Counteractive effect of mouse dermal fibroblasts during their adipogenic differentiation against Staphylococcus aureus infection and its mechanisms / 中华皮肤科杂志

Objective:To investigate the counteractive effect of mouse dermal fibroblasts (MdFBs) during their adipogenic differentiation against Staphylococcus aureus infection, and to explore its mechanisms. Methods:MdFBs were obtained from newborn C57BL/6 mice, and their adipogenic differentiation was induced by culture in an adipogenic medium for 48 hours. Real-time fluorescence-based quantitative PCR (RT-PCR) was performed to determine the mRNA expression of cathelicidin antimicrobial peptide (CAMP) on days 0-6 during the adipogenic differentiation of MdFBs, and Western blot analysis to determine the protein expression of CAMP in the culture supernatant of MdFBs during their adipogenic differentiation. MdFBs were divided into 4 groups: co-stimulation group stimulated by S. aureus suspensions and cultured in an adipogenic medium, adipogenic control group cultured in an adipogenic medium, S. aureus-stimulation group stimulated by S. aureus suspensions and cultured in a common medium, and control group stimulated by phosphate-buffered saline and cultured in a common medium; Western blot analysis and RT-PCR were conducted to determine the protein and mRNA expression of CAMP. S. aureus (5 × 10 4 CFU/ml) was cultured with the culture supernatant of MdFBs after 5-day adipogenic differentiation (adipogenic group), and the growth activity was evaluated every 2 hours during 10 - 24 hours after the start of co-culture; S. aureus cultured with the culture supernatant of MdFBs in a common medium served as the normal control group, and that cultured with cell-free culture supernatant served as the negative control group. Differences between groups were assessed using unpaired t-test or analysis of variance. Results:Significant differences were observed in the relative mRNA expression of CAMP among different time points (days 0, 1, 2, 4, and 6) during the adipogenic differentiation of MdFBs (1.14 ± 0.74, 68.04 ± 12.72, 683.12 ± 38.06, 1 390.68 ± 226.21, 454.57 ± 204.12, F = 50.08, P < 0.001) ; the CAMP mRNA expression was significantly higher on days 1, 2, 4, and 6 than on day 0 ( t = 9.09, 31.03, 10.63, 3.85, respectively, all P < 0.05), and showed an initial rise and subsequent fall during days 0 - 6. The CAMP protein expression in the culture supernatant of MdFBs peaked on days 2-5 and subsequently decreased. Significant differences were observed in the mRNA and protein expression of CAMP among the control group, S. aureus-stimulation group, adipogenic control group and co-stimulation group (mRNA: 0.08 ± 0.02, 0.38 ± 0.10, 0.49 ± 0.11, 0.80 ± 0.03, respectively, F = 43.25, P < 0.05; protein: 0.433 ± 0.176, 0.574 ± 0.176, 1.007 ± 0.176, 1.217 ± 0.176, respectively, F = 46.79, P < 0.05), and the relative mRNA and protein expression of CAMP was significantly higher in the co-stimulation group than in the adipogenic control group, S. aureus-stimulation group and control group (all P < 0.05). At 10 hours during culture, the growth activity of S. aureus was significantly lower in the adipogenic group (0.053 ± 0.015) than in the normal control group and negative control group (0.109 ± 0.015, 0.106 ± 0.015, t = 11.30, 13.26, respectively, both P < 0.05) ; during 10 - 24 hours, the growth activity of S. aureus also showed a significant decrease in the adipogenic group compared with the normal control group and negative control group (all P < 0.05) . Conclusion:MdFBs secreted CAMP during the adipogenic differentiation, and could inhibit the proliferation of S. aureus.

A preliminary study on Candida albicans-induced pyroptosis of murine bone marrow-derived macrophages / 中华皮肤科杂志

Objective:To investigate the effect of Candida albicans ( C. albicans) on pyroptosis of murine bone marrow-derived macrophages (BMDMs) . Methods:Live-cell imaging was used to observe morphologic changes of in vitro C. albicans-infected BMDMs (multiplicity of infection [MOI] = 50) so as to evaluate whether pyroptosis occurred. Cultured BMDMs were divided into a control group and a C. albicans group, which were treated with phosphate-buffered saline and C. albicans suspensions respectively for 6 hours; then, real-time fluorescence-based quantitative PCR was performed to determine the mRNA expression of NOD-like receptor pyrin domain containing 3 (NLRP3), interleukin (IL) -1β and IL-18, and Western blot analysis to determine the protein expression and cleavage levels of NLRP3, caspase-1 and gasdermin D (GSDMD). BMDMs were cultured with C. albicans suspensions for different durations (0, 10, 15, 20, and 25 hours), and enzyme-linked immunosorbent assay was conducted to detect secretion levels of IL-1β and IL-18. Cultured wild-type BMDMs and GSDMD-knockout BMDMs were treated with C. albicans suspensions for 15 minutes, and then rates of phagocytosis of C. albicans by wild-type BMDMs and GSDMD-knockout BMDMs were estimated by flow cytometry; after 6-hour treatment with C. albicans, flow cytometry and lactate dehydrogenase (LDH) release assay were performed to assess mortality rates of wild-type BMDMs and GSDMD-knockout BMDMs. In addition, some wild-type BMDMs and GSDMD-knockout BMDMs were separately divided into blank control group, control group, maximum enzyme activity-sample control group, IL-1β alone group, C. albicans alone group, and IL-1β + C. albicans group, and cell mortality rates were detected by the LDH release assay after treatment with IL-1β and/or C. albicans. Statistical analysis was carried out by using unpaired t test, Kruskal-Wallis test, analysis of variance, and other statistical methods. Results:After in vitro treatment with C. albicans, swelling and ballooning with large bubbles blowing from the plasma membrane occurred in BMDMs, suggesting the occurrence of cell pyroptosis; compared with the control group, the C. albicans group showed significantly increased mRNA expression levels of NLRP3 and IL-1β after 6-hour treatment with C. albicans ( t = 13.02, 17.51, respectively, P = or < 0.001), but no significant change in the IL-18 mRNA expression level ( P = 0.486), and Western blot analysis showed that C. albicans could increase the expression of NLRP3 inflammasomes, as well as cleaved caspase-1 and GSDMD. After the treatment with C. albicans for different durations (0, 10, 15, 20, and 25 hours), the secretion level of IL-1β by BMDMs gradually increased over time ( H = 12.90, P = 0.012), while the secretion level of IL-18 did not significantly change ( F = 0.48, P = 0.753), and the secretion level of IL-1β was significantly lower in the GSDMD-knockout BMDM group than in the wild-type BMDM group ( F = 24.22, P = 0.008). After 15-minute in vitro treatment with C. albicans, the phagocytosis rate of C. albicans was significantly lower in the GSDMD-knockout BMDM group (50.3% ± 1.10%) than in the wild-type BMDM group (58.53% ± 1.19%, t = 5.09, P = 0.007) ; after 6-hour treatment with C. albicans, the cell mortality rate was significantly higher in the GSDMD-knockout BMDM group than in the wild-type BMDM group (flow cytometry: 38.40% ± 0.50% vs. 34.37% ± 0.52%, t = 4.72, P = 0.009; LDH release assay: 22.52% ± 0.18% vs. 12.48% ± 0.15%, t = 42.36, P < 0.001) ; the cell mortality rates of wild-type BMDMs and GSDMD-knockout BMDMs both significantly decreased in the IL-1β + C. albicans groups compared with the C. albicans groups (both P < 0.001) . Conclusion:Pyroptosis could be induced in murine BMDMs after C. albicans infection, which promotes the release of IL-1β and may reduce the mortality rate of macrophages by improving their immune activity.

Cannabinoid alleviates anxiety- and depression-like behaviors in mice via inhibiting microglia activation / 中华微生物学和免疫学杂志

Objective:To investigate the effects and mechanism of cannabinoid (CBD) on the anxiety- and depression-like behaviors induced by lipopolysaccharide (LPS) in mice.Methods:C57BL/6J mice were intraperitoneally injected with LPS to establish the model of neuroinflammation. CBD was injected intraperitoneally 24 h after modeling. Behavioral tests were performed to evaluate the anxiety- and depression-like behaviors in mice. CBD-pretreated BV-2 microglia cells were stimulated with LPS in vitro. The levels of tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β) and CD86 in mouse cerebral cortex, hippocampus, prefrontal cortex and BV-2 cells were measured by qRT-PCR. The protein level of nuclear factor (NF-κB) in mouse brain and BV-2 cells was determined by Western blot. Results:CBD significantly increased the residence time and movement distance of LPS-treated mice in the central area in the open filed test (OFT), and reduced the immobility time in tail suspension test (TST) and force swimming test (FST). In addition, CBD alleviated the neuroinflammation and inhibited the activation of microglia in mouse brain. In vitro, CBD significantly inhibited the activation of BV-2 microglia cells. Both in vivo and in vitro experiments confirmed that CBD could inhibit NF-κB expression. Conclusions:LPS could induce the activation of BV-2 microglia cells and the expression of inflammatory factors in mouse brain accompanied with abnormal behaviors. CBD could inhibit the activation of microglia, alleviate the neuroinflammation in different regions of mouse brain and improve behavioral performance.

Radiotherapy is beneficial for long-term survival of patients with stage Ⅳ B thoracic esophageal squamous cell carcinoma at initial diagnosis / 中华放射肿瘤学杂志

Objective:To investigate the value of radiotherapy in patients with stage Ⅳ B thoracic esophageal squamous cell carcinoma (ESCC) at initial diagnosis. Methods:A total of 199 patients with stage Ⅳ B thoracic ESCC at initial diagnosis (according to UICC/AJCC Eighth Edition Esophageal and Esophagogastric Junction Cancer TNM Staging) who were treated in the Fourth Hospital of Hebei Medical University between January 2010 to December 2016 were recruited. Winthin the whole group, 130 patients (65.3%) had distant lymph node metastases alone, 51 cases (25.6%) of solid organ metastases alone and 18 cases (9.0%) of solid organ complicated with distant lymph node metastases. Among them, 16 patients (8.0%) were treated with chemotherapy alone, 50 cases (25.1%) of radiotherapy alone, 133 cases (66.8%) of radiochemotherapy (81 patients treated with concurrent radiochemotherapy and 52 patients treated with sequential radiochemotherapy). The survival rate was calculated by Kaplan-Meier method and the difference was analyzed by log-rank test. Clinical prognosis was assessed by multivariate Cox regression model. Results:The median overall survival (OS) of the entire cohort was 12.3 months (95% CI: 10.6-15.4m), and the 1-, 2-, 3-and 5-year OS rates were 52.1%, 25.2%, 19.1%, and 11.5%, respectively. Multivariate analysis showed that tumor length, the number of metastatic organs, and treatment modalities were the independent prognostic factors for OS. There was no significant difference in OS between concurrent radiochemotherapy and sequential radiochemotherapy ( P=0.955). The OS of patients in the radiotherapy dose of ≥6000 cGy group was significantly longer than that of their counterparts in the 4500-5039 cGy and 5040-6000 cGy groups (both P<0.001). Conclusions:For stage Ⅳ B thoracic ESCC patients at initial diagnosis, tumor length ≤3cm, single organ metastasis, and radiochemotherapy strategy are significantly correlated with longer OS. For stage Ⅳ ESCC patients with good physical status, radiotherapy can be supplemented on the basis of systemic chemotherapy. Concurrent or sequential radiochemotherapy needs to be individualized. If patients are tolerable, radiochemotherapy is recommended to the primary tumor or non-regional metastatic lymph nodes, aiming to prolong the OS of patients.

Observation on dynamic changes of gastrointestinal mucosal injury in rats with acute stage of cerebral infarction / 中国中西医结合急救杂志

Objective To investigate the dynamic characteristic changes of gastrointestinal mucosa and its relationship with disease progression in rats with acute cerebral infarction. Methods Fifty-six male Wistar rats were selected as the study subjects, and they were divided into three groups: normal control, sham operation and cerebral infarction model groups by random number table method. The middle cerebral artery occlusion (MCAO) model was prepared by the modified Longa thread embolic method. The levels of gastrin (GAS) were monitored in each group after modeling for 24 hours, 4 days and 7 days; after the rats were killed, the sections of gastric antrum and small intestine were taken and stained with hematoxylin-eosin (HE) staining method, the histopathological changes of gastric and small intestinal mucosa were observed under light microscope, in the mean time the gastric and small intestinal mucosal pathological scores were also performed, and the differences of pathological scores among the three groups were compared. Results There were no statistical significant differences in GAS, gastrointestinal mucosa and small intestinal mucosal pathological scores between the normal control group and sham operation group at each time point (all P > 0.05); the GAS level in cerebral infarction model group was decreased gradually with time prolongation, reaching the lowest level 7 days after modeling, but the GAS level in cerebral infarction model group was significantly higher than that in normal group and shamoperation group (ng/L: 205.02±7.68 vs. 130.51±8.03, 145.29±7.68, both P < 0.05). The pathological scores of gastrointestinal mucosa and small intestinal mucosa in the cerebral infarction model group were increased first and then decreased with time prolongation, peaked on 4th day and decreased significantly on 7th day, the pathological scores of gastrointestinal mucosa and small intestinal mucosa in the cerebral infarction model group at each time point were significantly higher than those in the normal control group and sham-operated group (gastric mucosal pathological score: 82.50±2.95 vs. 21.38±1.57, 36.10±3.41; small intestinal mucosal pathological score: 62.00±2.78 vs. 18.25±1.39, 25.55±1.75, all P < 0.05). Under light microscopy, the normal control group showed complete normal morphological appearance, normal structure, orderly arrangement of villi and no infiltration of inflammatory cells; in shamoperation group, inflammatory cells infiltrated the lamina propria at each time point, and there were villi slightly uneven, enlarged stroma, congestion, edema occasionally seen and no obvious ulcer; in cerebral infarction model group, the various layers of gastrointestinal mucosal were not very clear, the glands were arranged irregularly and the capillaries dilated, and in part of tissues, congestion, hemorrhage, edema and inflammatory cell infiltration were seen obviously. Conclusion The injury of gastrointestinal mucosa in acute stage of cerebral infarction should be related to the stress stimulation and disease progress of cerebral infarction itself, not due to the abnormal secretion of GAS.

Effect of electroacupuncture at different time on circadian rhythm and SCN epigenetic modification in mice with hepatocellular carcinoma / 中国针灸

<p><b>OBJECTIVE</b>To observe the effects of electroacupuncture (EA) on the circadian rhythm and suprachiasmatic nucleus (SCN) epigenetic modification in mice with hepatocellular carcinoma (HCC), and to explore the epigenetics mechanism of EA on circadian rhythm in patients with HCC.</p><p><b>METHODS</b>According to six zeitbeger time (ZT) of ZT0 (7:00), ZT4 (11:00), ZT8 (15:00), ZT12 (19:00), ZT16 (23:00) and ZT20 (3:00), a total of 108 eligible male C57BL/6J mice were divided into a blank group, a model group and an EA group at each ZT, 6 mice each group. Injection of H22 cancer cell suspension was used to establish the HCC model. After 11 days, EA (2 Hz/15 Hz, 0.2 mA) for 10 days was applied at "Ganshu" (BL 18) and "Zhiyang" (GV 9) in the EA group at each ZT, once a day, 15 min a time; the rats in the blank group and model group were treated with immobilization at the same time and under the same conditions. ClockLab (ACT-500) software was used to record the activity rhythm of mice. After 10 days intervention, MATLAB (R2007b) was used to export the circadian rhythm of mice, and the amplitude and peak phase of the mice were analyzed. The high-throughput epigenetics PCRarray array was applied to detect epigenetics-related gene expression in SCN.</p><p><b>RESULTS</b>(1) After modeling, compared with the blank group, the amplitude of activity was decreased and peak phase was delayed in the model group and EA group at each ZT (all <0.05), but the difference of rhythm parameters between the model group and EA group was not significant (all > 0.05). (2) After intervention, compared with the model group, the amplitude of activity in the EA group at ZT 8 was increased and peak phase was advanced (both <0.05); the difference of the activity amplitude and peak phase between the EA group and model group at ZT0, ZT4, ZT12, ZT16 and ZT20 was not significant (all >0.05); compared with the ZT0, ZT4, ZT12, ZT16 and ZT20, the amplitude of activity in the EA group at ZT 8 was increased and peak phase was advanced (all <0.05). (3) The results of epigenetic PCRarray array showed that after intervention at ZT 8, compared with the blank group, the expression of 48 epigenetic-related genes in SCN of HCC mice was up-regulated; compared with the model group, the relative expression of 49 epigenetic-related genes in the SCN was down-regulated in the EA group; there were 23 epigenetic-related genes differentially expressed among the three groups.</p><p><b>CONCLUSION</b>EA has benign regulation on circadian rhythm of HCC mice, and achieves the best efficacy at ZT 8. EA at ZT 8 could down-regulate the overexpression of epigenetic-related genes.</p>

Hydrogen sulfide protects intestinal mucosa in a neonatal rat model of necrotizing enterocolitis by upregulating the expression of HO-1 / 临床儿科杂志

Objective To explore the protective effects of GYY4137, a new hydrogen sulfide donor, on intestinal mucosa in a neonatal rat model of necrotizing enterocolitis (NEC), and its potential mechanism.Methods Sixty SD rats were randomly assigned into 4 groups: group A (control group), group B (NEC group), group C (NEC with GYY4137 treatment, H2S donor group), and group D (NEC with GYY4137 and Znpptreatment, HO-1 inhibitor group). The SD rat models of NEC were established using simulated milk feeding-hypoxia-cold stress-Lipopolysaccharides. The injury degree of intestinal mucosa was evaluated using HE-staining, and its mechanisms were investigated using biochemical indicators and Western blotting. Results Compared with control group, the pathology score and the total superoxide dismutase (T-SOD) in the NEC group was significantly higher, the concentrations of methane dicarboxylic aldehyde (MDA) and necrosis factor α (TNF-α) were lower(P<0.05). Compared with those in NEC group, the pathology score and the concentration of MDA and TNF-α in the H2S donor group were signiflcantly lower, the T-SOD, and the HO-1 expression was higher. The pathology score and the level of MDA and TNF-α were signiflcantly increased after treated with HO-1 inhibitor Znpp, and T-SOD was signiflcantly decreased.. Conclusions The GYY4137, as a new H2S donor, could attenuate the injury of intestinal mucosa in a neonatal rat model of NEC by upregulating the expression of HO-1.

Alizarin induced the function of beta subunits of BKCa channel relaxed interlobar renal artery and reduced SHR systolic pressure / 实用医学杂志

Objective To investigate the effect of alizarin on BKCa channel beta subunit and electrophysiological characteristics in interlobar renal artery of SHR.Methods Tail artery pressure measurement instrument,pressure myograph system,whole cell patch clamp technique and Western blot were used to measure the change of systolic pressure of SHR,renal interlobar arterial diameter,single vascular smooth muscle cell electrophysiological characteristics and expression of the BKCa channel beta subunit before and after alizarin intervention,respectively.Results (1) SHR systolic pressure was decreased after alizarin intervention (P ＜ 0.01).(2) ibTX inhibited alizarin-mediated renal interlobar arterial relaxation (P ＜ 0.01).(3) Alizarin enhanced BKCa channel-mediated outward currents (P ＜ 0.05).(4) Alizarin up-regulated BKCa channel beta subunits after alizarin intervention (P ＜ 0.01).Conclusion Alizarin reduced SHR systolic pressure and relaxed interlobar renal artery by enhancement BKCa currents via mediation of the function of beta subunits.

Comparison of PET/CT vs.EBUS-TBNA in Evaluating Lymph Node Metastasis in NSCLC / 华中科技大学学报(医学版)

Objective To compare positron emission tomography/computed tomography(PET/CT)vs.endobronchial ultra-sound-guided transbronchial needle aspiration(EBUS-TBNA)in evaluating lymph node metastasis in NSCLC.Methods We col-lected 79 NSCLC patients with enlarged mediastinal lymph nodes(diameter >1 cm by CT).The diagnostic values of PET/CT scanning and EBUS-TBNA for mediastinal staging were evaluated.Subgroup analysis according to histologic type was per-formed.Results There were 22 patients of N1 stage and 28 patients of N2 in these 79 cases.In the N1 patients,PET/CT's sen-sitivity and specificity was 59.1% and 75.4%,respectively.EBUS-TBNA's sensitivity and specificity was 86.4% and 100.0%,respectively.In the N2 patients,PET/CT's sensitivity and specificity was 67.9% and 76.5% and EBUS-TBNA's sensitivity and specificity was 89.3% and 100.0%.Conclusion EBUS-TBNA is more accurate than PET/CT in evaluating the metastatic condition of patients,and EBUS-TBNA can also benefit PET/CT(-)patients with adenocarcinoma.

CT pulmonary angiography using low tube voltage combination with different noise index, low concentration contrast agent: image quality and radiation dose / 中华放射学杂志

Objective To investigate the image quality,radiation dose and iodine intake of CT pulmonary angiography (CTPA) in patients using low tube voltage (100 kVp) combination of different noise indexes (NI) and low concentration contrast agent.Methods A total of 80 patients with suspected pulmonary embolism (PE) and other pulmonary diseases who had undergone CTPA were divided into four groups (A,B,C and D),with 20 patients in each group.Group A underwent 120 kVp CT scan protocol in combination with NI=25 and 370 mg iodine/ml contrast agent,while groups B,C and D underwent 100 kVp CT scan protocol in combination with NI=30,35,40,and 320 mg iodine/ml contrast agent,respectively.All images were restructured using 60％ adaptive statistical iterative algorithm 2.0.Objective image quality evaluation included CT values of pulmonary artery,noise values of pulmonary artery,signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR).Subjective image evaluation used a 5-point scoring method and was conducted by two independent radiologists.The CT dose index of volume (CTDIvol),dose-length product (DLP) and iodine intake was recorded,and the mean value was calculated.The DLP was converted to the effective dose (ED).Analysis of Variance or Kruskal-Wallis test was used to evaluate the differences among the four groups in terms of image quality,radiation dose and iodine intake.Results There was a significant difference in CT values of pulmonary artery among the groups A,B,C and D (P＜0.05),and the CT values of pulmonary artery of group A was the lowest.There was no significant differences in noises of pulmonary artery,SNR,CNR and subjective indexes scores among the groups A,B,C and D (P＞0.05).There was a significant difference in iodine intake among the groups A,B,C and D,iodine intake of the group A was the highest,iodine intake of the group D was the lowest.The iodine intake of groups B,C and D decreased by 12.4％ (42/340),13.2％ (45/340) and 15.0％ (51/340) relative to group A,respectively.There was a significant difference in radiation dose among the groups A,B,C and D,The CTDIvol,DLP and ED of group D decreased by 45.3％ (3.9/8.6),48.6％ (120/247) and 48.3％ (2.02/4.18) relative to group A,respectively (P＜0.01).Conclusion Low tube voltage combination with high NI value and low concentration contrast agent can more effectively reduce the radiation dose and iodine intake for CTPA while maintaining diagnostic image quality.

Feasibility of automatic spectral imaging protocol selection and adaptive statistical iterative reconstruction in reducing radiation and iodine contrast dose in abdominal CTA / 中国医学影像技术

Objective To investigate the feasibility of automatic spectral imaging protocol selection (ASIS) and adaptive statiatical iterative reconstruction (ASiR) technique to reduce radiation dose and dose of contrast agent.Methods Sixtyfour patients underwent routine abdominal examination were randomly divided into two groups.The test group used ASIS technique,with 30％ ASiR and 50％ ASiR reconstruction algorithm.The control group used 120 kVp tube voltage,FBP reconstruction method.The noise of liver,pancreas,sacrospinal muscle,CNR of liver and pancreas,subjective image score in arterial phase and portal venous phase were compared between the image of 70 keV+30％ ASIR and control group.CNR of abdominal aorta and its branchs,CNR of portal vein,and subjective image score were statistically analyzed between im age 55 keV+50％ ASiR and control group in the arterial phase and portal venous phase.Results Compared with control group,CT dose index volume for arterial phase and portal venous phase in test group decreased by 23.68％,23.57％ and dose length product decreased by 25.61％,18.45 ％,total contrast injection decreased 16.86 ％,the noise of liver,pancreas and sacrospinal muscle in 70 keV+30％ ASiR were lower than those of control group in abdominal arterial and portal phase (all P＜0.05).CNR of abdominal aorta,superior mesenteric artery,celiac axis and score in 55 keV+50％ ASiR were higher than those of control group in abdominal arterial phase (all P＜0.05),CNR of portal vein and score in portal phase had no statistically difference (all P＞ 0.05).Conclusion Combining of ASIS and ASiR including 70 keV + 30％ ASiR and 55 keV+50％ ASiR,images are superior to that of the conventional 120 kVp+FBP scan mode for abdominal CT image and vessel image quality,which can reduce the radiation dose and the dose of contrast agent.

Effects of zirconia/alumina ceramic composite on proliferation, apoptosis and bone induction activity of human periodontal ligament cells / 中国组织工程研究

BACKGROUND: Zirconia/alumina ceramic composite with excellent corrosion resistance, high strength and toughness has wide application prospect.OBJECTIVE: To investigate the effect of zirconia/alumina ceramic composite on proliferation, apoptosis and bone induction activity of human periodontal ligament cells.METHODS: Human periodontal ligament cells at passages 3-5 were cultured routinely as control group and co-cultured with zirconia/alumina ceramic composite as experimental group. After 1-5 days of culture, MTT,TUNEL, immunohistochemical detection, and alkaline phosphatase staining were used to detect cell proliferation,apoptosis, type I collagen expression and osteogenic activity, respectively.RESULTS AND CONCLUSION: Within 1-5 days of culture, there were no differences in cell proliferation, apoptosis and alkaline phosphatase activity between the two groups. At 5 days of culture, the type I collagen expression showed no difference between the two groups. Taken together, the zirconia/alumina ceramic composite has no effect on the proliferation, apoptosis and bone induction activity of human periodontal ligament cells.

Alizarin activates large-conductance Ca2+-activated K+ channels on renal interlobar arterial vascular smooth muscle cells in Wistar rat / 中国药理学通报

Aim To investage the regulatory effect of alizarin on renal interlobar arterial smooth muscle cells. Method The effect of alizarin on BKCa channels was observed by whole-cell patch clamp recording tech-nique. Results Selective BKCa channel blocker ibTX inhibited alizarin-induced outward current(P<0. 05);single smooth muscle cells were incubated in extracel-lular fluid with no Ca2+ 30 minutes, selective L-Ca2+channel blocker nimodipine, selective calcium ion che-lating agent BAPTA-AM and ryanodine inhibited the a-lizarin-induced BKCa channels current, too ( P <0. 05 ) . Conclusion Alizarin relaxes renales interlobar arterial smooth muscle via activation of L-Ca2+ channel firstly, which lead to Ca2+ flowed into cytoplam, and rising of Ca2+ in cytoplam ryanodine receptor indirect-ly, and BKCa is activated at last.

A meta-analysis of relationship between friendship quality and loneliness / 中国心理卫生杂志

Objective:To explore the relationship between friendship quality and loneliness and its influent factors like the measuring tool of friendship quality,the participants groups and cultural background. Methods:The studies on the association between the friendship quality and loneliness were collected through the databases of Chi-na National Knowledge Infrastructure,Wan Fang,Wiley Online library,Web of Science,Google Scholar and Duxiu from 1984 to 2014. Meta-analysis was conducted with the use of the CMA2. 0. Estimated value (r)and their 95%confidence interval (95%CI)was calculated to analyze the association between the friendship quality and loneliness from normal subjects except physiological disease (such as autism,etc. ). Results:Totally 46 literatures were met the requirements of meta-analysis with sample size of 19422,including 24 researches With 11846 participants who were from eastern countries,and 22 studies with 7676 western people. There was significant negative correlation of friendship quality and loneliness (r=-0. 28,95%CI:-0. 23--0. 32). The friendship quality measuring tool was able to adjust the relations of friendship quality and loneliness (Q=11. 95,P0. 05 ). Conclusion:It suggests that friendship quality has an influence on lone-liness. Measuring tools and subjects groups effect the relationship friendship quality and loneliness except cultural background.

Inhibiting effect of alcohol extract from Dioscore bulbifera on gastric cancer cells / 天津医药

Objective To investigate the inhibiting effects of alcohol extract from Dioscore bulbifera on proliferation, colony formation and migration of cancer cell lines. Methods Alcohol extract from Dioscore bulbifera was prepared using Soxhlet extraction. Human gastric cancer cell line MGC803 was treated with different concentrations(0, 60, 120 mg/L)of al?cohol extract from Dioscore bulbifera. In vitro, proliferation, colony formation and migration of gastric cancer cells were detect?ed by MTT, colony formation experiments and Transwell assay respectively. Results The proliferation(day2-day 6, F=29.130, 21.864, 67.826, 36.015, 43.656, P<0.01)and colony formation(F=11.918,P<0.01)of gastric cancer cells were significantly inhibited by administration of alcohol extract from Dioscore bulbifera at both 60 mg/L and 120 mg/L . The migra?tion(F=4.258,P<0.05)of gastric cancer cells were significantly suppressed after cells were treated with120 mg/L alcohol ex?tract from Dioscore bulbifera. Conclusion Alcohol extract from Dioscore bulbifera significantly inhibit proliferation, colony formation and migration of gastric cancer cells.

Outcomes of autologous osteo-periosteal cylinder graft transplantation for Hepple Ⅴ osteochondral lesions of the talus / 中华骨科杂志

Objective To study the outcomes of autologous osteo-periosteal cylinder graft transplantation for Hepple V osteochondral lesions of the talus (OLT) with large subchondral cyst.Methods The data of 27 consecutive patients of OLT with subchondral cyst was retrospectively analyzed who were treated by autologous osteo-periosteal cylinder graft transplantation from October 2007 to September 2011.There were 26 males and 1 female with an average age of 35.8 years (range,22-53 years).Visual analogue score (VAS) for pain during daily activities,the American Orthopaedic Foot and Ankle Society (AOFAS) ankle and hindfoot score,and subjective satisfaction were investigated.The plain radiographs,magnetic resonance imaging (MRI) of the ankle,and second look arthroscopy were analyzed.Results All the 26 patients were followed up for 22.4 months.At the last follow-up,the VAS score decreased from 5.4±1.0 points preoperatively to 0.8±0.8 points postoperatively,and the mean (50％) AOFAS score improved from 73.9±3.1 points preoperatively to 93.0±6.5 points postoperatively.In 26 cases,the radiolucent area of cysts disappeared on plain radiographs.The mean magnetic resonance observation of cartilage repair tissue (MOCART) score was 57.2,though small subchondral bone cyst was still found in 3 cases on postoperative MRI.The mean (50％) ICRS arthroscopic score of cartilage repair was 9.2 points according to second look arthroscopy of 18 cases.There were 16 cases receiving excellent effect,8 good and 2 fair.The excellent and good rate was 92.3％ (24/26).There were no major complications.Conclusion Autologous osteo-periosteal cylinder graft transplantation could repair the osteochondral defects.It yields satisfactory results,and is suitable for treating OLT with large subchondral cyst.

Role of insulin-like growth factor-1 in proliferation, migration and differentiation of neural stem cells in cerebral infarction rats / 中国组织工程研究

BACKGROUND: Insulin-like growth factor-1 (IGF-1) is a peptide hormone, it has been proved a promotion role on the proliferation of precursor cells. OBJECTIVE: To explore the intravenous injection of IGF-1 on the proliferation, migration and differentiation of neural stem cells in rats after cerebral ischemia. METHODS: Eight adult male SD rats were randomly divided into control group and experimental group, with 40 rats in each group. The rats in two groups were used to prepare models of focal cerebral ischemia using modified suture method, the rats in the experimental group were treated with tail vein injection of IGF-1, according to 100 μg/kg computation, the injection was given for 6 continuous days; in the control group, rats were given equal volume of saline. The rats were decapitated at 7, 14, 21, 28 days following intervention, respectively, and rats in each group were given intraperitoneal injection of the BrdU at 1 day before death. Immunohistochemistry and double staining were applied to detect the expressions of BrdU-positive cells, PSA-NCAM-positive cells, BrdU + PSA-NCAM double-positive cells, and BrdU + MAP2 double-positive cells. RESULTS AND CONCLUSION: The number of BrdU-positive cells and PSA-NCAM positive cells reached the peak at 7 days after ischemia; BrdU + PSA-NCAM double-labeled-positive cells could be detected in ischemic bilateral subependymal zone and dentate gyrus, the number was the most at 7 days, then followed by a gradual decrease; the BrdU + MAP2 double-positive cells began to increase from 14 days, and then gradually increased along with the decrease of BrdU + PSA-NCAM double-positive expression, showing a reverse trend. Intravenous injection of IGF-1 can induce the proliferation, differentiation and migration of neural stem cells in rats following ischemic brain injury.

The stable expression of human tissue-type plasminogen activator gene mediated by lipofectamine in human vein endothelial cell line cells / 生物医学工程学杂志

We have established a human umbilical vein endothelial cell (HUVEC) line monoclonal cells with the stable expression of human tissue-type plasminogen activator (t-PA) gene to provide a basis for further study on the vascular tissue engineering. Recombinant plasmid pcDNA3. 1-Myc-His B (-)/t-PA was constructed by insertion of t-PAcDNA originated from PBS/t-PA into eukaryotic expression vector pcDNA3. 1-Myc-His B(-) and transfected into hUVEC line cells mediated by lipofectamine. The positive clones were obtained by the screen of G418. The transcription and expression of t-PA gene were investigated by RT-PCR and Western blotting respectively. The t-PA activity was measured by chromogenic substrate assay. The positive clone cells which transcripted the mRNA of t-PA gene was obtained by RT-PCR. Immunoreactive human t-PA of the medium was significantly increased in the group of transfected gene when compared with that in the controlled and transfected plasmid without t-PA gene group. The biological activity of the protein of the t-PA in the media was increased significantly in the positive clone cells with t-PA gene transfected. The HUVEC line monoclonal cells with the stable expression of t-PA gene was established successfully.

Intrastriatal Gene Transfer of Vascular Endothelial Growth Factor Rescues Dopaminergic Neurons in a Rat Parkinson's Disease Model / 华中科技大学学报（医学）（英德文版）

To examine the ability of intrastriatal gene transfer of vascular endothelial growth factor 165 mediated by adenoviral vector to rescue dopaminergic neurons in a rat model of Parkinson's disease (PD), we constructed recombinant replication-deficent adenoviral vectors carrying the gene of VEGF165 (Ad-VEGF), and injected Ad-VEGF (or Ad-LacZ and PBS as controls) into the striatum of rats 7 days after the lesion by 6-hydroxydopamine. The rat rotational behavior analysis and tyrosine hydroxylase (TH) immunohistochemistry were performed to assess the change of dopaminergic neurons. Our results showed that the rats receiving Ad-VEGF injection displayed a significant improvement in apomorphine-induced rotational behavior and a significant preservation of TH-positive neurons and fibers compared with control animals. It is concluded that intrastriatal gene transfer by Ad-VEGF may rescue the dopaminergic neurons from degeneration in a rat model of PD.

Intrastriatal gene transfer of vascular endothelial growth factor rescues dopaminergic neurons in a rat Parkinson's disease model / 华中科技大学学报（医学）（英德文版）

To examine the ability of intrastriatal gene transfer of vascular endothelial growth factor 165 mediated by adenoviral vector to rescue dopaminergic neurons in a rat model of Parkinson's disease (PD), we constructed recombinant replication-deficent adenoviral vectors carrying the gene of VEGF165 (Ad-VEGF), and injected Ad-VEGF (or Ad-LacZ and PBS as controls) into the striatum of rats 7 days after the lesion by 6-hydroxydopamine. The rat rotational behavior analysis and tyrosine hydroxylase (TH) immunohistochemistry were performed to assess the change of dopaminergic neurons. Our results showed that the rats receiving Ad-VEGF injection displayed a significant improvement in apomorphine-induced rotational behavior and a significant preservation of TH-positive neurons and fibers compared with control animals. It is concluded that intrastriatal gene transfer by Ad-VEGF may rescue the dopaminergic neurons from degeneration in a rat model of PD.