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Cardiomyocyte loss and myocardial fibrosis are major determinants of myocardial infarction (MI) pathological changes. Mesenchymal stem cell (MSC)-derived exosomes (exos) and Danshen decoction (DSY) have been demonstrated to mediate cardiac repair following MI. BM-MSCs exos or BM-MSCsDSY exos were intramuscularly injected into post-MI rats. On the 7th, 14th and 28th days, serum CK, LDH, α-HBDH, ALT, and AST were measured and electrocardiogram changes were monitored to identify cardiac function; Triphenyltetrazolium chloride staining, Hematein&Eosin staining, Masson trichrome staining and Transmission Electron Microscope were adopted to analyze infarct area, cardiac morphology, histopathology, and fibrosis and cardiomyocyte ultrastructure; TUNEL assay, real-time PCR and western blot were performed to detect cardiomyocyte apoptosis and autophagy. As a result, BMMSCsDSY exos are superior to BM-MSCs-exos in improvement of cardiac function, morphology, histopathology and cardiomyocyte ultrastructure, as well as in reduction of infarction area and cardiac fibrosis by inhibiting apoptosis and promoting autophagy of cardiomyocytes., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

To investigate the dynamic homing process and characteristics of macrophages in different organs of immune-mediated aplastic anemia (AA) model mice. Macrophages in donor lymph nodes were sorted by magnetic beads and labeled with PKH67. After modeling according to the preparation method of the AA model, peripheral blood rountine analysis, bone marrow biopsy and HE staining results were analyzed to verify the modeling effect. On days 4, 8, and 12 of modeling, the bone marrow, spleen, and lymph node mononuclear cells were collected, and dynamic changes of PKH67-labeled macrophages in donor mice were analyzed by flow cytometry. In this study, dynamic changes in PKH67-labeled macrophages in the pathogenesis of AA model mice were explored. Macrophages in donor mice homed to the lymph nodes, expanding and differentiating in the lymph nodes, and finally transported to the bone marrow and spleen. Through proteomics mass spectrometry analysis, the related immune inflammatory response pathway of macrophages involved in the activation of the AA bone marrow microenvironment was preliminarily revealed, which provides a basis for the pathological macrophages involved in the pathogenesis of AA model mice.

Chemical constituents from the ethanol extract of Picrorhiza scrophulariiflora were isolated and purified by column chromatography. Their structures were identified by HR-MS, 1D and 2D-NMR, and their cytotoxicity was assessed by CCK-8 assay. Four compounds were isolated and identified as follows: 2β-D-glucosyloxy-3β,16α,20β-trihydroxy-9-methyl-19-norlanosterol-5,25-diene-22-one(1), 2β-D-glucosyloxy-3β,16α,20β-trihydroxy-9-methyl-19-norlanosta-5,24-diene-22-one(2), 25-acetoxy-2β-glucosyloxy-3β,16α,20β-trihydroxy-9-methyl-19-norlanosta-5-ene-22-one(3) and 25-acetoxy-2β-glucosyloxy-3β,16α,20β-trihydroxy-9-methyl-19-norlanosta-5,23-(E)-diene-22-one(4). Compound 1 represents a new cucurbitane glycoside. The half inhibitory concentrations of the 4 compounds exceeded 100 μmol·L~(-1) against four tumor cell lines, indicating no significant cytotoxicity.

Objective: To study the anti-fibrotic effect of ghrelin on high-fat diet-induced non-alcoholic steatohepatitis (NASH) in mice. Methods: 24 male C57BL/6 mice were randomly divided into a normal diet group, a normal diet + ghrelin group, a high-fat diet group, and the high-fat diet + ghrelin group. The HFD and HFD+ghrelin groups were fed high-fat diet for 16 weeks to induce non-alcoholic steatohepatitis. Among them, the NCD+ghrelin group and HFD+ghrelin group were continuously given ghrelin intervention (11nmol·kg(-1)·d(-1)) for 2 weeks after feeding for 14 weeks. 16 mice were euthanized on weekends. The plasma levels of alanine aminotransferase (ALT) and hyaluronic acid (HA) were measured in mice. The content of hydroxyproline (Hyp) was determined in liver tissue. RT-qPCR was used to detect the mRNA expression levels of transforming growth factor β1 (TGF-β1) and collagen types I, III, and IV in liver tissue. A Western blot was used to detect the expression level of the α-smooth muscle actin (α-SMA) protein in liver tissue. HE staining was used to observe the morphological changes in liver tissue. VG staining was used to observe the fibrotic condition in liver tissue. Results: Compared with the NCD group, plasma ALT (266.80±146.80)U/L, HA (219.00±39.47) ng/ml levels, Hyp content (0.35±0.05)μg/mg prot ( P < 0.05), mRNA expression levels of transforming growth factor β1 (TGF-β1) and collagen types I, III, IV ( P < 0.05), and the expression level of α-SMA protein in the HFD group were significantly increased ( P < 0.05), with congestion in the hepatic central lobular veins, hepatocytes swelling, and deposition of a large amount of collagen fibers in liver tissue. Compared with the HFD group, plasma ALT (57.17±20.88)U/L, HA (75.68±8.40)μg/mg levels, Hyp content (0.19±0.07)μg/mg prot, mRNA expression levels of transforming growth factor β1 (TGF-β1) and collagen types I, III, IV ( P < 0.05), and the expression level of α-SMA protein in the HFD+ghrelin mice group was significantly reduced ( P < 0.05), with only mild sinusoidal congestion in the liver tissue but significant improvement and reduction in liver injury and collagen fiber deposition. Conclusion: Ghrelin has a significant improvement effect on liver fibrosis in NASH mice.

To characterize the chemical constituents of Huanbei Zhike Prescription by ultra-high performance liquid chromatography-time of flight mass spectrometry( UPLC-Q-TOF-MS/MS). A Thermo Syncronls C18 column( 2. 1 mm×100 mm,1. 7 μm) was used with methanol( A)-0. 1% formic acid solution( B) as the mobile phase for gradient elution. The injection volume was 2 μL; the column temperature was 40 ℃; the flow rate was 0. 3 m L·min-1; and electrospray ionization( ESI) source was used to collect data in positive and negative ion modes. The ion scanning range was m/z 50-1 200,with capillary voltage of 3 000 V,ion source temperature of100 ℃,atomization gas flow rate of 50 L·h-1,desolvent gas flow rate of 800 L·h-1,desolvent temperature of 400 ℃,cone hole voltage of 40 V,with argon as the collision gas and the collision energy was 20-35 V. The excimer ion peak information was analyzed by Waters UNIFI data processing software. The molecular formula with error within 1×10-5 was compared with the data in database to identify the compounds. The secondary fragment ion information of the target compound was selected,and then compared with the retention time and fragmentation patterns provided by the database and the existing literature to further confirm the compositions and structures of the compounds. A total of 68 main compounds in Huanbei Zhike Prescription were identified,including 38 flavonoids,10 organic acids,6 terpenoids and 10 nitrogen-containing compounds,of which 12 compounds were verified by the control substances. This method is rapid and accurate,which provides a new strategy for the qualitative analysis of the chemical constituents of Huanbei Zhike Prescription,and lays a foundation for the further study and quality control of the compound pharmacodynamic substance.

Background: Melasma is the most common and distressing pigmentary disorder presenting to dermatology clinics. Various treatment protocols for melasma have been suggested in the previous literature and applied in various clinical settings. However, no satisfactory therapy has been widely accepted., Objective: To evaluate the efficiency and safety of a combination treatment with fractional Q-switched ruby laser (QSRL) and intense pulsed light (IPL) for melasma in Chinese population., Methods: Fifty-three Chinese melasma patients were enrolled in this study. Each patient underwent 2 courses of treatments at 2-week interval. One course was composed of 3 successive sessions of 694-nm fractional QSRL at intervals of two weeks followed by one IPL. The efficacy was evaluated by non-invasive measurements and subjective assessments. The adverse effects were recorded., Results: Mean melanin index (MI) and erythema index (EI) significantly decreased from 216.1 and 381.8 pre-treatment to 167.8 and 310.3 post-treatment, respectively. Mean melasma area and severity index (MASI) decreased dramatically from 14.66 before treatment to 5.70 after the final treatment. These values remained at low levels at 3-month follow-up. The percentage of patients who achieved moderate or significant improvements was 73.6%. Adverse effects of QSRL and IPL were minimal., Conclusion: The combination treatment of fractional QSRL and IPL would be a promising modality for managing melasma in Chinese patients.

In this study, five common buckwheats and nine tartary buckwheats grown at different locations were analyzed for the contents of rutin, quercetin, and amino acids by high-performance liquid chromatography and spectrophotometry. The rutin content was higher than quercetin in buckwheat seeds. Rutin content was in the range from 0.05 (0.05 g per 100 g dry seeds) to 1.35% of buckwheat seeds. Quercetin content varied from 0.01 to 0.17% and in some common buckwheats it was even difficult to detect. Comparatively, tartary buckwheat seeds contained more rutin and quercetin than common buckwheat seeds. Meanwhile, the bran has higher rutin content than the farina in tartary buckwheat seeds, with a respective content of 0.45 to 1.19% and 0.14 to 0.67%. It was found that amino acid contents were around 1.79 to 12.65% (farina) and 5.74 to 7.89% (bran) in common buckwheats, and 1.73 to 5.63% (farina) and 2.64 to 16.78% (bran) in tartary buckwheat seeds. The highest total rutin content was found to be 1.35% in tartary buckwheat seeds from Sichuan, China. The highest total amounts of amino acid were detected to be 20.13% in tartary buckwheat seeds from Changzhi, Shanxi Province (China). Our results suggested that food products made of whole-buckwheat flour are healthier than those made of fine white flour.

Background: Although pulsed dye laser (PDL) has long been regarded as the gold standard in treating port-wine stain (PWS), advanced PWS with deeper coloration may display resistance because of limited penetration depth of 585 or 595-nm light. Recently, a dual-wavelength laser system has been reported to achieve pronounced fading in many patients., Objective: The objective was to evaluate the efficacy and safety of a dual-wavelength laser device in treatment of neck and facial PWS in a direct side-by-side comparison., Methods: Sixteen Chinese patients with neck and/or facial PWSs were enrolled in the study. All lesions were randomly divided into two area, treated area and adjacent untreated area. Five successive treatments using a dual-wavelength laser system (595-nm PDL combined with 1,064-nm Nd:YAG laser) were delivered on treated areas at 4- to 6-week intervals. The adjacent area was not treated as self control. Two blinded dermatologists evaluated the clinical changes by comparing the before and after photos. Erythema index (EI) values were measured with a non-invasive instrument., Results: After five sessions of treatment, over 62.5% (10/16) patients achieved more than 50% (moderate or significant) improvement. The efficacy maintained at the 3-month follow-up visit. The values of EI on treated area showed a significant decrease. Adverse effects of treated area were limited., Conclusion: Using this split-face module, the dual-wavelength laser system is proved to be effective and well tolerated in treating neck and facial PWSs in Chinese patients. Adverse effects were minimal and acceptable.

Objective: To explore the protective effect of baicalin against rotenone-induced injury on PC12 cells, and the po-tential mechanism of action action was also explored., Method: PC12 cells were injured by rotenone and were treated with different concentrations (0.1, 1, 10 μmol x L(-1)) of baicalin at the same time. Cell viability was analyzed by MTT, and morphology was observed by phase-contrast microscopy. The cell apoptosis was detected by flow cytometry by Annexin V-FITC/PI staining. The intracellular ROS level was determined by fluorescence microscope with DCF-DA staining. The expression of Bcl-2, Bax and Caspase-3 was analyzed by Western blot., Result: The viability of PC12 cells exposure to rotenone for 24 hour was gradually decreased with dose escalating and 1.5 μmol x L was adopted to do the following experiment. Baicalin increased cell viability, improved cell morphology and decreased intracellular ROS level. Moreover, FACS indicated baicalin attenuated the apoptosis induced by rotenone significantly. Western blot showed that Bcl-2, Bax and Caspase-3 expression in rotenone-induced PC12 cells was reversed by baicalin., Conclusion: This study has demonstrated that baicalin protects PC12 cells against rotenone-induced apoptosis, at least in part, by scavenging excessive ROS and inhibiting the mitochondrion-dependent apoptotic pathway.

This study was purposed to investigate the expression of miR-16 in T lymphoblastic lymphoma/acute lymphoblastic leukemia (T-LBL/ALL) and its relation with target therapy and prognosis. The CD3, cCD3, CD10, CD20, CD34, CD43, CD99, TdT, PAX-5, BCL-2 and Ki67 in paraffin samples from 38 cases of T-LBL/ALL were detected by immunohistochemical labeling; the miR-16 expression level was detected by real-time RT-PCR. Fifteen cases of reactive hyperplasia of lymph nodes were selected as control. The results indicated that among 38 cases of T-LBL/ALL the positive rate of TdT was highest (94.7%), the positive rate of CD34 was lowest (22.1%), the PAX-5 and CD20 were found to be negative. The Ki67 expression level in 39.5% cases exceeded 80%. As compared with reactive hyperplasia of lymph node, the miR-16 expression in T-LBL/ALL was up-regulated, ant its expression level was 4.87-fold of reactive hyperplasia of lymph node (P < 0.05). The overall survival rate in group of miR-16 high expression decreased (P < 0.05). The prognosis of T-LBL/ALL patients with BCL-2 positive expression was better than that of patients with BCL-2 negative expression (P < 0.05). The miR-16 expression correlated with BCL-2 protein (r = 0.51, P < 0.05). It is concluded that the overall survival rate in miR-16 high expression group is higher than that in miR-16 low expression group, suggesting possible relation of miR-16 with prognosis. Moreover, the prognosis in BCL-2 positive expression group is better than that in negative expression group, which may be a factor influencing prognosis.

This study was aimed to investigate the expression of blood Th17 and CD4(+) CD25(+) regulatory T cells (Treg) in the patients with aplastic anemia (AA). Forty-five patients with AA were enrolled into this study, and were divided into mild aplastic anemia (MAA) group (n = 25) and severe aplastic anemia group (SAA) (n = 20), blood cell count was recorded. 15 healthy donors were enrolled as control. Proportions of blood Th17 and CD4(+) CD25(+) Treg cells were determined by flow cytometry. The serum levels of IL-17, IFN-γ and TNF-α, as well as their concentrations in culture supernatant of phytohemagglutinin (PHA) -stimulated peripheral blood mononuclear cells, were measured by ELISA. The results showed that the proportions of blood Th17 cells and concentration of blood serum IL-17 and IFN-γ increased in patients with SAA, compared with MAA and normal controls, but CD4(+) CD25(+) Foxp3(+) Treg cells obviously decreased in patients with SAA. The concentrations of IL-17 and IFN-γ significantly increased in culture supernatant of SAA group. Hemoglobin level in the patients with AA negatively correlated with the population of Th17 cells and serum IL-17 level, whereas positively correlated with the expression of CD4(+) CD25(+)Treg cells. It is concluded that the increased response of Th17 cells and deficiency of CD4(+) CD25(+) Treg cells present in severe aplastic anemia. The severity of anemia may be related with the imbalance between Th17 and CD4(+) CD25(+)Treg cell response.

Objective: To investigate the effects of activated AKT on murine myeloid precursor cells (32D cells), and the effects of IFN-γ on 32D cells and its mechanisms., Methods: Plasmid transduction was used to enhance the expression of AKT on 32D cells. After the transfected cells treated with IFN-γ for 24 hours, proliferation rate was tested by WST-1, apoptosis by flow cytometry, expression of phosphorylated Erk1/2, Stat3 and phosphorylated Stat3 was determined by Western blot., Results: (1) IFN-γ at low concentration (100 U/ml) enhanced the growth and proliferation of 32D cells, while at high concentration (1000 U/ml) suppressed them. (2) Compared with control groups, low concentration IFN-γ increased (1124 ± 13) Stat3 phosphorylation in 32D-cell, while it high concentration IFN-γ decreased (601 ± 13). 32D cells transfected with activated Akt grew rapidly (0.287 ± 0.010) and had a low apoptotic rate [(9.57 ± 0.17)% (P < 0.05)]. (3) The expression of p-Erk1/2 in transfected 32D-cell was significantly reduced (P < 0.05). (4) Apoptosis rate of IFN-γ treated group was significantly decreased in transfected 32D cells (P < 0.05)., Conclusions: IFN-γ has dual effects on 32D cells, namely, at low concentration enhanced the growth and proliferation of 32D cells, while at high concentration suppressed them. Its mechanisims is possibly through Stat3 pathway. Activated Akt can significantly promote the growth and proliferation of 32D cell and significantly inhibit apoptosis and IFN-γ can regulate cell proliferation and apoptosis through AKT. AKT activation can inhibit the Erk signal pathway, which may be affected by inhibition the modificaton of Raf1.

By using static chamber-gas chromatographic techniques, the CH4 and N2O emissions from the paddy soil in southeast Hubei were measured. Four treatments were installed, i.e., no-tillage plus no-fertilization (NT0), conventional tillage plus no-fertilization (CT0), no-tillage plus fertilization (NTC), and conventional tillage plus fertilization (CTC). In all treatments, the CH4 emission had a seasonal variation of increasing first and decreasing then, while the N2O emission had no significant seasonal variation. Fertilization increased the CH4 and N2O emissions significantly. NT0 increased the CH4 emission and decreased the N2O emission significantly, compared with CT0; NTC only decreased the CH4 emission and increased the N2O emission slightly, compared with CTC. The analysis on the integrated greenhouse effect of CH4 and N2O showed that NT0 increased the effect by 25.9%, compared with CT0, while NTC decreased the effect by 10.1%, compared with CTC. Therefore, a reasonable arrangement of fertilization and no-tillage could reduce the integrated greenhouse effect of CH4 and N2O from paddy field.