[The non-bacterial pathogenic and clinical characteristics of acute respiratory tract infection in children in a hospital of pediatric in Sichuan Province from 2019 to 2021].

Objective: To explore the non-bacterial pathogen distribution, epidemiological characteristics, and clinical features of acute respiratory infections in children in Sichuan Province. Methods: Using a retrospective cohort study method, this study selected hospitalized children diagnosed with acute respiratory infections at West China Second Hospital of Sichuan University from February 2019 to January 2021, and tested 13 pathogens using polymerase chain reaction (PCR)-fragment analysis. The children were divided into infant group (<1 year old), toddler group (1 year old ≤ age <3 years old), preschool group (3 years old ≤ age <6 years old) and school-age group (6 years old ≤ age <18 years old). The distribution of pathogen positive rates, seasonal epidemic characteristics, clinical characteristics, and some laboratory test indicators were analyzed in children. Statistical analysis was performed on the results using SPSS 22.0 software, with count data expressed as percentages and inter group comparisons using SPSS 22.0 software χ2 Inspection. Results: A total of 2 922 pediatric patients were included in this study, with 1 748 (59.8%) positive for pathogens detected. Among them, 1 391 (79.6%) were detected as a single pathogen, and 357 (20.4%) were detected as a mixture of two or more pathogens. The most commonly detected pathogens were rhinovirus (HRV) (39.7%), syncytial virus (RSV) (22.8%), and parainfluenza virus (PIV) (12.5%). Pathogen positivity is more common in children under 6 years old (χ2=146.59, P<0.001), with a slightly higher positivity rate in male children (61.3%, 1 047/1 707) than in female children (57.7%, 701/1 215) (χ2=3.91, P=0.048), and compared with pathogen negative children, positive children are more prone to symptoms such as cough, wheezing, and shortness of breath (χ2=259.15, 366.06, 12.48, P<0.001). The distribution of different pathogens varies among children of different age groups, and HRV is more common in children aged 1-3 and 3-6 years old (χ2=9.74, P<0.001), while RSV is more common in children under 1 year old (χ2=178.63, P<0.001), while mycoplasma pneumoniae (MP) and influenza virus (InfA/B) are less common in children under 1 year old (χ2=92.54, 12.90,22.21, P<0.01). The prevalence of multiple pathogens showed seasonal changes. HRV showed a high prevalence trend in spring and autumn, while the prevalence of RSV infection was mainly seen in autumn and winter festivals. The positive rate of different pathogens after the outbreak of novel coronavirus pneumonia was significantly lower than that before the outbreak (χ2=252.68, P<0.001). Conclusion: The detection rate of non-bacterial respiratory pathogens in children in Sichuan Province from 2019 to 2021 is high, which is prone to symptoms such as cough, wheezing, and shortness of breath, with HRV and RSV being the main types. The positive rate of respiratory pathogens varies among different age groups, genders, and seasons.

A novel double-variant RHAG allele leads to Rhmod phenotype.

AIMS/OBJECTIVES: We aimed to analyse the molecular backgrounds and red blood cell (RBC) antigen expression of a male blood donor with Rhmod phenotype and his family members. BACKGROUND: Rh deficiency phenotypes are rarely found worldwide and are characterised by the lack of Rh antigen expression on RBCs. During routine screening, we found a blood donor who seemingly lacked Rh antigens. Therefore, we recruited the donor and his family for further investigation. METHODS: RBC serotyping and antibody screening/identification were performed for each sample. A routine blood examination was also conducted. RHD, RHCE and RHAG were sequenced at the genomic DNA or RNA level. Eleven antigens or proteins associated with Rh complex were tested using flow cytometry analysis. RESULTS: The proband and one of his brothers showed extremely weak D antigen and Rh expression levels but did not manifest anaemia. Most of the expressed RBC antigens of the two Rh-deficient individuals were similar to the previously reported cases but with some exceptions. Molecular analyses demonstrated homozygous expression of a novel RHAG allele, namely, c.[572G>A;707A>C], both in the proband and one of his brothers. CONCLUSIONS: To our knowledge, we identified the second double-variant RHAG allele and the first one related to Rhmod phenotype. The novel allele was also confirmed to be heritable by family analyses.

[Application value of whole brain 3D artery spin labeling in diagnosis of intracranial tumors].

Objective: To investigate the perfusion characteristics of arterial spin labeling (ASL) in intracranial tumor and its application value in classification. Methods: The clinical, pathological and imaging data of 44 patients with gliomas confirmed by pathology were analyzed retrospectively, including 9 low grade gliomas, 15 high grade gliomas, 11 cases of meningiomas, 6 cases of neurilemmoma, 3 cases of metastatic tumors.Conventional plain scan, 3D- ASL and MRI dynamic enhanced imaging (DSC-MRI) were performed.The mean maximal cerebral blood flow (CBF) of the solid component of tumor was obtained based on the region of interest.Immunohistochemical staining was performed in 24 patients with glioma.The differences of cerebral blood flow map (CBF) and relative cerebral blood flow (rCBF) in 44 patients with intracranial tumors were compared. The results of paired t test between the tumor area and the contralateral mirror area were measured by the two methods. Results: Taken the normal control-lateral grey matter(GM) as reference to normalize the CBF of tumor, three normalized tumor blood flow (nTBF) acquired by ASL showed statistical difference between low grade and high grade gliomas respectively (P<0.05). While taken the mirror region (M) and normal control-lateral white matter (WM) as reference to normalize the CBF of tumor, it showed no statistical difference (P>0.05). There was no 1p deletion in the cases of ASL perfusion in low-grade glioma group.In the case of 1p deletion in high grade glioma group, ASL was low perfusion, and there was no 1p deletion in the cases of ASL perfusion. Conclusion: 3D ASL can be used to identify high-grade and low-grade gliomas which has important reference value in the qualitative diagnosis of brain tumors and preoperative grading of gliomas.A separate use of 3D-ASL might cause over-or underestimation of tumor diagnosis, therefore a comprehensive analysis is needed.

[Expression and clinical significance of moesin and E-cadherin in invasive carcinoma of breast, no specific type].

OBJECTIVE: To investigate the correlation of moesin and E-cadherin with biological behavior of breast cancer and its mechanism by comparing expression of moesin and E-cadherin in breast invasive carcinoma of no specific type(BIC-NST), breast ductal carcinoma in situ(BDCIS) and normal breast tissues adjacent to carcinoma. METHODS: Breast cancer cases of the Huizhou Municipal Center People Hospital were collected between Jan 2008 and Dec 2010, expression of moesin and E-cadherin in 104 cases of BIC-NST, 84 cases of BDCIS and 53 cases of normal breast tissues adjacent to carcinoma were detected by tissue-microarray and SP immunohistochemical staining. Western blot was used to detect moesin expression of 16 BIC-NST fresh tissues. RESULTS: Expression rate of moesin in BIC-NST and BDCIS were significantly higher than normal tissues(P<0.01), but the expression rate of E-cadherin in BIC-NST and BDCIS were significantly lower than those of normal tissues(P<0.01). Expression rate of moesin in BIC-NST grade Ⅲ group was significantly higher than that of the grade Ⅰ group.There was a significantly positive correlation between histological grade and moesin expression(P<0.05). However, E-cadherin expression rate in BIC-NST grade Ⅲ group was significantly lower than that in grade Ⅰ group , and there was a significantly negative correlation between histological grade and E-cadherin expression(P<0.05). Moreover, no significant correlation was observed between moesin and E-cadherin expression in BDCIS tissues. Expression of moesin in clinical stage Ⅱ + Ⅲ BIC-NST was significantly higher than that in stage Ⅰ(P<0.01) . Expression of moesin was significantly associated with lymph node metastasis (P<0.01). But no significant correlation was observed between moesin expression and age, tumor size and vascular invasion . However, expression of E-cadherin in clinical stage Ⅱ+ Ⅲ BIC-NST was significantly lower than that in stage Ⅰ(P<0.01). Expression of E-cadherin was significantly associated with lymph node metastasis and vascular invasion (P<0.01). But no significant correlation was observed between E-cadherin expression, age and tumor size. There was a negative correlation between expression of moesin and E-cadherin in BIC-NST(P=0.021)and BDCIS(P=0.032). CONCLUSION: Higher moesin and lower E-cadherin signal transduction is closely related to the recurrence and development of breast carcinoma, therefore moesin and E-cadherin might provide new targets for gene therapy in breast carcinoma.

Revisiting the relationship between environmental and financial performance in Chinese industry.

The debate on the relationship between corporate or industrial environmental performance (EP) and financial performance (FP) has yet to be resolved, and studies need to examine the possible moderating effects on the EP-FP link. We argue that industrial EP has a positive effect on FP and that industrial munificence and resource slack can moderate the EP-FP link. Using a dataset from Chinese industrial firms, we examine the direct effect of industrial EP on FP and the indirect effects of industrial munificence and resource slack on the EP-FP link. Our results show that improving corporate or industrial-level EP significantly influences FP and that slack resources play a significant role on the EP-FP link. However, we found no significant moderating effect of industrial munificence on the link.

[Activation of transcription factor NF-κB in a rat model of cardiac fibrosis induced by ß-adrenoceptor stimulation].

OBJECTIVE: To establish a model of cardiac fibrosis induced by isoproterenol (ISO), the non-selective ß adrenoceptor agonist, injected subcutaneously for 7 days in rats, and to observe changes of transcription factor NF-κB in the model. METHODS: Male SD rats weighing 280-320 g were injected with ISO (0.25 mg/kg/d) subcutaneously for 7 days to induce cardiac fibrosis. The collagen volume fraction was determined by quantitative morphometry of picrosirius red stained left ventricular sections. Collagen types I/III and IL-6 mRNA expressions were analyzed by real time PCR. The pathological changes of the heart were investigated by Hematoxylin and Eosin staining. NF-κB was localized by immunohistochemistry (IHC) and phosphorylated NF-κB levels were assessed by Western blot analysis. RESULTS: Compared with the controls, ISO significantly elevated the sirius red stained area and collagen volume fraction (12.01±1.644 vs. 0.95±0.067, P<0.001). Similarly, ISO increased the mRNA expressions of collagen Iand collagen III of the heart compared with the controls (10.51±0.47 vs. 0.98±0.02,P<0.001 for collagen I; 9.58±1.33 vs. 1.02±0.02, P<0.001 for collagen III). The number of nuclei was increased and nuclear accumulation was presented in myocardial tissue induced by ISO. The mRNA expression of IL-6 increased in ISO group (1.64±0.18 vs. 1.04±0.07, P<0.01). ISO induced NF-κB nuclear translocation, accompanied by an increase in phosphorylation of NF-κB (10.83±2.05 vs. 1.05±0.27, P<0.001). CONCLUSION: We conclude that the model of cardiac fibrosis can be successfully induced by ISO injected subcutaneously for 7 days in rats and the activation of nuclear factor NF-κB increased by ß-adrenoceptor stimulation.

MiR-296-3p promotes the development and progression of preeclampsia via targeting the CEMIP.

OBJECTIVE: Preeclampsia (PE) is one of common pregnancy diseases, which has seriously threatened the health of the gravidas. Although upregulated miR-269-3p has been found in the placentas of the patients with PE, the regulation mechanisms of miR-296-3p remain unclear. PATIENTS AND METHODS: In this study, the placentas of the patients and normal gravidas were used to observe the difference in miR-296-3p expression level, and HTR-8/Svneo and JAR cells were used to investigate the role of miR296-3p in trophoblast cells. Besides, qRT-PCR, Western blot, CCK-8 assay, Dual-Luciferase reporter gene assay and transwell assay were used to explore the functions and regulation mechanisms of miR-296-3p on PE. RESULTS: The results showed that miR-296-3p was upregulated in the PE-placentas, and increased miR-296-3p could inhibit the proliferation, invasion and migration of HTR-8/Svneo and JAR cells. Besides, miR-296-3p could directly target the 3'-UTR of CEMIP, and the phenomena induced by increased miR-296-3p, including decreased ß-catenin and p-AKT and weakened proliferation, invasion and migration abilities, could be reversed by upregulating the expression level of CEMIP. CONCLUSIONS: To summarize, this study suggests that miR-296-3p inactivates the Wnt/ß-catenin and PI3K/AKT pathways to promote the progression of PE via targeting the CEMIP.

[Research on feasibility of in vitro inflammatory wound microenvironment simulated by using inflammatory wound tissue homogenate of mice].

Objective: To investigate the feasibility of in vitro inflammatory wound microenvironment simulated by using inflammatory wound tissue homogenate of mice. Methods: (1) Ten eight-week-old C57BL/6 male mice were collected and full-thickness skin tissue with diameter of 1.0 cm on both sides of the midline of the back was taken with a perforator to make the normal skin tissue homogenate supernatant. At 48 h after the full-thickness skin defect wound was established, the wound tissue within 2 mm from the wound edge was taken to make inflammatory wound tissue homogenate supernatant. Two kinds of tissue homogenate supernatant were taken to adjust the total protein concentration to 1 mg/mL, and the tumor necrosis factor α (TNF-α) content was detected by enzyme-linked immunosorbent assay. The number of sample was 6. (2) The primary passage of human umbilical cord mesenchymal stem cells (hUCMSCs) were collected and cultured to the 3rd passage with the normal exosomes being extracted from the hUCMSCs after cultured for 48 h. Another batch of hUCMSCs in the 3rd passage was collected and stimulated with inflammatory wound tissue homogenate supernatant of 30, 50, and 100 µg/mL total protein and normal skin tissue homogenate supernatant of 30, 50, and 100 µg/mL total protein, respectively. After cultured for 48 h, the exosomes stimulated with normal protein of 30, 50, and 100 µg/mL and exosomes stimulated with inflammatory protein of 30, 50, and 100 µg/mL were extracted. Normal exosomes, exosomes stimulated with 30 µg/mL normal protein, and exosomes stimulated with 30 µg/mL inflammatory protein were collected, the morphology was observed by transmission electron microscope, the particle size was detected by nanoparticle tracking analyzer, and the expressions of CD9 and CD63 were detected by Western blotting. (3) Twenty one-day-old C57BL/6 mice were taken to isolate the primary passage of fibroblasts (Fbs) and the 3rd passage of Fbs, whose morphology was observed under the inverted phase contrast microscope. The Fbs of 3rd passage were collected to observe the expression of vimentin by cell crawling method combined with immunofluorescence method at culture hour (CH) 2. (4) The Fbs of 3rd passage were divided into control group, normal exosome group, 30, 50, 100 µg/mL normal protein stimulating exosome group, and 30, 50, 100 µg/mL inflammatory protein stimulating exosome group according to the random number table, with 4 wells in each group. Cells in control group received no treatment, and cells in the other 7 groups were respectively added with normal exosomes, exosomes stimulated with normal protein of 30, 50, and 100 µg/mL, and exosomes stimulated with inflammatory protein of 30, 50, and 100 µg/mL prepared in experiment (2). The final mass concentration of exosomes was adjusted to 10 µg/mL. The cell viability was detected by cell count kit 8 at CH 48. (5) Two batches of Fbs in the 3rd passage were divided and treated as those in experiment (4), with 4 wells in each group, and the final mass concentration of exosomes was adjusted to 1 and 10 µg/mL, respectively. The cell mobility was detected by cell scratch test at CH 6, 12, and 24. (6) Two batches of the Fbs of 3rd passage were collected, divided, and treated as those in experiment (4) except with no control group, with 3 wells in each group, and the final mass concentration of exosomes was respectively adjusted to 1 and 10 µg/mL. The mRNA expression levels of transforming growth factor ß(1) (TGF-ß(1)), TGF-ß(3), and α smooth muscle actin (α-SMA) were detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction at CH 48. Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, and Bonferroni method. Results: (1) The content of TNF-α in inflammatory wound tissue homogenate supernatant of mice was (116±3) pg/mL, significantly higher than (97±5) pg/mL in normal skin tissue homogenate supernatant at post injury hour 48 (t=3.306, P<0.05). (2) Normal exosomes, exosomes stimulated with 30 µg/mL normal protein, and exosomes stimulated with 30 µg/mL inflammatory protein of hUCMSCs showed the typical saucer-like shape. The particle sizes of the three exosomes of hUCMSCs were 30-150 nm, which were all within the normal particle size range of exosome. Three exosomes of hUCMSCs positively expressed CD9 and CD63. (3) The primary passage of cells were clearly defined and showed protruding spindle shape, irregular polygon shape, or slender strip shape. The morphology of the 3rd and the primary passage of cells is similar. At CH 2, vimentin in cells was positively expressed, and the cells were identified as Fbs. (4) At CH 48, the cell viability was (137.4±2.8)% in 30 µg/mL inflammatory protein stimulating exosome group, obviously higher than 100%, (107.5±2.4)%, (113.3±3.2)%, (104.0±2.0)%, and (101.9±1.5)% in control group, normal exosome group, 30 µg/mL normal protein stimulating exosome group, and 50 and 100 µg/mL inflammatory protein stimulating exosome groups, respectively (P<0.01), and cell viability in 30 µg/mL normal protein stimulating exosome group was obviously higher than that in control group, normal exosome group, and 50 and 100 µg/mL normal protein stimulating exosome groups [(103.4±2.2)% and (102.5±1.4)%], respectively (P<0.01). (5) At CH 6, 12, and 24, the mobility rate of cells in 30 µg/mL inflammatory protein stimulating exosome group was significantly higher than that in control group, normal exosome group, 30 µg/mL normal protein stimulating exosome group, and 50 and 100 µg/mL inflammatory protein stimulating exosome groups, respectively, when the final mass concentrations of exosome was 1 µg/mL (P<0.05) . At CH 12, the mobility rate of cells in 30 µg/mL normal protein stimulating exosome group was obviously higher than that in control group, normal exosome group, and 50 and 100 µg/mL normal protein stimulating exosome groups, respectively, when the final mass concentration of exosome was 1 µg/mL (P<0.05). At CH 6, the mobility rate of cells in 30 µg/mL inflammatory protein stimulating exosome group was significantly higher than that in control group and normal exosome group (P<0.05), and the mobility rate of cells in 30 µg/mL normal protein stimulating exosome group was significantly higher than that in 50 and 100 µg/mL normal protein stimulating exosome groups, respectively, when the final mass concentration of exosome was 10 µg/mL (P<0.05). At CH 12 and 24, the mobility rate of cells in 30 µg/mL inflammatory protein stimulating exosome group was significantly higher than that in control group, normal exosome group, and 50 and 100 µg/mL inflammatory protein stimulating exosome groups (P<0.05), and the mobility rate of cells in 30 µg/mL normal protein stimulating exosome group was significantly higher than that in control group, normal exosome group, and 50 and 100 µg/mL normal protein stimulating exosome groups, respectively, when the final mass concentration of exosome was 10 µg/mL (P<0.05). (6) There were no statistically significant differences in mRNA expression levels of TGF-ß(1), TGF-ß(3), and α-SMA of cells among the 7 groups at CH 48 when the final mass concentration of exosome was 1 µg/mL (F=1.123, 1.537, 1.653, P>0.05). There were no statistically significant differences in mRNA expression levels of TGF-ß(1) and α-SMA of cells among the 7 groups at CH 48 when the final mass concentration of exosome was 10 µg/mL (F=1.487, 1.308, P>0.05), and mRNA expression level of TGF-ß(3) of cells in 50 µg/mL inflammatory protein stimulating exosome group at CH 48 was significantly higher than that in normal exosome group, 50 µg/mL normal protein stimulating exosome group, and 30 and 100 µg/mL inflammatory protein stimulating exosome groups when the final mass concentration of exosome was 10 µg/mL (P<0.05). Conclusions: The pretreatment with inflammatory wound tissue homogenate supernatant of mice has no significant effect on the total protein of hUCMSCs exosomes. The hUCMSCs exosomes stimulated by low concentration inflammatory wound tissue homogenate supernatant can significantly promote the proliferation and migration ability of Fbs. The content of inflammatory mediators in the wound tissue homogenate supernatant during the inflammatory phase is extremely low, which may be the reason that the anti-inflammation and tissue repair paracrine effects of mesenchymal stem cell cannot be effectively started.

[Advances in the research of effects of epithelial-mesenchymal interaction on wound healing and scar formation].

Wound healing is a dynamic process which involves interaction of various types of cells, cytokines, and extracellular matrix. Among them, epithelial cells and mesenchymal cells are the key components which involve in wound healing and scar formation. Related scholars had done a great number of studies about the functions of epithelial cells and fibroblasts(Fbs) in wound healing and scar formation. The results showed that under the stimulation of complex microenvironment, epithelial cells would lose their epithelial characteristics and acquire the typical characteristics and migration ability of mesenchymal cells. At the same time, with the complex changes of cell structure and cell behavior, they would participate in the process of tissue wound repair, including normal or fibrotic repair, by covering the wound with migration. Fbs are the key cells for the wound fibrotic repair, and play important roles in the process of wound healing, including excessive wound healing or delayed wound healing. In the recent years, the researchers realized that the cross-talk between epithelial cells and Fbs in wound healing, which is referred to as epithelial-mesenchymal interaction, significantly changes the biological behaviors of these two cell types, which affects the dermal remodeling and re-epithelialization quality of wound. Epithelial-mesenchymal interaction plays an important role in skin morphogenesis during embryonic development and maintaining the structural integrity of adult skin. In the process of re-epithelialization, Fbs could promote the proliferation and migration of keratinocytes, meanwhile keratinocytes would receive the signals from Fbs to reconstruct functional epithelium, which has become a hot topic in the field of wound healing at present. In this paper, a comprehensive analysis of the literature on the role of epithelial-mesenchymal interaction in wound healing and scar formation at home and abroad in recent years is presented for the reference of relevant scholars.

[Influence of human amniotic mesenchymal stem cells on macrophage phenotypes and inflammatory factors in full-thickness skin wounds of mice].

Objective: To explore the influence of human amniotic mesenchymal stem cells (hAMSCs) on the in vivo and in vitro regulation of macrophage phenotypes and inflammatory factors associated with wound healing of full-thickness skin wounds in mice. Methods: Fresh amniotic membrane discarded from full-term delivery by 5 healthy pregnant women in the Department of Obstetrics and Gynecology of the Affiliated Hospital of Zunyi Medical University was used for the isolation and culture of hAMSCs by enzyme digestion method. The third passage of cells was used for identification of adipogenic and osteogenic differentiation. The fourth passage of cells was used for identification of hAMSCs surface markers. Ten C57BL/6 mice (all male, aged 6 to 8 weeks, the same gender and age below) were selected for extracting mouse peritoneal macrophages by intraperitoneal lavage, and M1-type macrophages were induced by Dulbecco's modified eagle medium (DMEM) medium containing interferon-γ. The M1-type macrophages were divided into hAMSCs+ macrophage group and macrophage alone group. Then 1×10(4) hAMSCs/per well of fourth passage were added to macrophage in hAMSCs+ macrophage group and cultured in 2 mL DMEM medium for routine culture. In macrophage alone group, each well was only added with 2 mL DMEM medium for routine culture. On day 1 and 7 in culture, the content of interleukin-12 (IL-12), arginase 1, and IL-10 in the cell culture supernatant of the 2 groups were detected by enzyme-linked immunosorbent assay with sample number of 6/per group. (2) Full-thickness skin wound model was reproduced in the back of 56 C57BL/6 mice, which were divided into hAMSCs group and phosphate buffer solution (PBS) group using the random number table, with 28 mice in each group. Mice in hAMSCs group were subcutaneously injected with 100 µL of cell suspension containing 1×10(7) hAMSCs per mL in PBS suspension along the wound edge. While mice in PBS group were only subcutaneously injected with 100 µL PBS along the wound edge. On post injection day (PID) 1, 3, 7, and 14, 7 mice in the two groups were sacrificed respectively. Histopathological observation was performed with hematoxylin-eosin staining. The expressions of macrophage surface markers [CD68 and inducible nitric oxide synthase (iNOS) double positive cells and CD68 and arginase 1 double positive] in the wounds were detected by immunofluorescent staining. The mRNA expressions of IL-10, macrophage inflammatory protein 1α (MIP-1α), and MIP-2 in the wounds were detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction. Data were statistically analyzed with analysis of variance for factorial design, t test, and Bonferroni correction. Results: (1) On day 1 in culture, the content of IL-12 and arginase 1 in the cell culture supernatant of the two groups were similar (t=0.448, 0.536, P>0.05), and the content of IL-10 in the cell culture supernatant of hAMSCs+ macrophage group was significantly lower than that in macrophage alone group (t=14.722, P<0.01). On day 7 in culture, the content of IL-12 in the cell culture supernatant of hAMSCs+ macrophage group was significantly lower than that in macrophage alone group (t=13.226, P<0.01), and the content of arginase 1 and IL-10 was significantly higher than that in macrophage alone group (t=30.172, 31.406, P<0.01). (2) On PID 1, a large number of inflammatory cells infiltration were observed in the skin wounds of both groups. On PID 3, the inflammatory cells infiltration in the skin wounds increased in both groups, and the inflammatory cells infiltration in hAMSCs group was less than that in the PBS group. On PID 7, the inflammatory cells infiltration in the wounds decreased in both groups, and the inflammatory cells infiltration in hAMSCs group was less than that in the PBS group. On PID 14, no obvious inflammatory cells infiltration was observed in the wounds in the two groups. (3) On PID 1 and 14, the percentages of CD68 and iNOS double positive cells and CD68 and arginase 1 double positive cells in the wounds were similar in the two groups (t(1 d)=0.134, 0.693, t(14 d)=1.146, 2.585, P>0.05). On PID 3 and 7, the percentages of CD68 and iNOS double positive cells in the wounds in hAMSCs group were significantly lower than those of PBS group (t=6.396, 4.787, P<0.01), while the percentages of CD68 and arginase 1 double positive cells were significantly higher than those of PBS group (t=3.928, 4.473, P<0.01). (4) On PID 1, the mRNA expressions of IL-10 in the wounds of mice in the two groups were similar (t=2.005, P>0.05). On PID 3, 7, and 14, the mRNA expressions of IL-10 in the wounds of mice in hAMSCs group were significantly higher than those of PBS group (t=7.758, 124.355, 80.823, P<0.01). On PID 1, 3, 7, and 14, the mRNA expressions of MIP-1α and MIP-2 in the wounds of mice in hAMSCs group (0.341±0.212, 0.648±0.004, 0.611±0.106, 0.763±0.049, 1.377±0.099, 1.841±0.042, 1.181±0.035, 0.553±0.028) were significantly lower than those of PBS group (3.853±0.035, 6.914±0.163, 3.648±0.113, 2.250±0.046, 11.119±0.495, 8.634±0.092, 5.722±0.021, 4.862±0.036, t=43.198, 101.904, 51.845, 58.231, 51.074, 177.501, 291.752, 251.614, P<0.01). Conclusions: hAMSCs demonstrates biological effects of promoting the transformation of M1-type macrophages into M2-type macrophages in full-thickness skin wounds of mice. They can up-regulate the expression of anti-inflammatory and anti-fibrotic factor IL-10, and down-regulate the expression of important inflammation mediated factors MIP-1α and MIP-2.

A Novel Magnetoelectric Biomagnetic Susceptometer on Iron Level Detection with Mice Tissue in Vitro.

Iron plays a vital role in human body. Liver Iron Concentration (LIC) is directly correlated to total body iron and can be an important indicator to a variety of pathologies. Non-invasive methods to quantitatively assess tissue iron with low cost and high sensitivity have drawn vast interests and investments. Among various methods, the magnetoelectric (ME) sensor based biomagnetic liver susceptometer (BLS) is of great promise because it operates at room temperature but with the same principle as that of the well-developed SQUID (Superconducting Quantum Interference Device). Here, we report a magnetoelectric (ME) sensor based BLS system exploiting the recently developed PIN-PMN-PT piezoelectric single crystal. The newly developed ME BLS, which employs the horizontal scanning mechanism with a water bath interface to automatically eliminate the diamagnetic background of the tissues and irregular shape of torso, exhibits an overall sensitivity advancement (300X) to the sensor system previously reported. A linear correlation (R2 = 0.97) found between the system measurements and the biopsy data demonstrates the validity of the system. The ability to detect signals from only 3cc of mouse liver tissue samples suggests a high spatial resolution which could be used for finer scanning and enable magnetic distribution image and profiling.

Upgraded flowing liquid lithium limiter for improving Li coverage uniformity and erosion resistance in EAST device.

We report on design and technology improvements for a flowing liquid lithium (FLiLi) limiter inserted into auxiliary heated discharges in the experimental advanced superconducting tokamak device. In order to enhance Li coverage uniformity and erosion resistance, a new liquid Li distributor with homogenous channels was implemented. In addition, two independent electromagnetic pumps and a new horizontal capillary structure contributed to an improvement in the observed Li flow uniformity (from 30% in the previous FLiLi design to >80% in this FLiLi design). To improve limiter surface erosion resistance, hot isostatic press technology was applied, which improved the thermal contact between thin stainless steel protective layers covering the Cu heat sink. The thickness of the stainless steel layer was increased from 0.1 mm to 0.5 mm, which also helped macroscopic erosion resilience. Despite the high auxiliary heating power up to 4.5 MW, no Li bursts were recorded from FLiLi, underscoring the improved performance of this new design.

Regional activation in the rat brain during visceral stimulation detected by c-fos expression and fMRI.

AIM: The aim of the study was to determine and compare the areas of brain activated in response to colorectal distention (CRD) using functional magnetic resonance imaging (fMRI) and c-fos protein expression. METHODS: For fMRI study (3.0 T magnet), anaesthetized rats underwent phasic CRD, synchronized with fMRI acquisition. Stimulation consisted of eight cycles of balloon deflation (90 s) and inflation (30 s), at 40, 60 or 80 mmHg of pressure. For c-fos study two sets of experiments were performed on anaesthetized rats: comparing (A) brain activation in rats with the inserted colorectal balloon (n = 5), to the rats without the balloon (n = 5); and (B) rats with inserted balloon (n = 10), to the rats with inserted and distended balloon (n = 10). The pressure of 80 mmHg was applied for 2 h of 30 s inflation and 90 s deflation, alternating cycles. RESULTS: Functional MRI revealed significant activation in the amygdala, hypothalamus, thalamus, cerebellum and hippocampus. Significant increase in c-fos expression was observed in amygdala and thalamus in the first set of experiments, and hypothalamus and parabrachial nuclei in the second. CONCLUSION: The two methods are not interchangeable but appeared to be complementary: fMRI was more sensitive, whereas c-fos had much greater resolution.

A flowing liquid lithium limiter for the Experimental Advanced Superconducting Tokamak.

A program involving the extensive and systematic use of lithium (Li) as a "first," or plasma-facing, surface in Tokamak fusion research devices located at Institute of Plasma Physics, Chinese Academy of Sciences, was started in 2009. Many remarkable results have been obtained by the application of Li coatings in Experimental Advanced Superconducting Tokamak (EAST) and liquid Li limiters in the HT-7 Tokamak-both located at the institute. In furtherance of the lithium program, a flowing liquid lithium (FLiLi) limiter system has been designed and manufactured for EAST. The design of the FLiLi limiter is based on the concept of a thin flowing film which was previously tested in HT-7. Exploiting the capabilities of the existing material and plasma evaluation system on EAST, the limiter will be pre-wetted with Li and mechanically translated to the edge of EAST during plasma discharges. The limiter will employ a novel electro-magnetic pump which is designed to drive liquid Li flow from a collector at the bottom of limiter into a distributor at its top, and thus supply a continuously flowing liquid Li film to the wetted plasma-facing surface. This paper focuses on the major design elements of the FLiLi limiter. In addition, a simulation of incoming heat flux has shown that the distribution of heat flux on the limiter surface is acceptable for a future test of power extraction on EAST.

15N NMR studies of the conformation of E. coli dihydrofolate reductase in complex with folate or methotrexate.

We have employed 15N NMR to characterize the conformations of Escherichia coli dihydrofolate reductase (ECDHFR) in complex with [5-15N]folate or [5-15N]methotrexate (MTX). Two 15N resonances were observed for DHFR/MTX binary complex. The relative population of these two conformations is pH dependent. Addition of NADP+ or NADPH results in the disappearance of the low field resonance. In contrast, only one conformation was observed for both the DHFR/folate and DHFR/folate/NADP+ complexes. However, the 15N chemical shift of [5-15N]folate in the binary DHFR/folate complex is 7.28 ppm upfield from that of the ternary complex, suggesting the possible loss of a hydrogen bonding to N5 of folate in the ternary complex.

The conformations of trimethoprim/E. coli dihydrofolate reductase complexes. A 15N and 31P NMR study.

We have employed 15N and 31P NMR techniques to characterize the conformations of trimethoprim (TMP)/E. coli dihydrofolate reductase (DHFR) complexes in the presence and absence of NADPH and NADP+. A single conformation was observed for TMP/DHFR, NADP+/DHFR, NADPH/DHFR, and TMP/NADPH/DHFR complexes. In the ternary complex of TMP/NADP+/DHFR both the 15N and 31P spectra revealed the presence of two conformations. However, the conformations of TMP and NADP+ in the ternary complex may not be correlated, resulting in the possible existence of four conformations for the protein ternary complex.

Sympathetic nerve activity during prolonged rhythmic forearm exercise.

Exercise is a potent stimulus to activate the sympathetic nervous system. Previous work suggests that metabolite-sensitive muscle afferents are activated near the point of fatigue, and, when activated, they determine the muscle sympathetic nerve activity (MSNA) response to isometric forearm exercise. Yet, studies using a more prolonged rhythmic exercise paradigm suggest that the sympathetic nervous system can be activated in a more graded fashion. The purpose of this study was to determine whether metaboreceptor stimulation would also be responsible for MSNA responses to prolonged rhythmic isotonic forearm exercise. Subjects (n = 16) performed rhythmic isotonic forearm exercise at 25% maximal voluntary contraction for 30 min as we measured MSNA (microneurography). We observed progressive increases in MSNA with a peak increase of 161 units from a baseline value of 180 units. We also performed posthandgrip circulatory arrest (PHG-CA) in nine of these subjects. This maneuver isolates the metaboreceptor contribution to MSNA. During PHG-CA, delta MSNA values were not different from those observed during a freely perfused recovery period (n = 7). We also compared MSNA responses during the rhythmic paradigm with those seen during a static protocol at 40% of maximal voluntary contraction in five subjects. The two types of exercise caused similar increases in MSNA, but only the static paradigm was associated with a sustained MSNA response during PHG-CA. Finally, 31P-nuclear magnetic resonance was used to evaluate muscle metabolic responses during rhythmic and static forearm exercise (n = 6). Static exercise caused muscle acidosis and an increase in H2PO4-, whereas rhythmic exercise had no effect on muscle metabolism.(ABSTRACT TRUNCATED AT 250 WORDS)

Ischemic exercise and the muscle metaboreflex.

In exercising muscle, interstitial metabolites accumulate and stimulate muscle afferents. This evokes the muscle metaboreflex and raises arterial blood pressure (BP). In this report, we examined the effects of tension generation on muscle metabolites and BP during ischemic forearm exercise in humans. Heart rate (HR), BP, P(i), H(2)PO(4)(-), and pH ((31)P-NMR spectroscopy) data were collected in 10 normal healthy men (age 23 +/- 1 yr) during rhythmic handgrip exercise. After baseline measurements, the subjects performed rhythmic handgrip for 2 min. At 2 min, a 250-mmHg occlusion cuff was inflated, and ischemic handgrip exercise was continued until near fatigue (Borg 19). Measurements were continued for an additional 30 s of ischemia. This protocol was performed at 15, 30, 45, and 60% of the subjects' maximum voluntary contraction (MVC) in random order. As tension increased, the time to fatigue decreased. In addition, mean arterial pressure and HR were higher at 60% MVC than at any of the other lower tensions. The NMR data showed significantly greater increases in H(2)PO(4)(-), P(i), and H(+) at 60% than at 15 and 30% MVC. Therefore, despite the subjects working to the same perceived effort level, a greater reflex response (represented by BP and HR data) was elicited at 60% MVC than at any of the other ischemic tensions. These data are consistent with the hypothesis that, as tension increases, factors aside from insufficient blood flow contribute to the work effect on muscle metabolites and the magnitude of the reflex response.

Augmented sympathetic tone alters muscle metabolism with exercise: lack of evidence for functional sympatholysis.

It is unclear whether sympathetic tone opposes dilator influences in exercising skeletal muscle. We examined high levels of sympathetic tone, evoked by lower body negative pressure (LBNP, -60 mmHg) on intramuscular pH and phosphocreatine (PCr) levels (31P-nuclear magnetic resonance spectroscopy) during graded rhythmic handgrip (30 contractions/min; approximately 17, 34, 52 and 69% maximal voluntary contraction). Exercise was performed with LBNP and without LBNP (Control). At the end of exercise, LBNP caused lower levels of muscle pH (6.59 +/- 0.09) compared with Control (6.78 +/- 0.05; P < 0.05). PCr recovery, an index of mitochondrial respiration, was less during the recovery phase of the LBNP trial. Exercise mean arterial pressure was not altered by LBNP. The protocols were repeated with measurements of forearm blood flow velocity and deep venous samples (active forearm) of hemoglobin (Hb) saturation, pH, and lactate. With LBNP, mean blood velocity was reduced at rest, during exercise, and during recovery compared with Control (P < 0.05). Also, venous Hb saturation and pH levels during exercise and recovery were lower with LBNP and lactate was higher compared with Control (P < 0.05). We conclude that LBNP enhanced sympathetic tone and reduced oxygen transport. At high workloads, there was a greater reliance on nonoxidative metabolism. In other words, sympatholysis did not occur.

Altered mechanisms of sympathetic activation during rhythmic forearm exercise in heart failure.

In congestive heart failure (CHF), the mechanisms of exercise-induced sympathoexcitation are poorly defined. We compared the responses of sympathetic nerve activity directed to muscle (MSNA) and to skin (SSNA, peroneal microneurography) during rhythmic handgrip (RHG) at 25% of maximal voluntary contraction and during posthandgrip circulatory arrest (PHG-CA) in CHF patients with those of an age-matched control group. During RHG, the CHF patients fatigued prematurely. At end exercise, the increase in MSNA was similar in both groups (CHF patients, n = 12; controls, n = 10). However, during PHG-CA, in the controls MSNA returned to baseline, whereas it remained elevated in CHF patients (P < 0.05). Similarly, at end exercise, the increase in SSNA was comparable in both groups (CHF patients, n = 11; controls, n = 12), whereas SSNA remained elevated during PHG-CA in CHF patients but not in the controls (P < 0.05). In a separate control group (n = 6), even high-intensity static handgrip was not accompanied by sustained elevation of SSNA during PHG-CA. 31P-nuclear magnetic resonance spectroscopy during RHG demonstrated significant muscle acidosis and accumulation of inorganic phosphate in CHF patients (n = 7) but not in controls (n = 9). We conclude that in CHF patients rhythmic forearm exercise leads to premature fatigue and accumulation of muscle metabolites. The prominent PHG-CA response of MSNA and SSNA in CHF patients suggests activation of the muscle metaboreflex. Because, in contrast to controls, in CHF patients both MSNA and SSNA appear to be under muscle metaboreflex control, the mechanisms and distribution of sympathetic outflow during exercise appear to be different from normal.