Alteration levels of IL-17 and IL-19 inflammatory factors among deadaptation personnel from the plateau / 军事医学

Objective To analyze the change in the concentration of IL-17 and IL-10 inflammatory factors among the deadaptation personnel who returned from the plateau.Methods A total 21 healthy males were investigated who averaged 25 years in age, lived permanently in the plains (200 m), and once stayed to the plateau (Lasha) for 6 months.Their venous blood was collected at three time points:the day before ascending to the plateau(control), the second day after return to the plains(d2) and the 30th day(d30), respectively.Their serum was seperated from the whole blood and the level of IL-17A and IL-10 was detected by ELISA method.Results The concentration of IL-17A and the IL-17A/IL-10 ratio were significant increased at d2 and d30, respectively, compared with control (P<0.05).Compared with d2, IL-17A and the IL-17A/IL-10 ratio were decreased obviously at d30(P <0.05).The level of IL-10 at d2 and d30 was significantly reduced compared with control ( P <0.05), but increased at d30 compared with d2.Correlative analysis showed that there was a negative correlation in the levels of IL-10 and IL-17A between control, d2 and d30, respectively (r1=0.948, P<0.05;r2=0.969, P<0.05;r3=0.972, P<0.05).A significant negative correlation was observed in the alteration levels of IL-10 and IL-17A between the three groups(r4=-0.793, P<0.05; r5=-0.756, P<0.05). Conclusion The concentration of inflammatory factors among the plateau deadaptation patients is imbalanced, but it is gradually reduced with time.The mechanism is still not clear.

Construction of bait plasmid containing SIGIRR cytoplasmic tail and detection of self-activation in yeast two hybrid system / 重庆医学

Objective To construct the bait plasmid of pSos-single immunoglobin IL-1 receptor related protein (SIGIRR) in Cy-toTrap yeast two hybrid system ,and to test its self-activation .Methods The cDNA fragments of SIGIRR(480 -1 230 bp) were amplified from pReceiver-LV19-SIGIRR and ligated into the bait plasmid pSos to generate the plasmid pSos-SIGIRR .The pSos-SI-GIRR was identified by DNA sequencing and dual-site endonuclease digestion .Then the recombinant plasmid and control plasmid were introduced into the yeast cell cdc25H .The transformants were inoculated on plates of 25 ℃ /SD/Glucose(-UL) ,25 ℃/SD/Ga-lactose(-UL) ,37 ℃ /SD/Glucose(-UL) and 37 ℃ /SD/Galactose ,respectively and the proliferation ability of transformant was ob-served for 6 d .The Western blot was adopted to detect the expression of target protein .Results The pSos-SIGIRR vector was cor-rectly constructed and proved of no self-activation and toxic action .The Western blot showed that the target protein was expressed in a form of fusion protein of 170KD .Conclusion The bait plasmid containing SIGIRR cytoplasmic tail can be applied to the yeast two-hybrid system and lays the important foundation for seeking the interacting protein with SIGRR from the human lung cDNA li-brary in .

Adenovirus-mediated protein-kinase-GIα suppresses the hypoxia-induced proliferation and phenotype-switching of pulmonary arterial smooth muscle cell / 中华内科杂志

Objective To observe the proliferation and phenotype-switching of pulmonary arterial smooth muscle cell (PASMC) induced by hypoxia and interfered by Ad-PKGIα. And to investigate the potential regulative role of PKGIα gene in the molecule mechanism of hypoxia pulmonary vessel remodeling (HPVR). Methods To establish the pure PASMC cultured by tissue-sticking methods. Semi-quantitative reverse transcription and polymerase chain reaction (RT-PCR) and Western blot were used to examine the PKGIα mRNA and protein expression after PASMC were transfected by Ad-PKG. The mRNA and protein expressive change of smooth muscle α actin(SM-α-actin) determined the degree of cell phenotype-switching. The changes of PASMC proliferation were determined by flow cytometry and ~3H-TdR incorporated way. Results Ad-PKGIα could transfect into PASMC and highly express. Hypoxia down-regulated the expression of SM-α-actin protein (44. 25±5.34 in normoxia, 32. 18±4. 19 in 12 h hypoxia condition, 21.90 ±2. 44 in 24 h hypoxia condition, P < 0. 05), that could be blocked by the transfeetion of Ad-PKGIα. Hypoxia could push PASMC mitosis and proliferating(~3H-TdR incorporated way: 7570 ± 371 in normoxia,12 020± 831 in 12 h hypoxia condition,14 924 ± 1491 in 24 h hypoxia condition, P <0. 05), that could be blocked by the transfection of Ad-PKGIα, too. Conclusions The results suggested that PKGIα signaling pathway might play an important role in the molecule mechanism of HPVR. And PKGIα gene might be a target point of gene therapy.

Negative regulation of cells proliferation by Gax gene and its mechanisms / 生物医学工程学杂志

Gax gene is a newly found negative transcriptional regulator of cells proliferation. This paper introduces the detection and structural features of Gax, details the inhibitory effect of Gax on the proliferation of vascular smooth muscle cells, vascular endothelial cells and cancer cells, and explains the putative mechanisms therein involved. The potential for providing therapeutic insights into human diseases by modulating Gax activity is prospected.

Construction of eukaryotic expression vector of Der p2 gene and its expression in mouse dendritic cells / 中国组织工程研究

BACKGROUND: Dendritic cells, the most potent antigen presenting cells known at present, have been extensively used in the immunotherapy as adjuvant. OBJECTIVE: The present study was to construct Der p 2 eukaryotic expression vector and validate its expression in the mouse bone marrow-derived dendritic cells. DESIGN, TIME AND SETTING: A single sample observation was performed at the Institute of Respiratory Disease,Xinqiao Hospital, Third Military Medical University of Chinese PLA between May and December 2005. MATERIALS: C57BL/6 mice were included. Plambd-Der p 2 was the product of Heska Company, USA.pCI-neo plasmid was provided by the Institute of Respiratory Disease, Xinqiao Hospital, Third Military Medical University of Chinese PLA.METHODS: Mouse bone marrow-derived dendritic cells were in vitro isolated and cultured.Complete Der p 2 cDNA was spliced from prokaryotic expression vector plambd-Der p 2, and then cloned into eukaryotic expression vector pCI-neo (pCI-neo-Der p 2).The positive recombinants pCl-neo-Der p 2 transfected into dendritic cells.Non-transfected and blank vector pCI-neo-transfected dendritic cells were used as controls. MAIN OUTCOME MEASURES: ①Identification of pCI-neo-Der p 2 recombinant plasmid.②Detection of Der p 2 mRNA and protein expression by reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot techniques. RESULTS: Sequencing results showed Der p 2 cDNA in pCI-neo-Der p 2 was in coincidence with the sequence registrated in Gene Bank.RT-PCR and Western Blot results showed that expression of Der p 2 mRNA and protein could be detectable in the pCI-neo-Der p 2-transfected dendritic cells. CONCLUSION: The Der p 2 cDNA was successfully constructed into the eukaryotic expression vector, and Der p 2 gene and protein could be expressed efficiently in dendritic cells.

Effect of Ad-Gax transfection on apoptosis of human lung adenocarcinoma A549 cells and its mechanism / 中国肺癌杂志

<p><b>BACKGROUND</b>Apoptosis is closely related to development of lung cancer. It is a strategy of lung cancer therapy to induce apoptosis. The aim of this study is to explore the effects of growth arrest-specific homeobox (Gax) transfection on apoptosis and expression of Bcl-2 and Bax proteins of human lung adenocarcinoma A549 cells.</p><p><b>METHODS</b>A549 cells were transfected with Gax gene by a replication-deficient adenovirus expressing the hemagglutinin-tagged Gax cDNA (Ad-Gax). Apoptosis of A549 cells was observed by transmission electronic microscope and terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) positive staining. Apoptotic rate of A549 cells was evaluated by flow cytometry. Expressions of Bcl-2 and Bax proteins in A549 cells were detected by immunocytochemistry.</p><p><b>RESULTS</b>Before Ad-Gax transfection, none or few of TUNEL-positive A549 cells were detected. After Ad-Gax transfection, a marked increase in TUNEL-positive staining occurred, especially at 24 h later. The ratio of apoptosis of A549 cellsin non-transfection group and transfection groups at 12 h, 24 h, 48 h were 0.25%, 12.57%, 17.29%, 15.03%, respectively. Compared with non-transfection group, the apoptotic rates of transfection groups increased significantly (Chi-square value was 7.357, 11.126 and 9.943 respectively, P < 0.01). The average optical density (AOD) of Bcl-2 protein in A549 cells in non-transfection group and transfection groups at 12 h, 24 h, 48 h were 2.02±0.07, 1.79±0.02, 1.25±0.51 and 1.21±0.24 respectively. Compared with non-transfection group, AOD of Bcl-2 protein in A549 cells in transfection groups decreased significantly (t value was 6.651, 7.089 and 7.438 respectively, P < 0.01). On the other hand, Bax protein expression in transfection groups increased, the AODs of Bax were 4.49±0.61, 4.24±0.37 and 3.95±0.43, respectively. Compared with non-transfection group (3.12±0.42), AOD of Bax protein in A549 celle in transfection groups increased significantly (t value was 7.469, 7.287 and 6.473 respectively, P < 0.01). In the Ad-Gax transfection groups the lower Bcl-2/Bax ratio was, the higher the apoptotic rate of A549 cells was (r=-0.49, P < 0.01).</p><p><b>CONCLUSIONS</b>Ad-Gax transfection can induce A549 cells apoptosis. Possible mechanism is that Gax can downregulate Bcl-2 protein expression and upregulate Bax protein expression, and A549 cells apoptosis is related to the Bcl-2/Bax ratio.</p>

Expression of canstatin gene in human lymphocytes and its inhibitory effect on growth and metastasis of Lewis lung carcinoma / 中国肺癌杂志

<p><b>BACKGROUND</b>In recent years, many progresses have been made in molecular target therapy for lung cancer, in which anti-angiogenic target therapy is a hot spot drawing researchers' attention. The aim of this study is to explore the expression of canstatin gene transfected into human lymphocytes and its inhibitory effect on growth and metastasis of Lewis lung carcinoma.</p><p><b>METHODS</b>The eukaryotic expression vector of pCMV-Script and the recombinant pCMV-Script/Canstatin vector were separately transfected into lymphocytes by electroporation. The expression of canstatin protein in supernatant of lymphocyues was examined by SDS-PAGE assay. Furthermore, Lewis lung carcinoma cells were subcutaneously inoculated to C57BL mice to make animal model of tumor. When the transplanted tumors on the mice developed to 1cm³, the 30 mice were randomized into 3 groups, which were injected with 0.2mL supernatant of lymphocytes transfected with recombinant vector or naked vector, or 0.2mL NS respectively. After the treatment for 14 days, the size and pathological section of subcutaneous tumors were observed, and the number of pulmonary metastatic node was calculated.</p><p><b>RESULTS</b>Canstatin protein was found in supernatant of the lymphocytes in the recombinant vector group by SDS-PAGE assay. After the treatment, the tumor size in the recombinant vector group (1.49cm³±0.18cm³) was significantly smaller than that in the naked vector group (2.44cm³± 0.19cm³) and NS group (2.53cm³±0.18cm³) (P=0.000). The numbers of pulmonary metastatic node were 3.40±1.14, 7.60±2.61 and 7.60±2.41 in the recombinant vector group, naked vector group and NS group respectively (recombinant vector group vs the other two groups, P=0.013).</p><p><b>CONCLUSIONS</b>The pCMV-Script/Canstatin vector can express canstatin protein in human lymphocytes. Canstatin has strongly inhibitory effect on growth and metastasis of mouse Lewis lung carcinoma.</p>

Effects of Gax gene transfection on proliferation and expression of proto-oncogenes in A549 cells / 中国肺癌杂志

<p><b>BACKGROUND</b>Lung cancer is the leading cancer of malignant tumor in China.It is the direction that poeple make efforts to seek gene therapy of lung cancer. The aim of this study is to explore the effects of transfected growth arrest-specific homeobox gene (Gax gene) on the proliferation and expressions of c-fos and c-jun mRNA in A549 cells.</p><p><b>METHODS</b>A549 cells were transfected with Gax gene by adenovirus. Expressions of Gax mRNA and protein were detected by RT-PCR and immunocytochemistry. The expressions of c-fos and c-jun mRNA were evaluated by RT-PCR. The proliferation inhibition effect of Gax transfection on A549 cells was evaluated by MTT assay.</p><p><b>RESULTS</b>Only in the A549 cells transfected with Gax gene the Gax expression was confirmed by RT-PCR and immunocytochemistry. Compared with that in the control group, c-fos and c-jun mRNA level decreased significantly in Gax-transfected A549 cells (t=7.755, P < 0.01; t= 5.938 , P < 0.01). MTT assay showed that the proliferation inhibition rates of A549 cells transfected by Ad-Gax for 24h, 48h and 72h were (47.35±5.36)%, (54.96±1.78)%, and (65.39±5.11)% respectively. And these proliferation inhibition rates were significantly higher than those in the control group (Chi-Square=7.152, 9.431 and 12.847, P < 0.01).</p><p><b>CONCLUSIONS</b>Gax gene can inhibit the proliferation of A549 cells. Its molecular mechanism may be through down-regulating the expressions of c-fos and c-jun.</p>

New Clinical Teaching Mode To Cultivate Practitioner / 中华医学教育探索杂志

The number of taking the Examination of Doctor Qualification is rising,while the pass rate is dropping each year.This status has undoubtedly made a big challenge for clinical teaching,and how to improve the effects of clinical lessons has become an important topic for the clinical teachers.The Affiliated Xinqiao Hospital of the Third Military Medical University has made some new changes in clinical teaching mode,which has a study clue of organ and system.Those methods give much help for students to pass the Examination of Doctor Qualification.

Effects of canstatin gene transfection on growth and apoptosis of lung cancer A549 cells and HUV-ECC cells / 中国肺癌杂志

<p><b>BACKGROUND</b>Angiogenesis is essential for tumor's growth and metastasis. Canstatin, a newly found potent endogenous angiogenesis inhibitor, has drawn researcher's attention to its powerful biological activities on endothelial cells. The aim of this experiment is to explore the expression and effects of canstatin gene in lung cancer A549 cells and HUV-ECC cells.</p><p><b>METHODS</b>Expression vector of pCMV- Script/Canstatin was transfected into A549 and ECC cells by electroporation, and the positive clone was screened by G418. Growth characteristics of the two cell lines were compared before and after transfection. Expression of canstatin protein in supernatant was examined by SDS-PAGE assay, and cell cycles of the two cell lines were analysed by flow cytometry.</p><p><b>RESULTS</b>The expression of canstatin gene was found in supernatant of the transfected A549 cells and ECC cells. The apoptotic rate in the transfected ECC cells (16.04%) was significantly increased compared with that of the naked plasmid control group (0.43%) and parental cell group (2.92%) (P < 0.01). The growth of the transfected ECC cells was significantly inhibited (P < 0.01). The apoptotic rate in the transfected A549 cells (0.19%) showed no marked difference from the naked plasmid control group (0.13%) and parental cell group (0.07%) (P > 0.05). No significant difference in cell growth was found among the transfected A549 cell, naked plasmid control and parental cell groups.</p><p><b>CONCLUSIONS</b>The results indicate that canstatin gene can express in lung cancer A549 cell line and HUV-ECC cell line, and it can specifically inhibit proliferation of endothelial cell and induce its apoptosis.</p>

Dynamic changes of apoptotic rate of peripheral blood polymorphonuclear neutrophils in the patients with chronic obstructive pulmonary disease / 中国组织工程研究

BACKGROUND: Chronic obstructive pulmonary disease(COPD) is characterized of chronic inflammation in airway, pulmonary parenchyma and pulmonary vessels. The mechanism of the increment and activity changes of these inflammatory cells is unclear at present.OBJECTIVE: To study the apoptotic character of peripheral blood polymorphonuclear neutrophils(PMNs) and its relationship with COPD to provide a reference for early intervention and function surveillance for COPD patients.DESIGN: An observatory comparative study based on COPD patients and healthy population as controls.SETTING: Department of pulmonary medicine in a military medical university of Chinese PLA affiliated hospital.PARTICIPANTS: Totally 98 COPD patients were admitted by the Department of Pulmonary Medicine, Research Institute of Surgery, Daping Hospital, Third Military Medical University of Chinese PLA between February 2003 and December 2003 due to COPD acute attack. Eighteen patients including 12 males and 6 females aged between 48 and 70 years old [mean of(56 ± 7) years old]were randomly selected into COPD group according to random number table.Totally 14 healthy adults including 10 males and 4 females aged between 50 and 70 years old [mean of (59 ± 8) years old] who were individuals came for physical check up in our hospital were selected in control group.METHODS: PMNs were separated from peripheral blood by density gradient centrifugation. The dynamic changes of PMNs apoptosis in peripheral blood was observed by flow cytometer and TUNEL method.MAIN OUTCOME MEASURES: Comparison of PMNs apoptotic rate in peripheral blood among groupsRESULTS: As indicated by flow cytometric analysis, PMNs apoptotic rate at early apoptotic phase in COPD patients at paracmasis was(8.5 ± 1.3)%,which was significantly lower than(12.5 ± 1.8)% of normal control group( t=6.25, P ＜ 0. 01); PMNs apoptotic rate was(5.1 ±0.6)% at acute aggravation stage, which was significantly lower than that of paracmasis group ( t =5.66, P ＜001). As indicated by TUNEL analysis, PMNs apoptotic rate at paracmasis was(12.42 ±2.7)% , which was significantly lower than (21.5±4.8)% of normal control group(t=5.76, P ＜ 0.01); PMNs apoptotic rate was(4. 9 ±0.4)% at acute aggravation stage, which was significantly lower than that of paracmasis group( t = 6. 12, P ＜ 0. 01 ) . PMNs changes at the late phase of apoptosis/necrosis had a contrary tendency, i. e.,PMNs rate at late apoptotic phase/necrosis was(2. 8 ± 0.5)% in COPD patients at paracmasis, which was significantly higher than(1. 3 ±0.4)% of normal control group ( t= 6. 37, P ＜ 0. 01 ); PMNs rate was (3.7 ± 0. 3) % at acute aggravation stage, which was significantly higher than that of paracmasis group(t=5.81, P ＜0.01).CONCLUSION: PMNs abnormal apoptosis might be an important reason that induces PMNs aggregation in airway and lung tissues in COPD. This process might have an important significance in the generation and development of chronic airway inflammation, which provides an etiologic basis for the primary rehabilitative intervention of COPD.

Summarization in the research of canstatin / 生物医学工程学杂志

Canstatin protein, a newly found angiogenesis inhibitor, has powerful anti-angiogenesis effect. Canstatin is the N terminal fragment of collagen IV alpha2 chain NC1 domain. Its distinct anti-cancer effect in mouse model and low toxicity has not only made it a promising new anti-cancer drug candidate, but also drawn much attention of researchers. The detection, denomination, biological characters and mechanism of canstatin protein, and also the prospect of its application were summarized.

Bcl-2-associated athanogene 1 regulates the glucocorticoid receptor activity in rat alveolar macrophages / 医学研究生学报

Objective: To investigate the expression changes of Bcl-2-associated athanogene 1(BAG-1) and its regulatory effect on the glucocorticoid receptor(GR) activity in rat alveolar macrophages in conditions of cell inflammation and glucocorticoid therapy.Methods: The expression changes of BAG-1 were detected by Western blot after lipopolysaccharide(LPS) and Dexamethasone(Dex) treatment of rat alveolar macrophages(AMs),the interaction between BAG-1 and GR determined by immune coprecipitation experiment,and the transcriptional activation of GR measured by relative luciferase activity assay.Results:After LPS and Dex treatment,the expression of BAG-1L in total protein increased but that of BAG-1S remained changed,BAG-1L rather than BAG-1S was detected in nuclear protein and its expression increased gradually within 24 hours,the interaction between BAG-1L and GR was observed in nucleoli,and the transcriptional activation of GR decreased,with a negative correlation between BAG-1L expression and GR activity.Conclusion:LPS and Dex acting on rat alveolar macrophages,the expression of BAG-1L increases,which,coupled with GR,translocates into nucleoli and inhibits GR activity.This might be the important mechanism that underlies glucocorticoid resistance in inflammation.

Changes of glucocorticoid receptor expression and activity in rat alveolar macrophages induced by lipopolysaccharide / 第三军医大学学报

Objective To investigate changes of glucocorticoid receptor (GR) expression and activity in rat alveolar macrophages (AMs) induced by lipopolysaccharide (LPS) within 24 h. Methods Primary cultured AMs were divided randomly into LPS treatment group and LPS+Dex treatment group. The expressions of GR mRNA and GR protein in AMs at different time points were detected by RT-PCR and Western blot. Electrophoretic mobility shift assay (EMSA) was used to measure the GR activity in AMs. Results The expression level of GR mRNA decreased after LPS treatment, but returned to the normal level at 24 h after LPS treatment. The expression level of GR also decreased and reached the lowest level at 4 h after LPS treatment. GR activity also decreased, reached the lowest level at 1 h after LPS treatment, but was still lower than that in the normal control group at 24 h after treatment. Dexamethasone treatment had no obvious effect on GR activity during the late period of treatment. Conclusion LPS treatment (100 ng/ml) down-regulates the protein expression of GR, which maybe associated with the decreased expression of mRNA and accelerate degradation of GR protein. The activity of GR is inhibited sharply, resulting in glucocorticoid resistance.

Mechanism of 11?-HSD2 gene expression up-regulated by endotoxin in vascular endothelial cells / 第三军医大学学报

Objective To investigate effects of endotoxin on 11?-HSD2 gene transcription in vascular endothelial cells to observe the role of p38 mitogen-activated protein kinase (p38 MAPK) signaling pathway. Methods The effects of endotoxin in the presence or absence of p38 MAPK specific inhibitor SB203580 on the transcription of 11?-HSD2 in vascular endothelial cells was evaluated by reverse transcription DNA polymerase chain reaction. Results Treatments of endotoxin (1.0, 10, 20, 50, 100 ?g/ L) for 24 h increased the ratios of 11?-HSD2mRNA/?-actin mRNA in vascular endothelial cells. The induction of 11?-HSD2 mRNA by endotoxin could be inhibited partially by 10 mmol/ L SB203580. Conclusion Endotoxin stimulated the transcription of 11?-HSD2 gene in vascular endothelial cells. The activation of p38 MAPK might be an important mechanism of 11?-HSD2 gene induced by endotoxin.

Repressive effects of RNA interference technique on expression and function of glucocorticoid receptor in human macrophage cell line U937 / 第三军医大学学报

Objective To establish a glucocorticoid receptor knockdown model of human macrophage cell line U937 with RNA interference technique. Methods Two RNAi recombinant plasmids (named pSilencer 3.1-GR 1 and pSilencer 3.1-GR 2) targeting to GR gene were constructed. After RNAi recombinant plasmids were transfected into human macrophage cell line U937, the expressions of GR mRNA and GR protein were evaluated with RT-PCR and Western blotting respectively. The transcriptional activation function of GR was evaluated through the detection of relative luciferase activity after dexamethasone treatment. Results Two RNAi recombinant expression plamids were constructed and identified by sequencing. pSilencer 3.1-GR 2 transfection could inhibit not only GR mRNA and protein expressions, but also transcriptional activation function of GR specially; pSilencer 3.1-GR 1 transfection had no significant changes as compared to normal control. Conclusion A glucocorticoid receptor knockdown model has been established successfully, which offers a new method for the further research of GR biological functions.

Asthmatic epidemiological study of Han nationality in Chongqing / 第三军医大学学报

Objective To investigate the genetic faction influence on asthmatic patients in Chongqing, China. Methods Case-control study was employed for genetic epidemiological survey. Results The heritability of asthma in Chongqing was (80.56?5.68)% in first-degree relatives of asthma probands. The segregation ratio was 0.18. The relative risk of first-degree relatives and siblings of asthma probands were 7.38 and 4.47, respectively. Conclusion The inheritance pattern of asthma in Chongqing coincided with polygenic inheritance pattern. Genetic factor is a major risk factor of asthma. The relative risk in siblings is high. We can localize the susceptible gene of asthma with positional cloning.

Biologic characteristics and chromosome karyotype analysis of lung adenocarcinoma cell strain induced by cis-diaminodichloroplatin / 第三军医大学学报

Objective To establish lung adenocarcinoma drug resistance cell strain induced by cis-diaminodichloroplatin and investigate the biologic characteristics and chromosome karyotype of cell strain.Methods Large dosage impact and step by step inducing method were used to establish A549 and SPC-A-1 drug resistance cells: A549/CDDP and SPC-A-1/CDDP.MTT was used to detect the cell drug resistance index. Bioluminescence was used to detect the energy metabolism.Cell cycle was analysed by flow cytometer.The differential staining technique and SKY were used to analyze the chromatosome of A549/CDDP and SPC-A-1/CDDP.Results The drug resistance index of A549/CDDP and SPC-A-1/CDDP were 14.0 and 12.0. The content of ATP,ADP and AMP decreased significantly.The cell cycle of A549/CDDP and SPC-A-1/CDDP were of no notable changes.The results of the differential staining technique and SKY showed that the chromatosome of A549/CDDP and SPC-A-1/CDDP had several derivative chromosomes.Both cells had the same derivative chromosome: der(21)t(21;22).Conclusion CDDP can induce lung adenocarcinoma cell strain drug resistance.The derivative chromosome,including der(21)t(21;22) may have relationship with the drug resistance of cell strains.

Comparing the characteristics of rat pulmonary arterial smooth muscle cell cultured by two methods / 第三军医大学学报

Objective To compare the growth curve, protein content, calcium ion content of pulmonary arterial smooth muscle cells (PASMCs) cultured by enzyme digestion and tissue-sticking methods. To investigate whether PASMCs cultured by two ways could be offered to one series of signaling pathway studies. Methods The protein content was determined by upgraded-lowry, the calcium ion content by fluorescence indicator. The growth of PASMCs was observed by light microscope. Results There existed certain difference in protein content, growth curve between PASMCs cultured by the two methods, especially in calcium ion content (P

Genetic analysis of bronchial hyperresponsiveness and serum total IgE of asthma pedigree / 第三军医大学学报

Objective To study the genetics pattern of bronchial hyperresponsiveness and serum total IgE in asthma pedigree. Methods Twenty-eight asthma families were collected, including 124 individuals, 72 with asthma and 52 without. Forty-five normal subjects of three generations without consanguinity among one another, 22 male and 23 female, were chosen as controls, excluding those with family history of asthma and hypersensitivity. Serum total IgE by ELISA and bronchial hyperresponsiveness (BHR) were detected in asthma pedigree members. Genetics pattern of BHR and serum total IgE was analysed. Results The frequency distribution of BHR was bimodal. Individuals with higher level of total IgE occupied 40 of BHR-affected. Conclusion BHR may be controlled by a single major gene. BHR and total IgE may be under separate genetic control.