Objective To investigate the expression and clinical significance of FoxA 2(hepatocyte nuclear factor 3β)in human non-small cell lung cancer(NSCLC)tissues and adjacent normal lung tissue.Methods The expressions of FoxA 2 protein in 80 cases with NSCLC tissues and adjacent normal lung tissue were tested by immunohistochemistry,the expressions of mRNA was detected by RT-PCR.Results The expressions of FoxA 2 both on protein and mRNA in NSCLC lung tissue were significantly less than those of adjacent normal lung tissue(protein expression comparison t value was 187.51,186.43,178.24 respectively;mRNA expression comparison t value was 236.70,260.13,126.27,respectively,P ＜ 0.05).However there was no difference on histopathological variability.F value equal to 1.86 and 1.69,P ＞ 0.05).In low differentiated group,the group of Ⅲ stage,with lymph node metastasis,the expression of FoxA 2 protein and mRNA was less than that in the group of high differentiated,the group of Ⅰ-Ⅱ stage,the group of non lymph node metastasis(pretein expression t values were 4.31,3.92,341,mRNA expression t values were 5.44,3.28,5.07,P＜0.05).Conclusion FoxA 2 might inhibit progression of human NSCLC.It may participate in the process of tumor tissue differentiation and lymph node metastasis in NSCLC.

Objective:This study aimed to observe the synergistic effect of a new tumor vaccine combined with metronomic che-motherapy in vivo on breast cancer. This study was also conducted to investigate the mechanism of this combination. Methods:Balb/c mice inoculated with 4T1 mouse breast cancer cell were used as tumor models. High-mobility group nucleosome-binding protein 1 (HMGN1) gene was used to transfect 4T1 cell lines as cancer vaccines. After 4T1 cell was inoculated, the mice were randomized into four groups:normal saline (NS);metronomic gemcitabine (GEM) alone;cancer vaccine alone;and combination therapy group. Tumor growth and potential toxicities of these regimens were observed. The Foxp3 expression of regulatory T cells (Tregs) was detected by western blot and immunohistochemical staining. The microvessel density (MVD) of the tumor was also detected by immunohistochemi-cal staining. Results:The tumor volume of the mice was significantly lower in the combination group than in the MET group or cancer vaccine group (P<0.05). This result exhibited a higher significant difference than the tumor volume of the mice in the NS group (P<0.01). Foxp3 expression was significantly lower in the mice treated with GEM (combination or MET group). MVD was significantly lower in these two groups than in the cancer vaccine group or NS group (P<0.05). Furthermore, adverse reactions slightly occurred in each group. Conclusion: The combination of cancer vaccines and metronomic GEM is a very active and well-tolerated regimen for breast cancer in mice.

Objective To investigate the mechanism of soluble β-1, 3-D-glucan in G-test positive serum in inhibiting ROS-dependent killing of Candida albicans ( C.albicans ) mediated by neutrophil Dectin-1.Methods The expression and distribution of internalized Dectin-1 and triggered ROS in human neutrophils were detected by using confocal/two-photon laser scanning microscopy upon stimulation with C.albicans (MOI=10) which was pretreated with β-1, 3-D-glucanase (10 U/ml) or not.Abrogation test was used to analyze whether intracellular Dectin-1 was involved in C.albicans-triggered ROS production in human neutrophils.Furthermore, flow cytometry analysis was performed to detect the expression of intracel-lular Dectin-1 and ROS in neutrophils which were pretreated respectively with G-test positive serum at differ-ent dilutions for 60 min and then stimulated with C.albicans for another 60 min at 37℃.Results After stimulated with C.albicans (MOI=10) for 60 min, the expression of Dectin-1 in neutrophils was recruited to the spores of opsonophagocytized C.albicans, and partly co-localized with the triggered ROS production . However, the expression of intracellular Dectin-1 was not observed in neutrophils when stimulated with β-1, 3-D-glucanase pretreated C.albicans for 60 min at 37℃.Abrogation test further showed that C.albicans-trig-gered ROS production in neutrophils was partly and irreversibly inhibited by adding Dectin -1 blocking mAb of 5 μg/ml.In addition , both the triggered expression of intracellular Dectin-1 and ROS production in neu-trophils stimulated with C.albicans ( MOI=10 ) in the presence of G-test positive serum were significantly lower than those of neutrophils stimulated only with C.albicans (LSD-t test, P<0.01).Linear regression a-nalysis suggested that the triggered intracellular Dectin-1 and ROS production in neutrophils upon stimulation with C.albicans were both inhibited by soluble β-1, 3-D-glucan in a dose-dependent manner (Dectin-1,R2=0.702,P<0.01;ROS,R2=0.588,P<0.01 ).Conclusion Taken together, these results demonstrated that the soluble β-1, 3-D-glucan in G-test positive serum may play a role in inhibiting the ROS-dependent killing of C.albicans by interfering with internalized expression of neutrophil Dectin-1.

OBJECTIVE: The aim of this study was to determine the expression of GATA-3 and the level of Th1 and Th2 cytokines upon repeated exposure to staphylococcal enterotoxin B(SEB) of different concentrations in the maxillary sinus mucosa of rabbits. METHOD: The rabbits were randomly divided into 2 groups (24 rabbits per group): low-dose SEB group and high-dose SEB group. The low-dose SEB group and high-dose SEB group received daily injections of 0.6 ng of SEB (2 ml) and 60 ng of SEB (2 ml) into the left maxillary sinus of rabbits for 28 days, respectively. Concurrent treatment of the right maxillary sinus with normal saline was used as a control. Six rabbits chosen randomly in two groups were killed on days 3, 7, 14, and 28, and to obtain the sinus mucosa from the two-side maxillary sinuses for measurement. Mucosal levels of IL-2, IL-4, IL-5, and IFN-γ were measured using ABC-ELISA. Tissue expression of GATA-3 were examined using Real-time PCR and immunohistochemistry. RESULT: IFN-γ and IL-2 levels were significantly elevated in the high-dose SEB group compared with the low-dose SEB and control groups on days 7, 14, and 28 (P < 0.05). However, IL-4 and IL-5 levels were markedly enhanced in the low-dose SEB group compared with the high-dose SEB and control groups on days 14 and 28 (P < 0.05). Real-time PCR showed that the expression of GATA-3 mRNA in the low-dose SEB group was markedly enhanced, and immunohistochemical staining illustrated that the number of GATA-3 positive cells was markedly increased in the low-dose SEB group as compared with the high-dose SEB group (P < 0.05). No significant differences were observed in GATA-3 expression between the high-dose SEB and the control groups (P > 0.05). CONCLUSION SEB promoted Th1 cytokines production at high concentrations, and enhanced Th2 cytokines expression and Th2 immune response at low concentrations.
Animals ; Cytokines ; metabolism ; Enterotoxins ; administration & dosage ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Interferon-gamma ; metabolism ; Interleukin-2 ; metabolism ; Interleukin-4 ; metabolism ; Interleukin-5 ; metabolism ; Male ; Maxillary Sinus ; drug effects ; metabolism ; Nasal Mucosa ; metabolism ; RNA, Messenger ; metabolism ; Rabbits ; Th1 Cells ; Th2 Cells ; Transcription Factors ; metabolism

OBJECTIVE:To study the effect of different dosage of grassleaf sweetflag rhizome on central nervous system.METHODS:The effects of different dosage of grassleaf sweetflag rhizome on the hours of sleep and hypoxia live time of mice were observed,and the effects of which on water contents of brain tissues,apoptosis,endothelin(ET),malonaldehyde(MDA),erythrocuprein(SOD),glutathione peroxidase(GSH-Px),etc.in rats with cerebral ischemia were observed as well.RESULTS:Compared with the control group,different dosage of grassleaf sweetflag rhizome could shorten the sleeping time of mice and prolong the live time of hypoxia mice;high dosage of grassleaf sweetflag rhizome could remarkably decrease the water content of brain tissue and MDA level and inhibit the apoptosis of brain tissue cells while increase the activity of SOD and GSH-Px.CONCLUSION:High dosage of grassleaf sweetflag rhizome has the strongest protective effects to central nervous system,the effects of middle and low dosage of grassleaf sweetflag rhizome are lower than that of the high dosage.

Objective To construct and identify a novel IL-10 delivery system by transforming a hIL-10-containing plasmid into B.longum (BL -hIL -10).Methods A plasmid vector pBADs -GFP was selected which had been built by previous test and biosynthetic hIL-10 plasmid,through double enzyme digestion and enzyme reaction,to construct and identify PBADs-hIL-10 shuttle plasmid,then to synthesis BL-hIL-10.hIL-10 was expressed and secreted into the culture supernatant of BL-hIL-10 after 0.2% L-arabinose induction in vitro as examined by Western blot,enzyme-linked immunosorbent assay (ELISA)and RT-PCR;Culture supernatants and bacterium pellets were collected after continuous culture for 12,24 and 36h,respectively.hIL-10 was expressed and secreted into the culture supernatant of BL-hIL-10 after 0.2% L-arabinose induction in vitro as examined by Western blot,enzyme -linked immunosorbent assay (ELISA)and RT -PCR;Culture supernatants and bacterium pellets were collected after continuous culture for 12,24 and 36h,respectively.Results The BL-hIL-10 bacterial strain that can stably express hIL-10 factor was successfully screened out,and the levels of hIL-10 in both superna-tant and cell pellet were similarly reached maximum at 24h of culture.Conclusion BL-hIL-10 as a novel oral hIL-10 delivery system has been successfully established,which established a basis for the treatment of IBS with transgenic Bifidobacterium.

Objective To explore the diagnosis rate.pathology types and positive rate of cancer cell in spu-tum of early central pulmonary carcinoma in three obstructive signs on chest X ray screened by fiberbromchoscope.Methods 326 cases of three obstructive signs with high risk of lung cancer were screened for central pulmonarycarcinoma by spiral CT.biopsy by fiberbronchoscope and cytological examination of sputum.Results 32 patients were diagnosed with central pulmonary carcinoma,with morbidity of 9.8%.In these patients,21 were confirmed with obstructive pneumonia(65.6%),7 with obstructive atelectasis(21.9%),4 with obstructive emphysema(12.5%);In terms of pathology type,16 cases were defined as squamous cell carcinoma(50.0%),9 as small cell carcinoma(28.1%).3 were as large cell carcinoma(9.4%).2 were as adenocarcinoma(6.3%),1 as admosquamous carci-noma(3.1%),1 as bronchial gland carcinoma(3.1%);cancer cell could be found in sputum of 5 patients of 32 cases,among them,it was found in 3 of 21 patients with obstructive pneumonia(14.3%),1 in 7 patients with ob-structive atelectasis(14.3%),1 in 4 patients with obstructive emphysema(25.0%).Conclusion The prevelance of early central pulmonary carcinoma in three obstructive signs on chest X-ray is 9.815%,in which squamous carci-noma and small-cell carcinoma are common in pathology type.Screening can increase the detection rate of early pul-monary carcinoma.

Objective To investigate the effects of simultaneous multi-level surgery for moderate to severe obstructive sleep apnea hypopnea syndrome (OSAHS). Methods A retrospective analysis was made on surgical cases of one hundred and thirty seven patients with moderate to severe OSAHS diagnosed by polysomnography (PSG). They were divided into multi-level group (n = 95) and UPPP group (n = 42). The two groups were compared in terms of postoperative complications as well as the related indicators of PSG , calgary sleep apnea quality of life index (SAQLI), epworth sleepiness scale (ESS), snore scales (SS) before operation and after operation. Results Just one patient in the multi-level group had difficulties in respiration and was rescued by timely tracheotomy. The AHI, LSaO2, TS90%, the total score and the scores on the four dimensions of SAQLI, ESS score, SS score in the multi-level group were significantly improved as compared both to the results after operation (P < 0.01) and to the UPPP group (P < 0.05). But only the AHI, LSaO2 and TS90% in the UPPP group were improved (P < 0.05). Conclusions The multi-level surgery is a safe and feasible therapy or moderate to severe OSAHS. The evaluation in subjective and objective ways can be more accurate in comprehensive reflecting the surgical efficacy and effects of OSAHS on patients′ of life quality.

Objective To investigate the effect of curcumin on proliferation and apoptosis of human nasopharyngeal carcinoma (NPC) cell line CNE-2.Methods The CNE-2 cells were treated by by different concentrations (0,10,20,40,60 μmol/L) of curcumin.The proliferation activity of CNE-2 cells was detected by MTT assay,the cell cycle and apoptosis rate of CNE-2 were detected by using the flow cytometry (FCM),and the apoptosis was observed by Hoechest33258 fluorescence staining.Results Curcumin could significantly inhibit the proliferation of CNE-2 cells,moreover which was increased with curcumin concentration increase,the inhibitory rate of CNE-2 cells showed an increasing trend (P＜0.05),the half inhibitory concentrations (IC50) of curcumin acting on CNE-2 cells at 24,48,72 h were (23.54 ± 0.36),(18.31 ± 0.42) and (8.56 ± 0.37) μmol/L respectively.Curcumin could significantly inhibit the proliferation effect of CNE-2 cells,showing the apparent concentration and time dependence.The FCM detection results showed that in treating CNE-2 cells by 0,10,20,40,60 μmol/L of curcumin,the apoptosis rate was increased with the curcumin concentration increase;the fluorescence staining results showed that CNE-2 cells without curcumin treatment were round or oval,the cell nucleis were uniform in size,chromatin distribution showed the homogeneous light blue fluorescence;after 24 h of 10 μmol/L curcumin treatment,the CNE-2 cell body was shrunk and cell nuclear chromatin was condensed,showing granular bright blue fluorescence;after 24 h of 20 μmol/L curcumin treatment,the cell body was shrunk,nuclear was condensed,chromatin was uneven,apoptotic bodies appeared,and even the nuclear fragmentation appeared;after 24 h of 40 μmol/L and 60 μmol/L curcumin treatment,the number of apoptotic cells was increased,a large number of nuclear fragmentation appeared.Conclusion Curcumin has a significant inhibitory effect on the proliferation of NPC cell line CNE-2,moreover promotes the apoptosis of CNE-2 cells.

Objective To investigate the ultrastructure of ciliated epithelia and inflammatory changes upon repeated exposure to staphylococcal enterotoxin A (SEA) of different concentrations in the maxillary sinus mucosa of rabbits.Methods The rabbits were randomly divided into 2 groups (24 rabbits per group):low-dose SEA group and high-dose SEA group.The low-dose SEA group and high-dose SEA group received daily injections of 0.6 ng of SEA (2 ml) and 60 ng of SEA (2 ml) into the left maxillary sinus of rabbits for 28 days,respectively.Concurrent treatment of the right maxillary sinus with normal saline was used as control.Six rabbits chosen randomly in two groups were examined by computed tomography (CT) scans and then sacrificed to obtain thc sinus mucosa from the two-side of maxillary sinuses for histological assessment on days 3,7,14 and 28.To characterize the inflammatory changes of the sinus mucosa examined using light microscope,hematoxylin and eosin (HE) and toluidine blue staining was performed.Scanning and transmission electron microscopy were performed to observe ultrastructure of ciliated epithelia in the maxillary sinus mucosa.SPSS 13.0 software was used to analyze the data.Results On days 14 and 28,CT images showed opacification of the left maxillary sinus in the high-dose SEA group.The percentage of epithelial disruption was (22.73 ± 5.72) ％ and (30.79 ± 4.30)％ in the high-dose SEA group respectively,and were significantly greater than those in the low-dose SEA group (5.12％ ± 1.98％ and 5.38％ ± 1.64％,q value was 10.079 and 19.132) and control group (4.08％ ± 1.29％ and 4.81％ ± 1.62％,q value was 11.016 and 19.592,respectively,all P ＜ 0.01).The subepithelial thickness in the high-dose SEA group was (113.34 ± 14.81) μm and (120.86 ± 12.35) μm respectively,and were significantly different from those of the low-dose SEA group[(71.08 ± 10.39) μm and (81.63 ±9.32) μm,q value was 8.090 and 8.782] and control group [(37.45 ± 7.67) μm and (38.79 ± 7.68) μm,q value was 15.759 and 19.541,all P ＜0.01].Viewed under the electron microscope,loss of cilia was observed,a few compound cilia and cytoplasmic protrusion were found,an obvious stretching of the endoplasmic reticulum and an obvious turgescence of the mitochondria was also observed.However,in the low-dose SEA group on days 14 and 28,CT scan of the left maxillary sinus showed transparency; light microscopy observations of the maxillary sinus mucosa showed the number of eosinophils was markedly increased as compared with the high-dose SEA and control groups,the differences were significant (q value was 5.871 and 6.766 on day 14,and q value was 7.572 and 8.970 on day 28,respectively,all P ＜ 0.05).But no significant differences were observed in epithelial disruption between the low-dose SEA and the control groups on days 14 and 28 (q value was 1.512 and 0.859 respectively,all P ＞0.05) ; inordinate array and adhesion of cilia was observed,but cilia loss,compound cilia,cytoplasmic protrusions,mitochondrial swelling and endoplasmic reticulum stretching were not found.Conclusions SEA may induce allergic inflammation of the sinus mucosa without damaging the structure of ciliated epithelia at low concentration.Whereas SEA impairs the structure of mitochondria and endoplasmic reticulum in ciliated epithelial cells at high concentration,and results in cilia loss and epithelial disruption,which may be one of the main reasons to induce acute sinusitis.