Ring finger protein 125, as a potential highly aggressive and unfavorable prognostic biomarker, promotes the invasion and metastasis of human gallbladder cancers via activating the TGF- ß1-SMAD3-ID1 signaling pathway.

Human gallbladder cancer (GBC) is a lethal aggressive malignant neoplasm. Identification of potential molecular biomarkers and development of targeted therapeutics for GBC patients is very necessary. In this study, we firstly investigated the correlation between ring finger protein 125 (RNF125) expression and the metastasis and prognosis of GBC, and the underlying molecular mechanism. RNF125 expression in a cohort of GBC tissues was examined; its correlation with clinicopathological and prognostic factors of GBC patients was analyzed. Moreover, the metastasis-related difference expressed genes in highly and lowly aggressive GBC cell lines were identified; and the influence of RNF125 knockdown on the metastatic phenotypes and characteristic EMT markers in highly aggressive GBC NOZ cells was detected. Furthermore, the underlying molecular mechanism of RNF125 effect was explored. The results showed that RNF125 was highly expressed in GBC tissues and related with aggressive characteristics such as Nevin stage (P = 0.041) etc. and unfavorable prognosis of GBC patients (P = 0.023, log-rank test). And, RNF125 was proved to a positive metastasis-related gene in vitro. RNF125 knockdown inhibited the invasion and migration, enhanced the adhesion, upregulated E-cadherin and ß-catenin expression, and downregulated vimentin and N-cadherin expression (all P < 0.001) of NOZ cells in vitro. RNF125 promoting effect on GBC tumor progression was identified to relate with the activation of TGF-ß1-SMAD3-ID1 signaling pathway. These findings firstly confirm that high RNF125 expression is related with aggressive characteristics and unfavorable prognosis of GBC patients; RNF125 promotes the invasion and metastasis of human GBCs via activating the TGF-ß1-SMAD3-ID1 signaling pathway.

Establishment and characterization of a novel highly aggressive gallbladder cancer cell line, TJ-GBC2.

BACKGROUND: Human gallbladder cancer (GBC) is an aggressive malignant neoplasm with a poor prognosis. The development of ideal tools for example tumor cell lines for investigating biological behavior, metastatic mechanism and potential treatment in GBCs is essential. In present study, we established and characterized a GBC cell line derived from primary tumor. METHODS: Primary culture method was used to establish this cell line from a primary GBC. Light and electron microscopes, flow cytometry, chromosome analysis, heterotransplantation and immunohistochemistry were used to characterize the epidemic tumor characteristics and phenotypes of this cell line. RESULTS: A novel GBC cell line, named TJ-GBC2, was successfully established from primary GBC. This cell line had characteristic epithelial tumor morphology and phenotypes in consistent with primary GBC, such as polygon and irregular cell shape, increased CA19-9 and AFP levels, and positive expression of CK7, CK8, CK19 and E-cadherin with negative vimentin. Moreover, about 25% of the cells were in the S-G2/M phase; abnormity in structure and number of chromosome with a peak number of 90-105 and 80% hypertetraploid were observed. Furthermore, this cell line had higher invasion and highest migration abilities compared to other GBC cell lines; and metastatic-related marker MMP9 and nm23 were positively expressed. CONCLUSIONS: A novel highly aggressive GBC cell line TJ-GBC2 was successfully established from primary GBC. TJ-GBC2 cell line may be efficient tool for further investigating the biological behaviors, metastatic mechanism and potential targeted therapy of human GBC.

A potential small-molecule synthetic antilymphangiogenic agent norcantharidin inhibits tumor growth and lymphangiogenesis of human colonic adenocarcinomas through blocking VEGF-A,-C,-D/VEGFR-2,-3 "multi-points priming" mechanisms in vitro and in vivo.

BACKGROUND: Tumor lymphangiogenesis plays an important role in promoting growth and metastasis of tumors, but no antilymphangiogenic agent is used clinically. Based on the effect of norcantharidin (NCTD) on lymphangiogenesis of human lymphatic endothelial cells (LECs), we firstly investigated the antilymphangiogenic activity of NCTD as a tumor lymphangiogenic inhibitor for human colonic adenocarcinomas (HCACs). METHODS: In vivo and in vitro experiments to determine the effects of NCTD on tumor growth and lymphangiogenesis of the in-situ colonic xenografts in nude mice, and lymphatic tube formation of the three-dimensional (3-D) of the co-culture system of HCAC HT-29 cells and LECs were done. Proliferation, apoptosis, migration, invasion, Ki-67, Bcl-2 and cell cycle of LECs and the co-culture system in vitro were respectively determined. Streparidin-peroxidase staining, SABC, western blotting and RT-PCR were respectively used to examine the expression of LYVE-1, D2-40, CK20 (including their LMVD), and VEGF-A, VEGF-C, VEGF-D, VEGFR-2 and VEGFR-3 in vitro and in vivo. RESULTS: NCTD inhibited tumor growth and lymphangiogenesis of the in-situ colonic xenografts in vivo, and these observations were confirmed by facts that lymphatic tube formation, proliferation, apoptosis, migration, invasion, S-phase cell cycle, and Ki-67 and Bcl-2 expression in vitro, and LYVE-1, D2-40, CK20 expression and their LMVD in vitro and in vivo were inhibited and affected. Furthermore, the expression of VEGF-A, VEGF-C, VEGF-D, VEGFR-2 and VEGFR-3 at protein/mRNA levels in the process of lymphatic tube formation in vitro and tumor lymphangiogenesis in vivo was downregulated; NCTD in combination with mF4-31C1 or Sorafenib enhanced these effects. CONCLUSIONS: NCTD inhibits tumor growth and lymphangiogenesis of HCACs through "multi-points priming" mechanisms i.e. affecting related malignant phenotypes, inhibiting Ki-67 and Bcl-2 expression, inducing S-phase cell cycle arrest, and directly or indirectly downregulating VEGF-A,-C,-D/VEGFR-2,-3 signaling pathways. The present finding strongly suggests that NCTD could serve as a potential antilymphangiogenic agent for tumor lymphangiogenesis and is of importance to explore NCTD is used for antitumor metastatic comprehensive therapy for HCACs.

Norcantharidin enhances TIMP­2 anti­vasculogenic mimicry activity for human gallbladder cancers through downregulating MMP­2 and MT1­MMP.

Vasculogenic mimicry (VM) is a tumor microcirculation pattern in highly aggressive gallbladder cancers (GBCs). We recently reported the anti­VM activity of norcantharidin (NCTD) in highly aggressive GBC­SD cells and xenografts. In this study, we further investigated that NCTD enhanced tissue inhibitor of matrix metalloproteinase­2 (TIMP­2) anti­VM activity for GBCs and the underlying mechanisms. In vivo and in vitro experiments were performed to determine the effects of NCTD in combination with TIMP­2 on tumor growth, host survival, VM formation, hemodynamic of GBC­SD xenografts, and VM­like networks and malignant phenotypes of GBC­SD cells. Expression of matrix metalloproteinase (MMP)­2 and membrane type 1­MMP (MT1­MMP) among human GBCs, GBC­SD cells and xenografts were determined, respectively. The results showed that expression of MMP­2 and MT1­MMP in human GBCs, GBC­SD cells and xenografts was significantly related to VM in GBCs; a shorter survival time of VM­positive patients with high expression of MMP­2 or MT1­MMP compared to that of the patients with low expression. After treatment with NCTD+TIMP­2, tumor growth, VM formation, VM hemodynamic of the xenografts in vivo were significantly inhibited as compared to control, NCTD or TIMP­2 group, with a prolonged survival time of the xenograft mice (log­rank test, P=0.0115); and these observations were confirmed by VM­like networks by 3­D matrices and showed that proliferation, apoptosis, invasion, migration of GBC­SD cells in vitro were markedly affected. Furthermore, expression of MMP­2 and MT1­MMP in VM formation of the xenografts in vivo and GBC­SD cells in vitro was downregulated as compared to control, NCTD or TIMP­2 group. Thus, we concluded that NCTD enhances TIMP­2 antitumor and anti­VM activities in GBCs through downregulating MMP­2 and MT1­MMP.

Inhibition of tumor vasculogenic mimicry and prolongation of host survival in highly aggressive gallbladder cancers by norcantharidin via blocking the ephrin type a receptor 2/focal adhesion kinase/paxillin signaling pathway.

Vasculogenic mimicry (VM) is a newly-defined tumor microcirculation pattern in highly aggressive malignant tumors. We recently reported tumor growth and VM formation of gallbladder cancers through the contribution of the ephrin type a receptor 2 (EphA2)/focal adhesion kinase (FAK)/Paxillin signaling pathways. In this study, we further investigated the anti-VM activity of norcantharidin (NCTD) as a VM inhibitor for gallbladder cancers and the underlying mechanisms. In vivo and in vitro experiments to determine the effects of NCTD on tumor growth, host survival, VM formation of GBC-SD nude mouse xenografts, and vasculogenic-like networks, malignant phenotypes i.e., proliferation, apoptosis, invasion and migration of GBC-SD cells. Expression of VM signaling-related markers EphA2, FAK and Paxillin in vivo and in vitro were examined by immunofluorescence, western blotting and real-time polymerase chain reaction (RT-PCR), respectively. The results showed that after treatment with NCTD, GBC-SD cells were unable to form VM structures when injecting into nude mouse, growth of the xenograft was inhibited and these observations were confirmed by facts that VM formation by three-dimensional (3-D) matrix, proliferation, apoptosis, invasion, migration of GBC-SD cells were affected; and survival time of the xenograft mice was prolonged. Furthermore, expression of EphA2, FAK and Paxillin proteins/mRNAs of the xenografts was downregulated. Thus, we concluded that NCTD has potential anti-VM activity against human gallbladder cancers; one of the underlying mechanisms may be via blocking the EphA2/FAK/Paxillin signaling pathway.

Norcantharidin inhibits tumor growth and vasculogenic mimicry of human gallbladder carcinomas by suppression of the PI3-K/MMPs/Ln-5γ2 signaling pathway.

BACKGROUND: Vasculogenic mimicry (VM) is a novel tumor blood supply in some highly aggressive malignant tumors. Recently, we reported VM existed in gallbladder carcinomas (GBCs) and the formation of the special passage through the activation of the PI3K/MMPs/Ln-5γ2 signaling pathway. GBC is a highly aggressive malignant tumor with disappointing treatments and a poor prognosis. Norcantharidin (NCTD) has shown to have multiple antitumor activities against GBCs, etc; however the exact mechanism is not thoroughly elucidated. In this study, we firstly investigated the anti-VM activity of NCTD as a VM inhibitor for GBCs and its underlying mechanisms. METHODS: In vitro and in vivo experiments to determine the effects of NCTD on proliferation, invasion, migration, VM formation, hemodynamic and tumor growth of GBC-SD cells and xenografts were respectively done by proliferation, invasion, migration assays, H&E staining and CD31-PAS double stainings, optic/electron microscopy, tumor assay, and dynamic micro-MRA. Further, immunohistochemistry, immunofluorescence, Western blotting and RT-PCR were respectively used to examine expression of VM signaling-related markers PI3-K, MMP-2, MT1-MMP and Ln-5γ2 in GBC-SD cells and xenografts in vitro and in vivo. RESULTS: After treatment with NCTD, proliferation, invasion, migration of GBC-SD cells were inhibited; GBC-SD cells and xenografts were unable to form VM-like structures; tumor center-VM region of the xenografts exhibited a decreased signal in intensity; then cell or xenograft growth was inhibited. Whereas all of untreated GBC-SD cells and xenografts formed VM-like structures with the same conditions; the xenograft center-VM region exhibited a gradually increased signal; and facilitated cell or xenograft growth. Furthermore, expression of MMP-2 and MT1-MMP products from sections/supernates of 3-D matrices and the xenografts, and expression of PI3-K, MMP-2, MM1-MMP and Ln-5γ2 proteins/mRNAs of the xenografts were all decreased in NCTD or TIMP-2 group; (all P < 0.01, vs. control group); NCTD down-regulated expression of these VM signaling-related markers in vitro and in vivo. CONCLUSIONS: NCTD inhibited tumor growth and VM of human GBCs in vitro and in vivo by suppression of the PI3-K/MMPs/Ln-5γ2 signaling pathway. It is firstly concluded that NCTD may be a potential anti-VM agent for human GBCs.

[Re-optimized technology of protective ileostomy with no need of reversal].

OBJECTIVE: To explore the clinical application of aoptimizedtechniquebased onpreviouslyreported protecting stoma with no need forreversal. METHODS: Thetechniquealso used "the assembly of drainage device" to performprotecting ileostomy. The original method includes enterotomy at the terminal ileum to placedrainage device, which was optimized as follows: two intestinal pursestring with 0.5 cm distance were placed 5 cm away from the ileocecal valve. Transverse enterotomy was performed in the anti-mesenteric side. The assembly was placed at the root of the appendix between two pursestring, and then the intestine purse suture was tighten. Ligation of the small intestine anastomosis between the anastomosis ring at both ends was carried out, and theanastomosis ring was deployed. From the root of the appendix in the cecum wall, the assembly was embedded about 2 cm and pulled out of abdominal cavitythough the Trocar hole. RESULTS: Seventeen cases of ultra-low rectal cancer completed protecting stoma, including 11 cases through ileocecal protective stoma. All the anastomosis healed well. Defecation drainage tube was removed 3-5 weeks after anastomosis ring degradation. Drainage nozzle healed after 3 to 5 days, and no complications occurred. CONCLUSION: The optimized ileocecal protective ileostomy has the following advantages: (1)wound healing time is significantly shorter. (2)secondary intestinal fistula can be prevented. (3)no need to fix ileum and less chance of subsequent volvulus, intestinal obstruction.

Suppression of lymphangiogenesis in human lymphatic endothelial cells by simultaneously blocking VEGF-C and VEGF-D/VEGFR-3 with norcantharidin.

Lymph node metastasis of tumors is a crucial early step in the metastatic process. Tumor lymphangiogenesis plays an important role in promoting tumor metastasis to regional lymph nodes. Norcantharidin (NCTD) has been reported to possess potent anti-angiogenesis and antitumor properties in several cell lines and xenograft tumor models. However, its role in tumor-associated lymphangiogenesis and lymphatic metastasis remains unclear. Here, we investigated the effect of NCTD on proliferation, apoptosis, migration, invasion and the lymphatic tube formation, lymphangiogenesis, of human lymphatic endothelial cells (HLECs) in vitro by MTT, proliferation assay, Hoechst staining and flow cytometry, scraping line method, Matrigel invasion assay, inverted or fluorescence microscope and transmission electron microscope. Moreover, the underlying mechanisms, such as VEGF-C, VEGF-D, VEGFR-3 at protein and mRNA levels in lymphangiogenesis using 3-dimensional (3-D) culture of HLECs were measured by immunohistochemistry, western blotting and real-time polymerase chain reaction (RT-PCR). It was shown that NCTD inhibited proliferation, migration, invasion and lymphatic tube formation (forming-lymphatic and/or formed-lymphatic) of HLECs, induced HLEC apoptosis (all P<0.01) significantly, in a dose- and time-dependent manner (IC50 6.8 µg/ml); and downregulated the expression of VEGF-C, VEGF-D and VEGFR-3 at protein or/and mRNA levels (P<0.01) in HLEC lymphatic tube formation. Thus, we identified for the first time that NCTD inhibited HLEC lymphangiogenesis by simultaneously blocking VEGF-C and VEGF-D/VEGFR-3 in vitro. The present findings may be of importance to explore the therapeutical target or strategy of NCTD for tumor lymphangiogenesis and lymphatic metastasis.

CD146 expression correlates with epithelial-mesenchymal transition markers and a poor prognosis in gastric cancer.

CD146 has been regarded as a novel potential therapeutic target for multiple cancers. The aim of the study was to investigate the expression of CD146 in gastric cancer and evaluate its clinical-pathological and prognostic significance. The expression of CD146 and three epithelial-mesenchymal transition (EMT)-related proteins (E-cadherin, ß-catenin and vimentin) was examined in 144 gastric cancers by immunohistochemistry. Fifty-nine cases (41.0%) were defined as positive for CD146 expression. We found that CD146 expression correlated positively with lymph node involvement and a poor prognosis, and retained an independent prognostic factor for gastric cancer patients. Furthermore, positive expression of CD146 was strongly associated with loss of the epithelial marker E-cadherin and acquisition of the expression of the mesenchymal markers nuclear ß-catenin and vimentin. These findings suggest that CD146 might promote EMT and progression in gastric cancer, and thus may be a potential therapeutic target for patients with gastric cancers.

[Evaluation of nasal patency in patients with vasomotor rhinitis].

OBJECTIVE: To measure the nasal patency in patients with vasomotor rhinitis (VMR) and healthy controls and to assess its correlation with visual analogue scale (VAS). METHODS: A total of 105 patients with VMR and 71 healthy controls were included in this study. By using nasal rhinomanometry, the pressure-flow curve and got total nasal resistances of 75 Pa and 150 Pa were measured. By means of acoustic rhinometry, the area-distance curve before and after using nasal vasoconstrictor substance was obtained, got the nasal minimum cross-sectional area (MCA), then calculated nasal congestion index (NCI). The outcomes of nasal resistance and acoustic rhinometry in two groups were compared. The correlation between VAS and nasal patency of VMR was evaluated. RESULTS: The correlation between the outcomes with nasal resistance and acoustic rhinometry and VAS of nasal symptom showed no statistical significance in VMR patients (all P > 0.05). MCA before and after decongestion showed no difference (Z value were -1.541 and -0.626, each P > 0.05), NCI had statistic differences in two groups (Z = -2.707, P < 0.05). Nasal resistance of 75 Pa had statistic differences in two groups (Z = -4.334, P < 0.05), 150 Pa showed no difference (Z = -1.314, P > 0.05). CONCLUSIONS: Vasomotor rhinitis is one of the most common non-allergic rhinitis. Subjective symptoms has no correlation with objective nasal patency tests. In clinical practice, comprehensive evaluation of subjective symptoms and objective test results of the patient is required.

[Impact of olfactory nerve transection on the apoptosis of mice olfactory receptor neurons].

OBJECTIVE: To analyze the impact of olfactory nerve transection on the apoptosis of mice olfactory receptor neurons (ORN), and discuss the reliability of this experimental model. METHODS: After olfactory nerve transection of mice, anterograde horseradish peroxidase (HRP) tracing was performed to confirm the completion of nerve transection. On 8 h, 2 d, 3 d and 5 d after surgery, TdT mediated deoxyuridine triphosphate-biotin nick end labelling (TUNEL) was used to observe the apoptosis of ORN, while relative semi-quantitative RT-PCR and immunohistochemistry were used to reflect the expression of olfactory marker protein (OMP, special marker of mature ORN) in olfactory epithelium. RESULTS: No HRP label was observed in olfactory bulb after olfactory nerve transaction. Both TUNEL-positive and OMP-positive cells were ORN. After the surgery, TUNEL-positive cells increased remarkably and peaked on 2 d after the surgery. Meanwhile OMP mRNA in olfactory epithelium began to decrease markedly till 5 d after the surgery, and the olfactory epithelium got thinner accordingly. CONCLUSIONS: This experimental model can be used reliably to sever mice olfactory nerve and manipulate simultaneous apoptosis of mice ORN.

[Effect of intranasal interleukin-12 gene therapy for allergic rhinitis in murine model].

OBJECTIVE: To investigate whether the local application of IL-12 gene with EBV-plasmid vector to nasal mucosa could prevent allergic inflammation in murine allergic rhinitis model. METHODS: Thirty-six BALB/C mice were randomly divided into allergic rhinitis group gene therapy group and control group. In mice with OVA-induced allergic rhinitis, the EBV/lipoplex (a novel cationic lipid combined with EBV-plasmid vector, pGEG. mIL-12) was locally administered into nasal mucosa before OVA challenge. The expression of IL-12 mRNA and protein, the change of eosinophilia and mast cell, and Th2 cytokine production in the nasal mucosa were measured. RESULTS: The amounts of IL-12 mRNA positive cells and IL-12 positive cells in nasal mucosa of gene therapy group were significantly higher than that of allergic rhinitis group (P < 0.01 and P < 0.05). The amount of eosinophils, mast cells, and the level of IL-5 expression in nasal mucosa in allergic rhinitis group were significantly higher than those in gene therapy group and control group (P < 0.01). The level of total IgE of peripheral blood in allergic rhinitis group was significantly higher than that in gene therapy group and control group (F = 1216.21, P < 0.01). CONCLUSIONS: These findings indicated that IL-12 mRNA and protein were expressed effectively after the local administration of pGEG. mIL-12 in the nasal mucosa. The local application of pGEG. mIL-12 is effective in modulating nasal allergic response and may be a convenient method for future approach to allergic rhinitis.

[Anatomical and computed tomographic analysis of the interaction between uncinate process and agger nasi cells].

OBJECTIVE: To investigate the anatomical interaction between uncinate process and agger nasi cell to better understand the anatomy of the frontal sinus drainage pathway by endoscopy, spiral computed tomography (CT) and sectioning. METHODS: Twenty-one skeletal skulls (forty-two sides) and one cadaver head (two sides) were studied by spiral CT together with endoscopy and collodion embedded thin sectioning at coronal plane. The sections with the thickness of 100 microm were stained with hemotoxylin and eosin. RESULTS: Under endoscopy, a leaflet of bone to the middle turbinate, which is given off by uncinate process, forms the anterior insertion of the middle turbinate onto the lateral nasal wall. The middle portion of the uncinate process attached to the frontal process of the maxilla in all of the skeletal nasal cavities, as well as the lacrimal bone in 78.6% of the skeletal nasal cavities. On CT scans, the agger nasi cell is present in 90.5% of the skeletal nasal cavities. While the lateral wall of the agger nasi cell is formed by lacrimal bone, the medial wall of the agger nasi cell is formed by uncinate process. And the anterior wall is formed by the frontal process of the maxilla. The superior portion of the uncinate process forms the medial, posterior and top wall of the agger nasi cells. The superior portion of the uncinate extends into the frontal recess and may insert into lamina papyracea (33.3%), skull base (9.5%), middle turbinate, combination of these (57.2%). CONCLUSIONS: The agger nasi cell is the key that unlocks the frontal recess.

[Relationship between GATA-3 and IL-4 and IL-5 in patients with allergic rhinitis].

OBJECTIVE: To evaluate the relationship between GATA-3 and IL-4, IL-5 in patients with allergic rhinitis. METHODS: The expression of GATA-3 was detected by immunochemistry and reverse transcriptase polymerase chain reaction (RT-PCR) in 23 patients with allergic rhinitis. IL-4, IL-5 were detected by enzyme-linked immunosorbent assay (ELISA). RESULTS: The positive rate of GATA-3 was 86.9% (20/23) and the relative density ratio of GATA-3 and glyceraldehyde phosphate dehydrogenase (GAPDH) were 0.602 +/- 0.11 in patients with allergic rhinitis. The concentrations of IL-4, IL-5 were (135.5 +/- 66.4) pg/mg, (77.5 +/- 29.4) pg/mg, respectively. The expressions of GATA-3 and IL-4, IL-5 had positive relationship (r = 0.45 and 0.62, P < 0.05). CONCLUSIONS: GATA-3 contributed to the production of IL-4, IL-5 in patients with allergic rhinitis.

[Effect of intranasal glucosteroid on the gene expression of interleukin-5 in nasal polyps].

OBJECTIVE: To investigate the effect of intranasal glucosteroid treatment on the expression of interleukin (IL)-5 gene in nasal polyps. METHODS: Nasal polyps from topical steroid treated patients (n = 20) and untreated patients (n = 20) were investigated with the technique of mRNA in situ hybridization. RESULTS: The majority of IL-5 mRNA positive cells in nasal polyps were lymphocytes or eosinophils. No statistical significance was found in the densities of IL-5 mRNA positive cells between allergic patients [(12.6 +/- 4.6)/0.25mm2] and nonallergic patients [(14.3 +/- 4.1)/0.25mm2] (t = -0.775, P > 0.05). Compared with the control group [(13.9 +/- 4.2)/0.25mm2], the density of IL-5 mRNA positive cells was decreased in the steroid-treated group [(10. 2 +/- 3.1)/0.25mm2], and the difference reached statistical significance (t = 3.114, P < 0.01). CONCLUSIONS: These findings indicate that topical steroid treatment may suppress the IL-5 gene expression, and steroids may inhibit eosinophil functions.

[Expression of chloride channel in nasal mucosa with allergic rhinitis].

OBJECTIVE: To evaluate the role of chloride channel ClC-3 in perennial allergic rhinitis. METHODS: The nasal mucosa from 19 patients with perennial allergic rhinitis were studied by immunohistochemistric staining and reverse transcriptase polymerase chain reaction (RT-PCR) for the expression of ClC-3. RESULTS: ClC-3 was positively stained in 17 cases of 19 patients, with a rate of 89.6%. It mainly distributed in cytoplasm of epithelium and submucosal gland. All tissues displayed expected DNA length after PCR. The relative density of ClC-3 in mucosa with allergic rhinitis was 0.685 +/- 0.114 and 0.140 +/- 0.075 in normal. CONCLUSION: ClC-3 played an important role in the over-secretion of liquor in allergic rhinitis.

[Relationship between expressions of GATA-3 and interleukin-5 in nasal polyps].

OBJECTIVE: To evaluate the role of transcription factor GATA-3 in the pathogenesis of nasal polyps. METHODS: The expression of GATA-3 was detected by immunohistochemistry and reverse transcriptase polymerase chain reaction (RT-PCR) in 28 cases of nasal polyps and 17 specimens of normal nasal epithelium. The expression of interleukin-5 (IL-5) in these specimens was detected by enzyme-linked immunosorbent assay (ELISA) meanwhile. RESULTS: Immunohistochemistry showed, as confirmed by RT-PCR, a positive rate of GATA-3 of 89.3% in the nasal polyp (25/28) and 29.4% in the normal nasal mucosa (5/17), P < 0.05. GATA-3 was mainly distributed in submucous inflammatory cells. The relative density ratio of GATA-3 to GAPDH was 0.618 +/- 0.137 in nasal polyp and 0.21 +/- 0.11 in normal nasal mucosa (P < 0.05) as indicated by RT-PCR and agarose electrophoresis. The concentrations of IL-5 were 69.4 +/- 15.1 pg/ml and 25.7 +/- 13.0 pg/mg in the two groups respectively. The expression of GATA-3 was positively correlated to the expression of IL-5 in the nasal polyp. CONCLUSION: Contributing to the overexpression of such cytokines as IL-5, GATA-3 may play an important role in the pathogenesis of nasal polyps.

[A study of the link between upper and lower respiratory inflammation].

OBJECTIVE: To evaluate the correlation between upper and lower respiratory inflammation. METHODS: Thirty rats were randomly divided into allergic rhinitis group (AR), asthma group (AS), and control group (Con). The pulmonary function was measured by pulmonary function instrument of animal model, the infiltration of eosinophils and mast cells was detected by hematotoxylin-eosin staining (HE staining) and toluidine blue staining respectively, the expression of VCAM-I and IL-13 was examined by immunohistochemistry. ELISA was used to measure the concentration of IL-5 in the peripheral serum. RESULTS: The numbers of eosinophils and mast cells in nasal mucosa and lung tissue of AR were both significantly higher than those of Con, and similar results were observed between AS and Con. The forced expiratory volume in 0.3 second (FEV0.3)/forced vital capacity (FVC) of AR and AS was both significantly lower than that of Con. The numbers of VCAM-I and IL-13 positive vessels in lung tissue of AR were both significantly higher than those of Con, and similar results were observed between AS and Con. The concentration of IL-5 in the serum of AR and AS was both significantly higher than that of Con. The concentration of IL-5 in serum was positively correlated with the number of eosinophils in peripheral blood of AR and AS. CONCLUSION: The inflammation is similar between AR and AS, and repeatedly challenge upper respiratory tract can impact on lower respiratory response.

[Interleukin-5 regulate the bone marrow response in the model of allergic rhinitis].

OBJECTIVE: To elucidate the mechanism of bone marrow response regulated by interleukin-5 (IL-5) in the model of allergic rhinitis. METHODS: Twenty SD rats were randomly divided into allergic rhinitis (AR) group and control group. The leucocytes in the smears of bone marrow and peripheral blood were counted, and the expression of IL-5 was detected by immunohistochemistry. RESULTS: The ratio of eosinophils to white cells in bone marrow smears of AR group was significantly higher than that of control group(P < 0.01). The ratio of basophils to white cells in bone marrow smears of AR group was significantly higher than that of control group(P < 0.01). The ratio of eosinophils to white cells in peripheral blood smears of AR group was significantly higher than that of control group(P < 0.01). The ratio of IL-5 positive cells to white cells in bone marrow smears of AR group was significantly higher than that of control group (P < 0.05). The ratio of IL-5 positive cells to white cells was significantly positively correlated with the ratio of eosinophils to white cells in bone marrow smears of AR group (R = 0.85, P < 0.01). CONCLUSION: IL-5 regulates the proliferation and differentiation of eosinophils in bone marrow.