Effects of microplastics and attached heavy metals on growth, immunity, and heavy metal accumulation in the yellow seahorse, Hippocampus kuda Bleeker.

Microplastics represent a new kind of environmental pollutant that has recently attracted extensive attention and become a research hotspot. Microplastics are similar in size to the food items of many marine organisms and are thus, often consumed by them, with potentially harmful and toxic effects. We examined the effects of microplastics on the growth of the yellow seahorse Hippocampus kuda. Seahorses were split into three groups fed Mysis + microplastics + heavy metals (group A), Mysis + microplastics (group B), and Mysis alone (group C). We analyzed and compared the accumulations of microplastics and heavy metals among the groups and monitored seahorse growth following the different treatments. Body length, body weight, condition factor, specific growth rate, and survival rate were all lower in group A compared with the other groups, but there was no significant difference in any of the parameters between groups B and C. The accumulation of microplastics was similar in groups A and B, and the accumulation of heavy metals was similar in groups B and C. These results suggest that the effect of microplastics on seahorse growth is caused by the accumulation of heavy metals, rather than by the microplastics themselves.

Visual structure and functional development of marine medaka, oryzias melastigma / 中国实验动物学报

Objective To observe the structure of the visual organ and functional development of marine medaka and to provide a theoretical basis for further studying its behavior. Methods The structure of visual organ of marine medaka was examined, with emphasis on the variations of its eyeball and retina. Results The medaka visual organ developed very rapidly. At the embryonic period, the pigment layer of the eyeball appeared, and there already had a significant eye area. At the first day,the structural development of the eyeball and retina were basically completed. At the 6th day, there was a significant difference of pigment layer and photoreceptor layer between light and dark vision. At the 23th day,the retinal structure was greatly changed, which can explain the reason about the change of medaka from the floating stage to the benthic stage. During from 9 to 14 month-old, the thickness of all layers in the retina decreased, indicating that the vision was degraded. Meanwhile,the kernel layer differentiation was obvious,as vertebral cells showed photoreceptor layer structure closely inlaid with rod cells, that medaka has good light sensitivity and strong visual acuity. The rapid development of medaka visual organ can be recognized as it needs to adapt the surroundings and to keep the life events such as feeding. Conclusions The development process of the visual organ of marine medaka is obvious and can provide a theoretical basis for further study of its behavior, and it may be a good experimental method for studying other related species.

The correlation of vitamin D level and vitamin D-binding protein gene polymorphism in chronic obstructive pulmonary disease / 中华内科杂志

Objective To assess the correlation of serum 25-hydroxyvitamin D (25-OHD) levels with vitamin D-binding protein (the group-specific component,GC) gene polymorphism in chronic obstructive pulmonary disease (COPD).Methods In a cross-sectional case-control study,250participants,including 116 COPD patients with smoking history and 134 healthy smokers,were investigated.A questionnaire about smoking history,vitamin D intake and comorbidities was collected.General pulmonary function was done by routine.Serum 25-OHD levels were detected by ELISA.The genetic variants (rs4588and rs7041) were genotyped by real time fluorescence polymerase chain reaction (RT-PCR) with TaqMan probe technology.Results The COPD patients had lower serum vitamin D level than the smoker subjects (36.58 nmol/L vs 43.80 nmol/L,P ＜0.001).In the COPD patients,vitamin D level was 39.43 nmol/L in those with percentage of predicted forced expiratory volume in 1 second (FEV1 ％ pred) greater than or equal to 80％.In other groups with FEV1 ％ pred 50％-80％,30％-50％ and lower than 30％,vitamin D levels were 35.32 nmol/L,32.21 nmol/L,26.25 nmol/L respectively (P ＜ 0.01).Moreover,there was a significant relevance of 25-OHD levels with FEV1 ％ pred in both COPD patients and healthy smokers (r2 =1.911; P ＜0.000 1).The mean 25-OHD concentration had a negative correlation with Global Initiative for Obstructive Lung Disease (GOLD) stages.Homozygous carriers of vitamin D-binding protein gene rs7041 T allele were independently related to 25-OHD levels and susceptibility of COPD (P ＜ 0.01 ; OR =2.140,95％ CI 1.157-3.959,P =0.015 respectively).Conclusions Patients with COPD are at high risk of vitamin D deficiency and the severity of COPD is inversely correlated with vitamin D levels.Furthermore,homozygous carrier of rs7041 T allele influences 25-OHD serum levels and is related to susceptibility of COPD,which may be a potential candidate gene for screening COPD.

Surfactant protein a polymorphism is associated with susceptibility to chronic obstructive pulmonary disease in Chinese Uighur population / 华中科技大学学报（医学）（英德文版）

This study investigated the correlation between surfactant protein-A (SP-A) polymorphism and the susceptibility of chronic obstructive pulmonary disease (COPD) in Xinjiang Uighurs. Genomic DNA was extracted from peripheral blood of 194 COPD smokers and 201 healthy smokers of Uighur who were hospitalized in or paid a visit to one of the four Xinjiang-based hospitals involved in the study, from March 2009 to December 2010. Single nucleotide polymorphisms (SNPs) were studied at aa62 (CCA/CCG rs1136451) and aa219 (CGG/TGG, rs4253527) in SP-A. Genotypes were determined by using the TaqMan polymerase chain reaction (PCR). Our results showed that genotype frequencies were different between the COPD and normal smokers in aa62 (x (2)=6.852, P=0.033). There were also significant differences in allele genotype frequencies between the COPD and the control and allele G might decrease the risk COPD (x (2)=6.545, P=0.011; OR=0.663; 95% CI: 0.484-0.909). The result suggested that polymorphism of aa62 (CCA/CCG, rs1136451) of SP-A may be associated with the susceptibility to COPD in Xinjiang Uighurs.

The expression of macrophage migration inhibition factor in pulmonary tissues of smokers with or without chronic obstructive pulmonary disease / 中华内科杂志

Objective To investigate the expression of macrophage migration inhibition factor (MIF) in pulmonary tissues of the smokers with and without chronic obstructive pulmonary disease (COPD).Methods The subjects were assigned into three groups:non-smokers without COPD (control group,n =12),smokers without COPD (smoker group,n =13) and smokers with COPD (COPD group,n =16).The specimens were obtained from lung tissues as far away from cancer focus as possible (＞ 5cm).Real-time quantitative PCR and immunohistochemistry were used to investigate the expression and distribution of MIF in pulmonary tissues.The relationship between the severity of airflow obstruction and the differential expressions of MIF in lung tissues of the smokers with or without COPD was analyzed.Results (1) MIF mRNA expression in COPD group (4.87 ± 1.79) was higher than that in the smoker group (2.16 ±0.72;P＜0.01),which was higher than that in the control group (1.09 ±0.48;P ＜0.01).(2)Immunohistochemistry analysis showed that MIF protein expression in lung tissues of the COPD group (0.277±0.025) was higher than that in the smokers group (0.199 ±0.034;P ＜0.01),which was significantly higher than that in control group (0.130 ±0.021 ;P ＜0.01).(3) Correlation analysis of MIF mRNA expression in the lung tissues and pulmonary function parameters of forced expired volume in one second (FEV1) percentage of predicted (FEV1 pred) and ratio of FEV1 to forced vital capacity (FEV1/FVC) suggested that MIF mRNA expression in the lung tissues was negatively related with FEV1 pred (r=-0.578,P ＜ 0.01) and FEV1/FVC (r =-0.607,P ＜ 0.01).Conclusions MIF expression significantly increases in the smokers with COPD,and MIF level in the lung is positively correlated with airflow limitation.The results suggest that MIF may play an important role in the pathogenesis of smokinginduced COPD.

Comparative study on the efficacy of tiotropium bromide inhalation and oral doxofylline treatment of moderate to severe stable chronic obstructive pulmonary disease / 华中科技大学学报（医学）（英德文版）

This study compared the efficacy and safety of tiotropium bromide inhalation powder (spiriva) and doxofylline oral tablet (doxofylline) in the treatment of chronic obstructive pulmonary disease (COPD). A multi-center, randomized, double-blind, double-dummy, parallel-controlled study involved 127 eligible stable moderate to severe COPD patients treated with inhaled tiotropium dry powder (18 μg/day) or oral doxofylline tablets (0.2 g/time, 2 times a day) for 12 and 24 weeks. Before and after treatment for 12 weeks and 24 weeks, respectively, pulmonary function, 6-min walking distance and dyspnea index were recorded. The results showed that in both tiotropium group and doxofylline groups, after 12-week treatment, FEV(1), FEV(1)/FVC% and 6-min walk distance were significantly higher than those before the medication, while dyspnea index decreased as compared with that before treatment. After 24-week treatment, a slight improvement in the measures was observed as compared with that of 12-weeks treatment, but the difference was not statistically significant. With both 12-week and 24-week treatment, the effect of tiotropium was slightly better than that of doxofylline tablets, with the difference being statistically insignificant. The major adverse events in the tiotropium group and doxofylline group were observed in 9 cases (9.9%) and 12 cases (12.9%), respectively, and no statistically significant difference was found between them. We are led to conclude that both tiotropium at 18 μg a day and doxofylline tablets at 0.2 g/day (two times a day) are effective and safe for the treatment of COPD.

Changes of protein kinase Calpha and cyclin D1 expressions in pulmonary arteries from smokers with and without chronic obstructive pulmonary disease / 华中科技大学学报（医学）（英德文版）

The purpose of this study was to investigate the changes of protein kinase Calpha (PKCalpha) and cyclin D1 expressions in pulmonary arteries from smokers with normal lung function and smokers with mild to moderate chronic obstructive pulmonary disease (COPD). The peripheral lung tissues were obtained from 10 non-smokers with normal lung function (non-smoker group), 14 smokers with normal lung function (smoker group), 11 smokers with mild to moderate COPD (COPD group). The morphological changes of pulmonary arteries were observed by HE-staining. The expressions of alpha-smooth muscle actin (alpha-SMA), proliferating cell nuclear antigen (PCNA), PKCalpha and cyclin D1 proteins in pulmonary artery smooth muscle cells (PASMCs) were immunohistochemically determined. The percentages of PCNA-positive cells were taken as the smooth muscle cells proliferation index (PI). The mRNA expressions of PKCalpha and cyclin D1 in PASMCs were evaluated by real-time fluorescence PCR. Morphometrical analysis showed that the ratio of pulmonary artery wall area to total area (WA%) in smoker group and COPD group was significantly greater than that in non-smoker group (P<0.01). The PASMCs proliferation index in smoker group and COPD group was significantly higher than that in nonsmoker group (P<0.01). The protein levels of PKCalpha and cyclin D1 in PASMCs were significantly increased in smoker group and COPD group as compared with non-smoker group (P<0.01). The mRNA expressions of PKCalpha and cyclin D1 in PASMCs were significantly elevated in smoker group and COPD group as compared with non-smoker group (P<0.01). Significant correlations were found between PKCalpha protein and WA% or PI (P<0.01). Correlations between cyclin D1 protein and WA% or PI also existed (P<0.01). The expression of PKCalpha was positively correlated with the expression of cyclin D1 at both protein and mRNA levels (P<0.01). In conclusion, increased expressions of PKCalpha and cyclin D1 might be involved in the pathogenesis of abnormal proliferation of PASMCs in smokers with normal lung function and smokers with mild to moderate COPD.

Conditioned medium from lung carcinoma cell line A549 increases the viability of human umbilical vein endothelial cells by activating the PI3K-Akt1 pathway / 肿瘤

Objective:To study the effects of the conditioned medium (CM) from human lung adenocarcinoma cell line A549 on the viability and apoptosis of human umbilical vein endothelial cells (HUVECs) and the role of PI3K-Akt signaling pathway in the process. Methods:HUVECs were cultured with CM of A549 cells. Cell viability was detected by XTT assay. The morphological changes of HUVECs were analyzed by Hoechst 33258 staining and fluorescence microscopy. The apoptosis was detected by flow cytometry. Expression levels of total Akt and phosphorylated Akt were assessed by Western blotting. PI3K inhibitors wortmannin(WT)and Akt1 siRNA(siAkt1)were used to block PI3K-Akt signaling pathway. The mRNA transcription of Akt subtype was determined by RT-PCR.Results:A549 CM significantly increased cell viability after 24 h treatment (P=0.037) and inhibited apoptosis (P=0.001) of HUVECs. CM time-dependently activated phosphorylation of Akt. Akt was phosphorylated at 15 min after CM treatment and reached the peak at 30 min and then tended to decline. Both WT and siAkt1 blocked the effects of CM. Conclusion:The CM of A549 cells increased the survival and inhibit the apoptosis of HUVECs. Akt1 played a significant role in the process.

XRCC1 Arg194Trp polymorphism and risk of chronic obstructive pulmonary disease / 华中科技大学学报（医学）（英德文版）

The DNA damage, caused by cigarette smoking, can cause airway cell apoptosis and death, which may be associated with the development of chronic obstructive pulmonary disease (COPD). However, just 20%-30% smokers develop COPD, which suggests that different degrees of DNA repair cause different outcomes in smokers. X-ray repair cross-complementing group 1 (XRCC1), a base excision repair protein, has multiple roles in repairing ROS-mediated, basal DNA damage and single-strand DNA breaks. The present study investigated the association between polymorphism in XRCC1 (Arg399Gln) and susceptibility of COPD. A total of 201 COPD cases and 309 controls were recruited and frequency-matched on age and sex. XRCC1 genotype was determined by PCR-restriction fragment length polymorphism analysis. Overall, compared with those with the XRCC1 Arg/Arg genotype, the risk for COPD had no significant difference among individuals with Trp/Trp genotype. However, after stratifying by smoking status, in former smokers, compared with those with the XRCC1 Arg/Arg genotype, the risk for COPD was significantly reduced among individuals with Trp/Trp genotype (adjusted OR=0.22, 95% CI 0.06-0.85, P=0.028); after stratifying by smoking exposure, in light smokers, compared with those with the XRCC1 Arg/Arg genotype, the risk for COPD was significantly reduced among individuals with Arg/Trp genotype and Trp/Trp genotype (adjusted OR=0.39, 95% CI 0.16-0.94, P=0.036; 0.24, 95% CI 0.07-0.79, P=0.019, respectively). In conclusion, XRCC1 Arg194Trp genotype is associated with a reduced risk of developing COPD among former and light smokers.

The role of PDK in regulating cytokine secretion by extracellular matrixes / 中华微生物学和免疫学杂志

Objective To investigate the regulatory effects of extracellular matrixes, including fi-bronectin(FN), and collagen Ⅰ (ColⅠ ) on the immunologic function of human airway smooth muscle cells (HASMCs) passively sensitized with asthmatic serum, and the role of phosphoinosifide 3-kinase (PI3K). Methods Primarily cultured HASMCs were inoculated on the blank plates or on the plates coated with difference matrix proteins, added 10% asthmatic serum to passively sensitized non-asthmatic HASMCs and 10% non-asthmatic serum treated HASMCs as control, cell pretreated with PI3K inhibitor LY294002 for 30 min. The expressions of RANTES, Eotaxin, TGF-β1 mRNA were observed by RT-PCR and RANTES, Eotaxin, TGF-β1 protein in the ceil culture supernatunts was detected by enzyme-linked immtmosorbent assay (ELISA). Results Compared with the control serum group, the expressions of RANTES, Eotaxin, TGF-β1 mRNA of HASMCs and those protein in HASMCs culture supematants were significantly increased in the asthmatic serum group and the control serum + FN group and the control serum + Col Ⅰ group ( P < 0.05 ). The expressions of RANTES, Eotaxin, TGF-β1 mRNA of HASMCs and protein in HASMCs culture superna-tants were significantly increased in the asthmatic serum + FN group and the asthmatic serum + Col Ⅰ group. 50 μmoL/L LY294002 could significantly inhibit the expressions of RANTES, Eotaxin, TGF-β1 mRNA of HASMCs and protein in HASMCs culture supematants(P < 0.05). Conclusion These results suggest ex-tracellular matrixe may regulate immunomodulatory function of HASMCs passively sensitized with asthmatic serum and PI3K signaling pathway may play an important role in the process.

The alteration and significance of surfactant protein A in rats chronically exposed to cigarette smoke / 华中科技大学学报（医学）（英德文版）

In order to confirm the alteration and significance of cigarette smoke exposure on SP-A in rats, 20 Wistar rats were assigned randomly to two groups: an N group (n=10), and an S group (n=10). The ultra-structural change was observed by electron microscopy. The number of cells positive for SPA was by immunohistochemically measured. The mRNA expression in the lung tissues was determined by reverse transcription polymerase chain reaction (RT-PCR). The number of cells positive for SPA of the S group (0.52 +/- 0.05) was lower than that of the N group (0.72+/-0.06) (P<0.05). The levels of mRNA of SPA in the lung tissues of the S group (0.3522+/-0.0512) was significantly lower than that of the N group (0.4432+/-0.05628) (P<0.05). It is concluded that cigarette smoke alone decreased the level of SP-A and that might have an important effect on surfactant metabolism and the host defense functions of surfactant in the peripheral airways, which might play a crucial role in the development of chronic obstructive lung disease.

Effects of puerarin on pulmonary vascular remodeling and protein kinase C-alpha in chronic cigarette smoke exposure smoke-exposed rats / 华中科技大学学报（医学）（英德文版）

In order to investigate the effects of puerarin on pulmonary vascular remodeling and protein kinase C-alpha (PKC-alpha) in chronic exposure smoke rats, 54 male Wistar rats were randomly divided into 7 groups: control group (C group), smoke exposure groups (S(4w) group, S(8w) group), puerarin groups (P(4w) group, P(8w) group), propylene glycol control groups (PC(4w) group, PC(8w) group). Rats were exposed to cigarette smoke or air for 4 to 8 weeks. Rats in puerarin groups also received puerarin. To evaluate vascular remodeling, alpha-smooth muscle actin (alpha-SM-actin) staining was used to count the percentage of completely muscularised vessels to intraacinar pulmonary arteries (CMA/IAPA) which was determined by morphometric analysis of histological sections. Pulmonary artery smooth muscle cell (PASMC) apoptosis was detected by in situ end labeling technique (TUNEL), and proliferation by proliferating cell nuclear antigen (PCNA) staining. Reverse transcription-polymerase chain reaction (RT-PCR), immunofluorescence staining and Western blot analysis were done to detect the PKC-alpha mRNA and protein expression in pulmonary arteries. The results showed that in cigarette smoke-exposed rats the percentage of CMA/IAPA and alpha-SM-actin expression were increased greatly, PASMC apoptosis was increased and proliferation was markedly increased; Apoptosis indices (AI) and proliferation indices (PI) were higher than in C group; AI and PI were correlated with vascular remodeling indices; The expression of PKC-alpha mRNA and protein in pulmonary arteries was significantly higher than in C group. In rats treated with puerarin, the percentage of CMA/IAPA and cell proliferation was reduced, whereas PASMC apoptosis was increased; The expression levels of PKC-alpha mRNA and protein were lower than in smoke exposure rats. There was no difference among all these data between S groups and PC groups. These findings suggested that cigarette smoke-induced pulmonary vascular remodeling was most likely an effect of the imbalance of PASMC proliferation and apoptosis. Puerarin appears to be able to reduce cell proliferation and vascular remodeling possibly through PKC signaling transduction pathway.

Expression of Leukemia Inhibitory Factor in Airway Epithelial Tissue of Asthmatic Rats / 华中科技大学学报（医学）（英德文版）

In order to investigate the expression of leukemia inhibitory factor (LIF) in airway epithelial tissues of normal and asthmatic rats, the influence of dexamethasone and the role of LIF in pathogenesis of asthma, 30 Sprague-Dawley (SD) rats were randomly divided into 3 groups (10 for each group): normal group, asthma model group, and dexamethasone-interfered group. In asthmamodel group and dexamethasone-interfered group, asthma rat models were established by intraperitoneal (i.p.) injection of 10% ovalbumin (OVA) and challenge with 1% OVA via inhalation. Rats in dexamethasone-interfered group were pretreated with dexamethasone (2 mg/kg, i.p) 30 min before each challenge. The expression of LIF protein in lung was detected by immunohistochemistry. The results showed that LIF protein was mainly expressed in cytoplasm of bronchial epithelial cells. The expression of LIF protein in the airway epithelial tissue of asthma model group was significantly higher than that in normal group and dexamethasone-interfered group (P＜0.01), but there was no significant difference between normal group and dexamethasone-interfered group (P＞0.05). It was concluded that the expression of LIF was increased significantly in the airway epithelial tissue of the asthma rats, and dexamethasone could down-regulate the expression of LIF. It was suggested that LIF might play an important role in the pathogenesis of asthma as an inflammation regulator.

The Effect of Ginkgo Biloba Extract on the Expression of PKCα in the Inflammatory Cells and the Level of IL-5 in Induced Sputum of Asthmatic Patients / 华中科技大学学报（医学）（英德文版）

To investigate the effect of the Ginkgo Biloba Extract (GBE) on the asthma and examine its possible mechanisms, 75 asthma patients were divided into 4 groups and the patients were respectively treated with fluticasone propionate for 2 weeks or 4 weeks, or treated with fluticasone propionate plus GBE for 2 weeks or 4 weeks. Fifteen healthy volunteers served as healthy controls. Sputum inhalation with inhaling hypertonic saline (4%-5%) was performed. Lung ventilatory function and forced expiratory volume in one second (FEV1) were measured. The numbers of different cells in induced sputum were calculated. The expression of PKCα in the cells was immunocytochemically detected and the percentages of positive cells in different cells were counted. Interleukin-5 (IL-5) in sputum supernatants was detected with enzyme-linked immunosorbent assay. The percentage of eosinophils, lymphocytes, PKCα positive inflammatory cells and the concentration of IL-5 in asthmatic patients were higher than those in the controls (P＜0.05), and the eosinophils, lymphocytes,positive expression of PKCα and the level of IL-5 were significantly decreased in asthmatic patients after they were treated with fluticasone propionate or fluticasone propionate plus GBE. However,they were still significantly higher than those of the controls. Compared to the group treated with glucocorticosteroid for 2 weeks, no significant decrease was found in the percentage of eosinophils,lymphocytes, PKCα positive inflammatory cells and the IL-5 in the supernatant of induced sputum.Compared with the group treated with glucocorticosteroid for 2 or 4 weeks, significant decrease in the same parameters was observed in the group treated with fluticasone propionate and GBE for 4 weeks. The IL-5 level in the supernatant of induced sputum was positively correlated with the percentage of PKCα-positive inflammatory cells and the percentage of eosinophils in the induced sputum in asthma patient groups respectively (n=150, r= 0.83, P＜0.01; n=150, r=0.76, P＜0.01). The FEV1 was negatively correlated with the percentage of PKCα-positive inflammatory cells and the IL-5 levels in supernatant of induced sputum in asthma patients respectively (n=150, r=-0.77,P＜0.01; n=150, r= -0.64, P＜0.01). It is concluded that GBE could significantly decrease the infiltration of inflammatory cells such as eosinophils and lymphocytes in the asthmatic airway and relieve the airway inflammation. GBE may decrease the activation of the PKCα in the inflammatory cells and thereby decrease the IL-5 level in induced sputum. GBE may be used as a complement to the glucocorticosteroid therapy for asthma.

Expression of Interleukin-17 in Lung and Peripheral Blood of Asthmatic Rats and the Influence of Dexamethasone / 华中科技大学学报（医学）（英德文版）

The expression of interleukin-17 (IL-17) in lung and peripheral blood of asthmatic rats and the influence of dexamethasone, and the role of IL-17 in the pathogenesis of asthma were inves-tigated. Thirty Sprague-Dawley (SD) adult rats were randomly divided into three groups (n=10 in each group): normal group, asthmatic group, and dexamethasone-interfered group. Rat asthmatic model was established by intraperitoneal (I.p.) injection of 10% ovalbumin (OVA) and challenge with 1% OVA via inhalation. Rats in dexamethasone-interfered group were pretreated with dexa-methasone (2 mg/kg, I.p.) 30 rain before each challenge. The expression of IL-17 protein in serum and bronchoalveolar lavage fluid (BALF) was detected by ELISA. The expression of IL-17 mRNA in peripheral blood mononuclear cells (PBMC) and BALF cells was semi-quantitatively detected by RT-PCR. The expression of IL-17 protein in serum and BALF of asthmatic rats was significantly elevated as compared with normal rats and dexamethsone-interfered rats (P<0.01), and there was sig- nificant difference between normal rats and dexamethsone-interfered rats (P<0.05). The expression of IL-17 mRNA in PBMC and BALF cells of asthmatic rats was markedly increased as compared with normal rats and dexamethsone-interfered rats (P<0.01), and significant difference was found between normal rats and dexamethsone-interfered rats (P<0.05). It was concluded that the expression of IL-17 was increased significantly in asthmatic rats and could be inhibited partly by dexamethasone, sug-gesting that IL-17 might play an important role in the pathogenesis of asthma as an inflammation regulation factor.

The Effect of Interleukin-18 on Airway Inflammation in Asthmatic Murine Models and Its Mechanisms / 华中科技大学学报（医学）（英德文版）

In order to investigate the effect of interleukin-18 (IL-18) on airway inflammation in asthmatic murine models and its mechanisms, BALB/C mice were randomly divided into three groups (n=10 in each group): group A (control group); group B (asthmatic model group); group C (IL-18-treated group). The asthmatic model was established in groups B and C by respiratory syncytial virus (RSV) killed by ultraviolet. Saline solution (0.1 mL) and IL-18 (0.1 mL, 1 μg) were intraperitoneally injected respectively in groups B and C at 7 time points (day 1, 2, 7, 8, 9, 21, 22). The number of eosinophils (EOS) and plasmacytes in the airway was observed. The levels of inter-feron gamma (IFN-γ) in bronchoalveolar lavage fluid (BALF) were measured by ELISA. The results showed that symptoms of asthma in group C were more severe than in groups A and B. In group A,there were no EOS and plasmacytes in the airway submucosa. The number of EOS [15±3 (average cell counts per microscopic visual field, the same below)] and plasmacytes (10±2) in group B were increased significantly. However, the number of EOS and plasmacytes in group C (6±2 and 2±1, re- spectively) was decreased significantly as compared with group B (both P<0.05). The levels of IFN-γ in groups A, B and C were 31±3, 40±5 and 63±5 pg/mL respectively, and those in group C were sig- nificantly higher than in groups A and B (both P<0.05). It was suggested that the mechanism by which IL-18 inhibited the airway inflammation in asthmatic mice might be contributed to the fact that IL-18 could induce the induction of IFN-γ.

Primary Culture of Alveolar Epithelial Type Ⅱ Cells and Its Bionomic Study / 华中科技大学学报（医学）（英德文版）

To establish a better method of primary culture for alveolar epithelial type Ⅱ cells (AEC Ⅱ) and to study its bionomics, alveolar epithelial type Ⅱ cells were isolated by digestion with tryp- sin and collagenase, which were then purified by plated into culture flask coated with rat immu- noglobulin G. The purified AEC Ⅱ were identified by alkaline phosphatase staining, electron mi-croscopy, immunocytochemical staining of pulmonary surfactant protein A (SPA). The SPA expres-sion and transfection characteristics were compared with those of A549 cell line. The results showed that AEC Ⅱ could be isolated by digestion with trysin and collagenase and purified by adhesive pu- rification by using IgG, with a yield of about 2-3×107, and a purity of about 75%-84 %. Cells could be quickly identified with AKP staining. AEC Ⅱ were different from A549 cell line in terms of SPA expression and transfection characteristics. It is concluded that adhesive purification with IgG can improve the purity of AEC Ⅱ, and AKP staining is simple in cell identification. AEC Ⅱ can not be completely replaced by A549 cells in some studies because the differences between them, such as SPA expression.

Increased Expression of PI-3K in Asthmatic Rat T Lymphocytes / 华中科技大学学报（医学）（英德文版）

In order to explore the expression of PI-3K in T lymphocytes of asthmatic rats and the relationship between PI-3K and activation of T lymphocytes, 24 Wistar rats were randomly divided into 4 groups: normal control group, asthmatic one-week group, asthmatic two-week group and asthmatic four-week group. T cells were purified from blood of each rat and the expression of PI-3K was observed by immunocytochemical fluorescence staining, the semiquantitative fluorescence intensity was measured by HPIAS-2000 analytic software, and the expression of IL-4 in supernatants was detected by ELISA. The results showed that the fluorescence intensity of T lymphocytes in asthmatic groups was significantly higher than that in normal control (P＜0.001), indicating that the expression of PI-3K in T lymphocytes of asthmatic rats was significantly higher than that in those of normal controls, and the difference between acute and chronic stage asthmatic groups was significant (P＜0.05). The expression levels of IL-4 protein in supernatants of asthmatic T lymphocytes were significantly higher than those in the normal controls (P＜0.05). There was a significant positive correlation between the expression of PI-3K in T lymphocytes and the IL-4 protein expression in supernatants (r=0.583, P＜0.01). It was suggested that PI-3K signal pathway may participate in the processes of activation and other cytological effects of asthmatic T lymphocytes, thus may play an important roles in the pathogenesis of asthma.

The Effects of Protein Kinase C (PKC) on the Tension of Normal and Passively Sensitized Human Airway Smooth Muscle and the Activity of Voltage-dependent Delayed Rectifier Potassium Channel (Kv) / 华中科技大学学报（医学）（英德文版）

The effects of protein kinase C (PKC) on the tension and the activity of voltage-dependent delayed rectifier potassium channel (Kv) were examined in normal and passively sensitized human airway smooth muscle (HASM), by measuring tones and whole-cell patch clamp techniques, and the Kv activities and membrane potential (Em) were also detected. The results showed that phorbol 12-myristate 13-acetate (PMA), a PKC activator, caused a concentration-dependent constriction in normal HASM rings. The constriction of the passively sensitized muscle in asthma serum group was significantly higher than that of the normal group (P＜0.05), and the constrictions of both groups were completely abolished by PKC inhibitor Ro31-8220 and calcium channel inhibitor nifedipine. Kv activities of HASM cells were significantly inhibited by PMA, and the Em became more positive, as compared with the DMSO (a PMA menstruum)-treated group (P＜0.01). This effect could be blocked by Ro31-8220 (P＜0.01). It was concluded that activation of PKC could increase the tones of HASM, which might be related to the reduced Kv activity. In passively sensitized HASM rings, this effect was more notable.

Serum Level of Vascular Endothelial Growth Factor in Patients with Obstructive Sleep Apnea Hypopnea Syndrome / 华中科技大学学报（医学）（英德文版）

To explore the relationship between the serum vascular endothelial growth factor (VEGF)level and the severity of obstructive sleep apnea hypopnea syndrome (OSAHS), the concentrations of serum VEGF in 40 OSAHS patients and 9 healthy controls were measured by using ELISA method.Meanwhile the correlation between the concentration of VEGF and parameters of polysomnography (PSG) was examined. Our results showed that the concentrations of VEGF were significantly higher in OSAHS patients with severe hypoxia (536.8±334.7 pg/mL) than in those with mild hypoxia (329.2±174.7 pg/mL) and healthy controls (272. 8±211.0 pg/mL) (P＜0.05 for both). The concentrations of VEGF were also significantly higher in OSAHS patients with hypertension (484.5±261.4 pg/mL) than in those without hypertension (311.0±158.4 pg/mL) and healthy controls (272. 8±211.0 pg/mL) (P＜0.05 for both). There was a positive correlation between the concentration of VEGF and the apnea hypopnea index (AHI) (γ=0.34, P＜0.05). It is concluded that the concentration of the serum VEGF is positively related to the severity of OSAHS. The elevated serum VEGF level may be involved in the pathogenesis of the complications of obstructive sleep apnea hypopnea syndrome.