[Mechanism of ultrafiltration extract of Angelicae Sinensis Radix and Hedysari Radix regulates HIF-1α signaling pathway mediated by renal hypoxia to ameliorate renal fibrosis in DKD rats].

This study explored the mechanism of the ultrafiltration extract of Angelicae Sinensis Radix and Hedysari Radix in ameliorating renal fibrosis in the rat model of diabetic kidney disease(DKD) based on the expression of hypoxia-inducible factor-1α(HIF-1α)/vascular endothelial growth factor(VEGF) and HIF-1α/platelet-derived growth factor(PDGF)/platelet-derived growth factor receptor(PDGFR) signaling pathways in the DKD rats. After 1 week of adaptive feeding, 50 male SPF-grade Wistar rats were randomized into a blank group(n=7) and a modeling group. After 24 h of fasting, the rats in the modeling group were subjected to intraperitoneal injection of streptozocin and fed with a high-sugar and high-fat diet to establish a DKD model. After modeling, the rats were randomly assigned into model(n=7), low-dose ultrafiltration extract(n=7), medium-dose ultrafiltration extract(n=7), irbesartan(n=8), and high-dose ultrafiltration extract(n=8) groups. After intervention by corresponding drugs for 12 weeks, the general conditions of the rats were observed. The body weights and blood glucose levels of the rats were measured weekly, and the 24 h urinary protein(24hUP) was measured at the 6th and 12th weeks of drug administration. After the last drug administration, the renal function indicators were determined. Masson staining was employed to observe the pathological changes of the renal tissue. The expression of prolyl hydroxylase domain 2(PHD2) and HIF-1α in the renal tissue was detected by immunohistochemistry(IHC). Real-time qPCR was employed to determine the mRNA levels of PHD2, VEGF, PDGF, and PDGFR in the renal tissue. Western blot was employed to determine the protein levels of HIF-1α, VEGF, PDGF, and PDGFR in the renal tissue. The results showed that compared with the model group, drug administration lowered the levels of glycosylated serum protein(GSP), aerum creatinine(Scr), and blood urea nitrogen(BUN) in a dose-dependent manner(P<0.05 or P<0.01) and mitigated the pathological changes in the renal tissue. Furthermore, drug administration up-regulated mRNA level of PHD2(P<0.05 or P<0.01), down-regulated the mRNA levels of VEGF, PDGF, and PDGFR(P<0.05 or P<0.01) and the protein levels of HIF-1α, VEGF, PDGF, and PDGFR(P<0.01) in the renal tissue, and increased the rate of PHD2-positive cells(P<0.01). In conclusion, the ultrafiltration extract of Angelicae Sinensis Radix and Hedysari Radix effectively alleviated the renal fibrosis in DKD rats by inhibiting the expression of key proteins in the HIF-1α signaling pathway mediated by renal hypoxia and reducing extracellular matrix(ECM) deposition.

Physiological feedback technology for real-time emotion regulation: a systematic review.

Background: Previous studies suggest that physiological feedback can be an effective method for emotion regulation (ER). However, studies on the specific effects of physiological feedback have shown conflicting results due to inconsistencies in study designs. Therefore, we present this systematic review to further validate the effectiveness of physiological feedback for ER, clarify its specific effects, as well as summarize the factors that influence its effectiveness. Method: This systematic review following PRISMA guidelines covers all studies using physiological feedback in emotions. A literature search was conducted in Web of Science, PubMed, PsychINFO, China National Knowledge Infrastructure, and WANFANG DATA. And a standardized quality assessment was performed. Results: We identified 27 relevant articles (25 studies), and the majority of these studies showed a significant regulatory effect of physiological feedback on different emotions. The feedback's content, explanation, authenticity, real-time capability, and modality were the key factors that influenced its effects, and this technology will achieve its optimal ER effect when these factors are considered comprehensively. Conclusions: These findings further confirmed the effectiveness of physiological feedback as an ER method, as well as providing key factors that should be addressed in its application. Meanwhile, due to the limitations of these studies, more well-designed studies are still needed.

Profiling GABA(A) Receptor Subunit Expression in the Hippocampus of PMDD Rat Models Based on TCM Theories.

γ-Aminobutyric acid type A receptors (GABAARs) play an important role in cognitive and emotional regulation and are related to the hippocampus. However, little is known regarding patterns of hippocampal GABAAR subunit expression in rat models of premenstrual dysphoric disorder (PMDD). This study investigated the above changes by establishing two PMDD rat models based on Traditional Chinese Medicine (TCM) theories, namely, PMDD liver-qi invasion syndrome (PMDD-LIS) and PMDD liver-qi depression syndrome (PMDD-LDS). Behavioral tests were used to detect depression and irritability emotion. Western blot analysis was used to investigate protein levels of GABAAR α1, α2, α4, α5, ß2, ß3, and δ subunits, whereas ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) analysis was performed to determine gamma-aminobutyric acid (GABA) and glutamate (Glu) levels in the hippocampus across each group. Concurrently, behavioral data indicated that the PMDD-LDS and PMDD-LIS rat models had been successfully established. GABAAR α2, α5, ß2, and δ subunit was significantly upregulated, whereas α4 was significantly downregulated (P < 0.05) in PMDD-LDS rat models relative to controls. On the other hand, GABAAR α1, α2, and ß3 were significantly downregulated while α4 and ß2 were significantly upregulated in PMDD-LIS rat models relative to the control group (P < 0.05). Moreover, GABA levels significantly decreased, while Glu and the ratio of glutamate to GABA increased in PMDD-LIS rat models (P < 0.05). Conversely, GABA and Glu levels significantly decreased, whereas the ratio of glutamate to GABA increased in PMDD-LIS rat models (P < 0.05). Conclusively, our results revealed differential expression of GABAAR α1, α2, α4, α5, ß2, ß3, and δ subunits between PMDD-LIS and PMDD-LDS rat models, suggesting that they may be biomarkers in the pathogenesis of PMDD.

[Observation on the efficacy of post-stroke dysphagia treated with He's santong acupuncture therapy through surface electromyography: a randomized controlled trial].

OBJECTIVE: To observe the clinical therapeutic effect on post-stroke dysphagia treated with He's santong (triple promotion) acupuncture therapy through surface electromyography (sEMG). METHODS: A total of 60 patients with post-stroke dysphagia were divided into a routine treatment group and a He's santong acupuncture therapy group, using blocked randomization, 30 cases in each one. In the routine treatment group, the secondary prevention and swallowing rehabilitation training were adopted. In the He's santong acupuncture therapy group, on the base of the treatment as the routine treatment group, weitong (mild promotion, routine acupuncture at bilateral Fengchi ï¼»GB20ï¼½, Fengfu ï¼»GV16ï¼½, Yifeng ï¼»TE17ï¼½, Lianquan ï¼»CV23ï¼½, Jia-lianquan ï¼»Extraï¼½, Fenglong ï¼»ST40ï¼½ and Tongli ï¼»HT5ï¼½, needle retaining for 30 min, 5 treatments a week), wentong (warm promotion, pricking with fire needle at bilateral GB20 and CV23, twice a week) and qiangtong (strong promotion, blood-letting with three-edge needle at Jinjin ï¼»EX-HN12ï¼½, Yuye ï¼»EX-HN13ï¼½ and Yanhoubi ï¼»Extraï¼½, twice a week) treatment was added. The therapy was given consecutively for 4 weeks in each group. The score of fiberoptic endoscopic examination of swallowing (FEES) and Rosenbek-penetration-aspiration scale (PAS), the score of swallowing grading scale, the score of the modified Mann assessment of swallowing ability (MMASA) and the peak amplitude of sEMG were recorded before and after treatment in patients. RESULTS: Compared with before treatment, the score of FEES and PAS after treatment was decreased (P<0.05), the scores of swallowing grading scale and MMASA after treatment were increased in both routine treatment group and He's santong acupuncture therapy group (P<0.05), the peak amplitude of sEMG was increased (P<0.05) in the He's santong acupuncture therapy group and decreased (P<0.05) in the routine treatment group. Compared with the routine treatment group, the score of FEES and PAS was decreased (P<0.05), the scores of swallowing grading scale and peak amplitude of sEMG were increased in the He's santong acupuncture therapy group (P<0.05). CONCLUSION: He's santong acupuncture therapy is obviously effective on post-stroke dysphagia, which may be related to its effects in increasing the contraction of swallowing-related muscles and improving the peak amplitude of hyoid muscle group.

Deciphering the spatial-temporal transcriptional landscape of human hypothalamus development.

The hypothalamus comprises various nuclei and neuronal subpopulations that control fundamental homeostasis and behaviors. However, spatiotemporal molecular characterization of hypothalamus development in humans is largely unexplored. Here, we revealed spatiotemporal transcriptome profiles and cell-type characteristics of human hypothalamus development and illustrated the molecular diversity of neural progenitors and the cell-fate decision, which is programmed by a combination of transcription factors. Different neuronal and glial fates are sequentially produced and showed spatial developmental asynchrony. Moreover, human hypothalamic gliogenesis occurs at an earlier stage of gestation and displays distinctive transcription profiles compared with those in mouse. Notably, early oligodendrocyte cells in humans exhibit different gene patterns and interact with neuronal cells to regulate neuronal maturation by Wnt, Hippo, and integrin signals. Overall, our study provides a comprehensive molecular landscape of human hypothalamus development at early- and mid-embryonic stages and a foundation for understanding its spatial and functional complexity.

Design, synthesis, and biological evaluation of tetrahydroquinolin derivatives as potent inhibitors of CBP bromodomain.

CREB-binding protein (CBP) is a large multi-domain protein containing a HAT domain catalyzing transacetylation and a bromodomain responsible for acetylated lysine recognition. CBPs could act as transcription co-activators to regulate gene expression and have been shown to play a significant role in the development and progression of many cancers. Herein, through in silico screening two hit compounds with tetrahydroquinolin methyl carbamate scaffold were discovered, among which DC-CPin7 showed an in vitro inhibitory activity with the TR-FRET IC50 value of 2.5 ± 0.3 µM. We obtained a high-resolution co-crystal structure of the CBP bromodomain in complex with DC-CPin7 to guide following structure-based rational drug design, which yielded over ten DC-CPin7 derivatives with much higher potency, among which DC-CPin711 showed approximately 40-fold potency compared with hit compound DC-CPin7 with an in vitro TR-FRET IC50 value of 63.3 ± 4.0 nM. Notably, DC-CPin711 showed over 150-fold selectivity against BRD4 bromodomains. Moreover, DC-CPin711 showed micromolar level of anti-leukemia proliferation through G1 phase cell cycle arrest and cell apoptosis. In summary, through a combination of computational and crystal-based structure optimization, DC-CPin711 showed potent in vitro inhibitory activities to CBP bromodomain with a decent selectivity towards BRD4 bromodomains and good cellular activity to leukemia cells, which could further be applied to related biological and translational studies as well as serve as a lead compound for future development of potent and selective CBP bromodomain inhibitors.

HuangqiGuizhiWuwu Decoction Prevents Vascular Dysfunction in Diabetes via Inhibition of Endothelial Arginase 1.

Hyperglycemia induces vascular endothelial dysfunction, which contributes to the development of vascular complication of diabetes. A classic prescription of traditional medicine, HuangqiGuizhiWuwu Decoction (HGWWD) has been used for the treatment of various cardiovascular and cerebrovascular diseases, which all are related with vascular pathology. The present study investigated the effect of HGWWD treatment in streptozocin (STZ)-induced vascular dysfunction in mouse models. In vivo studies were performed using wild type mice as well as arginase 1 knockout specific in endothelial cells (EC-A1-/-) of control mice, diabetes mice and diabetes mice treated with HGWWD (60 g crude drugs/kg/d) for 2 weeks. For in vitro studies, aortic tissues were treated with mice serum containing HGWWD with or without adenoviral arginase 1 (Ad-A1) transduction in high glucose (HG) medium. We found that HGWWD treatment restored STZ-induced impaired mean velocity and pulsatility index of mouse left femoral arteries, aortic pulse wave velocity and vascular endothelial relaxation accompanied by elevated NO production in the aorta and plasma, as well as reduced endothelial arginase activity and aortic arginase 1 expression. The protective effect of HGWWD is reversed by an inhibitor of nitric oxide synthesis. Meanwhile, the preventive effect of serum containing HGWWD in endothelial vascular dysfunction is completely blocked by Ad-A1 transduction in HG incubated aortas. HGWWD treatment further improved endothelial vascular dysfunction in STZ induced EC-A1-/- mice. This study demonstrates that HGWWD improved STZ-induced vascular dysfunction through arginase 1 - NO signaling, specifically targeting endothelial arginase 1.

Discovery of alkoxy benzamide derivatives as novel BPTF bromodomain inhibitors via structure-based virtual screening.

Bromodomain PHD finger transcription factor (BPTF), a bromodomain-containing protein, plays a crucial role in the regulation of downstream gene expression through the specific recognition of lysine acetylation on bulk histones. The dysfunction of BPTF is closely involved with the development and progression of many human diseases, especially cancer. Therefore, BPTF bromodomain has become a promising drug target for epigenetic cancer therapy. However, unlike BET family inhibitors, few BPTF bromodomain inhibitors have been reported. In this study, by integrating docking-based virtual screening with biochemical analysis, we identified a novel selective BPTF bromodomain inhibitor DCB29 with the IC50 value of 13.2 ±â€¯1.6 µM by homogenous time-resolved fluorescence resonance energy transfer (HTRF) assays. The binding between DCB29 and BPTF was confirmed by NMR and SPR. Molecular docking disclosed that DCB29 occupied the pocket of acetylated H4 peptide substrate and provided detailed SAR explanations for its derivatives. Collectively, DCB29 presented great potential as a powerful tool for BPTF-related biological research and further medicinal chemistry optimization.

Xiao-Shen-Formula, a Traditional Chinese Medicine, Improves Glomerular Hyper-Filtration in Diabetic Nephropathy via Inhibiting Arginase Activation and Heparanase Expression.

Hyperglycemia induces glomerular hyper-filtration, which contributes to the development of diabetic nephropathy (DN), a condition that remains a challenge for treatment. The present study investigated the effect of Xiao-Shen-Formula (XSF) used for treatment of renal injury in type 1 DN mice model induced by streptozotocin (STZ) and its underlying mechanism in cultured human glomerular endothelial cell (hGECs). Studies were performed using control, diabetic DN, DN treated with XSF groups (1 g/kg/d, LXSF or 3 g/kg/d, HXSF) for 6 weeks and hGECs were post-treated with mice serum containing HXSF (MS-HXSF) and arginase inhibitor (ABH, 100 µM) in high glucose medium. HXSF treatment restored STZ-induced renal hyper-filtration, glomerulosclerosis, renal microvascular remodeling and the increased levels of systemic reactive oxidative species and inflammatory cytokines, accompanied by preventing the decreased expression of glomerular heparin sulfate and the increased levels of cortical heparanase and argianse2 protein and arginase activity. In hGECs study, MS-HXSF ameliorated the enhancement in arginase activity, the protein/mRNA expression of heparanase, mRNA levels of vascular cell adhesion molecule-1, intercellular adhesion molecule-1, monocyte chemoattractant protein-1 and permeability of hGECs monolayers as well as the depression of nitric oxide production. Besides all these protective effects, XSF blunted the mRNA expression of TNF-α in vivo and vitro studies as well, which was not changed by the post-treatment of ABH or HXSF plus ABH. This study demonstrated that the protective effect of XSF might be related with vascular prevention, anti-inflammation and anti-oxidation through intervening multi-targets including glomerular endothelial arginase-heparanase signaling pathway in DN model.

The protective effects of acupoint gel embedding on rats with myocardial ischemia-reperfusion injury.

AIMS: Prevention and treatment of myocardial ischemia-reperfusion (I/R) injury has for many years been a hot topic in treating ischemic heart disease. As one of the most well-known methods of complementary and alternative medicine, acupuncture has attracted increasing interest in preventing myocardial I/R injury due to its remarkable effectiveness and minimal side effect. However, traditional acupuncture approaches are limited by cumbersome execution, high labor costs and inevitable pain caused by frequent stimulation. Therefore, in this work, we aimed to develop a novel acupoint gel embedding approach and investigated its role in protecting against myocardial I/R injury in rats. MAIN METHODS: Gels were embedded at bilateral Neiguan (PC6) points of rats and their protective effects against myocardial I/R injury evaluated in terms of changes in histomorphology, myocardial enzymology, antioxidant capacity, anti-inflammatory response, and anti-apoptosis of cells. KEY FINDINGS: We found that the approach of acupoint gel embedding could significantly reduce myocardial infarcted size, repair pathological changes, mitigate oxidative stress damage and inflammatory response, as well as inhibit apoptosis of cardiomyocytes. Such cardioprotective effects were found to be associated with Notch-1/Jagged-1 signaling pathway. SIGNIFICANCE: The proposed approach of acupoint gel embedding has advantages in continuous acupoint stimulation, dosing controls, and no side effects in the course of treatment, as well as in reducing the pain caused by frequent acupuncture. It can form an alternative therapy to not only protect against myocardial I/R injury but also hold great potential in treating other diseases in the future.

Synchronous front-face fluorescence spectroscopy for authentication of the adulteration of edible vegetable oil with refined used frying oil.

Synchronous front-face fluorescence spectroscopy has been developed for the discrimination of used frying oil (UFO) from edible vegetable oil (EVO), the estimation of the using time of UFO, and the determination of the adulteration of EVO with UFO. Both the heating time of laboratory prepared UFO and the adulteration of EVO with UFO could be determined by partial least squares regression (PLSR). To simulate the EVO adulteration with UFO, for each kind of oil, fifty adulterated samples at the adulterant amounts range of 1-50% were prepared. PLSR was then adopted to build the model and both full (leave-one-out) cross-validation and external validation were performed to evaluate the predictive ability. Under the optimum condition, the plots of observed versus predicted values exhibited high linearity (R(2)>0.96). The root mean square error of cross-validation (RMSECV) and root mean square error of prediction (RMSEP) were both lower than 3%.

Qualitative research on meaning in life in elderly patients with breast cancer / 中国实用护理杂志

Objective To explore the dimensions of meaning in life(ML) in elderly patients with breast cancer, and offer practical recommendations for holistic nursing in clinical practice. Methods Purposive and convenience sampling techniques from phenomenological methodology were used to and recruit twelve elderly patients (more than 65 years old) with breast cancer in the Third Department of Breast Cancer, Tianjin Medical University Cancer Institute and Hospital from February 2016 to October 2016. A depth interview was conducted for each patient to collect data. Results In breast cancer survivors′perspective,ML including 4 dimensions:focus on the current survival status,self-actualization, social support and participation in individualized treatment. Focus on the current survival status is the basic connotation of ML, self-actualization is the concentrative embodiment of ML, social support is an important path way for grabbing ML, and participation in individualized treatment is an important supplement to realize ML Conclusion The awareness of ML has its own characteristics in elderly patients with breast cancer. Focus on the current survival status, self-actualization, social support and participation in individualized treatment contribute to its main connotation. The truth of getting cancer affects their perceptions to ML. Thus, healthcare providers should pay more attention and take various measures to meet their demands to ML.

High-performance multiplexed fluorescence in situ hybridization in culture and tissue with matrix imprinting and clearing.

Highly multiplexed single-molecule FISH has emerged as a promising approach to spatially resolved single-cell transcriptomics because of its ability to directly image and profile numerous RNA species in their native cellular context. However, background-from off-target binding of FISH probes and cellular autofluorescence-can become limiting in a number of important applications, such as increasing the degree of multiplexing, imaging shorter RNAs, and imaging tissue samples. Here, we developed a sample clearing approach for FISH measurements. We identified off-target binding of FISH probes to cellular components other than RNA, such as proteins, as a major source of background. To remove this source of background, we embedded samples in polyacrylamide, anchored RNAs to this polyacrylamide matrix, and cleared cellular proteins and lipids, which are also sources of autofluorescence. To demonstrate the efficacy of this approach, we measured the copy number of 130 RNA species in cleared samples using multiplexed error-robust FISH (MERFISH). We observed a reduction both in the background because of off-target probe binding and in the cellular autofluorescence without detectable loss in RNA. This process led to an improved detection efficiency and detection limit of MERFISH, and an increased measurement throughput via extension of MERFISH into four color channels. We further demonstrated MERFISH measurements of complex tissue samples from the mouse brain using this matrix-imprinting and -clearing approach. We envision that this method will improve the performance of a wide range of in situ hybridization-based techniques in both cell culture and tissues.

[Effects of Jiangu granule containing serum on the cyclins in rat's osteoblast at G1 phase].

OBJECTIVE: To investigate the effects of Jiangu granule containing serum (JGG-serum) on the cyclins in rat's osteoblast at G1 phase. METHODS: Osteoblasts isolated by enzymatic digestion from SD rats were cultured and intervened with JGG-serum or normal saline (as control) respectively. Cell generation cycle was detected by flow cytometry, and expressions of Cyclin D1, cyclin-dependent kinase 4 (CDK4), oncogene protein (P21) in the osteoblast were detected dynamically using immuno-cytochemical and RT-PCR technique. RESULTS: As compared with the control, the cell generation cycle and cell proliferation were proceeding quicker in the JGG-serum (20%) intervention group; with higher protein and mRNA expressions of Cyclin D1 and CDK4, as well as much lowered expressions of P21 in nuclei of osteoblast detected at all time points (24 h, 48 h and 72 h after treatment, P < 0.05 or P < 0.01). CONCLUSION: JGG-serum can adjust the G1 phase cyclins in osteoblast cultured in vitro, increase the mRNA and protein expressions of Cyclin D1 and CDK4, and inhibit P21 expression, so as to accelerate the proliferation of osteoblast.

Production of SAM by recombinant Pichia pastoris / 生物工程学报

To utilize Pichia pastoris to produce S-adenosyl-L-methionine (SAM), an intracellular expression vector harboring S. cerevisiae SAM2 was transformed into GS115. A recombinant strain having 2 copies of expression cassette was obtained through G418 resistance screening. This strain had higher SAM synthetase activity and higher SAM production capacity than the original strain, when cultured in medium containing methanol and methionine. The carbon source and nitrogen source of medium was optimized. The results showed SAM production by this strain was closely related to carbon metabolism. With supplementation of 0.2% glycerol every day from the beginning of 3rd day, this strain produced 1.58 g/L SAM when cultured in a medium containing 0.75% L-methionine and optimized carbon and nitrogen source after 6 days.