HDL predominantly showed a negative association with elevated FPG in the second trimester (p < 0.05). CONCLUSIONS: Twin pregnancies with GDM have higher lipid levels. Increased TGs in the first and second trimesters are strongly associated with GDM, especially in elderly individuals, nonoverweight and ART groups. Lipid profiles varied among different GDM subtypes.

Association of Maternal Plasma Manganese with the Risk of Spontaneous Preterm Birth: A Nested Case-Control Study Based on the Beijing Birth Cohort Study (BBCS) in China.

We performed this study to clarify the dynamic changes in maternal manganese (Mn) concentration during pregnancy and its association with spontaneous preterm birth (SPB). A nested case-control study was conducted based on the Beijing Birth Cohort Study (BBCS) from 2018 to 2020. Singleton pregnancy women aged 18-44 (n = 488) were involved in the study, including 244 cases of SPB and 244 controls. All of the participants provided blood samples twice (in their first and third trimesters). Inductively coupled plasma mass spectrometry (ICP-MS) was used for the laboratory analysis, and unconditional logistic regression was used for the statistical analysis. We found that the maternal Mn levels were significantly higher in the third trimester than those in the first trimester (median: 1.23 vs. 0.81 ng/mL). The SPB risk was increased to 1.65 (95% CI: 1.04-2.62, p = 0.035) in the highest Mn level (third tertile) in the third trimester, especially in normal-weight women (OR: 2.07, 95% CI: 1.18-3.61, p = 0.011) or non-premature rupture of membrane (PROM) women (OR: 3.93, 95% CI: 2.00-7.74, p < 0.001). Moreover, a dose-dependent relationship exists between the SPB risk and maternal Mn concentration in non-PROM women (P trend < 0.001). In conclusion, dynamic monitoring of maternal Mn level during pregnancy would be helpful for SPB prevention, especially in normal-weight and non-PROM women.

Mutagenesis of the di-leucine motif in the cytoplasmic tail of newcastle disease virus fusion protein modulates the viral fusion ability and pathogenesis.

BACKGROUND: Newcastle disease virus (NDV) is a highly infectious viral disease, which can affect chickens and many other kinds of birds. The main virulence factor of NDV, the fusion (F) protein, is located on the viral envelope and plays a major role in the virus' ability to penetrate cells and cause host cell fusion during infection. Multiple highly conserved tyrosine and di-leucine (LL) motifs in the cytoplasmic tail (CT) of the virus may contribute to F protein functionality in the viral life cycle. METHODS: To examine the contribution of the LL motif in the biosynthesis, transport, and function of the F protein, we constructed and rescued a NDV mutant strain, rSG10*-F/L537A, with an L537A mutation using a reverse genetic system. Subsequently, we compared the differences in the syncytium formation ability, pathogenicity, and replication levels of wild-type rSG10* and the mutated strain. RESULTS: Compared with rSG10*, rSG10*-F/L537A had attenuated syncytial formation and pathogenicity, caused by a viral budding defect. Further studies showed that the LL-motif mutation did not affect the replication, transcription, or translation of the virus genome but affected the expression of the F protein at the cell surface. CONCLUSIONS: We concluded that the LL motif in the NDV F CT affected the regulation of F protein expression at the cell surface, thus modulating the viral fusion ability and pathogenic phenotype.

The triglyceride glucose index and CDKAL1 gene rs10946398 SNP are associated with NAFLD in Chinese adults.

BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) has a higher prevalence worldwide, and its pathogenesis is not clear. Genetic factors, dyslipidemia and dysglycemia have been proven to be associated with NAFLD. It has not been reported whether the triglyceride glucose index (TyG), which is estimated by triglyceride and fasting glucose, has a relationship with NAFLD in people from North China. Whether the CDKAL1 gene rs10946398 SNP, which has been found to be associated with BMI, has a relationship with TyG and NAFLD is not clear. METHODS: We recruited a total of 1760 subjects in this study, and we measured the clinical characteristics, abdominal ultrasound, and genotype of those participants. RESULTS: The results showed that 527 (29.9%) subjects suffered from NAFLD, the TyG index was associated with NAFLD (OR=5.456, 95% CI [3.526~8.442]), and the CDKAL1 gene rs10946398 SNP has a relationship with NAFLD (OR=1.509, 95% CI [1.046~2.178]). The distribution of the C allele of rs10946398 was statistically significant at different levels of the TyG index. CONCLUSIONS: We identified an association between the rs10946398 genotypes of CDKAL1 and NAFLD and the TyG index, and the TyG index was related to the risk of NAFLD.

MiR-130/SNAP-25 axis regulate presynaptic alteration in anterior cingulate cortex involved in lead induced attention deficits.

Brain volume decrease in the anterior cingulate cortex (ACC) after lead (Pb) exposure has been linked to persistent impairment of attention behavior. However, the precise structural change and molecular mechanism for the Pb-induced ACC alteration and its contribution to inattention have yet to be fully characterized. The present study determined the role of miRNA regulated synaptic structural and functional impairment in the ACC and its relationship to attention deficit disorder in Pb exposed mice. Results showed that Pb exposure induced presynaptic impairment and structural alterations in the ACC. Furthermore, we screened for critical miRNA targets responsible for the synaptic alteration. We found that miR-130, which regulates presynaptic vesicle releasing protein SNAP-25, was responsible for the presynaptic impairment in the ACC and attention deficits in mice. Blocking miR-130 function reversed the Pb-induced decrease in the expression of its presynaptic target SNAP-25, leading to the redistribution of presynaptic vesicles, as well as improved presynaptic function and attention in Pb exposed mice. We report, for the first time, that miR-130 regulating SNAP-25 mediates Pb-induced presynaptic structural and functional impairment in the ACC along with attention deficit disorder in mice.

Maternal and infant outcomes in women with and without gestational diabetes mellitus in the COVID-19 era in China: Lessons learned.

Aims: The global COVID-19 pandemic has required a drastic transformation of prenatal care services. Whether the reformulation of the antenatal care systems affects maternal and infant outcomes remains unknown. Particularly, women with gestational diabetes mellitus (GDM) are among those who bear the greatest brunt. Thus, this study aimed to evaluate the impact of COVID-19 lockdown during late pregnancy on maternal and infant outcomes in women stratified by the GDM status in China. Study design: The participants were women who experienced the COVID-19 lockdown during late pregnancy (3185 in the 2020 cohort) or not (2540 in the 2019 cohort) that were derived from the Beijing Birth Cohort Study. Maternal metabolic indicators, neonatal outcomes, and infant anthropometrics at 12 months of age were compared between the two cohorts, stratified by the GDM status. Results: Participants who experienced COVID-19 lockdown in late pregnancy showed lower gestational weight gain than those in the control cohort. Nevertheless, they displayed a worse metabolic profile. COVID-19 lockdown during pregnancy was associated with higher glycosylated hemoglobin (HbA1c) (ß= 0.11, 95% CI = 0.05-0.16, q-value = 0.002) and lower high density lipoprotein cholesterol level (HDL-C) level (ß=-0.09, 95% CI = -0.14 to -0.04, q-value = 0.004) in women with GDM, adjusted for potential confounders. In normoglycemic women, COVID-19 lockdown in late pregnancy was associated with higher fasting glucose level (ß= 0.10, 95% CI = 0.08-0.12, q-value <0.0001), lower HDL-C level (ß=-0.07, 95% CI = -0.08 to -0.04, q-value <0.0001), and increased risk of pregnancy-induced hypertension (adjusted OR=1.80, 95%CI=1.30-2.50, q-value=0.001). The fasting glucose level decreased less from early to late pregnancy in women who experienced COVID-19 lockdown than in the controls, regardless of the GDM status. The HDL-C has risen less with COVID-19 lockdown in the normoglycemic subgroup. In contrast, no significant differences regarding neonatal outcomes or infant weight were found between the two cohorts. Conclusion: Experiencing the COVID-19 lockdown in pregnancy was associated with worse maternal metabolic status but similar neonatal outcomes and infant weight.

Designed, synthesized and biological evaluation of proteolysis targeting chimeras (PROTACs) as AR degraders for prostate cancer treatment.

As a continuation of our research on developing potent and potentially safe androgen receptor (AR) degrader, a series of novel proteolysis targeting chimeras (PROTACs) containing the phthalimide degrons with different linker were designed, synthesized and evaluated for their AR degradation activity against LNCaP (AR+) cell line. Most of the synthesized compounds displayed moderate to satisfactory AR binding affinity and might lead to antagonist activity against AR. Among them, compound A16 exhibited the best AR binding affinity (85%) and degradation activity against AR. Due to the strong fluorescence properties of pomalidomide derivatives, B10 was found to be effectively internalized and visualized in LNCaP (AR + ) cells than PC-3 (AR-) cells. Moreover, the molecular docking of A16 with AR and the active site of DDB1-CRBN E3 ubiquitin ligase complex provides guidance to design new PROTAC degrons targeting AR for prostate cancer therapy. These results represent a step toward the development of novel and improved AR PROTACs.

Non-linear associations of body mass index with impaired fasting glucose, ß-cell dysfunction, and insulin resistance in nondiabetic Chinese individuals: a cross-sectional study.

INTRODUCTION: Identifying and managing patients with prediabetes is important. The study aims to investigate the association of body mass index (BMI) with impaired fasting glucose (IFG), ß-cell dysfunction, and insulin resistance in nondiabetic Chinese individuals. MATERIAL AND METHODS: This was a cross-sectional study of consecutive nondiabetic individuals enrolled between January 2014 and January 2015, divided into NFG [normal fasting glucose, fasting blood glucose (FBG) < 5.6 mmol/L) and IFG (n = 450; FBG ≥ 5.6 mmol/L) groups. Restricted cubic splines and piecewise-regression were used to model the association of IFG, impaired b-cell function, and insulin resistance with BMI. Stratified analyses were performed across sex and age. RESULTS: A total of 900 NFG and 450 IFG individuals were enrolled, with a median age of 41 (30-49) years and 1076 males (79.7%). After adjusting for age and sex, the restricted cubic splines showed that the risk of IFG was increasing rapidly until around 27.96 kg/m² of BMI and then started to plateau afterward (p for non-linearity = 0.010), which was similar in males and individuals ≤ 45 years old (p for nonlinearity < 0.001 and = 0.007, respectively). The risk of insulin resistance increased and ß-cell dysfunction decreased as the BMI increased in all participants (bothp for non-linearity > 0.05), consistent with the results in males, females, and ≤ 45 and > 45 year olds. CONCLUSIONS: The risk of IFG does not rise linearly as the BMI increases, and higher BMI seems to decelerate the rise of the risk.

The design, synthesis and anti-tumor mechanism study of new androgen receptor degrader.

Targeted protein degradation using small molecules is a novel strategy for drug development. In order to solve the problem of drug resistance in the treatment of prostate cancer, proteolysis-targeting chimeras (PROTAC) was introduced into the design of anti-prostate cancer derivatives. In this work, we synthesized two series of selective androgen receptor degraders (SARDs) containing the hydrophobic degrons with different linker, and then investigated the structure-activity relationships of these hybrid compounds. Most of the synthesized compounds exhibited moderate to good activity against all the cancer cell lines selected. Among them, compound A9 displayed potent inhibitory activity against LNCaP prostate cancer cell line with IC50 values of 1.75 µM, as well as excellent AR degradation activity. Primary mechanism studies elucidated compound A9 arrested cell cycle at G0/G1 phase and induced a mild apoptotic response in LNCaP cells. Further study indicated that the degradation of AR was mediated through proteasome-mediated process. For all these reasons, compound A9 held promising potential as anti-proliferative agent for the development of highly efficient SARDs for drug-resistance prostate cancer therapies.

In vitro treatment of 3 T3-L1 adipocytes with recombinant Calcium/calmodulin-dependent Protein Kinase IV (CaMKIV) limits ER stress and improves insulin sensitivity through inhibition of autophagy via the mTOR/CREB signaling pathway.

BACKGROUND: Recently, CaMKIV has been identified as a potential regulator of skeletal muscle glucose metabolism, it can also affect insulin gene expression in pancreas. However, its effects on adipose insulin resistance have yet to be explored. Autophagy has been shown as a potential therapeutic target for ER (endoplasmic reticulum) stress and insulin resistance. The purpose of this study is to investigate the effects of CaMKIV on ER stress, autophagic function and insulin signaling in tunicamycin-treated adipocytes. METHODS: In this study, mature 3 T3-L1 adipocytes were treated with tunicamycin to induce ER stress. Tunicamycin-treated 3 T3-L1 adipocytes were treated with recombinant CaMKIV in the presence or absence of targeted-siRNA mediated down-regulation of CREB and mTOR. The ER stress markers, autophagy activation, mTOR/CREB signaling and insulin sensitivity were analyzed by western blotting or electron microscopy. RESULTS: Treatment with CaMKIV significantly reversed tunicamycin-induced expression of p-PERK, cleaved-ATF6, Atg7 and LC3II. It also reduced p62 expression. In addition, levels of p-Akt and p-IRS-1 were increased. Moreover, CaMKIV inhibited activated ER stress and insulin resistance in Atg7 siRNA transfected adipocytes. However, the protective effects of CaMKIV on ER stress, insulin signaling, and autophagy function were nullified by suppression of mTOR or CREB in tunicamycin-treated adipocytes. CONCLUSION: This study proves recombinant CaMKIV inhibits tunicamycin-induced ER stress and insulin resistance by regulating autophagy. The protective effect of CaMKIV in adipocytes is affected at least partly through mTOR/CREB signaling. Our finding may offer novel opportunities for treating obesity and type 2 diabetes.

[Quality evaluation of Salvianolate for Injection by quantitative analysis of multi-components with single-marker].

Developing high-quality standard is useful for promoting the quality of traditional Chinese medicine injections, which could be evaluated by establishing the comprehensive quality control method. A method for simultaneous determination of salvianolic acid B, rosmarinic acid and lithospermic acid in Salvianolate for Injection was developed for quantitative analysis of multi-components with single-marker(QAMS). ZORBAX Eclipse Plus C_(18) chromatographic column was adopted, with 0.1% phosphoric acid and acetonitrile as mobile phase. The flow rate was set at 1 mL·min~(-1). The column temperature was set at 20 ℃, and the detection wavelength was 286 nm. Salvianolic acid B was used as internal reference. The relative correction factors of rosmarinic acid and lithospermic acid(f_(s/i)) were 0.58 and 0.94, respectively. About 85% of substances in Salvianolate for Injection were quantified by the established QAMS method. The analysis of different batches of intermediates and preparations during four years showed that the contents of salvianolic acid B were 77.1%-81.5% in intermediates and 70.5%-80.1% in preparations; The total content of rosmarinic acid and lithospermic acid was about 6%. The ratio of rosmarinic acid to lithospermic acid was(3.4∶1-10∶1) and(2.5∶1-5∶1), respectively, which showed that the ratio was more stable in preparation. The QAMS method established is feasible for comprehensive quality control of multiple components of in Salvianolate for Injection.

Dihydroartemisinin inhibits the tumorigenesis and metastasis of breast cancer via downregulating CIZ1 expression associated with TGF-ß1 signaling.

AIMS: Dihydroartemisinin (DHA) is currently considered as the promising cancer therapeutic drug. In this study, we aimed to investigate the anti-proliferative and anti-metastasis effects of DHA. MAIN METHODS: Utilizing breast cancer cells MCF-7, MDA-MB-231 and BT549, cell proliferation, migration and invasion were detected. RT-qPCR was performed to detect CIZ1, TGF-ß1 and Snail expression, and the interactions of these related molecules were analyzed by GeneMANIA database. Western blot detected CIZ1, TGF-ß1/Smads signaling and Snail expression in DHA-treated cells, in TGFß1-induced cells with enhanced metastatic capacity, and in cells treated with DHA plus TGFß1/TGFß1 inhibitor SD-208. KEY FINDINGS: Results indicated DHA inhibited breast cancer cell proliferation and migration, with more potent effects compared with that of artemisinin. RT-qPCR and Western blot showed DHA inhibited CIZ1, TGF-ß1 and Snail expression, and these molecules were shown to have protein-protein interactions by bioinformatics. Furthermore, TGFß1-treatment enhanced MCF-7 migration and invasion, and CIZ1, TGF-ß1/Smads signaling and snail activities; DHA, SD-208, combination of DHA and SD-208 reversed these conditions, preliminarily proving the cascade regulation between TGF-ß1 signaling and CIZ1. MCF-7 xenografts model demonstrated the inhibition of DHA on tumor burden, and its mechanisms and well-tolerance in vivo; combination of DHA and SD-208 tried by us for the first time showed better treatment effects, but possible liver impairment made its use still keep cautious. SIGNIFICANCE: DHA treatment inhibits the proliferation and metastasis of breast cancer, through suppressing TGF-ß1/Smad signaling and CIZ1, suggesting the promising potential of DHA as a well-tolerated antitumor TGF-ß1 pathway inhibitor.

Differentially expressed genes and key molecules of BRCA1/2-mutant breast cancer: evidence from bioinformatics analyses.

BACKGROUND: BRCA1 and BRCA2 genes are currently proven to be closely related to high lifetime risks of breast cancer. To date, the closely related genes to BRCA1/2 mutations in breast cancer remains to be fully elucidated. This study aims to identify the gene expression profiles and interaction networks influenced by BRCA1/2 mutations, so as to reflect underlying disease mechanisms and provide new biomarkers for breast cancer diagnosis or prognosis. METHODS: Gene expression profiles from The Cancer Genome Atlas (TCGA) database were downloaded and combined with cBioPortal website to identify exact breast cancer patients with BRCA1/2 mutations. Gene set enrichment analysis (GSEA) was used to analyze some enriched pathways and biological processes associated BRCA mutations. For BRCA1/2-mutant breast cancer, wild-type breast cancer and corresponding normal tissues, three independent differentially expressed genes (DEGs) analysis were performed to validate potential hub genes with each other. Protein-protein interaction (PPI) networks, survival analysis and diagnostic value assessment helped identify key genes associated with BRCA1/2 mutations. RESULTS: The regulation process of cell cycle was significantly enriched in mutant group compared with wild-type group. A total of 294 genes were identified after analysis of DEGs between mutant patients and wild-type patients. Interestingly, by the other two comparisons, we identified 43 overlapping genes that not only significantly expressed in wild-type breast cancer patients relative to normal tissues, but more significantly expressed in BRCA1/2-mutant breast patients. Based on the STRING database and cytoscape software, we constructed a PPI network using 294 DEGs. Through topological analysis scores of the PPI network and 43 overlapping genes, we sought to select some genes, thereby using survival analysis and diagnostic value assessment to identify key genes pertaining to BRCA1/2-mutant breast cancer. CCNE1, NPBWR1, A2ML1, EXO1 and TTK displayed good prognostic/diagnostic value for breast cancer and BRCA1/2-mutant breast cancer. CONCLUSION: Our research provides comprehensive and new insights for the identification of biomarkers connected with BRCA mutations, availing diagnosis and treatment of breast cancer and BRCA1/2-mutant breast cancer patients.