Imperatorin ameliorates pulmonary fibrosis via GDF15 expression.

Background: Pulmonary fibrosis features in damaged pulmonary structure or over-produced extracellular matrix and impaired lung function, leading to respiratory failure and eventually death. Fibrotic lungs are characterized by the secretion of pro-fibrotic factors, transformation of fibroblasts to myofibroblasts, and accumulation of matrix proteins. Hypothesis/purpose: Imperatorin shows anti-inflammatory effects on alveolar macrophages against acute lung injury. We attempt to evaluate the properties of imperatorin on the basis of fibroblasts. Methods: In in vitro, zymosan was introduced to provoke pro-fibrotic responses in NIH/3T3 or MRC-5 pulmonary fibroblasts. Imperatorin was given for examining its effects against fibrosis. The mice were stimulated by bleomycin, and imperatorin was administered to evaluate the prophylactic potential in vivo. Results: The upregulated expression of connective tissue growth factor (CTGF), α-smooth muscle actin (α-SMA), and collagen protein due to zymosan introduction was decreased by imperatorin in fibroblasts. Zymosan induced the activity of transglutaminase 2 (TGase2) and lysyl oxidase (LOX), which was also inhibited by the administration of imperatorin. Imperatorin alone enhanced sirtuin 1 (SIRT1) activity and growth differentiation factor 15 (GDF15) secretion in fibroblasts via LKB1/AMPK/CREB pathways. In addition, GDF15 exerted a beneficial effect by reducing the protein expression of CTGF, α-SMA, and collagen and the activities of TGase and LOX. Moreover, orally administered imperatorin showed prophylactic effects on bleomycin-induced pulmonary fibrosis in mice. Conclusion: Imperatorin reduces fibrotic marker expression in fibroblasts and also increases GDF15 secretion via the LKB1/AMPK/CREB pathway, attenuating pro-fibrotic responses in vitro. Imperatorin also alleviates pulmonary fibrosis induced by bleomycin in vivo.

[Epidemiological survey of malocclusion in 8-9 year-old children in Qingdao Laoshan district].

PURPOSE: To investigate the prevalence of malocclusion and the correlation between caries and malocclusion in mixed dentition of primary students in Laoshan district of Qingdao city, to provide a scientific basis for early orthodontic treatment. METHODS: According to WHO's oral health survey (basic methods), a survey concerning oral health was performed in 1803 children aged 8-9 years old. SPSS 17.0 software package was used for statistical analysis . RESULTS: 1322 of 1803 pupils suffered from malocclusion, the prevalence of malocclusion in children surveyed was (73.32%). There were 519 cases(39.26%) of Class I malocclusion, 374 cases(28.29%) of Class II malocclusion, 357(27.00%) cases of Class III malocclusion, 72 cases (5.45%) with asymmetric malformation. As regard to clinical manifestations, deep overbite accounted for(37.67%) ,followed by deep overjet(29.19%), dental crowding (16.94%) and crossbite(16.03%). The prevalence of caries was 64.23%. Among them, there were 625 males (34.66%) and 532 females(29.51%). CONCLUSIONS: The prevalence rate of malocclusion in mixed dentition of primary students in Laoshan district of Qingdao district is not optimistic, the incidence of malocclusion is closely related to caries.

Anti-inflammatory Property of Imperatorin on Alveolar Macrophages and Inflammatory Lung Injury.

Imperatorin is one of the furanocoumarin derivatives and exists in many medicinal herbs with anticancer, antiviral, antibacterial, and antihypertensive activities. In this study, we examined the anti-inflammatory effects of imperatorin on inflammation-associated lung diseases. Imperatorin reduced iNOS and COX-2 expression and also IL-6 and TNFα production enhanced by zymosan. Imperatorin also inhibited the signaling pathways of JAK/STAT and NF-κB. Moreover, in vivo study also revealed that zymosan-induced immune cell infiltration, pulmonary fibrosis, and edema were relieved by imperatorin in mice. We found that imperatorin exerts anti-inflammatory effects that are associated with amelioration of lung inflammation, edema, and rapid fibrosis. Studies on alveolar macrophages also reveal that imperatorin reduced the production of pro-inflammatory mediators and cytokines and inhibited pro-inflammatory JAK1/STAT3 and NF-κB signaling pathways. These results indicate that imperatorin may be a potential anti-inflammatory agent for inflammatory-associated lung diseases.

Circulating tight junction proteins mirror blood-brain barrier integrity in leukaemia central nervous system metastasis.

The aim of this study was to evaluate the clinical significance of circulating tight junction (TJ) proteins as biomarkers reflecting of leukaemia central nervous system (CNS) metastasis. TJs [claudin5 (CLDN5), occludin (OCLN) and ZO-1] concentrations were measured in serum and cerebrospinal fluid (CSF) samples obtained from 45 leukaemia patients. Serum ZO-1 was significantly higher (p < 0.05), but CSF ZO-1 levels were not significantly higher in the CNS leukaemia (CNSL) compared to the non-CNSL. The CNSL patients also had a lower CLDN5/ZO1 ratio in both serum and CSF than in non-CNSL patients (p < 0.05). The TJ index was negatively associated with WBCCSF , ALBCSF and BBB values in leukaemia patients. Among all of the parameters studied, CLDN5CSF had the highest specificity in discriminating between CNSL and non-CNSL patients. Therefore, analysing serum and CSF levels of CLDN5, OCLN and the CLDN5/ZO1 ratio is valuable in evaluating the potential of leukaemia CNS metastasis. Copyright © 2016 John Wiley & Sons, Ltd.

Diagnostic value of platelet indices and bone marrow megakaryocytic parameters in immune thrombocytopenic purpura.

Platelet indices could mirror megakaryopoietic activity in immune thrombocytopenic purpura (ITP), but its specificity and sensitivity need to be studied. The diagnostic performance of platelet indices was analyzed by receiver-operating characteristic curves, and the probability of true positive (sensitivity) and true negative (specificity) in predicting ITP, myelodysplasia, or controls was determined. Mean platelet volume (MPV) was higher, whereas plateletcrit (PCT) was significantly lower in ITP than in myelodysplasia and controls. The platelet distribution width in ITP patients was lower than in myelodysplasia, but higher than in controls. Increased megakaryocytes were only observed in ITP. A strong positive correlation was found between MPV and quantities of granular megakaryocytes, whereas a negative relationship existed between MPV and platelet-form megakaryocytes. In receiver-operating characteristic analysis, MPV and PCT gave a sensitivity of 70.3% (89.8%) and specificity of 74.8% (84.7%) at a cutoff of 9.35 (0.085) in diagnosis of ITP. Combined parallel test of MPV and PCT increased the sensitivity to 97.5 with 64.1% specificity, whereas series test increased the specificity to 94.7 with 62.7% sensitivity. Our results suggest that MPV, PCT, and platelet distribution width represent megakaryopoietic activity in bone marrow and may be reliable markers in ITP diagnosis.

Clinical and laboratory features parameters of human granulocytic anaplasmosis (HGA) in patients admitted to hospital in Guangdong Province, China.

BACKGROUND: Human granulocytic anaplasmosis (HGA) is an acute tick-borne infectious disease with increasing morbidity and mortality, but is rarely considered in clinical practice. Because human-to-human transfusion or nosocomial transmission can occur, diagnosis is difficult when the history of tick bites is not clear. METHODS: We present clinical features and laboratory data of HGA patients who had no clear tick bite history. RESULTS: All patients in the study presented with a high fever, petechiae, purpura, nose bleeding and leukopenia, and patients had abnormally high levels of serum ferritin and C-reactive protein. Morulae in leukocytes were observed in three patients. Foamy histiocytes and slight erythrophagocytic activity were only found in severely ill patients. CONCLUSION: In patients with fever and thrombocytopenia in whom no other diagnosis is evident on clinical assessment, HGA should be considered in the differential diagnosis, and tested for serologically if possible. For patients in whom the diagnosis of HGA is possible, and to whom tetracyclines can safely be given, it is apparent that these drugs hasten recovery and improve the prognosis.

The prognostic significance of serum and cerebrospinal fluid MMP-9, CCL2 and sVCAM-1 in leukemia CNS metastasis.

Metastasis to the central nervous system (CNS) is the primary obstacle in leukemia treatment. Matrix metalloproteinase-9 (MMP-9), chemokine ligand-2 (CCL2) and soluble vascular adhesion molecule-1 (sVCAM-1) play crucial roles in tumor cell adhesion, motivation and survival, but their roles in leukemia CNS metastasis remain to be elucidated. We investigated the prognostic significance of serum and cerebrospinal fluid (CSF) MMP-9, CCL2 and sVCAM-1 in leukemia patients to explore their potential as predictive biomarkers of the development of CNS leukemia (CNSL). MMP-9, CCL2 and sVCAM-1 were measured in paired CSF and serum samples collecting from 33 leukemia patients with or without CNS metastasis. Other risk factors related to CNSL prognosis were also analyzed. sVCAM-1Serum and CCL2Serum/CSF were significantly higher in the CNSL group than in the non-CNSL group and the controls (p < 0.05). MMP-9Serum was insignificantly lower in the CNSL group than in the non-CNSL group and the controls (p > 0.05). No differences were found for the sVCAM-1Serum, CCL2Serum, and MMP-9Serum levels between non-CNSL patients and controls (p > 0.05). MMP-9CSF was significantly higher in the CNSL group than both the non-CNSL and the control groups (p < 0.05). The indexes of sVCAM-1, CCL2, and MMP-9 in the CNSL group were lower than in the controls (p < 0.05). Positive correlations were determined between the MMP-9CSF and the ALBCSF/BBB value/WBCCSF, between sVCAM-1Serum and the WBCCSF/BBB value. Negative correlations existed between MMP-9Serum and the ALBCSF/BBB value/WBCCSF, and between the CCL2 index and ALBCSF. sVCAM-1Serum was positively associated with event-free survival (EFS), and patients with higher levels of ALBCSF, MMP-9CSF/Serum, CCL2CSF/Serum, and sVCAM-1CSF/Serum had shorter EFS. MMP-9CSF, CCL2CSF and sVCAM-1CSF are the first three principal components analyzed by cluster and principal component analysis. Our data suggest that MMP-9, CCL2 and sVCAM-1 in the CSF may be more potent than serum in predicting the possibility of leukemia metastatic CNS and the outcome of CNSL patients.

Association of miR-146a gene polymorphism with risk of nasopharyngeal carcinoma in the central-southern Chinese population.

This case-control study focused on estimating the association between miR-146a polymorphism and risk of nasopharyngeal carcinoma (NPC) in central-south China. In total, 160 patients with NPC and 200 healthy controls in central-south China were genotyped using polymerase chain reaction-restriction fragment length polymorphism assay. Chi-square test was used to assess the different distribution of miR-146a polymorphism between NPC patients and controls; and logistic regression analysis was applied to analyze the associations between miR-146a polymorphism with cancer risk in different contrast models. Significant differences between NPC patients and controls were found in genotype (P=0.033 for GG versus CG versus CC; and odds ratio (OR)=0.568, 95% confidence interval (CI)=0.354-0.912, P=0.019 for CG versus CC; and OR=0.503, 95% CI=0.261-0.971, P=0.041 for CG versus CC; and OR=0.564, 95% CI=0.360-0.884, P=0.012 for GG+CG versus CC, respectively) and allelic analysis (P=0.025 for G versus C). Our findings suggested that polymorphism of mir-146a was associated with NPC in the central-southern Chinese population.

Association study between miR-149 gene polymorphism and nasopharyngeal carcinoma.

Association studies between single-nucleotide polymorphism (SNP) rs2292832 on miR-149 gene and cancer risk have been previously analyzed in several types of cancer. The aim of this study was to evaluate the association between miR-149 polymorphism and risk of nasopharyngeal carcinoma (NPC). miR-149 gene polymorphism was genotyped using polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) in 158 patients with NPC and 242 healthy individuals. Associations with cancer risk and clinicopathological characteristics were analyzed by χ2 test. No significant difference was observed for miR-149 gene polymorphism in NPC patients and healthy controls in either genotype (P=0.427 for CC vs. CT vs. TT, P=0.247 for CT vs. TT and P=0.323 for CC vs. TT, respectively) or allelic analysis (P=0.216). No significant difference was noted between the genotypes and the clinicopathological parameters examined with the exception of clinical stage. A significantly higher CC distribution in clinical stage I-II compared with III-IV was observed under the dominant model (CC vs. CT vs. TT, P=0.026) and the co-dominant model (CC vs. TT, P=0.030). The results of this study suggested that the CC genotype of miR-149 contributes to the progression and development, rather than the initiation of NPC.

[Detection and significance of transcription factors and cytokines of Th17/Treg cells in peripheral blood in the gastric cancer patients].

OBJECTIVE: To detect the expression levels of transcription factors and associated cytokines of Th17 and Treg cells in peripheral blood mononuclear cells (PBMC) of patients with gastric cancer, and explore the possible pathological mechanism of these cells involved in the development of gastric cancer. METHODS: The mRNA levels of RORgammat, FoxP3 in PBMC were determined by quantitative real-time PCR (QRT-PCR) from 57 patients with gastric cancer, 31 patients with benign gastric illness and 40 healthy people. The concentration of IL-17, IL-23, TGF-beta, IL-10 in plasma were detected by enzyme linked immunosorbent assay (ELISA). RESULTS: Compared with healthy volunteers, patients with gastric cancer showed higher levels of RORgammat and FoxP3 in PBMC (P < 0.05). The ratio of FoxP3/RORgammat in gastric cancer group was higher than that in the volunteer group and benign gastric illness group (P < 0.05). The ratio of FoxP3/RORgammat was higher in advanced disease than early disease (P < 0.05). The expressions of IL-17, IL-23, TGF-beta and IL-10 were higher in patients with gastric cancer than that in healthy volunteers (P < 0.05). In addition, The expression of TGF-beta and IL-10 were significantly increased in the advanced disease group than that in the early group (P < 0.05), but IL-17 and IL-23 was not significantly changed between the two groups (P > 0.05). CONCLUSION: There are higher levels of Th17 and Treg cells in gastric cancer patients, and it also shows a persistent predominant tendency of Treg cells and a reduced tendency of Th17 cells in advanced disease. Detecting the expression of Th17/Treg transcription factor and related cytokines would contribute to the diagnosis and prediction of the disease development and prognosis.

Effects of estrogen and tibolone on bladder histology and estrogen receptors in rats.

BACKGROUND: Estrogen deficiency causes atrophic changes within the urogenital tract, and is associated with urinary symptoms. The purpose of this study was to investigate the effects of estrogen and tibolone on bladder histology, and the changes of estrogen receptor alpha and beta (ERalpha and beta) protein expression in the detrusor muscle. METHODS: Forty female rats were separated into four groups of ten each. They received a sham operation (Sham), ovariectomy (Ovx), ovariectomy plus estrogen replacement (Ovx + E), or ovariectomy plus tibolone treatment (Ovx + T). After 12 weeks each rat was anesthetized and the bladders were removed. The bladders' ultra structure, collagen fiber (CF) to smooth muscle (SM) ratio and ER subtypes were studied. Statistical analyses were performed using the one-way analysis of variance test. RESULTS: Ovx resulted in significant degeneration in bladder ultra structure; however, estrogen and tibolone reversed those changes. Ovx increased the CF/SM ratio, estrogen and tibolone resulted in an increase. Two estrogen receptors (ERs) were expressed in the bladder detrusor, with ERbeta the main subtype. Ovx resulted in up-regulation of ERalpha and down-regulation of ERbeta. With estrogen and tibolone treatment, ERbeta showed a significant increase but ERalpha showed no significant difference compared with Ovx. CONCLUSIONS: Estrogen deficiency deteriorates bladder ultra structure and histology. Supplementary estrogen can improve bladder function which may be due to inhibition of collagen hyperplasia and increased SM density. ERbeta has an important role in mediating estrogen function in the bladder. Tibolone has a mild estrogenic action and has an effect on bladder function and structure to some degree.

[Cloning and expression of human Runx3 gene obtained from human peripheral blood CD8(+) T lymphocyte].

AIM: Runx3, a type of Runt family member, plays an important role in immune regulation. In this study, we cloned and analyzed the cDNA encoding human Runx3 from T lymphocyte, expressed Runx3 protein in E.coli system, and studied the relation between Runx3 and some immune disorders or tumors. METHODS: The CD8(+) T were isolated from human peripheral blood with MACS, Runx3 cDNA was amplified by RT-PCR and cloned into pMD19-T vector, and recombinant was transformed into competent cells DH5alpha and recombinant sequencing were performed. The identical was subcloned into pQE30 vector and expressed in E.coli M15. The fusion protein was identified by Western blot. RESULTS: The 1,248 bp fragment amplified by RT-PCR was the same as the anticipated one in size and encodes 415 amino acids. Runx3 protein was gained. CONCLUSION: Human Runx3 gene was cloned and expressed in E.coli system successfully, which brought a foundation for further research on its biological function.

[Effect of ovariectomy and estrogen replacement on bladder function and structure in rats].

OBJECTIVE: To investigate the mechanism of estrogen action on bladder. METHODS: By examining the functional, histological and ultrastructural changes that occurred in the bladder of rats with estrogen depletion and supplementation, we studied the actions of estrogen on bladder. A total of 30 female adult Sprague-Dawley rats were divided into 3 groups, including control group, bilateral ovariectomy (OVX) group and bilateral ovariectomy plus estradiol valerate (OVX + E) group. After medication for 12 weeks, cystometry was performed. Masson stain was used to quantify the area densities of collagen fiber and smooth muscle and their ratios, and electron microscopy was used to evaluate ultrastructure of detrusor muscle. RESULTS: In OVX group, bladder maximal capacity, (0.32 +/- 0.20) ml, compliance, (0.012 +/- 0.006) ml/cm H2O (1 cm H2O = 0.098 kPa), and maximal contraction force, (1.4 +/- 0.4) cm H2O were obviously lower than those in control group, which were (1.11 +/- 0.09) ml, (0.026 +/- 0.003) ml/cm H2O, and (4.4 +/- 0.3) cm H2O, respectively (P < 0.05). In OVX + E group, bladder maximal capacity (0.83 +/- 0.10) ml was lower than that in control group (P < 0.05), but compliance, (0.029 +/- 0.003) ml/cm H2O and maximal contraction force, (4.8 +/- 1.4) cm H2O were similar to those of control group (P > 0.05). Collagen fiber area density and collagen fiber to smooth muscle area density ratio in OVX group (0.218 +/- 0.041 and 0.54 +/- 0.08) were higher than those in control group (0.160 +/- 0.039 and 0.32 +/- 0.09, P < 0.05); however, there were no significant differences between OVX + E group (0.178 +/- 0.027, 038 +/- 0.06) and control group (P > 0.05). In OVX group, ultrastructure of detrusor muscle showed degenerative change, while it was not found in the other groups. CONCLUSIONS: When the rat was ovariectomized, bladder function became poor and supplementary estrogen could improve bladder function. This effect may be due to inhibition of collagen hyperplasia and protection of cell organelles. Estrogen replacement should be beneficial for treating postmenopausal lower urinary tract symptoms.