Common 1H-MRS Characteristics in Patients With Alzheimer's Disease and Vascular Dementia Diagnosed With Kidney Essence Deficiency Syndrome: A Preliminary Study.

Context • Traditional Chinese medicine (TCM) indicates that both Alzheimer's disease (AD) and vascular dementia (VD) should be categorized as dementia and that they have a common etiology and pathogenesis under TCM classification of syndromes, such as with kidney essence deficiency syndrome (KEDS). The pathological location is mainly in the brain. However, it remains unclear whether AD and VD patients with KEDS exhibit a metabolic commonality in the same region of the brain. Objective • The study intended to investigate the metabolic characteristics of the brain using proton magnetic resonance spectroscopy (1H-MRS) in patients with AD and VD who had been diagnosed with KEDS. Design • The research team designed a pilot study, with the participants being allocated to 3 groups: (1) an AD group, (2) a VD group, and (3) a control group. All data analysis was carried out by a trained radiologist who was blinded to each participant's diagnosis. Setting • The study took place at the Tongde Hospital of Zhejiang Province (Zhejiang Sheng, China). Participants • Participants were patients at the Tongde Hospital with mild AD or VD who had been diagnosed with KEDS. The normal controls were patients' spouses or guardians with normal cognitive function. Outcome Measures • All participants underwent 1H-MRS. The N-acetyl aspartate (NAA)/myo-inositol (mI), NAA/creatine (Cr), choline (Cho)/Cr, and mI/Cr ratios were bilaterally measured in the posterior cingulate gyrus (PCG) and anterior cingulate gyrus (ACG) by the Syngo spectroscopy postprocessing package. Demographic characteristics and 1H-MRS data were assessed across the AD, VD, and normal control groups. Results • Thirteen patients with mild AD with KEDS, 15 patients with mild VD with KEDS, and 18 normal controls were recruited from May 2013 through May 2014. The AD and VD groups did not significantly differ in the NAA/mI, NAA/Cr, Cho/Cr, and mI/Cr ratios in either the PCG or the ACG, with the exception being the Cho/Cr ratio in the right ACG, for which the ratio of the AD group was significantly lower when compared with that of the VD group (P < .05). Conclusions • Mild AD and VD with KEDS showed similar patterns for the 1H-MRS on the cingulate cortex, providing evidence for a common pathogenesis for the KEDS that is associated with AD and VD, providing a modern biological basis for the philosophy of providing the same treatment for different diseases.

An improved method on isolation and serial passage of Chlamydia pneumoniae from human peripheral blood mononuclear cells.

BACKGROUND: Conventional method for Chlamydia pneumoniae (Cpn) isolation and propagation is technically challenging and time-consuming. Here, we developed a method to improve the isolation and passage of Cpn collected from human peripheral blood mononuclear cells (PBMCs). METHODS: PBMCs positive with Cpn antigen (Cpn-Ag) were isolated, then centrifuged and cultured with Hep-2 cells after being broken. Cells were broken again and put into new Hep-2 cells to finish totally four passages with isolated and imported Cpn. Microimmunofluorescence method was used to detect Cpn. Inclusion forming unit (IFU) number was counted for each passage. Polymerase chain reaction (PCR) method was used to detect Cpn DNA. Efficiency of different centrifugation modes was compared. RESULTS: Hep-2 cells of the first and second passages were strong positive with Cpn-Ag, the third passage was positive, and the fourth negative. Degeneration appeared in the fourth passage for isolated Cpn and third passage for imported strain. Centrifugation mode of 1,000 rpm for 2 h was the most efficient for Cpn propagation and passage. CONCLUSION: This simplified method achieved efficient isolation, propagation, and passage of Cpn from PBMCs, and isolated strain was superior to imported strain on propagating ability.

Effect of Pseudomonas aeruginosa on the expressions of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 in the airway of bronchiectasis patients.

OBJECTIVE: To study the expressions of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the airway of bronchiectasis (BE) patients, and evaluate the effect of pseudomonas aeruginosa (PAE) on the expression of TIMP-1 and MMP-9. METHODS: In this case-control study, subjects were divided into BE group and control group, and the BE group was further divided into PAE group and other bacteria group based on the culture results of bronchoalveolar lavage fluid (BALF). BALF was obtained by bronchoscopy, the expressions of MMP-9 and TIMP-1 were determined by ELISA, then the ratio of TIMP-1/MMP-9 was calculated. Furthermore, the tissue of bronchic endomembrane was obtained by transbronchial biopsy and the expressions of MMP-9 and TIMP-1 were determined using immunohistochemical method. RESULTS: The levels of MMP-9 in the BALF of PAE group and other bacteria group were significantly higher than that in control group (P=0.0000 both), and the expressions of MMP-9 in bronchic endomembrane of PAE group and other bacteria group were also significantly higher (P=0.0421 and 0.0003, respectively). The level of TIMP-1 in BALF of PAE group was significantly lower than that in other bacteria group (P=0.0324). The ratio of TIMP-1/MMP-9 in BALF of BE group was significantly lower than that in control group(P=0.0000), and this ratio of PAE group was significantly lower than those in both other bacteria group and control group (P=0.0026 and 0.0000, respectively). CONCLUSION: PAE infection in BE patients can suppress the expression of TIMP-1 and stimulate the expression of MMP-9, and thus make the disease even worse.

Study on the Effects of Kuanxiong Aerosol on the Isolated Artery and Rabbits Acute Myocardial Ischemia Model.

BACKGROUND: Kuan Xiong aerosol (KXA) is a Chinese herbal compound used to regulate qi-flowing to relieve pain and improve angina. However, only a few pharmacological studies on this traditional Chinese medicine preparation have been reported to confirm these activities. OBJECTIVES: This article aims to observe the effect of resisting acute myocardial ischemia (AMI) in vivo and dilating vessel in vitro of KXA. METHODS: The AMI model involves intravenously injecting the pituitary (2 U.kg-1) into the ear of rabbits. Electrocardiograph (ECG) T waves were then recorded after administration, and the falling range was calculated. Following this, the level of serum Cardiac troponin T (cTn-T) and the histopathology of the cardiac muscle tissue were evaluated. In vitro, the effect of KXA on vasodilation of isolated aortic rings that had been pre-contracted with KCl (30 mM) was observed. RESULTS: It was found that KXA reduced ECG ST-T waves and serum cTn-T in the rabbit AMI model, protecting myocardial tissue from fracturing and loss of myocardial fibers and inhibiting inflammatory cell infiltration, cavitation degeneration, and karyopyknosis of the myocardial matrix. Furthermore, the administration of 0.215, 1.075, and 2.150 mg.mL-1 of KXA resulted in significant relaxation of the aortic rings at a rate of 69.63 %, 90.14 %, and 118.72 % (p < 0.01) in the untreated ones, and a second shrinkage ratio of 20.17 %, 4.29 %, and 4.54 % (p < 0.01) in the untreated ones, respectively. CONCLUSION: These results suggest that KXA protects against AMI, contributes to the dilation of blood vessels, and has long-acting effectiveness.

Inhibition of miR-29b-1-5p Attenuates Inflammatory Response and Pulmonary Fibrosis in LPS-Induced Acute Lung Injury by Regulating RTN4 Expression.

Objective: Acute lung injury (ALI) is a severe respiratory disorder causing alveolar-capillary barrier, leading to a high rate of morbidity and death in critically ill individuals. microRNAs (miRNAs)-mediated mechanism in the pathogenesis of ALI has attracted much interest. Herein, we attempt to characterize a candidate miRNA and its downstream target that is linked to the pathogenesis of ALI. Methods: LPS-conditioned MH-S cells were treated with miR-29a-1-5p mimic, inhibitor, and RNT4 expression vector, and the ALI animal model was injected with agomir and antagomir of miR-29b-1-5p and RNT4 expression vector, in which the pro-inflammatory cytokine production, cell viability and apoptosis, myeloperoxidase (MPO) activity, wet/dry (W/D) ratio, and expression of TGF-ß1, α-smooth muscle actin (α-SMA), E-cadherin, and vimentin were examined. miR-29a-1-5p inhibition of RTN4 translation was confirmed by luciferase activity assays. Results: An elevated miR-29a-1-5p expression was demonstrated in LPS-conditioned MH-S cells. miR-29a-1-5p inhibitor transfection attenuated the production of pro-inflammatory cytokines and MH-S cell viability but enhanced the apoptosis. miR-29a-1-5p inhibition of RTN4 translation was demonstrated in the setting of LPS-induced ALI. LPS-induced murine models demonstrated upregulated miR-29a-1-5p. Intravenous injection of miR-29b-1-5p agomir attenuated mouse lung injury and pulmonary fibrosis. RTN4 overexpression resisting to miR-29a-1-5p overexpression was demonstrated in LPS-induced murine models. Conclusion: The findings obtained from the study that disturbing the action of miR-29a-1-5p may be a novel therapeutic strategy for preventing ALI.

High expression of hypoxia inducible factor 1α related with acquired resistant to EGFR tyrosine kinase inhibitors in NSCLC.

The acquired resistance of the first generation epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) is a main factor leading to poor prognosis of non-small cell lung cancer (NSCLC), so we researched whether the high expression of hypoxia-inducible factor-1α (HIF-1α) in EGFR-TKIs sensitive NSCLC tissue tends to induce the acquired resistance. We detected the HIF-1α in normal lung tissue, EGFR-TKIs sensitive NSCLC tissue, the first generation EGFR-TKIs acquired resistant NSCLC tissue and acquired EGFR T790M mutation NSCLC tissue with the method of immunohistochemistry. Then, we compared the expression of HIF-1α in these tissues, and evaluate the effect of HIF-1α expression to the occurrence of acquired resistance. The expression of HIF-1α was much higher in the EGFR-TKIs sensitive NSCLC tissue than that in normal lung tissue. HIF-1α level became higher after the occurrence acquired resistance. There was negative correlation between HIF-1α level before receiving treatment and the time of acquired resistance occurring as well as the acquired EGFR T790M mutation occurring. As the treatment going on, EGFR-TKIs sensitivity rate of low HIF-1α level group was much higher than that of high level group. The high expression of HIF-1α related with the acquired resistance of the first generation EGFR-TKIs, and HIF-1α can be a biomarker to predict the early occurrence of acquired resistance.

HIF-1 Inhibitor YC-1 Reverses the Acquired Resistance of EGFR-Mutant HCC827 Cell Line with MET Amplification to Gefitinib.

BACKGROUND: Acquired resistance occurred in the majority of nonsmall cell lung cancer (NSCLC) patients receiving epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) therapy, and this may be related to the activation of the HIF-1 pathway. Therefore, we examined the influence of the hypoxia-inducible factor-1 (HIF-1) pathway inhibition on the sensitivity of HCC827 gefitinib-resistant (HCC827 GR) cells with MET amplification to gefitinib. METHODS: We established HCC827 GR cell line with MET amplification and set four groups with different treatment. An MTT assay, a colony formation analysis, and a wound healing assay were performed to determine the sensitivity change of HCC827 GR cells after different treatments. HIF-1α, p-EGFR, and p-Met levels were detected with western blot. Correlations among HIF-1α, p-EGFR, and p-Met levels of HCC827 GR cells with different treatments were analyzed with Pearson's correlation analysis. RESULTS: HIF-1 inhibitor YC-1 enhanced the sensitivity of HCC827 GR cells to gefitinib. p-Met level was correlated with HIF-1α level, while there was no correlation between p-Met level and p-EGFR level. CONCLUSION: HIF-1 inhibitor YC-1 is able to reverse the acquired resistance of HCC827 GR to gefitinib, and the regulation of the HIF-1 pathway on MET may be one of the mechanisms.

Hypoxia-inducible factor-1 signaling pathway influences the sensitivity of HCC827 cells to gefitinib.

The majority of patients with non-small cell lung cancer (NSCLC) with activating epidermal growth factor receptor (EGFR) mutations inevitably progress in stage despite an initial substantial and rapid response to EGFR-tyrosine kinase inhibitors (EGFR-TKIs). Previous research indicates that hypoxia may be associated with resistance to EGFR-TKIs in EGFR mutation-positive NSCLC. Therefore, the present study regulated the activity of hypoxia-inducible factor-1 (HIF-1) signaling pathway to observe if it is able to alter the sensitivity of lung cancer cells to gefitinib. The present study selected 3-(5'-hydroxymethyl-2'-furyl)-1-benzylindazole (YC-1) and dimethyloxalylglycine (DMOG) as a HIF-1 signaling pathway inhibitor and activator, respectively, on HCC827 cells. Cells were incubated with different treatments for different durations: A blank control, DMOG, gefitinib, or DMOG and gefitinib combined, for 36 and 48 h; and then a blank control, YC-1, gefitinib, or YC-1 and gefitinib combined, for 16 and 28 h. A western blot analysis assay was performed to evaluate the protein expression levels of HIF-1α and phosphorylated hepatocyte growth factor receptor (p-MET), an MTT assay was used to determine cell proliferation, a colony formation assay was used to investigate the colony-forming ability and a wound healing assay was used to test the cell migration ability. Additionally, Pearson's correlation analysis was used to evaluate the correlation between p-Met and HIF-1α expression levels. Finally, it was identified that gefitinib and DMOG combined notably improve the growth and cell migration ability of HCC827 cells, compared with gefitinib alone. When gefitinib and YC-1 were combined, the inhibiting effect on the growth and cell migration ability of HCC827 cells was substantially enhanced, compared with the control cells. Pearson's correlation analysis revealed that the p-Met expression level had a strong positive correlation with HIF-1α expression levels. Thus, it was concluded that the HIF-1 signaling pathway influences the sensitivity of HCC827 cells to gefitinib. The positive correlation between p-Met and HIF-1α expression levels may be the underlying mechanism of the HIF-1 signaling pathway influencing the sensitivity of HCC827 cells to gefitinib.

Value of Serum Carbohydrate Antigen 19-9 and Carcinoembryonic Antigen in Evaluating Severity and Prognosis of Connective Tissue Disease-Associated Interstitial Lung Disease.

OBJECTIVES: This study aims to detect serum carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA 19-9) levels in connective tissue disease-associated interstitial lung disease (CTD-ILD) patients and to demonstrate their values in evaluating the severity and prognosis of CTD-ILD. PATIENTS AND METHODS: The study included 82 CTD-ILD patients (54 males, 28 females; mean age 67.9 years; range 29 to 91 years) and 82 controls (54 males, 28 females; mean age 68.1 years; range 30 to 92 years). Patients were followed-up for 12 months. Correlations of serum CEA and CA 19-9 with disease severity parameters (pulmonary function, oxygenation index and involvement score on high resolution computed tomography) were analyzed. Survival analysis was used to evaluate significance of serum CEA and CA 19-9 as prognosis predictors. RESULTS: Serum CEA and CA 19-9 levels were higher in CTD-ILD patients compared with controls (both p<0.05) and correlated with disease severity (p<0.05 for all R2). High levels of serum CEA and CA 19-9 were associated with poor survival (both p<0.05). Serum CEA level was indicated as a prognostic factor for cumulative survival (hazard ratio=1.685, 95% confidence interval: 1.405-2.021, p=0.001). CONCLUSION: In CTD-ILD patients, serum CEA and CA 19-9 are elevated and can be indicators of disease severity. Moreover, serum CEA is a significant and independent predictor of survival.