Research progress of lung aging in chronic respiratory diseases / 生理学报

Cell aging is an extremely complex process, which is characterized by mitochondrial structural dysfunction, telomere shortening, inflammatory microenvironment, protein homeostasis imbalance, epigenetic changes, abnormal DNA damage and repair, etc. Aging is usually accompanied by structural and functional damage of tissues and organs which further induces the occurrence and development of aging-related diseases. Aging includes physiological aging caused by increased age and pathological aging induced by a variety of factors. Noteworthy, as a target organ directly contacting with the outside air, lung is more prone to various stimuli, causing pathological premature aging which is lung aging. Studies have found that there is a certain proportion of senescent cells in the lungs of most chronic respiratory diseases. However, the underlying mechanism by which these senescent cells induce lung senescence and their role in chronic respiratory diseases is still obscure. This paper focuses on the causes and classification of lung aging, the internal mechanism of lung aging involved in chronic respiratory diseases, and the application of anti-aging treatments in chronic respiratory diseases. We hope to provide new research ideas and theoretical basis for the clinical prevention and treatment in chronic respiratory diseases.

Effects of GSK3β/eEF2K signaling pathway on pulmonary fibrosis in mice / 中国应用生理学杂志

Objective: To investigate the effects of glycogen synthase kinase-3β (GSK3β)/eukaryotic extension factor kinase 2 (eEF2K) signaling pathway on the process of pulmonary fibrosis through in vivo experiments, and find new ideas for clinical treatment of pulmonary fibrosis. Methods: The pulmonary fibrosis model of C57BL/6 male mice was induced by bleomycin with intratracheal injection at the dose of 2 mg/kg. After 14 days of modeling, animals were divided into model group, negative inhibition group and inhibition group (n=5 for each group), and control group was not processed. The inhibition group was treated with TDZD-8 (4 mg/kg) after modeling, the negative inhibition group was given DMSO solution after modeling, and the samples were collected after 28 days. Hematoxylin-eosin staining method was used to detect lung fibrosis in mice and scored according to Ashcroft scale. Expression levels of GSK3β, p-GSK3β, eEF2K, p-eEF2K (Ser70, Ser392, Ser470), precursor protein of matrix metalloproteinase-2 (pro-MMP-2), matrix metalloproteinase-2 (MMP-2), collagen I (Col I), collagen Ⅲ (Col Ⅲ) and α-smooth muscle actin (α-SMA) were detected by Western blot. Results: Compared with control group, the fibrosis score was up-regulated, the expression levels of GSK3β, p-GSK3β, p-eEF2K (Ser70, Ser392, Ser470), pro-MMP-2, MMP-2, Col I, Col Ⅲ and α-SMA were increased, while that of eEF2K was decreased in model group (P＜0.05). Compared with model group, the fibrosis score, expression levels of GSK3β, p-GSK3β, p-eEF2K (Ser70, Ser392, Ser470), pro-MMP-2, MMP-2, Col I, Col Ⅲ and α-SMA were decreased, but the expression level of eEF2K was increased in inhibition group (P＜0.05). Conclusion: GSK3β can activate eEF2K by phosphorylation at the sites of Ser70, Ser392 and Ser470, increase the contents of fibrosis indicators, promote the formation of pulmonary fibrosis, and aggravate lung tissue lesions.

Application of DNA Microarray in Genetic Mutation Detection in Patients with Thalassemia / 中国实验血液学杂志

OBJECTIVE@#To perform dried blood spots thalassemia gene detection in patients with positive blood phenotypes by microarray technology, and evaluate its value in clinical detection.@*METHODS@#DNA samples were extracted from dried blood spots of 410 patients. Microarray technology was used to detect 3 deletion and 3 non-deletion types of α-thalassemia and 19 β-thalassemia point mutations which were common gene mutions in China.@*RESULTS@#There were 357 positive cases in all the 410 tested samples with the positive rate 87.07%, among which 299 cases (72.93%) carried deletion or point mutations of α-thalassemia, 29 cases (7.07%) carried point mutations of β-thalassemia and 29 cases (7.07%) carried gene mutations of complex αβ-thalassemia syndrome. The mutations of α-thalassemia were involved with --@*CONCLUSION@#The most common genetic mutations are --

Methodological Evaluation of Microarray in the Detection of α-Thalassemia / 中国实验血液学杂志

OBJECTIVE@#To proceed the clinical evaluation of DNA microarray for thalassemia gene detection.@*METHODS@#Peripheral blood samples of 166 thalassemia gene test subjects were collected and tested for thalassemia genes by microarray chip method and Gap-PCR method combined with PCR-reverse dot blot hybridization method according to double-blind control test. The specificity, sensitivity, positive predictive value, negative predictive value, and total coincidence rate of the microarray chip method were evaluated. When the two methods were inconsistent, multiplex ligation dependent probe amplification (MLPA) was used to verify the deletional α-thalassemia.@*RESULTS@#Compared with Gap-PCR method, specificity, sensitivity, positive predictive value, negative predictive value, Youden index, and total coincidence rate of microarray chip method was 100% (70/70), 96.88% (93/96), 100% (93/93), 95.89% (70/73), 0.969, and 97.59% (162/166), respectively, while compared with PCR-reverse dot blot hybridization method was 100% (125/125), 100% (41/41), 100% (41/41), 100% (125/125), 1, and 100% (166/166), respectively.@*CONCLUSION@#The microarray chip method for α-thalassemia gene detection shows the advantages of high specificity, sensitivity, and throughput.

Research progress of respiratory syncytial virus (RSV) infection and respiratory diseases in infancy / 生理学报

Lower respiratory tract infection (LRTI) induced by respiratory syncytial virus (RSV) is an important cause of hospitalization for infants. Compared with adults, infants are more likely to cause serious respiratory diseases after RSV infection due to the specific immature airway structure and immune system. The balance of immune resistance and immune tolerance of the host is critical to effective virus clearance and disease control. This paper reviews the relationship between RSV infection and respiratory diseases in infancy, the influence factors of the high pathogenicity of RSV infection in early life, as well as the research progress of anti-RSV therapy, and expands the specific molecular events regulating immune resistance and immune tolerance. We expect to present new ideas for the prevention and treatment of RSV-related respiratory diseases in clinical practice.

Progress of epithelial-mesenchymal transition in respiratory system and the modulatory mechanism of cell adhesion / 生理学报

Epithelial-mesenchymal transition (EMT) plays an important role in the development and pathogenesis of respiratory system. Epithelial cells are characterized by well-developed, intercellular contacts, whereas EMT triggers the sequential destabilization of cell-cell adhesive junctions. The dynamic remodeling of the epithelial cell adhesion molecules is important for maintaining the integrity and normal function of epithelium. This paper reviews the research progress of EMT in lung development, lung injury repair and chronic lung diseases, and summarizes the effect of cell junctions and cell adhesion molecules on EMT molecular events.

COVID-19:Emergency response measures for GCP in Hunan province under the first level response of major public health emergencies(2.0 Version) / 中国临床药理学与治疗学

According to the epidemic situation in Hunan province, COVID-19: Emergency response measure for GCP in Hunan Province under the first-level response of major public health emergencies (version 2.0) as the consensus is reached by experts with years of rich experience in the field of drug clinical trials. We hope this paper can provide reference for GCP practitioners.

The research progress of integrin β4 / 生理学报

Integrin is a transmembrane receptor that mediates the connection between cells and their external environment, such as extracellular matrix (ECM). Integrin β4 (ITGβ4) plays a number of functions due to its special structures: forms α6β4 with ITGα6 subunit and participates in the formation of hemidesmosomes; mediates cell-to-cell matrix interaction and cell-to-cell interaction, cell proliferation and survival, as well as migration and invasion. Also, ITGβ4 participates in various disease processes by activating multiple signaling pathways. In this paper, the structure, physiological function and function of ITGβ4 in respiratory system, tumor, nervous system and other related diseases will be reviewed.

Effects comparison of gemstone energy spectrum CT atomic number method and infrared spectroscopy for analyzing composition of urinary calculus / 重庆医学

Objective To investigate the effects of gemstone energy spectrum CT atomic number method and infrared spec-troscopy for analyzing the composition of urinary calculi and to compare their values in qualitative diagnosis of urinary calculi .Meth-ods Two hundreds and sixty cases of urinary tract stones were performed the gemstone spectrum CT urinary scanning and the stone composition was identified by atomic number method .After removing stone ,the stone composition analyzed by infrared spec-troscopy served as the gold standard .Then the consistency identified by the two methods was analyzed .Results The Kappa consis-tency test results showed that the two kinds of method for identifying stone type had good consistency (Kappa=0 .787 ,P<0 .01) . The paired chi square test results showed that the difference of the two methods for identifying the stone type had no statistical sig-nificance(χ2 =6 .581 ,P=0 .254) .The stone crystal composition types measured by gemstone energy spectrum CT atomic number method were less than those measured by infrared spectroscopy .The precise quantification of the stones with different crystal struc-tures was not as accurate as that of infrared spectroscopy (calcium oxalate monohydrate and calcium oxalate dihydrate ) .Conclusion The two methods for analyzing theurinary stone composition all have clinical significance ,the stone analysis method should be se-lected according to the actual situation .

Allergens-induced sensitization alters airway epithelial adhesion molecules expression in mice / 生理学报

To explore the relationship between the epithelial adhesion molecules and immune responses of airway epithelium, we observed the expression of integrin β4 and intercellular adhesion molecule-1 (ICAM-1) in the mice airway epithelium after sensitization with allergens. BALB/c mice were sensitized with intraperitoneal injection of ovalbumin (OVA) or house dust mite (HDM) and then developed airway hyper-responsiveness as determined by barometric whole-body plethysmography. Both OVA and HDM sensitization led to increases of the number of peripheral leukocytes as well as inflammatory cells infiltration in lungs. OVA sensitized mice showed more severe inflammatory cells infiltration than HDM sensitized mice. Immunohistochemistry analysis of mice lung tissues revealed that sensitization with both allergens also led to a decrease of integrin β4 expression and an increase of ICAM-1 expression in airway epithelia. OVA sensitized mice showed a more significant increase of ICAM-1 expression compared with HDM sensitized mice. siRNA mediated silencing of integrin β4 gene in 16HBE cells resulted in an up-regulation of ICAM-1 expression. Our results indicate a possible role of airway epithelial adhesion molecules in allergen-induced airway immune responses.

MSCT measurement of anterior clinoid process and clinoid segment of internal carotid artery / 实用放射学杂志

Objective To observe the anatomy status of the anterior clinoid process (ACP)and the anterior clinoid segment of in-ternal carotid artery (ICA)respectively by multisliced computed tomography (MSCT),and to provide useful imaging information for ACP removal surgery.Methods A total of 100 patients (200 sides)had volume rendering reconstruction of skull.Cranium was removed along cranio-orbital bone in simulation.Then the anatomical structures of the ACP and its surrounding were observed in cephalad direction.The total length,medium length,basic width,medium width of the ACP and the sagittal view curve length of anterior clinoid segment of the ICA from both sides were measured.Results Total length of left ACP was (9.82±2.48)mm,basal width was (9.47±1.88)mm,medium length was (5.03±1.55)mm,medium width was (6.1 9 ±1.75)mm;for right side total length was (10.41±2.1 6)mm,basal width was (9.66 ±2.21)mm,medium length was (5.86 ±2.48)mm,medium width was (6.66±1.5 1)mm.Left anterior clinoid segment of ICA curve length was (6.74±2.25)mm;right was (8.54±3.00)mm.Paired sample t test showed no significant difference in total length,basal width and medium width of ACP in both sides (P >0.05);while the difference in medium length and curve length of the anterior clinoid segment of ICA were statistically significant respectively (P <0.05).Conclusion MSCT can clearly display the vivisection and variation status of the ACP and the anterior clinoid segment of the ICA and can provide useful imaging information for removal of ACP in operation.

Efficacy of oral fludarabine in patients with chronic lymphocytic leukemia/small lymphocytic lymphoma / 中南大学学报(医学版)

OBJECTIVE@#To investigate the efficacy and safety of oral fludarabine in patients with chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL).@*METHODS@#The patients received oral fludarabine 40 mg/(m2.d) for 5 consecutive days, each treatment lasting 4 weeks. The efficacy was assessed with National Comprehensive Cancer Network (NCCN) criteria for response.@*RESULTS@#Twenty-two patients received the treatment, a median of 4 cycles per patient. The rate of complete response (CR), partial response (PR), and overall response (OR) was 40.9% (9/22), 45.5% (10/22), and 86.4% (19/22), respectively. Among the 17 previously untreated patients, 7 (41.2%) achieved CR and 8 (47.0%) achieved PR. Two of the 5 pre-treated patients achieved CR and the other 2 achieved PR. During a median observation of 24 months, the overall survival rate was 81.8%. The main adverse reactions were myelosuppression and infection. Grade 1 to 3 granulocytopenia was found in 7 (31.8%) patients, and infection in 3 (13.6%) patients. Nonhematologic toxicity was mild. All the adverse reactions were reversible.@*CONCLUSION@#The oral fludarabine is effective, safe, and well-tolerated in the patients with CLL/ SLL.

Multislice Spiral CT-based Simulation in Enlarging Exposure Extent of Internal Carotid Artery Siphon Before and After Removing Anterior Clinoid / 中国医学影像学杂志

Purpose To study the exposure extent of internal carotid artery siphon (ICAS) before and after removing anterior clinoid process (ACP) using multislice spiral CT (MSCT) simulation, and to improve the tumor resection rate and ensure the operation effect. Materials and Methods MSCT three-dimensional images reconstruction simulating supraorbital keyhole approach of 100 patients (200 sides) were observed, the distance between the crotch of anterior cerebral artery and middle cerebral artery and ICAS before and after removing ACP (exposure extent) was measured. Results In 100 patients (200 sides ACP), the exposure extent before and after removing ACP were (14.3±3.9) mm and (30.5±4.2) mm, respectively on the left side with statistical difference (t=45.278, P0.05). Conclusion MSCT simulating supraorbital keyhole approach in removing ACP can effectively increase the exposure length of ICA, and enlarge the exposure extent of sella region, thus provide reliable imaging information for removing tumor and selecting surgical project in this region.

Cerebral hemodynamic changes in hyperlipemia patients with transcranial Doppler / 中南大学学报(医学版)

OBJECTIVE@#To analyze cerebral hemodynamic changes by means of transcranial Doppler (TCD) and generally to explore the clinical application of this method in hyperlipemia patients.@*METHODS@#Cerebral hemodynamics were detected by TCD in 63 patients with hyperlipidemia and compared with the hemodynamics of 64 health people.@*RESULTS@#No statistically significant changes were found in the cerebral artery blood flow velocity and pulsatility index between the hyperlipidemic and control groups (P>0.05). Spectral shape, however, was abnormal in 52 patients in the hyperlipemia group (82.54%), which was statistically different (P<0.005) from controls. These abnormalities were classed as follows: 22 patients had abnormal spectra of the vertebrobasilar system, 2 patients had abnormal spectrum of the internal carotid arterial system, and 28 patients had abnormal spectra of both systems. The incidence of the abnormal spectra in vertebrobasilar system was significantly higher than the internal carotidartery (P<0.005).@*CONCLUSION@#TCD examination can reveal abnormal spectral shape in the cerebralartery and vertebrobasilar arterial systems in hyperlipidemia patients, and thus has some clinical value in determining changes in the brain of patients with high cholesterol levels and atherosclerosis.

Effect of neferine combined with mdr-1shRNA on the expression of mdr-1/P-gp in K562/A02 cell line / 中南大学学报(医学版)

OBJECTIVE@#To determine the effect of neferine (Nef) combined with mdr-1shRNA on the expression of mdr/P-gp in K562/A02 cell line.@*METHODS@#MTT assay was used to observe the cell proliferation. The expression level of P-gp was determined by Western blot and the transcription of mdr-1 gene was detected by semi-quantitative RT-PCR.@*RESULTS@#After K562/A02 cells were treated by Nef or mdr-1shRNA alone or both for 24 h, the proliferation of K562/A02 cells was significantly higher in the Nef combined with mdr-1shRNA treatment group than that of Nef or mdr-1shRNA alone group (P<0.01).The expression of mdr-1/P-gp in the Nef with mdr-1 shRNA group was significantly lower than that of Nef or mdr-1shRNA alone group.@*CONCLUSION@#Nef enhances the inhibition of mdr-1shRNA expression vector on K562/A02 cell proliferation and on P-gp protein to effectively reverse multidrug resistance induced by mdr-1 gene encoding P-gp.

Influence of intrapulmonary regulatory peptides on the expressions of HLA-DR, CD80 and CD86 in human bronchial epithelial cells / 生理学报

Antigen presenting is the initial step of the immune responses. In order to verify that human bronchial epithelial cells (HBECs) can express antigen presentation molecules, which can be modulated by intrapulmonary regulatory peptides, the present study was designed to examine the expressions of human leukocyte antigen DR (HLA-DR), CD80 and CD86 in resting or ozone-stressed HBECs by using immunocytochemistry and flow cytometry analysis. The results showed that HBECs expressed HLA-DR, CD80 and the expressions of HLA-DR and CD80 molecules were down-regulated under ozone stress. While VIP, P3513 and CGRP upregulated the expression of HLA-DR in resting or ozone-stressed HBECs, they had different effects on CD80 expression. VIP did not influence the expression of CD80 under resting state, but increased the expression of CD80 under ozone stress. CGRP decreased CD80 expression in resting HBECs, but increased CD80 expression in ozone-stressed HBECs. P3513 increased CD80 expression in resting HBECs, but decreased CD80 expression in ozone-stressed HBECs. The expression of CD86 was absent in resting or ozone-stressed HBECs. The results obtained demonstrate that HBECs have the capability to act as antigen presenting cells and the expression of HLA-DR and costimulatory molecules can be modulated by intrapulmonary regulatory peptides.

Effect of intermittent hypoxia on leptin and leptin receptor expression in obesity mice / 生理学报

In order to explore the effect and underlying mechanism of hypoxia on body weight, the effect of intermittent moderate hypoxia on high-fat diet-induced obesity was observed in mice, and the role of leptin in hypoxic effect was identified. Healthy Kunming mice were divided randomly into 4 groups (n=20 in each group). The control group: the mice were fed normally under the normal oxygen pressure. Hypoxia group: the mice were fed normally, and given intermittent moderate hypoxia training. Obesity group: the mice were fed diet rich in fat and sugar under the normal oxygen pressure. Hypoxia + obesity group: the mice were fed diet rich in fat and sugar, and given intermittent moderate hypoxia training. After 40 d of feeding and training, the body weight of mice was determined, and the average increasing rate of body weight in each group was calculated and normalized with food intake. Meanwhile, plasma leptin level was measured with ELISA method, and fatty degeneration and leptin receptor expression in liver were observed by Sudan III staining and immunohistochemistry, respectively. The obesity mouse model was successfully established with increases in body weight, plasma leptin level and distribution of adipocytes in the liver. The average body weight and density of adipocytes in the liver in hypoxia and hypoxia + obesity groups decreased obviously, while plasma leptin level and leptin receptor expression in the liver were increased. It is suggested that intermittent moderate hypoxia reduces body weight through elevating plasma leptin level and/or enhancing leptin receptor expression in the liver.

The role of bronchial epithelial cells in airway hyperresponsiveness / 生理学报

It is commonly accepted that airway hyperresponsiveness (AHR) is a chronic airway inflammation although the exact mechanism of its pathogenesis is still unclear. In the past ten years, an epithelial defect hypothesis has gradually gained supports from the main stream. Airway epithelium is no longer considered only as a simple mechanic barrier but an active interface between the inner and outer environment. Bronchial epithelial cells play a critical role in maintenance of homeostasis in the airway local microenvironment through a wide range of physiologic functions including anti-oxidation, exocrine/endocrine secretions, mucus production and antigen presentation under health and stressed/inflamed/injured conditions. It is reasonably hypothesized that disruption of these functional processes or defects in airway epithelium integrity may be the initial steps leading to airway hyperresponsiveness such as in asthma and chronic obstructive pulmonary disease.

Experimental study of acute lung injury induced by glutamate in vivo / 中南大学学报(医学版)

OBJECTIVE@#To investigate the possible injury induced by glutamate in the lung.@*METHODS@#The lung wet weight/body weight (LW/BW), lung wet/dry weight (W/D), the content of cells and the total protein (TP) in bronchoalveolar lavage fluid (BALF) were determined together with the micromorphology observation.@*RESULTS@#(1) The LW/BW, W/D, the content of white blood cells, red blood cells and TP in BALF increased in a dose dependent manner 2 hours after the administration of the glutamate (0.50 - 0.75 g/kg). (2) Examination of histological sections showed the presence of lung inflammation charactered by neutrophils recruitment 2 hours after the glutamate administration. (3) The increase of W/D caused by glutamate (0.50 g/kg) was nearly abolished by pre-treatment with MK801 (a specific blocker of NMDA receptor, 0.1 mg/kg) for 30 minutes (P<0.05).@*CONCLUSION@#Glutamate can cause the acute lung injury through the activation of NMDA receptor in vivo.

Reversal of multidrug resistance by MDR1 shRNA expression vector in human leukemia K562/A02 cells / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To construct a short hairpin RNA (shRNA) eukaryotic expression vector specific to MDR1 gene in multidrug resistance (MDR) human leukemia cell line K562/A02 to observe its silencing effect on MDR1 and P-glycprotein (P-gp) expression.</p><p><b>METHODS</b>The shRNA expression vector was constructed by gene recombination, then transfected into the cultured K562/A02 cells. The transcription of MDR1 gene was detected by semi-quantitative RT-PCR and the expression level of P-gp was determined by Western blot. 50% inhibition concentration (IC50) of ADM in K562/A02 cells was determined by MTT method. The intracellular doxorubicin (ADM) concentration was determined by HPLC.</p><p><b>RESULTS</b>The introduction of pEGFP-C1/U6/MDR1-A or pEGFP-C1/U6/MDR1-B expression vector was shown to efficiently and specifically inhibit the expression of P-gp according to results of Western blot, with an inhibitory rate of 50.67%. Semi-quantitative RT-PCR showed that mRNA transcription of MDR1 gene was reduced by (48.2 +/- 2.5)%. On the contrast, the control plasmid did not exhibit inhibitory effect on the protein expression and mRNA transcription of MDR1. The relative efficiency of K562/A02 to ADM was 40.8% or 62.4%, respectively, and the intracellular accumulation of ADM increased after shRNA treatment.</p><p><b>CONCLUSION</b>The shRNA expression vector targeting MDR1 gene showed dramatic inhibition on RNA transcription and protein expression. It could effectively restore the sensitivity of K562/A02 cells to conventional chemotherapeutic agents.</p>