Chinese herbal medicine formula Gu-Ben-Fang-Xiao-Tang attenuates airway inflammation by modulating Th17/Treg balance in an ovalbumin-induced murine asthma model.

Gu-Ben-Fang-Xiao-Tang (GBFXT) is a traditional Chinese medicine formula consisting of 11 medicinal plants, which has been used in the treatment of asthma. The present study aimed to determine the protective effects and the underlying mechanisms of GBFXT on ovalbumin (OVA)-induced allergic inflammation in a mouse model of allergic asthma. A total of 50 mice were randomly assigned to the following five experimental groups: Normal, model, montelukast (2.6 mg/kg), 12 g/kg GBFXT and 36 g/kg GBFXT groups. Airway responsiveness was measured using the forced oscillation technique, while differential cell count in the bronchoalveolar lavage fluid (BALF) was measured by Wright-Giemsa staining. Histological assessment was performed by hematoxylin and eosin staining, while BALF levels of Th17/Treg cytokines were measured by enzyme-linked immunosorbent assay, and the proportions of Th17 and Treg cells were evaluated by flow cytometry. The results showed that GBFXT suppressed airway hyperresponsiveness during methacholine-induced constriction, reduced the percentage of leukocytes and eosinophils, and resulted in decreased absolute neutrophil infiltration in lung tissue. In addition, GBFXT treatment significantly decreased the IL-17A cytokine level and increased the IL-10 cytokine level in the BALF. Furthermore, GBFXT significantly suppressed Th17 cells and increased Treg cells in asthmatic mice. In conclusion, the current results demonstrated that GBFXT may effectively inhibit the progression of airway inflammation in allergic asthma, partially by modulating the Th17/Treg cell balance.

Imbalance of Peripheral Th17 and Regulatory T Cells in Children with Allergic Rhinitis and Bronchial Asthma.

The purpose of the present study is to investigate the prevalence of Th17 and regulatory T (Treg) cells in children with allergic rhinitis (AR) accompanying with bronchial asthma (BA). 24 children with AR, 22 children with BA, 18 children with AR accompanying with BA, and 20 healthy controls were recruited. The prevalence of peripheral blood Th17 and Treg cells were determined by flow cytometry. mRNA expression of retinoid-acid receptor-related orphan receptor (ROR)-γt and forkhead box P3 (Foxp3) were determined by realtime polymerase chain reaction. Cytokine expressions in plasma were determined by enzyme linked immunosorbent assay. The frequency of Th17 cells, ROR-γt mRNA expression, and the plasma levels of IL-17 were significantly higher, while Treg cells and Transforming growth factor (TGF)-ß1 were significantly lower in children with AR accompanying with BA compared with those in children with AR or BA alone or control subjects. In children with allergic airway disease, total IgE levels were positively correlated to the frequency of Th17 cells (r=0.607, p<0.01), plasma IL-17 levels, and negatively correlated to the frequency of Treg cells (r=-0.429, p<0.01) and TGF-ß1 levels (r=-0.224, p<0.01). While Forced expiratory volume in one second (FEV1) (% predicted) was negatively correlated to the frequency of Th17 cells (r=-0.602, p<0.01), plasma IL-17 levels (r=-0.577, p<0.01), and positively correlated to the frequency of Treg cells r=0.504, p<0.01) and TGF-ß1 levels (r=0.231, p<0.05). Our results demonstrate that the imbalance of peripheral Th17/Treg cells plays an important role in the pathogenesis of AR accompanying with BA.

[The microsurgical anatomic research of the internal auditory canal area on the retrosigmold approach].

OBJECTIVE: To evaluate the safety of the circular round window and discus anatomic landmarks of posterior wall of internal auditory canal by investigating the microscopic anatomy of internal auditory canal area of the retrosigmold approach, which can provide the anatomical basis for acoustic neutrinomas surgery. METHOD: Fifteen adult cadaver heads (30 sides) fixed with formalin were used in the study. The retrosigmold approach operations were imitated to dissect the blood vessels and nerves in internal auditory canal area by opening round bony window and removing posterior wall of internal auditory canal. RESULT: Fifteen specimens of 30 sides circular bone window were opened without injury with transverse sinus and sigmoid sinus. The vertical distance between the highest point of bone window margo superior and the lowest point of transverse sinus margo inferior was (4.02 ± 0.32) mm. The vertical distance from the most anterior point of bone window leading edge to the most posterior point of sigmoid sinus trailing edge was (6.31 ± 0.43) mm. The internal auditory canal tubercle located in the anterior superior position of internal auditory canal. The vertical distance from the highest point of internal auditory canal tubercle to the upper margin of internal auditory canal was (2.31 ± 0.32) mm. To expose the whole internal auditory canal, the length and width of the internal auditory canal posterior wall removal was (7.29 ± 0.32) mm, (4.12 ± 0.29) mm. Within this removal range, no case of cochlea, semicircular canal or venous was injured in 30 specimens. CONCLUSION: The method of opening round window through retrosigmold approach is simple, practial and convenient. With little variation and easiness of location, the sinternal auditory canal tubercle can be used in the identification of the internal auditory canal. When exposing the whole internal auditory canal, the removal scope of the posterior wall should be paid more attention to, in order to avoid the damage of cochlea, semicircular canal and jugular bulb.

Full-length spleen tyrosine kinase inhibits the invasion and metastasis of human laryngeal squamous cell carcinoma.

OBJECTIVE: This study aimed to investigate correlation between full-length spleen tyrosine kinase [SYK (L)] expression and clinical characteristics of laryngeal squamous cell carcinoma (LSCC), and explore effects of SYK (L) on invasion and metastasis of LSCC. METHODS: The human laryngeal cancer Hep-2 cells with low SYK (L) expression were transfected with pIRES2-EGFP-SYK (L) vector and empty vector pIRES2-EGFP to generate Hep-2-SYK (L) cells and Hep-2-neo cells. The cell invasion and migration abilities were determined. RESULTS: The SYK (L) positive expression rate in LSCC tissues was significantly lower than in vocal cord dysplasia tissues and normal laryngeal tissues (P < 0.05). There was a significant correlation between SYK (L) expression and LSCC T stage, histopathological grade and lymph node metastasis (P < 0.05). mRNA expression of SYK (L) in Hep-2-SYK (L) cells was significantly higher than in Hep-2-neo cells and Hep-2 cells (P < 0.01). The protein expression of SYK (L) in Hep-2-SYK (L) cells was markedly higher than in Hep-2-neo cells and Hep-2 cells (P < 0.01). The number of invasive cells was significantly lower in Hep-2-SYK (L) group than in Hep-2-neo group and Hep-2 group (P < 0.01). The average number of migrating cells in Hep-2-SYK (L) group also markedly reduced as compared to Hep-2-neo group and Hep-2 group (P < 0.01). CONCLUSION: The SYK (L) expression was down-regulated in LSCC, which was closely correlated with cancer growth and lymph node metastasis. SYK (L) up-regulation was able to inhibit the invasion and metastasis of LSCC, therefore suppressing tumor development. Thus, SYK (L) may be a potential target for the LSCC treatment.

[Inhibitory effect of full-length spleen tyrosine kinase on invasion and metastasis of human laryngeal squamous cells].

OBJECTIVE: To study the effect of gene transfection of full length spleen tyrosine kinase (Syk (L)) on the biological behavior of malignant cancer cells. METHODS: Eukaryotic expression vector pIRES2-EGFP-Syk (L) was constrauted and sequenc. Laryngeal carcinoma cell line Hep-2 were transfected with pIRES2-EGFP-Syk (L) vectors or blank vectors. The expressions of mRNA and protein were examined by real time fluorescence quantitative polymerase chain reaction (Q-RT-PCR) and Western blot analysis. CCK-8 method was used for evaluating cell proliferation, Transwell for cell invasion capacity in vitro, and tumor formation in nude mice for in vivo tumorigenicity. RESULTS: pIRES2-EGFP-Syk (L) vectors were successfully construct and transfected to Hep-2 cells. Q-PCR showed that mRNA expression level in Hep-2 cells transfected with Sky (L) (28.395 ± 0.067) was higher than those in Hep-2-neo cells transfected with blank vectors (3.891 ± 0.021) and Hep-2 cells with no transfection (1.005 ± 0.012), with statistically significant difference (F = 104.02, P < 0.01). Western blot showed that protein expression level of transfected-Sky (L) cells (0.821 ± 0.047) was significantly higher than those of Hep-2-neo cells (0.558 ± 0.031) and Hep-2 cells (0.468 ± 0.031), and the difference was statistically significant (F = 112.32, P < 0.01) ; CCK-8 assay showed OD value (1.390 ± 0.067) of transfected-Sky (L) cells was lower than those of Hep-2-neo cells (1.830 ± 0.067) and Hep-2 cells (1.920 ± 0.040), and the difference was statistically significant (F = 107.64, P < 0.01). Transwell assay showed average cell number per field of transfected-Sky (L) cells (176.04 ± 22.32) was higher than those of Hep-2-neo cells (301.02 ± 21.45) and Hep-2 cells (336.04 ± 26.01) with statistically significant difference (F = 123.46, P < 0.01). The volume (250.77 ± 34.83) mm(3) tumor formed from transfected-Sky (L) cells in nude mice, was less than those from Hep-2-neo cells (750.77 ± 40.83) mm(3) and Hep-2 cells (770.77 ± 30.83) mm(3), with statistically significant difference (F = 165.78, P < 0.01). CONCLUSION: Down-regulation of Syk in Hep-2 cells is associated with the malignant biological behaviors of the cells. Syk (L) may be a potential target in gene therapy for laryngeal squamous cell carcinoma.

5-aza-CdR induces the demethylation of Syk promoter in nasopharyngeal carcinoma cell.

The present study employed 5-aza-2'-deoxycytidine (5-aza-CdR) to treat nasopharyngeal carcinoma cell line CNE-1, CNE-2 and non-cancerous human nasopharyngeal epithelial cell line NP-69 to understand the effects on spleen tyrosine kinase (Syk) gene promoter methylation. The results showed that the methylation level of Syk gene is negatively associated with the differentiation level of the cell lines, and the 5-aza-CdR treatment decreased the methylation level in nasopharyngeal carcinoma cell lines. Additionally, the drug sensitivity of low-differentiated cell line was significantly higher than the high-differentiated cell line. In conclusion, the Syk gene promoter methylation reflects the cell differentiation status, and 5-aza-CdR treatment could induce the demethylation of Syk gene promoter.

The promoter methylation of the Syk gene in nasopharyngeal carcinoma cell lines.

The aim of this study was to investigate the mRNA and protein expression levels of the Syk gene as well as its promoter methylation in nasopharyngeal carcinoma (NPC) cell lines. The CNE-1 (highly differentiated), CNE-2 (poorly differentiated) and NP69 (non-cancerous human immortalized nasopharyngeal epithelial cells) cell lines were used in the present study. The MS-PCR, Q-RT-PCR and western blotting methods were used to examine the Syk gene promoter methylation levels and mRNA and protein expression in the three cell lines. The promoter methylation levels in CNE-1, CNE-2 and NP69 cells were 36%, 62% and 0, respectively. The mRNA levels in CNE-1 and CNE-2 cells were 42±3.5 and 28±2% of that in NP69, respectively; the protein levels in CNE-1 and CNE-2 cells were 36±4.5 and 16±2.5 of that in NP69, respectively; the statistical differences between groups were significant. The lower differentiation levels of the NPC cell lines correlate with lower levels of mRNA and protein expression of the Syk gene, as well as higher promoter methylation levels.

The expression of cyclin G in nasopharyngeal carcinoma and its significance.

To investigate the expression of cyclin G1, cyclin G2 in nasopharyngeal carcinoma (NPC) cell lines and its significance. The protein expression of cyclin G1, cyclin G2 in NPC cell lines of different differentiation degree (HNE2, CNE1) was detected by indirect immunofluorescence. The mRNA expression of cyclin G1, cyclin G2 in HNE2 and CNE1 was measured with RT-PCR. The cyclin G1 expression in HNE2 and CNE1 was weak, and cyclin G2 expression in the cytoplasm near cell membrane was strong, continuous, and homogeneous. The expression level of cyclin G1-mRNA in HNE2 was 2.097 ± 0.262, which was significantly higher than CNE1 (0.997 ± 0.286, P < 0.05); the expression level of cyclin G2-mRNA in HNE2 was 0.708 ± 0.107, which was significantly lower than CNE1 (1.216 ± 0.037, P < 0.05). Abnormal expression of cyclin G was closely related to tumor differentiation, the origin, and progression of NPC.

[Study of demonstrating main operative mark of transmastoid-epitympanum approach of the facial nerve using double oblique multi-planar reconstruction in multi-slice CT].

OBJECTIVE: To explore a method of demonstrating the facial nerve anatomical landmarks under transmastoid and epitympanum approach with multi-slice CT using double oblique multi-planar reconstruction (MPR). METHOD: Two temporal bone of a corpse were dissected, under transmastoid and epitympanum approach, to observe the anatomical landmarks of facial nerve. Based on that, the anatomical landmarks of facial nerve under transmastoid and epitympanum approach in 30 (60 ears) normal temporal bones of adult corpses were reconstructed using double oblique MPR in multi-slice CT. The achievement ratio was calculated and the differences among transverse plane, coronal plane, sagittal plane and double oblique were compared. RESULT: The different part of facial nerve, such as mastoid segment, tympanum segment, pyramid segment, geniculate ganglion and the outer labyrinthine segment could be exposed clearly with the main anatomical landmarks, such as horizontal semicircular canal, epitympanic recess and cochleariform process through transmastoid and epitympanum approach. The image of anatomical landmarks could be showed in the same sections by double oblique multi-planar reconstruction. The double oblique multi-planar reconstruction to show the landmarks of facial nerve displaying on the same imaging is better than transverse plane, coronal plane and sagittal plane. The achievement ratio of every section is 100%. CONCLUSION: Double oblique MPR is a new method to demonstrate anatomical landmarks through transmastoid and epitympanum approach in one slice. Combined with the operative approach and purpose, the reconstructive images with double oblique MPR can provide valuable information for operation.

[The value of induced hypotension in nasal endoscopic surgery].

OBJECTIVE: To observe the value of induced hypotension in nasal endoscopic surgery with local anesthesia. METHOD: Sixty Patients were randomly divided into two groups. Group I : induced hypotension group; Group II: control group. MAP, HR, SaO2 amount of blood loss, clear visual field of operation and the operation time were observed and compared between two groups. RESULT: The difference of MAP, HR, SaO2 data between two groups showed no statistic difference with anatomical measurement (P >0.05). The difference of amount of blood loss, the operation time and clear visual field of operation between two groups was significant (P <0.05). CONCLUSION: The induced hypotension with local anesthesia can produce time of operation and amount of blood loss decreased. It can safely applied to the in clinical practice.

[Three dimensional reconstruction study of multi-slice helical CT imaging on optic canal area].

OBJECTIVE: To investigate the accuracy and clinical application of multi-slice spiral CT imaging on optic canal area. METHOD: Forty intact specimens of human corpse head were tested with three dimensional reconstruction of multi-slice spiral CT imaging. The results of radiology and anatomy were compared on adjacent structures of optic canal interior wall and its developmental relationship with sphenoid sinus. RESULT: (1) The high spatial resolution of integrity could be obtained with dimensional processing technique. The anatomical structure of optic canal and its surrounding area could be demonstrated clearly. (2) With the technique of virtual endoscopy, the interior reconstruction of optic canal could demonstrate the inner topography of optic canal, the reconstruction image of which was identical to that of the control subjects. (3) There was no significant difference between the data of multi-slice spiral CT imaging and that of anatomy (P > 0.05). CONCLUSION: Associated application of dimensional processing technique and virtual endoscopy, not only can the subtle structures of optic canal be demonstrated and measured, but also the detailed information on spatial relationships of optic canal area can be provided for operation.