FHL1 has been recognized for a long time as a tumor suppressor protein that associates with both the actin cytoskeleton and the transcriptional machinery. We present in this study a paradigm that phosphorylated FHL1 functions as an oncogenic protein by promoting tumor cell proliferation. The cytosolic tyrosine kinase Src interacts with and phosphorylates FHL1 at Y149 and Y272, which switches FHL1 from a tumor suppressor to a cell growth accelerator. Phosphorylated FHL1 translocates into the nucleus, where it binds to the transcription factor BCLAF1 and promotes tumor cell growth. Importantly, the phosphorylation of FHL1 is increased in tissues from lung adenocarcinoma patients despite the down-regulation of total FHL1 expression. Kindlin-2 was found to interact with FHL1 and recruit FHL1 to focal adhesions. Kindlin-2 competes with Src for binding to FHL1 and suppresses Src-mediated FHL1 phosphorylation. Collectively, we demonstrate that FHL1 can either suppress or promote tumor cell growth depending on the status of the sites for phosphorylation by Src.

MicroRNAs (miRNAs/miRs) show great promise as novel cancer biomarkers. Several studies have revealed an association between abnormal miRNA expression and the risk of various cancer types. However, the diagnostic accuracy and reliability of miRNAs remains unclear. The present meta-analysis was performed to summarize the overall diagnostic performance of miR-195 for cancer. The PubMed, Cochrane Library, Wanfang and China National Knowledge Infrastructure databases were searched for associated literature published until December 10, 2017. Eligible studies were selected using multiple search strategies based on study selection criteria. Measures, including sensitivity and specificity, of the performance of miR-195 as a cancer diagnostic tool were pooled using bivariate meta-analysis models. All analyses were performed using Stata 14.0. The pooled analysis included 8 studies comprising 735 cases and 547 controls. The pooled diagnostic results calculated from all studies were as follows: Sensitivity, 0.79 [95% confidence interval (CI), 0.69-0.87]; specificity, 0.84 (95% CI, 0.68-0.93); positive likelihood ratio, 4.9 (95% CI, 2.50-9.50); negative likelihood ratio, 0.25 (95% CI, 0.18-0.35); diagnostic odds ratio, 20 (95% CI, 10.00-38.00); and area under the curve, 0.87 (95% CI, 0.84-0.90). Deeks' funnel plot asymmetry test suggested no potential publication bias (P=0.53). The present meta-analysis indicated that miR-195 could be a reliable non-invasive biomarker for the diagnosis of cancer. Further large-scale prospective studies are necessary to confirm the present findings and the clinical value of miR-195 for future diagnostics.

BACKGROUNDPatients with mild or moderate chronic obstructive pulmonary disease (COPD) rarely receive medications, because they have few symptoms. We hypothesized that long-term use of tiotropium would improve lung function and ameliorate the decline in lung function in patients with mild or moderate COPD.METHODSIn a multicenter, randomized, double-blind, placebo-controlled trial that was conducted in China, we randomly assigned 841 patients with COPD of Global Initiative for Chronic Obstructive Lung Disease (GOLD) stage 1 (mild) or 2 (moderate) severity to receive a once-daily inhaled dose (18 mu g) of tiotropium (419 patients) or matching placebo (422) for 2 years. The primary end point was the between-group difference in the change from baseline to 24 months in the forced expiratory volume in 1 second (FEV 1) before bronchodilator use. Secondary end points included the between-group difference in the change from baseline to 24 months in the FEV 1 after bronchodilator use and the between-group difference in the annual decline in the FEV 1 before and after bronchodilator use from day 30 to month 24.RESULTSOf 841 patients who underwent randomization, 388 patients in the tiotropium group and 383 in the placebo group were included in the full analysis set. The FEV 1 in patients who received tiotropium was higher than in those who received placebo throughout the trial (ranges of mean differences, 127 to 169 ml before bronchodilator use and 71 to 133 ml after bronchodilator use; P<0.001 for all comparisons). There was no significant amelioration of the mean (+/- SE) annual decline in the FEV 1 before bronchodilator use: the decline was 38 +/- 6 ml per year in the tiotropium group and 53 +/- 6 ml per year in the placebo group (difference, 15 ml per year; 95% confidence interval [CI], -1 to 31; P = 0.06). In contrast, the annual decline in the FEV 1 after bronchodilator use was significantly less in the tiotropium group than in the placebo group (29 +/- 5 ml per year vs. 51 +/- 6 ml per year; difference, 22 ml per year [95% CI, 6 to 37]; P = 0.006). The incidence of adverse events was generally similar in the two groups.CONCLUSIONSTiotropium resulted in a higher FEV 1 than placebo at 24 months and ameliorated the annual decline in the FEV 1 after bronchodilator use in patients with COPD of GOLD stage 1 or 2. (Funded by Boehringer Ingelheim and others; Tie-COPD ClinicalTrials.gov number, NCT01455129.)

In the past several years, the αv integrin subfamily has been repeatedly found to be involved in tumor progression and angiogenesis. The aim of this study was to investigate the expression of the integrin αv subfamily in laryngeal squamous cell carcinoma (LSCC), and to correlate the expression rate with tumor biological behavior and angiogenesis of the LSCC. The integrin αv subfamily, including αv, β1, β3, β5, β6 and β8 subunits, was immunohistochemically found to be expressed in 64 patients with LSCC, and we analyzed the relationship between the expression rate and the clinicopathological stage of this cancer. Immunohistochemical staining for CD105 was carried out in the same group of the patients. The intratumoral microvessel density (IMVD) of the LSCC was calculated by CD105 staining, and the correlation between the IMVD and αv subfamily expression was discussed. The results showed that all members of the integrin αv subfamily could be detected in the LSCC. The expression rate of integrin αv and β5 subunits in primary cancer was significantly higher than in normal tissue, and their expression rate in the group with lymphatic metastasis was significantly higher than in the group without metastasis. The IMVD of the group with positive expression of αv and β5 subunits was significantly higher than in the group with negative expression, but there were no significant effects on the β1, β3, β6 and β8 subunits in these biological processes. In conclusion, the expressions of integrin αv and β5 subunits were significantly associated with lymphatic metastasis and angiogenesis of the LSCC. Among the members of integrin αv subfamily, integrin αvβ5 might play an important role in invasion and metastases of the LSCC, and it may become a valuable marker for the evolution of the LSCC.

Objective: To study the effects of acanthopanax giraldii harms var hispidus hoo polysaccharides (AGP) on proliferation and expression of nuclear transcription factor p53 and c-myc in human breast cancer cell line MDA-. MB-231. Methods: The proliferation rate of human breast cancer cell line MDA-MB-231, which was incubated by different concentration of AGP (0.15625, 0.3125, 0.625, 1.25, 2.5g/L) in DMEM medium for 1, 3, 5, 7 days in vitro respectively, was studied by WST-1 cell proliferation and cytotoxicity assay kit. The changes of human breast cancer cell line MDA-MB-231 morphology were observed by an inverted microscopy. The expression of apoptosis-related proteins such as p53 and c-myc was detected by fluorescence immunoassay and Western blotting. Results: WST-1 assay showed that different concentration of AGP inhibited proliferation of human breast cancer cell line MDA-MB-231 treated for 3day, 5day, 7day when compared with the negative control group (P<0.05). The inhibition rate depended on AGP dose and reaction time. According to fluorescence immunoassay and Western blotting analysis, c-myc expression in cytoplasm reduced and p53 expression increased significantly after the treatment of AGP on human breast cancer cell line MDA-MB-231. Conclusion: AGP has the effect of inhibiting proliferation on human breast cancer cell line MDA-MB-231. Its role may be related to reduce c-myc expression and increase p53 expression of human breast cancer cell line MDA-MB-231.

OBJECTIVETo investigate the potential of human amniotic mesenchymal cells (hAMC) serving as seeding cells in bone tissue engineering.METHODShAMC were isolated and cultured. The third passage of hAMC was cultured in osteogenic induced media [DMEM supplemented with 10% (v/v) FBS, 0.1 mumol/L dexamethasone, 50 mg/L ascorbic acid and 10 mmol/L beta-glycerophosphate] for one week. Calcified nodules were shown by alizarin red staining and counted under light microscope. Immunofluorescence cytochemical staining was used to detect collagen I (COL I) and alkaline phosphatase (ALP). Expression of FasL was examined in the amnion and hAMC by immunohistochemistry or immunocytochemistry.RESULTSAfter osteoblast differentiation, calcified nodules were formed, on the average 18 per well. hAMC in calcified nodules showed positive expression of COL I and ALP. FasL was detected positive both in cells contained in amnion and in hAMC.CONCLUSIONhAMC are potential ideal candidates for seeding cells in bone tissue engineering.

OBJECTIVE:To investigate the alterations in structure and function of sternohyoid muscle of metabolic syndrome (MS) rats effects and the effects of taurine thereupon contractile properties.METHODS:Twenty-one healthy male SD rats were randomly assigned to three groups: control group (Group A, n=6) fed with normal food, MS group (Group B, n=8) fed with high-lipid forage for 9 weeks to induce MS and then fed with the same high-lipid forage for 12 weeks in addition, and taurine group (Group C, n=7), fed with high-lipid forage for 20 weeks and given gastric perfusion of taurine 50 mg x d(-1) x kg(-1) since the tenth week for 12 weeks. Venous blood samples were collected to undergo biochemical examination of plasma total cholesterol (TC), triglyceride (TG), high-density lipoprotein (HDL), low-density lipoprotein (LDL), and glucose (Glu). Radioimmunoassay was used to detect the plasma insulin. The sternohyoid muscles were collected. Muscle tissue homogenate was made. The superoxide dismutase (SOD) and malondialdehy (MDA) levels were measured. Myosin-ATPase histochemistry was used to assay the size and capillary density of the isolated sternohyoid muscles. Isometric contractile properties were determined by electrostimulating the strips of isolated sternohyoid muscles at different frequencies (from 10 Hz to 100 Hz) to observe the changes of the sternohyoid contractile properties in different conditions.RESULTS:(1) The levels of TG, LDL, Glu, PGI, and MDA of Group B were significantly higher than those of Groups A and C, while the HDL level of Group B was significantly lower than that of Group A (P < 0.01). The OD level of Group B was significantly lower than those of Groups A and C. (2) The capillary density, capillary-to-fiber ratio (C/F), and cross-sectional area of type I fiber in sternohyoid muscles of Group B were (140 +/- 5)/ nm2, 0.90 +/- 0.11, and (6119 +/- 165) microm2 respectively, all significantly higher than those of Group A [(278 +/- 17)/mm2, 1.43 +/- 0.05, and (9371 +/- 68) microm2 respectively, all P < 0.01] and Group C [(269 +/- 10)/mm2, 1.40 +/- 0.07, and (9007 +/- 136) microm2 respectively, all P < 0.01]. (3) The tensions of sternohyoid muscle of Group B at the frequencies from 10 Hz to 60 Hz were (29 +/- 6) g/cm2, (34 +/- 7) g/ cm2, (41 +/- 6)g/cm2, (47 +/- 6)g/cm2, (53 +/- 7)g/cm2, and (59 +/- 20)g/cm2 respectively, all significantly lower than those of Group A group[(40 +/- 3)g/cm2, (52 +/- 13)g/cm2, (60 +/- 17) g/cm2, (74 +/- 32) g/cm2, (80 +/- 29)g/cm2, and (96 +/- 24)g/cm2 respectively, all P < 0.05] and Group C [(43 +/- 26)g/cm2, (48 +/- 6)g/cm2,(55 +/- 9)g/cm2, (67 +/- 14)g/cm2, (75 +/- 15)g/cm2, and (80 +/- 15)g/cm2 respectively, all P < 0.05]. (4) In fatigue test, the tension percentages of sternohyoid muscle at the time intervals of 1 min, 2 min, 3 min, 4 min, and 5 min of Group B were (80.5 +/- 8.0)%, (64.1 +/- 1.2)%, (59.1 +/- 1.1)%, (56.4 +/- 10.9)%, and (53.5 +/- 9.1)% respectively, all significantly lower than those of Group A [(87.7 +/- 3.5)%, (78.5 +/- 1.5)%, (76.0 +/- 1.2)%, (72.3 +/- 15.0)%, and (68.7 +/- 17.2)% respectively, all P < 0.05] and Group C group [(87.4 +/- 2.4)%, (77.9 +/- 5.5)%, (73.6 +/- 1.1)%, (71.3 +/- 8.7)%, and (68.0 +/- 6.7) respectively, all P < 0.05].CONCLUSION:With the development of MS, the capillary density, C/F, cross-sectional area of type I fiber of upper airway muscles decrease, in part via oxidative stress, which leads to reduction of the contractile function of upper airway muscles, thus contributing to the onset of obstructive sleep apnea-hypopnea syndrome.

Objective To observe the distribution and the effect of the quantum dots(QDs) on mouse abdominal cavity macrophages.Methods The QDs were co-cultured with mouse abdominal cavity macrophages in vitro.The differentiation and effect of the QDs on macrophage ultrastructures were observed under electronic microscope. Results The QDs were enveloped with unit membrane and internalized in the cytoplasm of the macrophage under transmission electron microscope.And it formed vacuolelike structures in the macrophage.There were many lamellar processes on the surface of the macrophage under scanning electron microscope.Conclusion The QDs can promote macrophage activation,and make its surface projection increased.The QDs were internalized by the macrophage,distributed in the cytoplasm,and formed vacuolelike structures enveloped with unit membrane.

Objective To investigate the distribution and significance of CD20 positive cells in the early human embryo. Methods Distribution and morphology of CD20 positive cells in 10 specimens of human embryo aged from 6 to 7 weeks were brown,and studied with immunohistochemical method. Results 1.CD20 positive cells appeared in the liver of early human embryo.2.The immunohistochemical positive substances were brown,and found chiefly in the nuclei of B cells as unevenly-distributed granules,and were not detected in the cytoplasma and on the cell membrane.Conclusion The accuracy of location and distribution of CD20 positive cells in the nuclei of B cells of early human embryo liver may provide an important clue for further exploration of the functional mechanism of CD20 in the process of B cell proliferation and differentiation.

Objective Study the effect of quantum dots(QDs) on macrophage cytochemistry and enzyme activity of mice in vitro. Methods To observe the macrophage biocompatibility with QDs and effect of QDs on PAS reaction,Feulgen reaction,ATPase,acid phosphatase(AcP),alkaline phosphatase(ALP),ANAE,SDH and LDH using inverted phase contrast microscope,fluorescence microscope,and cytochemistry methods were used.Results QDs hadn't effect to macrophage structure in(3.125mg/L) density of QDs,but its cytochemistry and cell enzyme activity had changed differently,that was PAS reaction,Feulgen reaction,AcP,ALP,ANAE and Mg~(2+)-ATP showed positive,and SDH and LDH showed negative.Feulgen reaction,ANAE,ALP and Mg~(2+)-ATP had statistical significance with the control group(P0.05),PAS and AcP doesn't have statistical significance with the control group(P0.05).Conclusion In cell level,macrophage enzymes have changed differently by quantum dots,but QDs don't affect macrophage structure and phagocytosis. QDs can be applied to label living cell in invariably limited dose of QDs in biomedicine area,and cells doesn't have visible influence.

Objective To analyse the number of dendritic cells and macrophages and the size of macrophages in the spleen of SARS patients so as to provide evident for the study of pathology and pathogenesis of SARS. Methods Immunohistochemical method with four specific antibodies(S-100,CD68,HLA-DR,CD83) were used to detect the dendritic cells and macrophages in the spleens of six dead patients of SARS and six accidental deaths as the controls,The number or the size of these positive cells was analysed with image analysis system. Results In the spleens of SARS patients,the number of S-100~+ dendritic cells in the white pulp was reduced by 80.4% on average,and even disappeard.The number of CD68~+ macrophages in the red pulp was reduced by 39.48% in SARS spleens,and the average size of individual macrophages was increased by 2.21 times.HLA-DR~+ antigen presenting cells(APC) reduced remarkably in the SARS spleen white pulp.CD83~+ mature dendritic cells did not exist in either SARS spleens or the control spleens.Conclusion The function of antigen presentation had been damaged severely,which supports that SARS should be categorized as a viral immune deficiency disease.SARS virus doesn't induce the maturation of DC.The increase in the size of macrophages indicated that they were in an activated state and may play a role in the pathogenesis of SARS.

Objective To investigate the number,distribution and expression intensity of dendritic cells and T lymphocytes in human cervical intraepithelial neoplasia and invasive carcinoma for seeking biotherapeutic evidence of cervical carcinoma. Methods The number,distribution and expression intensity of CD-(1a),S-100,CD-3, CD-8 positive cells were determined in cervical intraepithelial neoplasia and invasive carcinoma and human normal cervix tissue as the control,by using immunohistochemical technique and computerized imaging analysis system. Results Compared with human normal cervical tissue,the number and the expression intensity of CD~+-(1a),S-100~+,CD~+-3,CD~+-8 cells in cervical intraepithelial neoplasia significantly increased(P0.05) and had a positive relation to histological grade(P0.05);the number and the expression intensity of four antibody positive cells in invasive carcinoma decreased and had a negative relation to the clinical stage(P0.05).Conclusion The number and the expression intensity of dendritic cells and T lymphocytes changed in the process of cervical carcinogenesis,and the results suggest that there are some changes in local immunologic function in human pathological cervix.

Objective To investigate the effect of macrophage-conditioned medium on the growth of mouse myoblasts. Methods After a 3-day intraperitoneal injection with a mixture of muscle homogenate and starch,macrophages were obtained from mouse peritoneal cavity and cultured for 48 hours in serum-free DMEM/F12.Then the cultured medium was harvested.Myoblasts were obtained from newborn mice skeletal muscle and grown in DMEM/F12 supplied with 10% FBS for 36-48 hours,and then exposed to 0.5% FBS medium with or without macrophage-conditioned medium.Cells were harvested at 72 hours later,their growth was observed with Wright staining,LDH cytochemistry and immunocytochemistry. Results Macrophage-conditioned medium can maintain the normal morphology and the activity of myoblasts grown in low concentration of serum and promote their proliferation.There was a significant difference between average absorbance and integral absorbance of LDH positive cells of the two groups(P0.01 and P0.000 1 respectively).The experiment group contained more positive cells compared with the control group,identified by immunocytochemistry for desmin.Conclusion Macrophage-conditioned medium contains factors that promote growth of mice myoblasts.

PURPOSE: To explore the characteristics of SARS Coronovirus-induced lung injury by comparing the protein expression of CD34,VEGF and AQP-1 in SARS lungs with those in non-SARS lung injury.

Objective To observe the expression of NF-κB p50 protein in the process of the human peripheral blood monocytes differentiating and maturing into dendritic cells. Methods Human peripheral blood monocytes were obtained by discontinuous density gradient centrifuged method. Monocytes of different groups (cytokine group, saikoside group, co-culture group)were cultured with different stimulating factors such as GM-CSF, IL-4, TNF-α and saikoside in vitro. The inverted phase contrast microscope, Wright's staining, immunocytochemical staining were used respectively to observe the changes of morphology, surface differentiation antigen including CD14, CD83, HLA-DR,S-100, and the expression of intracellular transcription factor NF-κB p50 protein, in the process of monocytes differentiating into dendritic cells. The image analysis and SPSS software were used to analyze the results. Results The obtained monocytes and dendritic cells possessed their respective cell shape and cell surface antigen marker by cell morphology observation and cell surface marker immunocytochemical staining such as CD14, HLA-DR, S-100 and CD83. The monocytes differentiated into immateure dendritic cells(imDC) when cultured with RPMI-1640 medium containing GM-CSF and IL-4 for 7 days. The imDC differentiatd into mature dendritic cells (mDC) when cultured with RPMI-1640 medium containing GM-CSF,IL-4 and TNF-α for another 3 days. The expresion of NF-κB p50 in the two cells showed a significant difference (P0.05). When Monocytes were cultured with RPMI-1640 medium containing saikoside a only for 7 days, NF-κB p50 staining in their nucleus was brown-yellow, but differentiated imDCs was not found. As monocytes were cultured with RPMI-1640 medium containing GM-CSF,IL-4 and saikoside for 7 days, the cells were identified as imDCs by immunocytochemical staining. Then the maturation promoting factor TNF-α was added, cultured imDC for another 3 days, the nucleus appeared obvious NF-κB p50 brown-yellow staining which was stronger than that of cytokine group. These cells were identified as mDC. The surface marker expression of co-culture group stronger than that in cytokine group.Conclusion NF-κB p50 quantity increased in the nucleus when monocytes differentiated into imDC and mDC low concentration of Saikoside a can not stimulate monocytes to differentiate into dendritic cells, but when combined with GM-CSF, IL-4 and TNF-α, the DCs grew much better and the surface antigen expression increased.

It has been proposed that immune injury is the central mechanism of pathogenesis of the infectious disease, severe acute respiratory syndrome (SARS). To gain a better understanding of immune injury in the spleen, we investigated the number and distribution of various immune cell types in the spleens of SARS patients. We performed autopsies on six confirmed SARS cases, with six normal subjects as controls; spleen samples from these autopsies were examined with hematoxylin and eosin (H&E) sections, in situ hybridization for SARS virus genomic sequences, and immunohistochemistry with seven monoclonal antibodies to five cell types. The number and distribution of these cells were measured and analyzed using an image analysis system. SARS genomic sequences were detected in all SARS spleens. The SARS spleens all had severe damage to the white pulp and showed an alteration of the normal distribution of various cell types. Immunocytes in the red pulp were decreased by 68.0-90.7% except for CD68+ macrophages and human leukocyte antigen (HLA)-DR positive antigen-presenting cells (APC), which were decreased to a lesser degree. On average, CD68+ macrophages were increased in size by 2.21-fold. We hypothesize that the collapse of the splenic immune system plays a key role in the clinical outcome of these patients.

Background Chemokine-like factor 1 (CKLF1) was recently identified as a novel cytokine. The full-length CKLF1 cDNA contains 530 bp encoding 99 amino acid residues with a CC motif similar to that of other CC family chemokines. Recombinant CKLF1 exhibits chemotactic activity on leucocytes and stimulates proliferation of murine skeletal muscle cells. We questioned whether CKLF1 could be involved in the pathogenesis of inflammation and proliferation in the lung. Therefore we used efficient in vivo gene delivery method to investigate the biological effect of CKLF1 in the murine lung.Methods CKLF1-expressing plasmid, pCDI-CKLF1, was constructed and injected into the skeletal muscles followed by electroporation. Lung tissues were obtained at the end of week 1,2,3 and 4 respectively after injection. The pathological changes in the lungs were observed by light microscope.Results A single intramuscular injection of CKLF1 plasmid DNA into BALB/c mice caused dramatic pathological changes in the lungs of treated mice. These changes included peribronchial leukocyte infiltration, epithelial shedding, collagen deposition, proliferation of bronchial smooth muscle cells and fibrosis of the lung. Conclusions The sustained morphological abnormalities of the bronchial and bronchiolar wall, the acute pneumonitis and interstitial pulmonary fibrosis induced by CKLF1 were similar to phenomena observed in chronic persistent asthma, acute respiratory distress syndrome and severe acute respiratory syndrome. These data suggest that CKLF1 may play an important role in the pathogenesis of these important diseases and the study also implies that gene electro-transfer in vivo could serve as a valuable approach for evaluating the function of a novel gene in animals.