AIMTo investigate the influence and mechanisms of 17beta-estradiol on the CTP: phosphorylcholine cytidylyltransferase (CCT) activity from cultured lung explants without serum.

METHODSWe detected the amount of [M-14C] choline incorporation into phosphatidylcholine so as to reflect CCT activity by liquid scintillation.

RESULTS(1) 17beta-estradiol increased the CCT activity in dose-dependence and time-dependence. (2) Both the protein kinase C inhibitor H-7 and calmodulin antagonist W-7 abolished the stimulatory effect of 17beta-estradiol (3 x 10(-6) mol/L) on the CCT activity.

CONCLUSION17beta-estradiol can increase CCT activity in cultured lung explants, its mechanism is related to protein kinase C and calmodulin.

Animals ; Calmodulin ; metabolism ; Choline-Phosphate Cytidylyltransferase ; metabolism ; Culture Media, Serum-Free ; Estradiol ; pharmacology ; In Vitro Techniques ; Lung ; drug effects ; enzymology ; Male ; Protein Kinase C ; metabolism ; Rats ; Rats, Wistar

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Manipulation, Orthopedic ; Middle Aged ; Temporomandibular Joint Disorders ; therapy

OBJECTIVETo compare the CC-chemokine ligand 18 (CCL18) expression in the serum and malignant pleural effusion (MPE) of NSCLC patients and explore its regulatory effect on differentiation of monocyte-derived dendritic cells (Mo-DC).

METHODSCCL18 levels in the serum and MPE from 62 NSCLC patients were quantitated by immunoassay. CCL18 in sera from 26 healthy individuals, 28 exudative pleural effusions from inflammatory pulmonary diseases and 17 transudative pleural effusions from non-inflammatory diseases were used as control. Mo-DC was generated by culturing NSCLC-derived monocytes with GM-CSF and IL-4 in the presence or absence of CCL18. The mean fluorescent intensity (MFI) of CD14, CD80, CD83, CD86 and HLA-DR were analyzed by flow cytometry (FCM). Mo-DC was then co-cultured with purified T cells and the percence of CD25(+)FoxP3(+) cells was assayed by FCM.

RESULTSCCL18 levels in the sera of NSCLC patients and healthy individuals were (132.70 ± 15.52) ng/ml and (18.44 ± 0.99) ng/ml, respectively (P < 0.001). The levels of CCL18 in MPE, exudative PE and transudative PE were (155.6 ± 13.58) ng/ml, (190.4 ± 22.33) ng/ml and (20.89 ± 3.03) ng/ml, respectively. CCL18 in the MPE was significantly higher than that in transudates (P < 0.001), however, no significant difference was observed between CCL18 expression in exudative PE and MPE (P = 0.172). Of note, a moderate positive correlation (r = 0.421, P < 0.01) was observed between CCL18 levels in the paired MPE and serum of NSCLC. In the healthy control group, Mo-DC cultured in the presence of CCL18 showed 31.4 ± 15.8 (MFI) of CD14 expression, which was significantly higher than that in Mo-DC cultured in the absence of CCL18 (18.5 ± 8.9, P < 0.05). In contrast, the expressions of MFI of CD80, CD83, CD86 and HLA-DR were significantly decreased upon CCL18 induction (P < 0.05). In the NSCLC group, GM-CSF+IL-4+CCL18 induced a MFI of 45.2 ± 13.8 of CD14 expression in Mo-DC, which was also significantly higher than that of GM-CSF+ IL-4 induction (22.6 ± 10.5, P < 0.01). Similarly, the expressions of MFI of CD80, CD83, CD86 and HLA-DR were significantly decreased in the presence of CCL18 (P < 0.05). Furthermore, the MFI of CD14, CD83, CD86 and HLA-DR had significant differences between GM-CSF/IL-4/CCL18-induced Mo-DC derived from NSCLC patients and healthy control (P < 0.05). Finally, CD4(+) T cells co-cultured with NSCLC-derived, GM-CSF/IL-4/CCL18-treated Mo-DC had significantly higher percent of CD25(+)FoxP3(+) cells compared with that of CD4(+) T cells stimulated with Mo-DC induced by GM-CSF/IL-4(P < 0.01).

CONCLUSIONSCCL18 is present at a high level in MPE and serum of NSCLC patients complicated with pleural effusion and a moderate positive correlation exists between CCL18 levels in the two fluids. CCL18 inhibits maturation of Mo-DC, which consequently stimulates T cells to differentiate into CD25(+)FoxP3(+) regulatory T cells.

Carcinoma, Non-Small-Cell Lung ; metabolism ; Cell Differentiation ; Chemokines ; Chemokines, CC ; metabolism ; Coculture Techniques ; Dendritic Cells ; metabolism ; Flow Cytometry ; Granulocyte-Macrophage Colony-Stimulating Factor ; metabolism ; Humans ; Interleukin-4 ; metabolism ; Ligands ; Lung Neoplasms ; Monocytes ; physiology ; Pleural Effusion ; T-Lymphocytes, Regulatory

OBJECTIVETo investigate the efficacy of the procedure for prolapse and hemorrhoids (PPH) combined with external hemorrhoids excision in the treatment of III or IV mixed hemorrhoids.

METHODSOne hundred and twelve patients with III or IV mixed hemorrhoids admitted for surgical treatment were randomly divided into three groups: PPH 1 group (34 cases), PPH2 group (36 cases), and Milligan-Morgan group (42 cases). PPH1 group received the standard PPH operation, PPH2 received PPH and external hemorrhoids excision, and Milligan-Morgan group received Milligan-Morgan hemorrhoidectomy. Postoperative 24 h-pain index, pain index when defecating, bleeding, anal discomfort feeling , wound edema, the ability of controlling feces, operating time, hospitalization time and charges were recorded. The change of anal dynamics was detected by anorectal manometry. All the patients were followed-up for 0.5-1 year.

RESULTSThere were no significant differences among the three groups in bleeding, anal discomfort feeling, the ability of controlling feces (P>0.05). The postoperative 24 h-pain index of PPH1 group was lower than those of the other two groups (P<0.05). PPH1 group and PPH2 group were better than Milligan-Morgan group in pain index when defecating, wound edema, operating time, and hospitalization time (P<0.05). Milligan-Morgan group was better than the other two groups in postoperative urinary retention and hospital charges (P<0.05). The change of anal duct pressure of Milligan-Morgan group was less than those of the other two groups (P<0.05). Within 0.5-1.0 year follow-up, 3 patients got thrombosed external hemorrhoid in PPH1 group, 2 patients recurred and 1 patient got thrombosed external hemorrhoid in Milligan-Morgan group, no recurred patients in PPH2 group.

CONCLUSIONPPH combined with external hemorrhoid excision is a safe and effective treatment for mixed hemorrhoids, which is suitable for mixed hemorrhoids with severe external hemorrhoids.

Adult ; Aged ; Anal Canal ; surgery ; Female ; Follow-Up Studies ; Hemorrhoids ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Prolapse ; Surgical Stapling

Background:Despite the growing epidemic of heart failure (HF),there is limited data available to systematically compare non-cardiac comorbidities in the young-old,old-old,and oldest-old patients hospitalized for HF.The precise differences will add valuable information for better management of HF in elderly patients.Methods:A total of 1053 patients aged 65 years or older hospitalized with HF were included in this study.Patients were compared among three age groups:(1) young-old:65 to 74 years,(2) old-old:75 to 84 years,and (3) oldest-old:≥85 years.Clinical details of presentation,comorbidities,and prescribed medications were recorded.Results:The mean age was 76.7 years and 12.7％ were 85 years or older.Most elderly patients with HF (97.5％) had at least one of the non-cardiac comorbidities.The patterns of common non-cardiac comorbidities were different between the young-old and oldestold group.The three most common non-cardiac comorbidities were anemia (53.6％),hyperlipidemia (45.9％),and diabetes (42.4％) in the young-old group,while anemia (73.1％),infection (58.2％),and chronic kidney disease (44.0％) in the oldest-old group.Polypharmacy was observed in 93.0％ elderly patients with HF.Additionally,29.2％ patients were diagnosed with infection,and 67.0％ patients were prescribed antibiotics.However,60.4％ patients were diagnosed with anemia with only 8.9％ of them receiving iron repletion.Conclusions:Non-cardiac comorbidities are nearly universal in three groups but obviously differ by age,and inappropriate medications are very common in elderly patients with HF.Further treatment strategies should be focused on providing optimal medications for age-specific non-cardiac conditions.

We have previously shown that the binding of integrins with extracellular matrix component fibronectin (Fn) can improve the ability of bronchial epithelial cells (BECs) in resisting oxidant injury by up-regulating the activity of catalase and increasing the content of GSH. However, the molecular mechanism or its signaling pathway of this protection is still unclear. In order to examine the intracellular signaling mechanism activated by Fn-integrin binding reaction, the present study investigated the mRNA expression of catalase in primary cultured rabbit BECs using RT-PCR based on a cell-injury model made with ozone exposure. The product bands of target gene CAT were checked with Southern blot and oligonucleotide probe hybridization. The results showed that Fn (10 microg/ml) promoted the catalase mRNA transcription (P<0.01). This effect was abolished either by protein-tyrosine kinase inhibitor genistein or calmodulin inhibitor W(7) (P<0.01). These results indicate that the promotion of catalase activity induced by Fn-integrin reaction is partly due to the elevation of catalase mRNA transcription, and that its signalling are possibly relevant to tyrosine phosphorylation or calmodulin pathway.
Animals ; Bronchi ; cytology ; metabolism ; Calmodulin ; metabolism ; Catalase ; biosynthesis ; genetics ; Cells, Cultured ; Epithelial Cells ; cytology ; metabolism ; Female ; Fibronectins ; physiology ; Integrins ; physiology ; Male ; Protein-Tyrosine Kinases ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Rabbits ; Signal Transduction ; Up-Regulation ; drug effects

To explore the role of intrapulmonary neuropeptides in the development of airway hyperresponsiveness, we established an animal model of airway hyperresponsiveness (AHR) in rabbits by using ozone exposure. With the model, after test of the mechanics of respiration and bronchoalveolar lavage assay, the levels of vasoactive intestinal peptide (VIP) and calcitonin gene-related peptide (CGRP) in the lungs were determined by radioimmunoassay, and the expression of mRNA coding receptors of these two neuropeptides was evaluated by reverse transcriptional-polymerase chain reaction (RT-PCR). At the same time, the distribution of VIP receptor-1 (VIPR1) and CGRP receptor-1 (CGRPR1) in lung tissues and its time-course were examined by in situ hybridization. The results showed: (1) in ozone-stressing groups, airway resistance increased significantly and typical inflammatory pathological changes were observed in pulmonary tissue slides, including neutrophil and eosinophil infiltration, mucus exudation and bronchial epithelial cells (BECs) shedding; (2) with elongation of ozone exposure, the levels of VIP and CGRP in the lungs increased at first, reaching a peak on d 2 to 4, then decreased slowly, and CGRP peaked somewhat earlier than VIP; (3) mRNA expression of the two neuropeptide receptors in the lungs changed in a similar manner like VIP and CGRP, but the high level of mRNA expression of VIPR1 lasted longer than that of CGRPR1; and (4) in situ hybridization for neuropeptide receptors demonstrated that, in unstressed control, VIPR1 and CGRPR1 positive cells appeared in the airway epithelium, pulmonary interstitial and focal areas of airway and vascular smooth muscles. With the elongation of ozone exposure, hybridization stained deeper and the majority of positive cells were located around the vessels and bronchus except a few in the alveoli. At 8 d, only a small number of positive cells were seen in the lungs. From the results, it is concluded that ozone-stressing can induce the development of AHR, in which VIP and CGRP may play important roles. That implies, through binding to CGRPR1, CGRP stimulates an early inflammation response which contributes in cleaning up of irritants, while VIP exerts a later dampening of pulmonary inflammation response. These two neuropeptides may play sequential and complementary roles in the development of AHR.
Animals ; Bronchi ; pathology ; Bronchial Hyperreactivity ; chemically induced ; metabolism ; Bronchoalveolar Lavage Fluid ; Calcitonin Gene-Related Peptide ; metabolism ; Epithelium ; metabolism ; Lung ; metabolism ; Ozone ; Rabbits ; Receptors, Calcitonin Gene-Related Peptide ; metabolism ; Receptors, Vasoactive Intestinal Peptide ; metabolism ; Vasoactive Intestinal Peptide ; metabolism

Intercellular adhesion molecule-1 (ICAM-1) is an important adhesion molecule leading to adhesion between cells; NF-kappaB, being universally distributed in the organism, is an important nuclear transcription factor leading to a rapid response to the stimuli. Line of evidence have shown that ICAM-1 transcription and NF-kappaB activation is an important step of inflammatory reaction. To testify that intrapulmonary regulatory peptides modulate inflammatory lesion of bronchial epithelial cells (BECs) through their effect on ICAM-1 expression and nuclear factor kappaB (NF-kappaB) activation, we used immunocytochemistry, RT-PCR, and electrophoretic mobility-shift assay (EMSA) to determine the ICAM-1 expression and NF-kappaB activity in BECs. The effects of NF-kappaB inhibitor MG-132 on ICAM-1 expression were also observed. The results showed that vasoactive intestinal peptide (VIP) and epidermal growth factor (EGF) decreased ICAM-1 expression in O(3)-stressed BECs, while endothelin-1 (ET-1) and calcitonin gene-related peptides (CGRP) increased ICAM-1 expression in resting BECs. MG-132 blocked ICAM-1 expression induced by O(3), ET-1 and CGRP. The results obtained by using EMSA confirmed that VIP and EGF restrained the activation of NF-kappaB in O(3)-stressed BECs; CGRP and ET-1 promoted activation of NF-kappaB. These observations indicate that VIP and EGF abated the injury by means of down-regulatory effects on ICAM-1 transcription and NF-kappaB activation, while ET-1 and CGRP enhanced the inflammation reaction by an up-regulatory effect. It is suggested that a developing and intensive airway inflammation correlates closely with a persistent expression of ICAM-1 and repeated activation of NF-kappaB.
Animals ; Bronchi ; cytology ; Cell Adhesion ; physiology ; Cells, Cultured ; Endothelin-1 ; metabolism ; Epithelial Cells ; cytology ; metabolism ; Humans ; Inflammation ; metabolism ; Intercellular Adhesion Molecule-1 ; metabolism ; NF-kappa B ; metabolism ; Peptides ; physiology ; Rabbits ; Vasoactive Intestinal Peptide ; physiology

To investigate the influence of vasoactive intestinal peptide (VIP) on chemotaxis of bronchial epithelial cells (BECs). Rabbit chemotactic migration of primary BEC was assessed in a blind-well Boyden chamber. Radioimmunoassay and radio-ligand affinity analysis were used for determining VIP secretion and vasoactive intestinal peptide receptor (VIPR) expression. The results showed: (1) the method for determining chemotaxis of BECs by using insulin as chemotactic factor was stable and reproducible (r=0.9703, P<0.01). (2) VIP (0.001-1 micromol/L) elicited chemotaxis of BECs which was substantial and concentration-dependent. The effects of VIP were inhibited by W-7 and H-7 (P<0.01). (3) Heat stress enhanced the secretion of VIP (P<0.01) and upregulated the expression of VIPR on BECs (P<0.05). These results indicate that VIP in the lungs may play an important role in the repair of damaged epithelium, accelerating restoration of the airway to its normal state. Calmodulin and protein kinase C may be involved in the signal transduction of VIP effects.
Animals ; Bronchi ; cytology ; Cells, Cultured ; Chemotaxis ; drug effects ; physiology ; Epithelial Cells ; drug effects ; physiology ; Female ; Insulin ; pharmacology ; Male ; Rabbits ; Receptors, Vasoactive Intestinal Peptide ; biosynthesis ; Vasoactive Intestinal Peptide ; pharmacology

Objective: This study was designed to improve complete remission(CR) rate in the patients with advanced lymphoblastic lymphoma by using early extensive induction chemotherapy. Method:A total of 11 cases of untreated lymphoblastic lymphoma in Stage Ⅲ /Ⅳ were received CVDLP regimen, including cytoxan(CTX) 1000 mg/m2 d1， vincristine(VCR) 1.5 mg/m2 d1,d8,d15,d21， Adriamycin(ADR) 40 mg/m2 d1， d2， d21， L-asparaginase(L-ASP) 10000 U/m2 d15～24， Prednison 60 mg/m2 d1～28， gradually decreased dosage at d15. methotrexate＋ Ara-C IT qw× 4. Efficacy were evaluated at d28～35. Simultaneously,retrospective analysis for 9 cases of untreated lymphoblastic lymphoma in Stage Ⅲ /Ⅳ treated with 2 cycles of CHOP were made. Efficacy were evaluated at d35. Results: CVDLP group: 10/11 cases of patients achieved CR, and 1/11 case had PR, rate of complete remission was 90.9％ ;10/11 cases had Grade Ⅳ hematological toxicity,1/11 cases had Grade Ⅲ hematological toxicity(WHO). CHOP group:3/9 got CR;5/9 got PR;1/9 had MR,rate of complete remission was 33％ . 3/9 had Grade Ⅲ hematology toxicity;6/9 had GradeⅡ hematological toxicity. Conclusion:CVDLP regimen can induce higher CR rate than CHOP regimen in untreated lymphoblastic lymphoma with Stage Ⅲ /Ⅳ , but hematology toxicity was also higher than CHOP regimen. However this induction regimen is safe and viable with strengthening supportive care.