Objective To investigate the aetiology of acute exacerbations of COPD (AECOPD),and the effects of moxifloxacin in the treatment of AECOPD.Methods Patients with stable COPD based on GOLD criteria were included in the study.Sputum collected at first exacerbation was analyzed for bacteria count and culture.IL-6,IL-8 and TNF-?were measured by enzyme- linked immunosorbent assay (ELISA).Eligible patients were randomized to receive moxifloxacin (400 mg qd for 5 days) or ce- faclor (250 mg q8h for 7 days).Efficacy parameters were evaluated at 7 and 14 days after treatment initiation and 1 year later. Results Of the 46 patients with moderate or severe COPD (male 38,moderate 24),21 (45.65%) were microbiologically evalu- able at baseline.The main pathogen was Haemophilus influenzae (10/21).Clinical efficacy rate was 87.0% in moxifloxacin group and 82.6% in cefaclor group.Bacterial eradication rate was 80.0% and 72.7% respectively.The difference between groups was not statistically significant in terms of clinical or microbiological efficacy.In moxifloxacin arm,the frequency of ex- acerbation was 2.6?1.0,significantly lower than control arm (3.5?1.4,P

Cell viability of Pichia pastoris was detected by flow cytometry (FCM) with two reagents fluorescein diacetate (FDA) and propidium iodide (PI). Compared with FDA/PI double-stained dot plots and PI single-stained dot plots,the latter could divide dead and living cells into two separate zones,and get the correct proportion. Then PI single-stained method was used to detect the change of cell viability in Pichia patoris fermentation. At glycerol batch and fed-batch phase,little dead cells were detected. At methanol fed-batch phase,cell viability decreased when cell weight increased,and was only 73.8% at 88 h.

Animals ; Endothelins ; physiology ; Humans ; Mice ; Pulmonary Fibrosis ; etiology ; Rats

Objective To explore the expression of TLR4/MyD88/NF-κB signaling and its correlation with the progression of acute intestinal graft-versus-host disease (iGVHD) after allogeneic bone marrow transplantation in mice.Method Recipient BALB/c female mice were lethally irradiated and were reconstituted within 4-6 h with a transplant of bone marrow cells (1 × 107) and different amounts of splenocytes (1 × 107,n =12 or 2 × 107,n =12) from MHC-mismatched C57BL/6 donors to induce iGVHD,and 6 healthy BALB/c mice served as controls.A globe survey observation of GVHD by survival,clinical manifestation,and histological detection was performed.RT-PCR,immunohistochemistry,and Western blotting technology were used to detect the mRNA and protein levels of TLR4,MyD88 and NF-κB p65 in the small intestine tissue.Result The exacerbation of iGVHD was associated with the increasing dose of allogeneic spleen lymphocytes.The mRNA expression of TLR4,MyD88 and NF-κBp65 was increased as the iGVHD progressed.All of them in severe iGVHD model were significantly increased as compared with the healthy controls (P＜0.05).The expression of corresponding proteins had the same tendency as mRNA.All of the three genes expression was not only positively correlated with each other,but also with the clinical GVHD score:TLR4 (R =0.814,P＜0.001),MyD88 (R=0.828,P＜0.001),and NF-κB p65 (R=0.568,P =0.034).Conclusion Excessive activation of TLR4/MyD88/NF-κB signaling pathway does exist in iGVHD,and the enhanced levels of gene transcription and translation are positively correlated with the deterioration of iGVHD.

Objective To investigate the effects of acupuncture combined with conventional treatment on swallowing function and hemorheology in patients with pseudobulbar palsy following stroke. Methods One hundred thirty-one patients with laughing sickness after cerebral stroke from March 2010 to November 2013 were selected and divided into two group by random number table method:a treatment group (66 patients) and a control group (65 patients). The control group was treated with aspirin and rosuvastatin, while the treatment group was additional treated with acupuncture for benefiting vital energy and eliminating phlegm for 4 weeks. The swallowing function was evaluated by Kubota Water Swallow Test and the hemorheological measurements were performed. Results The total improvement rate of swallowing function in the treatment group was significantly higher than that in the control group (86.36% vs. 64.61%; χ2=8.391, P=0.004). After the treatment, the whole blood viscosities at high shear rate (4.96 ± 0.53 mPa?s vs. 5.32 ± 0.63 mPa?s;t=3.541, P=0.001) and low shear rate (23.23 ± 0.94 mPa?s vs. 23.81 ± 1.01 mPa?s;t=3.403, P=0.001), and the plasma viscosity (1.52 ± 0.24 mPa?s vs. 1.61 ± 0.28 mPa?s;t=1.976, P=0.050) in the treatment group were significantly lower than those in the control group. Conclusion Acupuncture combined with conventional treatment can improve the swallowing function and the hemorheological parameters in patients with pseudobulbar palsy following stroke.

Purpose To explore the clinicopathological characteristics of primary non-Hodgkin lymphoma ( PNHL) of lung. Methods The clinical features, morphology and immunohistopathological phenotypes were retrospectively studied in 28 cases of PNHL and re-viewed of the literature. Results The composition of this group of cases is 18 cases of male, 10 cases female. The a median age of pa-tients was 57 years old. According to the WHO classification, all of the 28 cases of in our PNHL series were diagnosed as non-Hodgkin lymphoma ( NHL) , including 17 mucosa-associated lymphoid tissue extranodal marginal zone lymphomas ( MALT) , 5 diffuse large B cell lymphomas ( DLBLC) , 2 NK/T cell lymphomas and 2 anaplastic large cell lymphomas, 1 mantle cell lymphoma ( MCL) and 1 pe-ripheral T cell lymphomas, unspecified, respectively. One-third PNHL patients presented with specific clinical symptoms such as cough, chest pain, dyspnea, and fatigue, Imaging examination showed unilateral or bilateral pulmonary infiltrate, single lesions or multiple nodules, and the lesions always involved with the trachea, bronchus and lung. Follow-up was completed in 16 patients ( range, 3 to 38 months) . 3 cases were conducted with pneumonectomy, among which 2 cases were given postoperative adjuvant chem-otherapy. Seven patients were given chemotherapy alone, and 5 patients did not give any treatment following initial diagnosis. At the time of last follow-up, 13 patients were alive with disease, 2 patients were died. The tumor metastasis in the left inguinal lymph node was found in one patient after 2 years by surgery. Conclusions It was shown that there was no specific clinical manifestations and fea-tures of pulmonary PNHL. Among of them, the MALT is the most common diseases, and the highly aggressive lymphomas many be oc-cur, such as the DLBLC, NK/T cell lymphoma. The diagnosis of PNHL depends on pathological examination. The immunohistochemi-cal staining and molecular pathological technology may helpful for the diagnosis and differential diagnosis of the tumors.

Proteolytic degradation has been a severe problem when Pichia pastoris is employed to express recombinant proteins.One alternative method to circumvent this problem is to construct protease gene disruptant.However,the main study of gene disruption is focused on nonrecombinant Pichia pastoris rather than recombinant strain.In our study,we established two different methods to directly disrupt PRC1 and KEX1 gene in recombinant Pichia pastoris.On the basis of this,we further discussed and compared the application and advantages of both methods.

The kinase domain of receptor tyrosine kinase(RTK) ErbB2 was expressed fused with GFP in Pichia pastoris. Recombinant expression vector pPIC3.5K was constructed in Escherichia coli TOP10. The right P. pastoris transformants were screened on his-deficient plates and YPD-G418 plates by turns after electroporation of recombinant vector, and then induced by methanol in baffled shake bottles. The strain with highest protein yield was scaled up in a 5 L fermentor. Recombinant protein was analyzed with tyrosine kinase assay after Ni2+ affinity chromatograph. Results showed that the 100 kD recombinant protein with tyrosine kinase activity was successfully expressed in P. pastoris.

The bioreactor production of recombinant Lateolabrax japonicus growth hormone (rljGH) expressed intracellularly by Pichia pastoris was investigated. A strategy of feeding methanol at the exponential rate was established and the effect of specific growth rate on the rljGH production was examined. The results indicated that the average specific production rate increased and the rljGH production duration decreased as the specific growth rate increased. The maximum specific rljGH production (0.58 mg/g WCW) was achieved at a specific growth rate of 0.029/h. The effect of supplementing ammonium sulfate, peptone and yeast ex- tract on the rljGH production was further investigated. The results indicated that the effects of ammonium sulfate and peptone were not significant. Supplementing yeast extract of 2.5 g/L was advantageous for the rljGH production. The duration of the rljGH production was increased to 23 h from 17 h and the fermenta- tion stability of run-to-run could be improved.

A fusion expression vector pPIC3.5K-PDGFR? was constructed to express recombinant receptor tyrosine kinase PDGFR? and the right Pichia pastoris transformants were screened on his-deficient plates and YPD-G418 plates by turns after electroporation of strain GS115, a high yield strain named M3 was screened. The strain M3 was cultured in a 5 L fermentor and His-GFP-PDGFR? fusion protein was purified by Ni2+ chelating affinity chromatography. One distinct peak was obtained after elution with 250 mmol/L imidazole. Fusion protein was proved to be 90.08 kD by western blotting, and have tyrosine kinase activity by ELISA. Results showed that the receptor tyrosine kinase PDGFR? was successfully expressed in P. pastoris and could be used as a target for small molecule selective inhibitors screening.