[Clinical features of vestibular syncope associated with tumarkin attacks in delayed endolymphatic hydrops].

Objective: To analyze the clinical characteristics of vestibular syncope (VS) associated with drop attacks (DA) in delayed endolymphatic hydrops (DEH). Methods: DEH cases with complete data were retrospectively analyzed, including three DEH cases with DA and VS (VS group), and six DEH cases without DA or VS (control group). The clinical profile, the results of neurotological examinations [such as pure tone audiometry, electrocochleography (EcochG), caloric test, vestibular evoked myogenic potentials (VEMP), and video head impulse test (vHIT)] and treatment outcomes were analyzed. Results: (1) In the VS group, there were three cases of ipsilateral DEH; in the control group, there were six cases of ipsilateral type. One case in each group had a history of migraine. (2) The prevalence of abnormal results in caloric test, vHIT, cervical VEMP, and ocular VEMP in the VS group was 3/3, 1/3, 2/2, and 2/2, respectively, and in the control group was 3/6, 0/3, 1/6, and 4/6, respectively. Two cases in each group underwent EcochG, and no identifiable waveform was elicited on the affected side, and-SP/AP ratio of unaffected side was less than 0.4. (3) Patients in both groups were initially treated with conservative medication. Two cases in the VS group subsequently received intratympanic injections of dexamethasone. No DA or VS occurred during a follow-up period lasting over one year. All patients achieved good control of vertigo during the follow-up period. Conclusions: VS may occur in the patients with DEH. The differential diagnosis of syncope in patients with otogenic vertiginous disease can help improve clinical diagnosis and treatment.

[On-pump total arterial revascularization in coronary artery disease patients with left ventricular dysfunction: a multi-center retrospective study].

Objective: To evaluate the clinical outcomes of on-pump total arterial revascularization with bilateral radial artery (BRA) and left internal mammary artery (LIMA) as conduits in coronary artery bypass grafting (CABG) patients with left ventricular dysfunction (LVD). Methods: All the perioperative medical records and follow-up results of coronary artery disease patients with left ventricular ejection fraction (LVEF) ≤ 40% undergoing CABG from 24 heart centers of 15 provinces and autonomous regions in China between July 2015 and December 2019 were retrospectively analyzed. Results: A total of 87 consecutive patients (55 males and 32 females) underwent on-pump CABG with BRA and LIMA, with a mean age of (57.5±9.1) years old. There were 22 patients complicated with primary hypertension, 12 with diabetes mellitus, 8 with peripheral vascular disease, 7 with chronic obstructive lung disease, 12 with mild renal injury and 3 with partial aortic calcification. There were 43 cases with in-stent stenosis, and 21 had left main disease. The mean LVEF and left ventricular end-diastolic diameter (LVEDD) was (35.5±7.3)% and (65.5±2.6) mm, respectively. The mean graft number, aortic cross-clamp time and cardiopulmonary bypass duration was 3.2±0.9, (90.5±22.7) min and (113.4±19.2) min, respectively. There were 32 mitral and 9 aortic valve replacements, and 5 tricuspid annuloplasties. Prophylactic intra-aortic balloon pumps were implanted in 27 patients. There were 2 operative deaths from acute heart failure. After surgery, there were 15 cases of atrial fibrillation, 1 case of acute kidney injury, 1 case of acute myocardial infarction, and 1 cases of stroke. All the patients fulfilled the follow-up, with a mean time of (39.5±7.7) months. At 3 months after surgery, LVEDD was decreased and LVEF was improved significantly compared with pre-operative indicators [(53.0±1.5) mm vs (65.5±2.6) mm, t=9.51 P=0.02; (45.2±3.3)% vs (35.5±7.3)%, t=13.79, P=0.001]. No major cardiac events were reported during the follow-up. At (30.5±7.4) months after surgery, 62.4% of patients (53/85) underwent coronary CT angiography examination, and the results indicated that the graft patency was 98.8%, with only one case of RA occlusion occurred. Conclusion: In selected patients of LVD, on-pump total arterial revascularization with BRA and LIMA conduits was proved to be safe and effective.

The extra domain A of fibronectin facilitates osteoclastogenesis in radicular cysts through vascular endothelial growth factor.

AIM: To analyse the effects of the alternatively spliced fibronectin (FN) gene and its isoforms on osteoclastogenesis in radicular cysts. METHODOLOGY: Specimens of radicular cysts were collected surgically from 22 patients whose radiolucent periapical areas were measured on digital panoramic radiographs before surgery. The associations between the radiolucent areas and FN isoforms, vascular endothelial growth factor (VEGF) expression or micro-vessel density, as well as the relationships amongst them, were analysed by immunohistochemical staining using the antibodies IST-9, BC-1, P1F11, VEGF and CD34. Fibroblasts isolated from those specimens were used to induce Trap + MNCs, and the effects of induction were assessed by blocking FN containing extra domain A (EDA + FN), COX-2 or VEGF in vitro. The effects of EDA exon knockout using CRISPR/Cas system were also assessed. Quantitative PCR was used to analyse relative expression of FN isoforms and osteoclastogenic genes. Data were analysed using linear regression, Spearman's rank correlation analysis, chi-square test and Student's t-test; P < 0.05 was considered significant. RESULTS: Micro-vessel density and EDA + FN staining were positively associated with the size of radiolucent periapical areas (mm2 ; P < 0.05), consistent with a positive association between Trap + MNCs and VEGF expression in fibroblasts (P < 0.05). Blocking the interaction between EDA + FN and fibroblasts inhibited Trap + MNC formation. In addition, EDA exon knockout decreased VEGF expression and inhibited Trap + MNC formation to the extent of blocking VEGF by bevacizumab, but osteoclastogenic induction was restored by recombinant VEGF. Using retrospective clinicopathological data, VEGF staining was shown to be positively associated with EDA + FN staining, micro-vessel density and the size of radiolucent areas (P < 0.05). CONCLUSION: In fibrous capsules of radicular cysts, the alternatively spliced isoform EDA + FN generated by fibroblasts stimulated VEGF expression via an autocrine effect and then facilitated osteoclastogenesis. Both blockage of VEGF and EDA exon knockout could be used to inhibit bone destruction.

BDNF activates TrkB/PLCγ1 signaling pathway to promote proliferation and invasion of ovarian cancer cells through inhibition of apoptosis.

OBJECTIVE: Abnormal expression and activation of tropomyosin-related kinase receptor B (TrkB) are observed in many pathological conditions, including many types of cancer. We try to explore the relationship between ovarian cancer and Brain-derived neurotrophic factor (BDNF), a ligand of TrkB. MATERIALS AND METHODS: Human ovarian cancer cell line SKOV-3 was used in this study. qPCR, immunohistochemistry, and immunoblot were used to assay BDNF and TrkB expression level. Scratch assay was used to test the cell motility, and transwell assay was used to test the cell migration ability. RESULTS: We found that BDNF promotes the proliferation and invasion of human ovarian cancer SKOV-3 cells depend on the activation of TrkB. To illuminate the downstream pathway of BDNF/TrkB, we silenced AKT1 and PLCγ1 by siRNA. The functional assay showed that activated PLCγ1 signaling pathway is necessary for the proliferation and invasion of cancer cells other than the AKT pathway. Further study showed that PLCγ1 could inhibit the apoptosis of cancer cells. CONCLUSIONS: BDNF triggers TrkB/PLCγ1 signaling pathway to promote proliferation and invasion of ovarian cancer cells through inhibition of apoptosis.

Acute nutrient treatment causes alterations in intra-follicular antioxidation and AKT signaling.

This study aimed to determine if short-term nutrient alteration affects (1) ovarian morphology, (2) plasma and ovarian antioxidant capability and (3) cell apoptosis and AKT signaling within the ovary. After estrus synchronization, 24 Hu sheep were assigned to three groups based on the nutrient requirement recommended for maintenance (M): 1 × M (Control), 1.5 × M (S) and 0.5 × M (R) during days 7-14 of their estrous cycle. The results indicated that undernourishment significantly increased the counts and volume of follicles <2.5 mm and decreased the counts and volume of follicles ≥2.5 mm (P < 0.05). Feed restriction altered the plasma and follicular redox balance within follicles ≥2.5 mm by inhibiting total antioxidant capacity, increasing malondialdehyde concentration (P < 0.05) and reducing the mRNA expression levels of superoxide dismutase 2 (SOD2) and glutathione peroxidase (GSH-PX), as well as the activities of total SOD and GSH-PX. Feed restriction also attenuated B-cell lymphoma-2 (BCL2) but enhanced Bcl-2-associated X protein (BAX) and BAX/BCL2 transcription and translation levels in granulosa cells (P < 0.05). Uniform staining intensities of AKT and P-AKT-Ser473 were observed in each follicle stage, whereas weaker P-AKT-Thr308 staining in the antral follicle than in the pre-antral follicle suggested possible involvement of P-AKT-Thr308 during the beginning of follicle development. P-AKT-Ser473 levels in follicles ≥2.5 mm was significantly reduced in the R group (P < 0.05). The results presented in this study demonstrate that suppressed folliculogenesis caused by feed restriction might be associated with attenuated AKT signaling, reduced follicular antioxidant capacity and enhanced granulosa cells apoptosis.

[The effect of monrustet on the exhalation of nitric oxide and inflammatory cytokines in patients with asthma combined with OSAHS].

Objective:To study the effect of monrustet on fractional exhaled nitric oxide and inflammatory cytokines in the elderly with bronchial asthma combined obstructive sleep apnea hypopnea syndrome.Method:150 cases diagnosed as asthma were enrolled in this study. The patients were divided into bronchial asthma combined with OSAHS (n=72)and bronchial asthma (n=78). According to the different treatment methods , patients were divided into observation group and control group. Two groups were given common treatment with inhaled corticosteroids, and the observation group was taken orally montrast 10mg/night additionally. The fractional exhaled nitric oxide was detected before and after treatment. Serum were collected for detection of tumor necrosis factor alpha (TNR-α), Creactive protein (CRP) and interleukin 6 (IL-6), according to the sleep monitoring results, the sleep apnea low ventilation index (AHI) was calculated, and the changes of FeNO level,CRP,TNR-α,IL-6 and AHI were analyzed in the two groups.Result:Before treatment, the gender, age, degree of disease, FeNO level,TNR-α, CRP,IL-6 and AHI differences were not statistically significant in the two groups (P>0.05).After treatment, both groups of FeNO, TNR-α,CRP,IL-6 and AHI decreased before treatment (P < 0.05).The observation group was lower than the control group, and the difference was statistically significant (FeNO level, AHIP< 0.01,TNR-α, CRP,IL-6 P< 0.05).Conclusion::Montelukast can reduce the FeNO level, the expression of TNR-α, CRP, and IL-6 of patients of bronchial asthma combined with OSAHS. Further improvement of the patient's condition, and provide reference value for the clinical medicine of patients with asthma.

[Effects of ketotifen on fractional exhaled nitric oxide in patients with combined allergic rhinitis and asthma syndrome].

Objective:To study effects of ketotifen on fractional exhaled nitric oxide in patients with combined allergic rhinitis and asthma syndrome.Method:One hundred and twelve patients with asthma were selected from department of respiration, Affiliated Hospital of North China University of science and technology from January 2016 to 2017 in March. Eighty-four patients with allergic rhinitis were selected as the subjects, Randomly divided into two groups,including observed group(n=42) and control group(n=42).The control group was treated with antitussive,expectorant and inhaled corticosteroids, while the observed group in the control group combined with ketotifen 1 mg 2/ day oral treatment,the changes of FeNO and IL-17 in the two groups were compared after one month treament.Result:Before treatment, two groups of patients in general IL-17 and fractional exhaled nitric oxide ware no significant difference(P> 0.05).After treatment, compared to the control group, the observed group was significantly more effective,the difference was statistically significant(P <0.05);fractional exhaled nitric oxide in treatment group were lower than those in control group, the difference was statistically significant(P<0.05,P< 0.01).And the observed group was lower than the control group, and the differences were statistically significant,t(P<0.05).Conclusion:Ketotifen significantly reduce the FeNO of allergic rhinitis and asthma syndrome, reduce airway inflammation.

Additional chromosomal abnormalities in core-binding factor acute myeloid leukemia.

Despite sharing a similar genetic abnormality, patients with core binding factor acute myeloid leukemia (CBF-AML), which is characterized by the presence of t(8;21) or inv(16)/t(16;16), show heterogeneous survival. Other molecular or cytogenetic factors are supposed to have an impact on the prognosis. We enrolled 24 CBF-AML patients to determine the impact of cytogenetic abnormality, and c-KIT, FLT3, NPM1, and CEBPA mutations on the prognosis. Only three patients had the c-KIT mutation (3/24, 12.5%) and one had the FLT3 mutation. However, over half of the patients (14/24) harbored additional cytogenetic changes, including ten with loss of sexual chromosomes (LOS) [all in the t(8;21) group], and six had additional abnormalities (two cases had both LOS and additional abnormalities). From this small-number study, no association was found between c-KIT mutation and survival and relapse rate. However, additional chromosome abnormalities had a significant association with relapse of the disease (P = 0.027). Stem cell transplant had a trend of benefitting patients after relapse (P = 0.065). This implies that chromosome abnormalities occur in CBF-AML and might take part in the heterogeneous nature of CBF-AML.

The CRISPR/Cas system inhibited the pro-oncogenic effects of alternatively spliced fibronectin extra domain A via editing the genome in salivary adenoid cystic carcinoma cells.

OBJECTIVES: To identify the association of fibronectin (FN) extra domain A (EDA) with the progression of salivary adenoid cystic carcinoma (SACC). Accordingly, the exclusion of EDA exon through the CRISPR/Cas9 system was investigated as the rescue for such pro-oncogenic splicing. MATERIALS AND METHODS: SACC-83 cells were transiently transfected with plasmids containing recombinant EDA, and the cellular growth and motility were then accessed in vitro. Epithelial-mesenchymal transition (EMT) was investigated with immunohistochemistry, Western blot, and real-time PCR analysis. SACC tissues from 81 patients were used to access the associations between EDA+FN and clinical-pathological parameters. CRISPR/Cas9 plasmids containing sgRNA were designed and co-transfected into SACC-83 cells; the effects of EDA knockout on cellular growth and motility were then accessed. RESULTS: The recombinant EDA exhibited little effect on the proliferation of SACC cells, but significantly promoted the migration and invasion of the cells (P < 0.05), accompanied with upregulated EMT (P < 0.05); consistently, the expression of EDA+FN was positively associated with the metastasis, nerve invasion and recurrence of SACC (P < 0.05). Furthermore, the EDA knockout from the FN gene in most SACC cells resulted in a decrease in cell motility and invasion, as well as prolonged population doubling time, compared with untreated SACC-83 cells (P < 0.05). CONCLUSION: The EDA domain significantly promoted the motility of SACC cells, and positively associated with the tumor progression in patients with SACC. Thus, it is a potential risk factor and also a therapeutic target for SACC. The CRISPR/Cas9 system may control a pro-oncogenic splicing process through the exclusion of EDA exon from the FN gene, leading to inhibition of motility, invasion and proliferation of cancer cells.

Fibroblasts isolated from a keratocystic odontogenic tumor promote osteoclastogenesis in vitro via interaction with epithelial cells.

OBJECTIVES: Investigate the role of the epithelial-mesenchymal interaction of keratocystic odontogenic tumor (KCOT) in influencing osteoclastogenesis. MATERIALS AND METHODS: Fibroblasts isolated from KCOT fibrous capsule and normal gingival mucosa were, respectively, co-cultured with human immortalized oral epithelial cells (HIOECs), and the supernatant was collected to make conditioned medium, in which the osteoclastogenesis of osteoclast precursor cell line Raw 264.7 was observed. Genes related to bone resorption (RANKL, OPG, COX-2, and M-CSF) were analyzed by real-time PCR. Antibodies against human sRANKL and inhibitor of COX-2: NS398 were added to conditioned medium to investigate the inhibitory effect on osteoclastogenesis. RESULTS: Compared with co-cultured gingival fibroblasts and HIOECs (GE-CM), the conditioned medium from co-cultured KCOT fibroblasts and HIOECs (KE-CM) induced more osteoclast-like cell formation and increased NFATC1 mRNA in Raw264.7 cells (P < 0.05). Co-cultured KCOT fibroblasts (KF) and HIOECs, respectively, expressed more COX-2 mRNA than the co-cultured gingival fibroblasts (GF) and HIOECs (P < 0.05). While the ratio of RANKL/OPG in HIOECs co-cultured with KF was also significantly higher than that co-cultured with GF (P < 0.05). The anti-human sRANKL antibody in KE-CM inhibited osteoclastogenesis of Raw264.7 cells; however, NS398 displayed little inhibition. CONCLUSION: An interesting phenomenon of osteoclastogenic effect of KE-CM in vitro was investigated, which suggested an indispensable role of epithelial-mesenchymal interaction of KCOT in its bone destruction. It could be at least partly attributed to the up-regulated ratio of RANKL/OPG in epithelium induced by KCOT fibroblasts, the aggressiveness of tumor as result of epithelial-mesenchymal interaction deserves exploration further.

Hepatitis C transmission from viremic donors in hematopoietic stem cell transplant.

Transmission of hepatitis C virus (HCV) to recipients of hematopoietic stem cell transplant (HSCT) occurs frequently from HCV viremic donors and causes complications. Here, we report the outcomes of 3 cases from our 265 allogeneic HSCTs, whose donors had HCV infections. Successful prevention of HCV transmission was noted in 1 recipient by pretreatment of the donor with peginterferon/ribavirin to undetectable levels of HCV viremia before stem cell harvest. This case stressed the important role of effective antiviral therapy and HCV RNA seronegativity before cell harvest for prevention of HCV transmission in HSCT.

Effects of metoprolol and small intestine RNA on marrow-derived endothelial progenitor cells applied for autograft transplantation in heart disease.

AIMS: The objective of this project was to improve the effect of EPC autograft transplantation and observe the tolerance of EPCs to I/R injury affected by metoprolol and small intestine RNA. MATERIALS AND METHODS: We isolated bone marrow-derived EPCs and examined the effects of metoprolol and small intestine RNA on EPCs to ischemia at different time points after reperfusion. EPCs growth curve, secretion, apoptosis and mortality were also analyzed. RESULTS: EPCs will be better protected if the blood can be recovered within 4 hours after ischemia for cardiac muscle cells and pretreatment of EPCs with metoprolol or small intestine RNA could protect and promote EPCs proliferation. CONCLUSIONS: Our study demonstated that pretreatment of EPCs with metoprolol or small intestine RNA will increase the EPCs proliferation and may improve the EPCs autograft transplantation ability.

Association between migraine and irritable bowel syndrome: a population-based retrospective cohort study.

BACKGROUND AND PURPOSE: Migraine and irritable bowel syndrome (IBS) share many similarities characterized by their epidemiology, periodic pain, lack of definable organic causes, trigger factors, comorbidities and proposed pathophysiology. In this retrospective case-control study, the association between migraine and IBS was investigated using a nationwide population-based database in Taiwan. METHODS: The data were retrieved from the National Health Insurance Research Database (NHIRD) of Taiwan. In all, 14 117 newly diagnosed migraine cases were identified in a subset of the NHIRD and 56 468 migraine-free individuals were randomly selected as the comparison cohort. The multivariate Cox proportional hazards regression model was used to explore the risk of IBS in migraine sufferers after adjusting for demographic characteristics and comorbidities. RESULTS: After adjusting for the covariates, the incidence of IBS was 1.95-fold higher in the migraine cohort than in the comparison cohort (73.87 vs. 30.14 per 10 000 person-years). The adjusted cumulative incidence of IBS was also higher in the migraine group than in the control group in the follow-up years (log-rank test, P < 0.0001). In addition, the risk was most prominent in the youngest group (<30 years old), exhibiting a 3.36-fold increased risk (95% confidence interval 2.44-4.63) of IBS compared with the migraine-free cohort. Moreover, the incidence of IBS in migraine sufferers tended to increase with the frequency of migraine diagnoses. CONCLUSION: The current population-based study demonstrated that migraine is associated with an increased risk of IBS after adjusting for comorbidities, particularly in the young population.

Cladosporium cladosporioides Identified in China on Tobacco Seeds.

Tobacco (Nicotiana tabacum L.) is a leafy, annual, solanaceous plant grown commercially for its leaves. China is the biggest single tobacco market and accounts for more than 40% of the global tobacco consumption (1). Tobacco seed harvested in Guiyang, Guizhou Province, China, are commonly contaminated or infected by various fungal pathogens, which can cause abnormal seedlings with dark brown lesions and stunting of roots and decayed seeds. In 2013, five samples of 500 seeds from tobacco cv. Guiyan 4 were tested for germination on moistened paper on petri dishes. On average, 35% of the seeds from all five samples developed into abnormal seedlings or were decayed and were plated onto potato dextrose agar media and grown for 5 days at 25°C in darkness to confirm the presence of a pathogen. However, one fungus was isolated from an average of 10% of the 500 seeds sampled. It was identified morphologically as Cladosporium cladosporioides (Fresen.) de Vries based on the velvety olive-brown with almost black reverse colony color and dimensions and color of conidia and conidiophores. Conidia formed in long branched chains that readily disarticulate, single celled, elliptical to limoniform, 2 to 8 (avg. 4.3) × 2 to 3 (avg. 2.1) µm. Conidia were pale to olive brown and smooth to verruculose. Ramoconidia were 0 to 1 septate, 7 to 14 (avg. 9.2) × 2 to 4 (avg. 2.6) µm, smooth or sometimes minutely verruculose. Conidiophores were pale to olive brown, macro- and micronemateus, smooth or sometimes verruculose, and of various lengths up to 320 µm long and 2 to 5 µm wide. Primer pair ITS1 and ITS4 was employed to amplify the regions of ITS1-5.8s-ITS2 of the pathogens. Sequences of all three isolates (G3, G10, and G18) (Accession Nos. KF841547, KF841554, and KF841560) were identical to each other and to four sequences in GenBank (JX230994.1, JQ768317.1, JQ768322.1, and AB763555.1). Pathogenicity of the three isolates of C. cladosporioides was verified on tobacco seedlings of 3-week-old grown on wet filter paper in the petri dishes (9 cm in diameter). For each isolate, 20 seedlings incubated in one plate were inoculated with 0.5 ml of a suspension of 105 conidia/ml. Twenty seedlings were treated with sterile water as control treatment. After inoculation, the petri dishes were incubated at 25°C, 100 to 120 µEm-2 S-1, RH > 80%, and 16 h light per day for disease development. At 96 h after inoculation, symptoms comprising medium brown to black lesions on the roots were clearly visible on inoculated plants but not on the control plants. All seedlings inoculated died 9 days after inoculation whereas control seedlings remained symptomless. Re-isolation attempts on PDA from roots demonstrated C. cladosporioides to be present in symptomatic seedlings but not in roots of the control plants. Moreover, the characteristics of the cultured fungi were exactly the same as those originally isolated. Isolates G3, G10, and G18 (KF841547, KF841554, and KF841560) were deposited with the Tobacco Diseased Fungi, Guizhou Academy of Tobacco Sciences, Guizhou, China. Previously, C. cladosporioides has also been isolated from macadamia (Macadamia integrifolia Maiden & Betche) racemes in South Africa (4), from diseased papaya (Carica papaya L.) in Taiwan province of China (2), and from seeds of Amaranthus spp. in Poland (3). To the best of our knowledge, this is the first report of C. cladosporioides causing seed disease on tobacco in China and the disease should be considered in existing disease management practices. References: (1) British American Tobacco Annual Report, 8, 2012. (2) R. S. Chen, et al. Plant Dis. 93:426, 2009. (3) W. Pusz. Phytopathologia 54:15, 2009. (4) N. van den Berg et al. Plant Dis. 92:484, 2008.

Isolated familial pneumothorax in a Taiwanese family with Birt-Hogg-Dubé syndrome.

Primary spontaneous pneumothorax usually occurs as a sporadic event, but may be clustered in certain families with an underlying inherited disorder. Birt-Hogg-Dubι (BHD) syndrome is a rare autosomal dominant disease accounting for familial pneumothorax. BHD syndrome, caused by mutation of the folliculin gene, is characterized by skin fibrofolliculoma, pulmonary cysts, pneumothorax, and renal cancer. We describe a BHD-affected Taiwanese family with clinical and genetic study. A rare mutation of the folliculin gene was detected in the patient and members with pulmonary cysts or pneumothorax, but no skin or renal lesions were found. This mutation was reported in a Taiwanese family and might indicate a pneumothorax-predominant phenotype. Isolated pneumothorax is an uncommon initial presentation of BHD syndrome. Family history should be carefully reviewed when managing a patient with pneumothorax.

Elastase induced lung epithelial cell apoptosis and emphysema through placenta growth factor.

Chronic pulmonary obstructive disease (COPD) is the fourth leading cause of death worldwide, however, the pathogenic factors and mechanisms are not fully understood. Pulmonary emphysema is one of the major components of COPD and is thought to result from oxidative stress, chronic inflammation, protease-antiprotease imbalance and lung epithelial (LE) cell apoptosis. In our previous studies, COPD patients were noted to have higher levels of placenta growth factor (PlGF) in serum and bronchoalveolar lavage fluid than controls. In addition, transgenic mice overexpressing PlGF developed pulmonary emphysema and exposure to PlGF in LE cells induced apoptosis. Furthermore, intratracheal instillation of porcine pancreatic elastase (PPE) on to PlGF wild type mice induced emphysema, but not in PlGF knockout mice. Therefore, we hypothesized that PPE generates pulmonary emphysema through the upregulation of PlGF expression in LE cells. The elevation of PlGF then leads to LE cell apoptosis. In the present study, we investigated whether PPE induces PlGF expression, whether PlGF induces apoptosis and whether the downstream mechanisms of PlGF are related to LE cell apoptosis. We found that PPE increased PlGF secretion and expression both in vivo and in vitro. Moreover, PlGF-induced LE cell apoptosis and PPE-induced emphysema in the mice were mediated by c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (p38 MAPK) pathways. Given these findings, we suggest that the increase in PlGF and PlGF-induced JNK and p38 MAPK pathways contribute to PPE-induced LE cell apoptosis and emphysema. Regulatory control of PlGF and agents against its downstream signals may be potential therapeutic targets for COPD.

Epigenetic silencing of PTPRR activates MAPK signaling, promotes metastasis and serves as a biomarker of invasive cervical cancer.

Epigenetic modifications are a driving force in carcinogenesis. However, their role in cancer metastasis remains poorly understood. The present study investigated the role of DNA methylation in the cervical cancer metastasis. Here, we report evidence of the overexpression of DNA methyltransferases 3B (DNMT3B) in invasive cervical cancer and of the inhibition of metastasis by DNMT3B interference. Using methyl-DNA immunoprecipitation coupled with microarray analysis, we found that the protein tyrosine phosphatase receptor type R (PTPRR) was silenced through DNMT3B-mediated methylation in the cervical cancer. PTPRR inhibited p44/42 MAPK signaling, the expression of the transcription factor AP1, human papillomavirus (HPV) oncogenes E6/E7 and DNMTs. The methylation status of PTPRR increased in cervical scrapings (n=358) in accordance with disease severity, especially in invasive cancer. Methylation of the PTPRR promoter has an important role in the metastasis and may be a biomarker of invasive cervical cancer.

The growth and osteoclastogenic effects of fibroblasts isolated from keratocystic odontogenic tumor.

OBJECTIVES: To investigate the growth characteristics and effects on osteoclastogenesis in fibroblasts isolated from keratocystic odontogenic tumor (KCOT) fibrous capsule. MATERIALS AND METHODS: Fibroblasts isolated from KCOT fibrous capsule and normal gingival mucosa were cultured in vitro. Their colony-forming units and proliferative activity were investigated, and the osteoclastogenic effects were also observed by a co-culture system with osteoclast precursor cell line Raw264.7. The mRNA of several genes related to bone resorption (IL-6, VEGF, COX-2, and M-CSF) was analyzed by real-time PCR. RESULTS: Keratocystic odontogenic tumor fibroblasts developed fewer CFU and had longer population doubling time than gingival fibroblasts (P < 0.05). In contrast to gingival fibroblasts, KCOT fibroblasts expressed less IL-6, COX-2, and M-CSF (P < 0.05); however, the Raw264.7 co-cultured with KCOT fibroblasts developed more osteoclast-like cells and expressed higher level of nfatc1 than that co-cultured with gingival fibroblasts. Increased COX-2 expression and VEGF expression were detected in KCOT fibroblasts and Raw264.7 co-culture system (P < 0.05). CONCLUSION: Although KCOT fibroblasts showed lower level of cell proliferation than gingival fibroblasts, higher osteoclastogenic ability was detected when co-cultured with Raw264.7. These results suggest that the cell-cell interaction in the co-culture system, possibly by increasing COX-2 and VEGF expression, may be responsible for the increased osteoclastogenic effects of KCOT fibroblasts.

First Report of Fusarium Wilt of Tobacco Caused by Fusarium kyushuense in China.

Tobacco (Nicotiana tabacum L.) is a leafy, annual, solanaceous plant grown commercially for its leaves. China accounts for more than 39.6% of total global tobacco production (3). In May 2012, seedlings of tobacco cv. Honghuadajinyuan in a Guiyang tobacco commercial field (Guizhou, China, 26.35° N, 106.42° E) developed symptoms of severe wilting, chlorosis, and stunting. The main stem and taproot exhibited reddish to light brown vascular discoloration; further progression of these symptoms eventually caused mortality of infected seedlings. To isolate the causal agent, necrotic tissues from the symptomatic root were placed on potato dextrose agar (PDA) and incubated at 25°C in darkness. Colonies with white to rose mycelia and red-brown colony colors developed on PDA after 5 days of incubation. Microconidia were abundant, straight or slightly curved, clavate, 0- to 3-septate, and 7.5 to 20.0 × 2.5 to 5.0 µm. Macroconidia were straight or slightly curved, slender, 3- to 5-septate, and 25.0 to 45.0 × 3.3 to 5.0 µm. Based on the observed colony attributes, growth patterns, absence of chlamydospores, micro- and macro-spore attributes (1), and PCR amplification (using primers ITS1/4) combined with translation elongation factor primers (EF1/2) (2), the fungus was identified as F. kyushuense O'Donnell & T. Aoki. Sequence of ITS1-5.8s-ITS2 region of rDNA (GenBank Accession No. JX235957) exactly matched the sequences of F. kyushuense accession AB587020.1 (100% similarity). Analysis of the elongation factor (EF-1alpha) gene of the fungus (JX658565) resulted in a 99% match for F. kyushuense accession AB674297.1. Pathogenicity of the fungus was confirmed by performing Koch's postulate as follows. Pure cultures of the fungus F. kyushuense obtained from symptomatic tissues of tobacco seedlings were grown on PDA for 6 days. Tobacco plants to be used in pathogenicity tests were germinated and grown on potting soils in a plastic container. Additional fertilization was supplied by adding 0.2 g/L of 20-20-20 (N-P-K) in the float water. When seedlings got 6-leaf stage, they were ready for pathogenicity tests. Spores harvested from these culture plates were suspended in sterile distilled water, adjusted to a concentration of 1 × 104 conidia/ml, and inoculated by irrigating 10 ml of the conidia suspension onto roots of each of the 12 tobacco seedlings with 6-leaf stage. A group of 12 seedlings of the same age treated with sterile water served as control. Inoculated seedlings were maintained at 25°C, 100 µE m-2.s-1, relative humidity >70%, and 16 h light per day, and monitored for 9 days for symptom development. Seedlings inoculated with conidia developed disease symptoms with roots with vascular discoloration of roots whereas control seedlings remained symptomless. F. kyushuense was reisolated from the symptomatic seedlings 9 days after inoculation. F. kyushuense has also been isolated from rice seeds in China (4), and from diseased wheat in Japan (1). The common tobacco Fusarium disease reported in China was caused by F. oxysporium f. sp. nicotianae. However, to the best of our knowledge, this is the first report of F. kyushuense causing wilt on tobacco in China and the disease must be considered in existing disease management practices. References: (1) T. Aoki and K. O'Donnell. Mycoscience. 39:1, 1998. (2) D. M. Geiser et al. Eur. J. Plant Pathol. 110:473, 2004. (3) US Census Bureau. Foreign Trade Statistics. Washington DC, 2005. (4) Z. H. Zhao and G. Z. Lu. Mycotaxon. 102:119, 2007.