Exploration of potential active ingredients and mechanism of action of Xihuang pill-medicated serum against glioma based on HPLC-Q-TOF-MS/MS, network pharmacology and experimental verification / 药学学报

Qualitative analysis of the ingredients absorbed into blood and their metabolites of Xihuang pill (XHP) were conducted using high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (HPLC-Q-TOF-MS/MS) technology. Network pharmacology was used to explore the potential anticancer mechanisms of the ingredients against glioma, and their specific mechanisms were validated through molecular docking and experimental verification. SD rats were intragastrically administered with XHP, and rat serum samples were collected. Ingredients absorbed into blood and their metabolites were identified based on the retention time of chromatographic peaks, accurate molecular mass, characteristic fragment ions, and comparisons with reference substances and literature data. PharmMapper and SwissTarget Prediction databases were used to obtain the targets of the XHP-medicated serum, while GeneCards, OMIM, PharmGKB, TTD, and DrugBank databases were used to obtain glioma disease targets. The "component-target" network relationship diagram was constructed using Cytoscape 3.9.1 software. The protein-protein interaction (PPI) network diagram was constructed using the STRING database, and the targets were analyzed using GO and KEGG analyses. Molecular docking was used to verify the binding ability of core targets with their corresponding compounds in XHP-medicated serum. The potential mechanism of the anti-glioma effect of 11-keto-β-boswellic acid (KBA), a representative component of XHP-medicated serum, was verified using CCK-8 and Western blot assays. A total of 40 compounds were identified in the XHP-medicated serum, including 28 prototype components and 12 metabolites. The network pharmacology results showed that elemonic acid, 3-acetyl-β-boswellic acid, KBA, α-boswellic acid, and other 5 compounds might be the active ingredients of XHP-medicated serum in the treatment of glioma. Glutathione reductase (GSR), glucose-6-phosphate dehydrogenase (G6PD), ATP-citrate lyase (ACLY), aldo-keto reductase family 1 member B1 (AKR1B1) and glutaredoxin (GLRX) were identified as key targets, involving pathways such as glutathione metabolism and the pentose phosphate pathway. Further cell experiments showed that KBA significantly inhibited the proliferation of T98G cells with an IC50 of 30.96 μmol·L-1, and KBA (30 μmol·L-1) significantly downregulated the protein expression levels of GSR in T98G cells. In summary, XHP-medicated serum may exert its anti-glioma effect by regulating GSR and G6PD-targeted pathways involved in glutathione metabolism. These results provide valuable evidence for further investigating the mechanism of XHP in treating glioma. The animal welfare and experimental procedures were approved by the Ethical Committee of Laboratory Animals at Nanjing University of Chinese Medicine (approval No. ACU221001).

The effect and mechanism of Xihuang pill inhibits the formation of vasculogenic mimicry in human glioblastoma cells by down-regulating HIF-1α/VEGFA/VEGFR2 signaling pathway / 药学学报

Our studies were aimed to explore the effect and mechanism of the inhibition of the formation of vasculogenic mimicry (VM) in human glioblastoma cells by Xihuang pill (XHP) medicated serum through regulating the hypoxia inducible factor-1α (HIF-1α)/vascular endothelial growth factor A (VEGFA)/vascular endothelial growth factor receptor 2 (VEGFR2) signaling pathway. The medicated serum of XHP was prepared by gavage for 7 days to male SD rats (approval number of animal experiment ethics: 202105A051). The hypoxia model of U251 cells was established using 200 μmol·L-1 of CoCl2. After treatment with XHP-medicated serum, cell viability and proliferation of U251 cells were detected by CCK-8 and cell cloning experiment. Cell apoptosis and cell cycle of U251 cells were determined by flow cytometry. Cell migration and invasion were evaluated by wound healing and Transwell invasion assay. The formation of VM was assessed by three-dimensional cell culture of U251 cells. The protein expression levels of HIF-1α, VEGFA, VEGFR2, phosphorylated-VEGFR2 (p-VEGFR2), vascular endothelial-cadherin (VE-cadherin), Eph receptor tyrosine kinases A2 (EphA2), matrix metalloproteinase 2 (MMP2), matrix metalloproteinase 14 (MMP14) and laminin γ2 in U251 cells were detected by Western blot. The results showed that 10% XHP-medicated serum had little effect on the cell viability, proliferation, apoptosis and cell cycle of U251 cells under hypoxia. Compared with the model group, 10% XHP-medicated serum at 1.0, 1.5 and 2.0 h significantly decreased the migration rate (P < 0.01) and the number of invading U251 cells (P < 0.01). 10% XHP-medicated serum at 2.0 h significantly suppressed the formation of VM tubular structures in U251 cells under the condition of hypoxia (P < 0.01). Western blot experiment showed that 10% XHP-medicated serum significantly down-regulated the expression of HIF-1α, VEGFA, phospho-VEGFR2, VE-cadherin, EphA2 and MMP14 proteins (P < 0.05). In conclusion, XHP could inhibit the formation of VM in human glioblastoma U251 cells to suppress the angiogenesis by down-regulating the HIF-1α/VEGFA/VEGFR2 signaling pathway.

Multimodal deep learning model for staging diabetic retinopathy based on ultra-widefield fluorescence angiography / 中华眼底病杂志

Objective:To apply the multi-modal deep learning model to automatically classify the ultra-widefield fluorescein angiography (UWFA) images of diabetic retinopathy (DR).Methods:A retrospective study. From 2015 to 2020, 798 images of 297 DR patients with 399 eyes who were admitted to Eye Center of Renmin Hospital of Wuhan University and were examined by UWFA were used as the training set and test set of the model. Among them, 119, 171, and 109 eyes had no retinopathy, non-proliferative DR (NPDR), and proliferative DR (PDR), respectively. Localization and assessment of fluorescein leakage and non-perfusion regions in early and late orthotopic images of UWFA in DR-affected eyes by jointly optimizing CycleGAN and a convolutional neural network (CNN) classifier, an image-level supervised deep learning model. The abnormal images with lesions were converted into normal images with lesions removed using the improved CycleGAN, and the difference images containing the lesion areas were obtained; the difference images were classified by the CNN classifier to obtain the prediction results. A five-fold cross-test was used to evaluate the classification accuracy of the model. Quantitative analysis of the marker area displayed by the differential images was performed to observe the correlation between the ischemia index and leakage index and the severity of DR.Results:The generated fake normal image basically removed all the lesion areas while retaining the normal vascular structure; the difference images intuitively revealed the distribution of biomarkers; the heat icon showed the leakage area, and the location was basically the same as the lesion area in the original image. The results of the five-fold cross-check showed that the average classification accuracy of the model was 0.983. Further quantitative analysis of the marker area showed that the ischemia index and leakage index were significantly positively correlated with the severity of DR ( β=6.088, 10.850; P<0.001). Conclusion:The constructed multimodal joint optimization model can accurately classify NPDR and PDR and precisely locate potential biomarkers.

Changes of subfoveal choroidal thickness and choroidal volume in different stages of diabetic retinopathy through automated choroid segmentation / 中华实验眼科杂志

Objective:To analyze the changes of subfoveal choroidal thickness (SFCT) and choroidal volume in type 2 diabetes patients with diabetic retinopathy (DR) through automated choroid segmentation.Methods:A cross-sectional study was conducted.Sixty-four patients (88 eyes) with type 2 diabetes diagnosed in The First Affiliated Hospital with Nanjing Medical University from May 2016 to May 2017 were enrolled, including 27 males (32 eyes) and 37 females (56 eyes), with an average age of (62.7±7.4) years and an average diabetes duration of (13.7±7.2) years.Best corrected visual acuity, slit lamp ophthalmoscopy, indirect ophthalmoscopy and spectral domain optical coherence tomography (SD-OCT) examination were carried out among all the patients.The patients were divided into non-DR (NDR) group (28 eyes), nonproliferative DR (NPDR) group (36 eyes) and proliferative DR (PDR) group (24 eyes) according to the Early Treatment Diabetic Retinopathy Study.Nineteen age-matched normal subjects (32 eyes) were enrolled as normal control group.SFCT and choroidal volume were measured through automated choroid segmentation based on SD-OCT with enhanced depth imaging.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of The First Affiliated Hospital with Nanjing Medical University (No.2017-SR-176).Results:Mean SFCT and choroidal volume of normal control group, NDR group, NPDR group and PDR group were (277.8±45.3)μm and (8.8±1.0)mm 3, (220.9±38.7)μm and (7.6±1.3)mm 3, (231.8±26.7)μm and (8.2±0.9)mm 3, (252.0±42.2)μm and (8.7±1.0)mm 3, respectively, with significant overall differences among the groups ( F=14.615, P<0.001; F=7.711, P<0.001). Mean SFCT and choroidal volume were significantly smaller in NDR and NPDR group than those in normal control group (both at P<0.05). Mean SFCT and choroidal volume of PDR group was greater than those of NDR group (both at P<0.05). Conclusions:SFCT and choroidal volume decrease during the early course of diabetes and increase significantly as DR worsens from NDR to PDR.

Research progress in ecological prevention of dental caries / 中华口腔医学杂志

Dental caries is one of the most common oral diseases around the world. Dental plaque attached to the surfaces of teeth is the main biological factor leading to caries. Although fluoride is still one of the most commonly used methods to prevent caries, with the change of epidemiological characteristics of caries and the update of the understanding of caries etiology, it is necessary to use other ecological methods such as antimicrobial peptides, arginine, probiotics and natural products, etc. to enhance the effect of fluoride in preventing dental caries. The present article reviews the research progress on the ecological approaches for caries prevention in recent years.

Evolution Characteristics and Clinical Significance of Blood Separation in Patients with Multiple Myeloma / 中国实验血液学杂志

OBJECTIVE@#To investigate the evolution of blood separation results by gel extraction of multiple myeloma (MM) patients, and to evaluate the clinical value of abnormal blood separation results for the evaluation of disease and prognosis.@*METHODS@#The clinical data of 5 patients diagnosed newly MM patients with abnormal blood separation of gel collection vessels in our hospital were retrospectively analyzed, and the changes of blood separation results and blood index levels were followed up with the improvement of treatment effect, and the correlation of different blood index levels was analyzed.@*RESULTS@#In 5 patients with newly diagnosed MM, the blood separation result showed floating phenomenon after centrifugation, which divided into three layers and the order from top to bottom is separator gel, serum, and red blood cells(RBC). With partial remission of clinical symptoms, the blood separation results were still abnormal, which were divided into three layers from top to bottom: serum, RBC and separator gel. Finally, with complete remission of the disease, blood separation results returned to normal, from top to bottom: serum, separator gel, RBC. With the blood separation results from abnormal to normal, the blood routine indicators: Hb, Hct levels gradually increased, neutrophil/lymphocyte ratio (NLR), monocyte/lymphocyte ratio (MLR) gradually decreased; biochemical indexes: TP, GLB, Ig and β2-MG levels gradually decreased. Tumor load related indicators: serum IL-6, TNF-α, IL-17 levels gradually decreased, and IL-35 levels gradually increased; and the differences were statistically significant (P<0.05). Pearson correlation analysis showed that serum β2-MG was positively correlated with IL-6, TNF-α and IL-17 levels (r=0.710, 0.756, 0.581, P<0.05), and negatively correlated with IL-35 level (r=-0565, P<0.05).@*CONCLUSION@#Abnormal blood separation exists in MM patients, and there are significant differences in blood, tumor load and immune balance related indexes in patients with different blood separation results, which provides partial experimental basis for evaluation of disease, efficacy and prognosis with different blood separation results.

Detecção de áreas vulneráveis ​​ao escorpionismo e sua associação com fatores ambientais em São Paulo, Brasil / Detection of areas vulnerable to scorpionism and its association with environmental factors in São Paulo, Brazil

Accidents caused by scorpions are considered a neglected condition and represent a major health problem in most tropical countries, especially for children and elderly people. In Brazil, scorpionism is recurrent in the southeast region, mainly in the state of Sao ˜ Paulo, due to the progressive increase in scorpions found in urban habitats. Thus, our study aimed to provide better insights into the geographic and epidemiological characteristics of scorpion envenomation in Sao ˜ Paulo state and identify the environmental factors that are associated with these accidents. This is an ecological and retrospective study with secondary data on scorpion accidents in the state of Sao ˜ Paulo from 2008 to 2018 obtained from the Notifiable Disease Information System. The SatScan software was used to identify the higher- and lower-risk spatiotemporal clusters. A total of 145,464 scorpion sting cases were recorded in the state of Sao Paulo, between 2008 and 2018; there was a four-fold increase in the incidence rate. Accidents occurred more frequently in the spring season, wherein higher-risk clusters were in the north and northwest regions of the state. High temperatures, low precipitation, and poor natural vegetation are associated with higher risk areas. Our study mapped vulnerable areas for scorpion accidents that can aid in the design ofefficient public health policies, which should be intensified during the spring season.

Regulation of LncRNA00067110 on Proliferation, Apoptosis and Melanins Production of B16-F10 Cells by Targeting Cabyr / 中国生物化学与分子生物学报

Long non-coding RNA (lncRNA) is a type of non-coding RNA with the more than 200 nucleotides. Several lncRNAs have been identified as the potential targets for cancer therapy. LncRNA00067110 is one of the differentially expressed genes in the transcriptome profiles of melanoma B16-F10 cells compared to normal mice melanocytes. To investigate whether lncRNA00067110 regulates the proliferation, apoptosis and melanogenesis of B16-F10 cells, the calcium-binding tyrosine phosphorylation regulated protein (Cabyr) target gene was predicted by LncTar and verified by dual luciferase activities. The regulating function of lncRNA00067110 was investigated by the analysis of transcriptome profiles and to detect the proliferation, apoptosis and melanin production of B16-F10 cells transfected by the overexpression plasmids of lncRNA00067110. The results showed that the relationship of lncRNA00067110 targeting Cabyr, the mRNA and protein levels of proliferation (MEK/ERK/MNK/CREB) and melanogenesis-related genes (TYR family and CREB) were significantly down-regulated, while the mRNA and protein levels of apoptosis-related genes (AKT and Bcl-2) were up-regulated in B16-F10 cells with lncRNA00067110 overexpression. The transcriptome profile of B16-F10 cells with lncRNA00067110 overexpression showed that 17 genes were differentially expressed, among which Cabyr was up-regulated. Furthermore, the effect of lncRNA00067110 on the phenotypes of cell proliferation and apoptosis were verified. The results suggested that lncRNA00067110 might be a novel target for the treatment of melanoma by targeting Cabyr, which regulate the expression of related genes to inhibit the proliferation and melanogenesis, as well as to induce the apoptosis of B16-F10 cells.

The anti-neoplastic activities of aloperine in HeLa cervical cancer cells are associated with inhibition of the IL-6-JAK1-STAT3 feedback loop / 中国天然药物

Cervical cancer (CC) is recognized as the most common neoplasm in the female reproductive system worldwide. The lack of chemotherapeutic agents with outstanding effectiveness and safety severely compromises the anti-cipated prognosis of patients. Aloperine (ALO) is a natural quinolizidine alkaloid with marked anti-cancer effects on multiple malignancies as well as favorable activity in relieving inflammation, allergies and infection. However, its therapeutic efficacy and underlying mechanism in CC are still unclear. In the current study, MTT assay was employed to evaluate the viability of HeLa cells exposed to ALO to preliminarily estimate the effectiveness of ALO in CC. Then, the effects of ALO on the proliferation and apoptosis of HeLa cells were further investigated by plate colony formation and flow cytometry, respectively, while the migration and invasion of ALO-treated HeLa cells were evaluated using Transwell assay. Moreover, nude mice were subcutaneously inoculated with HeLa cells to demonstrate the anti-CC properties of ALO in vivo. The molecular mechanisms underlying these effects of ALO were evaluated by Western blot and immunohistochemical analysis. This study experimentally demonstrated that ALO inhibited the proliferation of HeLa cells via G2 phase cell cycle arrest. Simultaneously, ALO promoted an increase in the percentage of apoptotic HeLa cells by increasing the Bax/Bcl-2 ratio. Additionally, the migration and invasion of HeLa cells were attenuated by ALO treatment, which was considered to result from inhibition of epithelial-to-mesenchymal transition. For molecular mechanisms, the expression and activation of the IL-6-JAK1-STAT3 feedback loop were markedly suppressed by ALO treatment. This study indicated that ALO markedly suppresses the proliferation, migration and invasion and enhances the apoptosis of HeLa cells. In addition, these prominent anti-CC properties of ALO are associated with repression of the IL-6-JAK1-STAT3 feedback loop.

Association between circulating CD39+CD8+ T cells pre-chemoradiotherapy and prognosis in patients with nasopharyngeal carcinoma / 中华医学杂志(英文版)

BACKGROUND@#The mortality rate among patients with nasopharyngeal carcinoma (NPC) has improved significantly with the advent of chemoradiotherapy strategies. However, distant metastasis remains problematic. Tumor-specific reactivity in cancer patients has been detected exclusively in CD39+ T cells, particularly in CD39+CD103+ T cells. Circulating cancer-specific T cells are important for protecting against metastasis. This study aimed to evaluate the predictive value of circulating CD39+CD8+ T cells for metastasis in patients with NPC.@*METHODS@#We performed a cross-sectional, longitudinal study of 55 patients with newly diagnosed NPC of stage III-IVa. All patients were initially treated with standard combined chemoradiotherapy. Blood samples were obtained from 24 patients before and at 1 month and 6 months after treatment. T cell expression of CD39 and CD103, together with the markers of T cell exhaustion programmed death-1 (PD-1)/T cell immunoglobulin and mucin domain-containing protein 3 (Tim-3) and markers of cell differentiation CD27/CC-chemokine receptor 7/CD45RA, was examined by flow cytometry. The Wilcoxon rank-sum test analysis was used to analyze the differences between two groups. Kaplan-Meier analysis was used for analysis of progression-free survival (PFS).@*RESULTS@#The expression of circulating CD39+CD8+ and CD39+CD103+ CD8+ T cells was significantly higher in patients without distant metastasis (CD39+CD8+: 6.52% [1.24%, 12.58%] vs. 2.41% [0.58%, 5.31%], Z=-2.073, P=0.038 and CD39+CD103+CD8+: 0.72% [0.26%, 2.05%] vs. 0.26% [0.12%, 0.64%], Z=-2.313, P = 0.021). Most CD39+ T cells did not express PD-1 or Tim-3. Patients with high expression of CD39+CD103+CD8+ T cells had better PFS than patients with low expression (log rank value = 4.854, P = 0.028). CD39+CD8+ T cells were significantly elevated at 1-month post-treatment (10.02% [0.98%, 17.42%] vs. 5.91% [0.61%, 10.23%], Z = -2.943, P = 0.003). The percentage of advanced differentiated CD8+ T cells also increased at 1-month post-treatment compared with pre-treatment (33.10% [21.60%, 43.05%] vs. 21.00% [11.65%, 43.00%], Z = -2.155, P = 0.031). There was a significant correlation between elevated CD39+CD8+ T cells and increased effector memory T cells (intermediate stage: r = 0.469, P = 0.031; advanced stage: r = 0.508, P = 0.019).@*CONCLUSIONS@#CD39+CD8+ circulating T cells have preserved effector function, contributing to an improved prognosis and a reduced risk of metastasis among NPC patients. These cells may thus be a useful predictive marker for a better prognosis in patients with NPC.

MicroRNA-100-5p Regulates Melanogenesis in Alpaca Melanocytes by Targeting FGF21 / 中国生物化学与分子生物学报

Although many microRNAs (miRNAs) are known to function as regulators of coat color and melanogenesis, the underlying molecular mechanisms of miR-100-5p governing melanogenesis were not completely known.The goal of this study was to determine the effect of miR-l()()-5p on melanogenesis in alpaca melanocytes.Fibroblast growth factor 21 (FGF21) is a predicted target gene of miR-100-5p and the luciferase reporter assay demonstrated that miR-100-5p regulates FGF21 by binding to its 3' untranslated region (3'UTR).In this study, alpaca melanocytes were transfected with miR-100-5p, inhibitor and negative control plasmid.Results showed that miR-100-5p overexpression significantly decreased mRNA and protein expression of FGF2\.Meanwhile, the ERK signal pathway was inhibited, with subsequent up-regulation of microphthalmia-associated transcription factor (MITF) , tyrosinase (TYR) and tyrosinase-related protein 2 (TYRP2), which increased melanin production.The results suggest that miR-100-5p may regulate melanogenesis by targeting FGF21 via extracellular regulated MAP kinase (ERK) signaling pathway.

DKK3 Regulates Melanogenesis in Melanocytes of Alpaca / 中国生物化学与分子生物学报

Dickkopf-3 (DKK3) , as a critical inhibitor of the Wnt/p-catenin signaling pathway, may he involved in melanogenesis.In the current study, we investigated the effects of DKK3 on melanogenesis in melanocytes of alpaca.Overexpression of DKK3 in alpaca melanocytes, the expression of Wntl, Lefl , Myc and the major target genes termed microphthalmia-associated transcription factor (M1TF) and its downstream genes, including tyrosinase (TYR), tyrosinase-related protein 1 (TYRP1) and tyrosinase- related protein 2 (TYRP2) were significantly decreased at both mRNA and protein levels (P<0.05); total alkali melanin, pheomelanin and eumelanin were decreased by 80.30%, 72.17% and 64.60% (P <0.05), respectively.In contrast, in the melanocytes transfected with siRNA-DKK3 (a small interference RNA targeting DKK3) , the expression of Wntl, Lefl, Myc, MITF, TYR, TYRPl and TYRP2 were significantly increased at both mRNA and protein levels (P<0.05) ; total alkali melanin, pheomelanin and eumelanin were significantly increased by 1.65 folds, 1.25 folds and 1.21 folds (P< 0.05) , respectively.These results indicate that DKK3 regulates melanogenesis in alpaca melanocytes via the Wnt/p-catenin signaling pathway and down-regulates MITF.

SGOLl Promotes Proliferation and Invasion of Lung Cancer Cells Through PKM2 / 中国生物化学与分子生物学报

Shugoshin-1 (SGOLl) is one of the human homologues of yeast Shugoshin that locates in the centromeric region. It prevents premature division of the eentromerie cohesin complex and maintaining chromosome stability. Very recently, the role of SGOLl in tumors has emerged, but its role in lung cancer is unclear. In this study, we identify that SGOLl was upregulated in lung cancer samples from the TCGA database (n=529, P< 0.00001), which was correlated to the overall survival time of lung cancer patients (P = 0.0049). Detected by qRT-PCR and Western blotting, we demonstrate that SGOLl was consistently upregulated in lung cancer cell lines than normal epithelial cell line. Regulatory effects of SGOLl on cell viability, clonality, migration and invasion of lung cancer cells A549 and NCI-H2405 were examined by Cell Counting Kit-8 (CCK8), Colony formation, Scratch and Transwell assays, respectively. Compared with the control group, knockdown of SGOLl significantly inhibited proliferation, migration and invasion in A549 and NCI-H2405 cells (P<0. 05). A positive correlation was identified between expression levels of pyruvate kinase muscle isoenzyme 2 (PKM2) and SGOLl in the TCGA data-base (r=0. 38, P = 0). Western blotting results showed that knockdown of SGOLl in A549 and NCI-H2405 cells down-regulated PKM2 as well. In addition, co-interventions of SGOLl and PKM2 were performed in A549 and NCI-H2405 cells to clarify the interaction mechanism between them. The results showed that overexpression of PKM2 in lung cancer cells could partially reverse the regulatory effects of SGOLl knockdown on proliferation, migration and invasion (P<0. 05). Therefore, our study showed that SGOLl promoted the proliferation, migration and invasion of lung cancer cells by regulating PKM2. This study provided a theoretical basis for the mechanism of SGOLl in lung cancer.

TMEM106B Induces Melanogenesis by Regulating ERK/CREB Signaling Pathway / 中国生物化学与分子生物学报

The TMEM106B protein is a type-II transmembrane protein, which localizes in the endosome and lysosome of dendrites in primary neurons. TMEM106B is essential for maintaining and branching of dendrites, and thus regulates retrograde lysosomal trafficking of dendrites in primary neurons. Mammalian melanocytes are derived from neural cells, while melanosomes are originated from early endosome. However, the function of TMEM106B protein in melanocytes and its potential molecular mechanism in melanogenesis still remain unknown. Recently it was reported that transcription factor EB (TFEB) was the regulator of lysosome synthesis and TMEM106B protein overexpression promoted TFEB translocation into the nucleus. However, MITF (microphthalmia-associated transcription factor) and TFEB regulate each other in melanoma cells in vitro. Here in, plasmid containing gene for TMEM106B overexpression was transfected into melanocytes to investigate the regulation of TMEM106B on melanogenesis. The results showed that TMEM106B protein was localized in the cytoplasm of melanocytes. Compared with the negative control (NC), the mRNA levels of cyclic AMP-responsive element-binding protein (CREB) and MITF, especially CREB, were significantly increased in melanocytes with TMEM106B overexpression P< 0. 001). Western blot analysis showed that the expression of phosphorylated MAP kinase (p-ERK) was apparently increased (P<0.001) and resulted in the up-regulation of melanogenic regulatory proteins, including MITF, tyrosinase (TYR), tyrosinase-related protein-1 (TYRP1) and 2 (TYRP2). Masson-Fontana method showed that TMEM106B influenced the production of melanin in melanocytes. The spectrophotometry assay indicated that the amount of total melanin (ASM) (P<0. 001) and eumelanin (EM) (P<0. 05) were increased in alpaca melanocytes transfected with TMEM106B, while pheomelanin (PM) (P<0. 001) was decreased. These results demonstrated that TMEM106B played a vital role in melanogenesis in melanocytes by regulating ERK/CREB signaling pathway.

Effect of Kangquan Recipe () on BAMBI Expression in Hypothalamic-Pituitary-Prostate in Rats with Benign Prostatic Hyperplasia / 中国结合医学杂志

OBJECTIVE@#To investigate the effect of Kangquan Recipe (, KQR) on bone morphogenetic protein and activin membrane-bound inhibitor (BAMBI) expression and its mechanism in rats with benign prostatic hyperplasia (BPH).@*METHODS@#Forty-eight male Sprague-Dawley rats were divided into 6 groups using a random number table, with 8 in each group: the normal group (normal saline 10 mL/kg), the model group (normal saline 10 mL/kg), the finasteride group (0.5 mg/kg), the low-dose KQR group (3.5 g/kg), the middle-dose KQR group (7 g/kg), and the high-dose KQR group (14 g/kg). The 40 rats were subcutaneously injected with testosterone propionate after castration for 30 days to establish the BPH rat model except for those in the normal group. At the same time, the corresponding drugs were administered by gavage for 30 consecutive days. The effects of different doses of KQR on the protate wet weight, prostate volume and prostate index (PI) were observed. The changes in histopathology were monitored with hematoxylin-eosin staining. BAMBI protein and mRNA expression contents were determined by Western blot and quantitative real-time polymerase chain reaction, respectively.@*RESULTS@#All doses of KQR could decrease prostatic epithelial tissue proliferation. Compared to the model group, the high and middle-dose KQR significantly reduced prostate wet weight, prostate volume and PI; increased BAMBI protein expression in the hypothalamus, pituitary and prostate tissue; all doses of KQR up-regulated BAMBI mRNA expression in serum, prostatic fluid and prostate tissue (P<0.05 or P<0.01).@*CONCLUSIONS@#KQR could inhibit the proliferation of rat prostatic tissue, promote BAMBI protein expression in the hypothalamic-pituitary-prostate of rats with BPH; and increase BAMBI mRNA expression in the blood, prostatic fluid and prostate tissue of rats with BPH, showing a dose-effect relationship. KQR can be used as a potential drug for the treatment of BPH.

Characteristics of Electroencephalogram Power Spectrum Network on Uninjured Side of Post-stroke Aphasia / 中国康复理论与实践

Objective:To clarify the characteristics of electroencephalogram (EEG) power spectrum network of right hemisphere in patients with post-stroke aphasia (PSA) after left brain damaged. Methods:From December, 2018 to June, 2019, twelve PSA patients with left hemisphere injury were recruited and twelve healthy adults were matched as control group. China Rehabilitation Research Center aphasia examination (CRRCAE) was used to evaluated the linguistic function, and the EEG was collected. The functional connection characteristics of Alpha and Theta power spectrum were compared, and the correlation with language items was analyzed. Results:In PSA patients, the functional connection enhanced in Alpha and Theta frequency bands of central area, frontal, parietal and frontal-parietal joint areas, and decreased in Theta frequency bands of temporoparietal, parietal-occipital area, frontal, fronta-parietal and frontal central areas. The enhancement of alpha frequency functional connection from the right parietal occipital region to the central region was significantly correlated to the reduction of the expression ability (P < 0.05), while the weakening of theta frequency functional connection between the right parietal occipital region and the forehead and the center of the forehead was significantly correlated to the ability of speaking, reading, copying, dictation and calculation (r = -0.676~-0.717, P < 0.05). Conclusion:EEG power spectrum network can reflect the reorganization of right brain function network, and the change of right frontal parietal central network function connection may be closely related to PSA language injury and recovery.

Stimulation Mode and Model of Word-picture Language Task in Functional Magnetic Resonance Imaging Test for Post-stroke Aphasia (review) / 中国康复理论与实践

The cognitive neuroscience researches about post-stroke aphasia provide the interpretation of all aspects of linguistics. The word-picture research paradigm can be applied to assess different types of aphasia, in various ways of stimulation modes and models. It is more helpful combining functional magenetic resonance imaging to research the mechanism of brain damage and recovery objectively. The interactive application of language task and imaging has also become a new direction in the mechanism study of aphasia.

Functional Connectivities in Right Hemisphere for Post-stroke Aphasia: Based on Dual Stream Model / 中国康复理论与实践

Objective:To study the effect of right brain language network in post-stroke aphasia (PSA) patients with left hemisphere injury. Methods:From December, 2018 to June, 2019, twelve PSA patients with left hemisphere injury, and twelve matched healthy controls were recruited to accept rest-state functional magnetic resonance imaging (f-MRI) scan, and analyzed the characteristics of right brain function network with Dual Stream model. Results:There were two patients lost. Compared with the controls, for dorsolateral lingual pathway, the functional connections increased from superior marginal gyrus to middle frontal gyrus and inferior frontal gyrus of trigone in the patients, while those decreased from posterior central gyrus to inferior frontal gyrus of insula. For ventral lingual pathway, the functional connection increased from angular gyrus to orbital inferior frontal gyrus. For ventral and dorsolateral double-pathway, the functional connections increased from temporal lobe to lenticular pallidum and angular gyrus, from caudate nucleus to inferior frontal gyrus of insula, from lenticular putamen nucleus to middle frontal gyrus and trigonometry, while it decreased from superior marginal gyrus and temporal lobe to inferior frontal gyrus of insula. There was a negative correlation between the functional connection from inferior frontal gyrus to lenticular putamen and repeating (r = -0.720, P < 0.05), between the functional connection from inferior frontal gyrus to the caudate nucleus to speaking and repeating (r < -0.696, P < 0.05). In terms of network index, there were significant differences between the patients and the controls in both local and global indexes for language key brain area in right brain (|t| > 2.143, P < 0.05). Conclusion:The functional network has reorganized in right hemisphere of PSA patients. However, the increase of connection between language critical cortex and subcortical nuclei may play a role in improvement of language function.

Protective effect of bisoprolol against hypoxia/reoxygenation injury of cardiac microvascular endothelial cells / 医学研究生学报

Objective　To observe the protective effect of bisoprolol against hypoxia/reoxygenation injury of cardiac microvascular endothelial cells and explore the mechanism.　Methods　Left ventricular of cardiac microvascular endothelial cells (CMECs) were isolated from 8-week-old male C57BL/6N mice. CMECs were randomized into four groups: control group, vehicle group, hypoxia/reoxygenation group (H/R group), hypoxia/reoxygenation + bisoprolol group. The level of cell proliferation, apoptosis, superoxide anion, Cleaved caspase-3 and Nox2 expression were measured in each group.　Results　Compared with control group, H/R group had lower cell proliferation, higher apoptotic level, more superoxide anion level and the expression of Cleaved caspase-3 and Nox2 (P < 0.05). Furthermore, bisoprolol reversed hypoxia/reoxygenation-induced the decreased cell proliferation, the increased apoptosis, superoxide anion level, Cleaved caspase-3 and Nox2 expression (P < 0.05).　Conclusion　Bisoprolol can protect CMECs against hypoxia/reoxygenation injury by reducing the expression of Nox 2 that decreases oxidative stress.