Adult ; Asthma ; metabolism ; physiopathology ; Basic-Leucine Zipper Transcription Factors ; genetics ; physiology ; Fanconi Anemia Complementation Group Proteins ; genetics ; physiology ; Female ; Glutamate-Cysteine Ligase ; metabolism ; Humans ; Inflammation ; metabolism ; pathology ; Male ; NF-E2-Related Factor 2 ; genetics ; physiology ; RNA, Messenger ; genetics ; metabolism ; Sputum ; cytology

AIMTo investigate the effects of protein tyrosine kinase on the inflammation and airway remodeling in lung of guinea pigs with bronchial asthma.

METHODS30 adult male guinea pigs were randomly divided into 3 groups (n=3): control group (C group), asthmatic group(A group)and genistein group (B group). Asthmatic model was established by ovalbumin intraperitoneal injection and ovalbumin inhalation. The total cell and the proportion of inflammatory cell in bronchial alveolar lavage fluid(BALF), inflammatory cell infiltration and index of remodeling of bronchiole were measured, respectively. The expression of p-tyrosine in lung tissue was examined by immunohistochemistry.

RESULTSThe total cell and proportion of eosinophil in BALF of A group were significantly higher than that of C group (P < 0.01), but compared with A group, the total cell and proportion of eosinophil in BALF of B group were much lower (P < 0.01). The number of eosinophile and lymphocyte of bronchiole in A group were significantly higher than that of C group (P < 0.01), but compared with A group, the number of eosinophile and lymphocyte in bronchiole of B group were much lower (P < 0.01). Compared with A group, the remodeling of bronchiole of B group was significantly relieved (P <0.01), there was no difference between B and C group (P > 0.05). Immunohistochemistry indicated that in A group the p-tyrosine was more positively expressed at the bronchial smooth muscle, bronchial epithelium, smooth muscle of vessel and inflammatory cell, especially at smooth muscle of bronchi and vessel and inflammatory cell than that of C group (P <0.01), there was no difference between B group and C group (P > 0.05).

CONCLUSIONPTK played a key role in inflammation and bronchial remodeling in lung of guinea pigs with bronchial asthma. The Protein tyrosine kinase inhibitor genistein could prevent and inhibit the inflammation and bronchial remodeling in lung of guinea pigs with bronchial asthma.

Airway Remodeling ; physiology ; Animals ; Asthma ; physiopathology ; prevention & control ; Genistein ; pharmacology ; Guinea Pigs ; Inflammation ; prevention & control ; Male ; Ovalbumin ; Protein-Tyrosine Kinases ; antagonists & inhibitors ; physiology ; Random Allocation

Objective To investigate the clinical effect of Ethibond suture and anchor fixation by arthroscopic technique for the treatment of avulsion fracture of the anterior cruciate ligament (ACL) tibial insertion.Methods Twenty patients with avulsion fracture of the ACL tibial insertion hospitalized between July 2013 and June 2014 were collected retrospectively.There were 12 males and 8 females,aged 18-41 years (mean,25.3 years).All patients were identified with type Ⅱ and type Ⅲ fractures according to the Meyers-McKeever classification.Results of Lachman test and anterior drawer test were both positive.All patients accepted the operation that avulsion fracture of the ACL tibial insertion was treated with the Ethibond suture and anchor fixation by arthroscopic technique within 3 weeks after injury.Follow-up X-ray examinations were carried out to evaluate the bone union.Lysholm and international knee documentation committee (IKDC) scores were used to evaluate the function of knee joint postoperatively.Results Operation time was 45-70 min (mean,50 min).Blood loss was 5-15 ml (mean,10 ml).Follow-up was conducted for 12-24 months (mean,16.3 months).Postoperative X-ray showed the reduction was satisfactory.Lachman test and anterior drawer test were both negative after operation.Knee functions were recovered to normal.Lysholm and IKDC scores were 89-96 points [(93.5 ± 2.3) points]and 84-96 points [(91.0 ± 3.9)points] respectively at the final follow-up,improved compared to the preoperative 42-61 points [(51.1 ± 6.2) points] and 46-68 points [(55.2 ± 7.0) points] (both P ＜0.05).Conclusion Arthroscopic Ethibond suture and anchor fixation for avulsion fracture of the ACL tibial insertion is an effective procedure with the advantages of small trauma,reliable fixation,good functional recovery and satisfactory clinical effect.

Objective To seek the curative effect of stereotactic radiation therapy (SRT) combined with gemcitabine for unresectable advanced pancreatic carcinoma.Methods 24 patients were treated by SRT of 6MV X-ray.Patients were fixed with the rack of stereotactic localization and heated plastics mould.The CT scanning results were put into the treatment planning system.According to the target area of tumor,sensitive organs and moving error drew GTV,CTV and PTV.The best plan was selected by the dose-volume histogram (DVH).5 to 7 beams of non-coplanar radiation ray were chosen.PTV was surrounded by ≥95 ％ isodose curves.Conventional fraction,5 fractions per week,1.8-2 Gy per fraction was used.All patients received a total dose of 50-60 Gy.Gemcitabine was performed 1000 mg/m2,once a week,iv gtt,in the 1st,2nd and 4th,5 th weeks.Results In chemo-radiotherapy,50.0 ％ (12/24) patients showed light nausea,41.7 ％ (10/24) patients showed leucopenia or thrombocytopenia of grade 1 or 2,after symptomatic treatment,all patients completed the planned treatment.In a period of one month to three months,after SRT combined with chemotherapy,appetite improvement was 83.3 ％ (20/24),jaundice disappeared in 6 of 6 patients (100.0 ％),abdominal pain was relieved in 21 of 24 patients (87.5 ％),3 patients were relieved completely among them.The complete remission (CR) rate was 16.7 ％ (4/24) and partial remission (PR) rate was 66.7 ％ (16/24),with CR+PR rate of 83.4 ％ (20/24).One-year survival rate was 70.8 ％ (17/24).As a consequence of cachexia,intestinal obstruction or bleeding,7 patients died within one year.Nobody survived more than 2 years.Conclusions SRT combined with gemcitabine for advanced pancreatic carcinoma may relieve symptoms.It is an effective approach to improve life quality and prolong survival time for advanced pancreatic carcinoma,especially lod and weak patients are more suitable to select SRT combined with gemcitabine.

Objective To investigate the relationship of the1675A/G polymorphism of AT2 gene with the therapeutic effect of indapamide sustained release tablets in female patients with primary hypertension.Methods Two hundred and twenty female patients with primary hypertension were treated with Indapamide Sustained Release Tablets ( 1.5 mg · qd) for 8 weeks.The blood samples from the patients were collect to determine AT2 gene polymorphism by PCR combined with HRM and sequencing.Results Two hundred and five patients completed the test.In female patients,the therapeutic efficacy of indapamide sustained telease tablets among different AT2R genotypes( AA:70.6％,AG:71.6％,GG:71.4％ ) showed no significant difference ( x2=2.53,P=0.49 ),neither do the decline of BP after therapy ( F=0.39 and 0.19 respecrively,P ＞ 0.05).Conclusion The AT2 genotype was assumed to be not correlated to the blood pressure lowering response to Indapamide Sustained Release Tablets in female primary hypertension patients.

Objective To explore the synergetic effect of HBX protein and M2 macrophages in inflammatory microenvironment on invasion and metastasis of hepatocellular carcinoma cells.Methods Hep3B cells were infected with recombinant lentivirus carrying HBx gene,following co-culture with THP-1 original M2 macrophages.The cells were divided into six groups:two infected groups (Hep3B +and Hep3B + + M2),four non-infected groups (Hep3B-,Hep3B-+ LV5,Hep3B-+ M2,Hep3B-+LV5 + M2).Western blot (WB) was used to assess the expression changes of E-cadherin and N-cadherin,markers of epithelial-mesenchymal transition (EMT).The cellular location of EMT markers was observed by immunofluorescence confocal microscopy.Transwell assay was used to evaluate the invasion ability of Hep3B cells.Results HBX protein overexpressed in Hep3B cells by lentivirus infection.After 72 h co-culture with M2 macrophages,WB results showed that E-cadherin descreased significantly in Hep3B+ (0.42 ±0.11) when compared with Hep3B-(1.00 ±0.18) (t =4.762,P ＜0.05),while N-cadherin was significantly higher in Hep3B + (2.85 ± 0.44) than in Hep3B-(1.00 ± 0.17) (t =4.762,P ＜ 0.05).M2macrophages decreased E-cadherin expression in Hep3 B + + M2 (0.1 ± 0.13) compared with Hep3 B + (t =3.255,P ＜0.05),while N-cadherin expression increased in Hep3B+ + M2 (4.18 ± 0.52) (t=10.009,P ＜ 0.05).Non-Infected groups didn't change the markers of E-cadherin and N-cadherin.It was suggested that invasion ability of Hep3B increased by HBx overexpression.Conclusions HBX protein and M2 macrophages synergetically mediated the invasion and metastasis of hepatocellular carcinoma cells by EMT.

In order to construct a eukaryotic co-expression plasmid containing membrane-anchored Sjcl4FABP and Sjc26GST genes and identify their expression in vitro, Sj14 and Sj26 genes were obtained by RT-PCR with total RNA of Schistosoma japonicum adult worms as the template and cloned into eukaryotic expression plasmid pVAC to construct recombinant plasmids pVAC-Sj14 and pVAC-Sj26. Then a 23 amino-acid signal peptide of human interleukin-2 (IL-2) upstream Sj14 or Sj26 gene and a membrane-anchored sequence containing 32 amino-acids of carboxyl-terminal of human placental alkaline phosphatase (PLAP) downstream were amplified by PCR as the template of plasmid pVAC-Sj14 or pVAC-Sj26 only to get two gene fragments including Sj14 gene and Sj26 gene. The two modified genes were altogether cloned into a eukaryotic co-expression plasmid pIRES, resulting in another new recombinant plasmid pIRES-Sj26-Sj14. The expression of Sj14 and Sj26 genes was detected by RT-PCR and indirect immunofluorescent assays (IFA) when the plasmid pIRES-Sj26-Sj14 was transfected into eukaryotic Hela cells. Restriction enzyme analysis, PCR and sequencing results revealed that the recombinant plasmids pVAC-Sj14, pVAC-Sj26 and plRES-Sj26-Sj14 were successfully constructed and the expression of modified Sj14 and Sj26 genes could be detected by RT-PCR and IFA. A bivalent membrane-anchored DNA vaccine encoding Sj14 and Sj26 genes was acquired and expressed proteins were proved to be mostly anchored in cellular membranes.

Objective To establish a real-time quantitative polymerase chain reaction (qPCR) assay for B-cell lymphoblastic leukemia according to individualized and specific immunoglobulin gene rearrangements in leukemia cells, and to use it for the monitoring of minimal residual disease (MRD) of B-cell lymphocytic leukemia. Methods The immunoglobulin gene rearrangements of bone marrow samples from 15 cases of B-cell lymphoblastic leukemia were analyzed with a validated European BIOMED-2 system, and the individualized and specific qPCR-based quantification of leukemic immunoglobulin gene rearrangements was established. Results Unique and specific gene rearrangements of immunoglobulin light and heavy chains were identified in 14 cases and Ig-qPCR based on these gene rearrangements had a sensitivity of 10-5 and high specificity which met the international criteria in 10 patients. Leukemia MRD quantification with immunoglobulin gene rearrangement-based qPCR was similar as compared with other MRD detection methods. Conclusion Immunoglobulin gene rearrangement-based leukemia MRD quantification is feasible, sensitive, specific, precise and much valuable for clinical decision of treatments in B-cell lymphoblastic leukemia.

OBJECTIVETo investigate the dynamic expression and role of SENP1 (SUMO-specific proteases-1) in the pulmonary vascular wall of rat during the development of hypoxic pulmonary hypertension (HPH).

METHODSForty adult male Wistar rats were randomly divided into 5 groups (n = 8), and exposed to normoxia (Control group) or exposed to hypoxia for 3, 7, 14 or 21 d, respectively. The HPH models were established by normobaric intermittent hypoxia. Mean pulmonary arterial pressure (mPAP), right ventricle hypertrophy index (RVHI), and vessel morphometry were measured. Reverse transcriptase-polymerase chain reaction(RT-PCR) and in situ hybridization were used to determine the mRNA expression of SENP1. Immunohistochemistry and Western blot were used to determine the protein expression of SENP1.

RESULTSThe hypoxic rats developed pulmonary vascular remodeling in pulmonary arterioles after 7 d of hypoxia exposure. Pulmonary vascular remodeling in pulmonary arterioles significantly increased after 14 d of hypoxia. The level of mPAP in hypoxic rats increased significantly after 7 d of hypoxia, reached its peak after 14 d of hypoxic exposure. RVHI was markedly increased after 14 d of hypoxia. In situ hybridization and immunohistochemical analysis showed that SENP1 mRNA and protein were positively stained in control. SENP1 mRNA expression had little changes after exposure to hypoxia compared with the control, however, SENP1 protein expression was declined gradually after 7 d of hypoxia. The results of RT-PCR and Western blot showed that the same dynamic expression of SENP1 mRNA and protein in lung tissues of rats. Linear correlation analysis showed that SENP1 protein were negatively correlated with mPAP, pulmonary vascular remodeling index and RVHI.

CONCLUSIONUnder chronic hypoxia, SENP1 protein can be degradated. The dynamic expression of SENP1 protein may play a role in implicating in the development of HPH.

Animals ; Endopeptidases ; metabolism ; Hypertension, Pulmonary ; etiology ; metabolism ; Hypoxia ; complications ; metabolism ; Male ; Pulmonary Artery ; metabolism ; Rats ; Rats, Wistar

AIMTo investigate the expression and relationship of gamma-glutamylcysteine synthetase (gamma-GCS) and NF-E2-related factor2 (NRR2) in lung of rat with chronic obstructive pulmonary disease (COPD)in order to elucidate the possible important role of gamma-GCS and NRF2 in pathogenesis of COPD.

METHODSThe rat COPD model was established by intratracheal instillation of lipopolysaccharide twice and exposed to cigarette smoke daily. The gamma-GCS activity was measured, the expression of gamma-GCS mRNA in lung was examined by in situ hybridization (ISH) and reverse transcription-polymerase chain reaction (RT-PCR), the protein expressions of NRF2, gamma-GCS in lung were detected by immunohistochemical (IH) and Western blot respectively.

RESULTSThe gamma-GCS activity was higher in COPD group than that in control group. The expressions of gamma-GCS mRNA in COPD group was stronger than those in control group. ISH showed that the gamma-GCS mRNA was expressed in alveolar epithelium and bronchial smooth muscle cell in COPD. The protein expressions of NRF2, gamma-GCS were significantly higher than the control group. IH showed that NRF2, gamma-GCS proteins were expressed in alveolar and bronchial epithelium in the COPD group. There was a positive correlation between NRF2 and gamma-GCS and gamma-GCS mRNA.

CONCLUSIONNRF2 may play an important role in the mechanism of COPD oxidative stress vis up-regulation of gamma-GCS.

Animals ; Glutamate-Cysteine Ligase ; metabolism ; Lung ; metabolism ; Male ; NF-E2-Related Factor 2 ; metabolism ; Oxidative Stress ; Pulmonary Disease, Chronic Obstructive ; metabolism ; physiopathology ; Random Allocation ; Rats ; Rats, Wistar