BACKGROUND:Studieshavesuggestedthat oncogene expression in liver cancer stem cels has a certain relationship withtheoccurrence and development of liver cancer, but there is stil a lack of research on bioinformatics and mechanisms.
OBJECTIVE:To identify differentialy expressed genes in liver cancer stem cels and to analyze these genes by bioinformatics.
METHODS:The human hepatoma cel line HepG2 was the cel tool of liver cancer stem cels to prepare total RNA, andfluorescent labelingexperiment was conducted.Usinggene chipshybridization, mRNA expression profiles were obtained and were screened, and then differentialy expressed mRNAs were obtained,GO and Pathway annotationswere analyzedusing bioinformatics.
RESULTS AND CONCLUSION:The detection rate was 73.21% for hybridization experiment, indicating the hybridization experiment is successful. In this study, a total of 38342 mRNA werefound, and after further analysis, 1236 differentialy expressed genes werescreened (P< 0.05, fold change≥2), including599up-regulated and637down-regulated genes, respectively (P< 0.05). Biological functions of differentialy expressed geneswere mainly involved inthe histone H4 acetylation, cel mitosis and proliferation, synthesis and decomposition of cel-associated proteins, chromosome segregation, cel differentiation and apoptosis, signaltransduction, as wel as nutrienttransportation and transcription. Pathway annotationsweremainly related to cytokine-mediated inflammatory signaling pathway, Wnt signaling pathway, Hedgehog and Notch signaling pathways, TGF-Beta, Jak-STAT and so on. These results demonstrate that differentialy expressed genes in liver cancer stem celsarerelatedto the occurrence of liver cancer, and may be involved in the regulation of signaling pathways, which provides a new target for the treatment of liver cancer.

The vagus nerve innervates most visceral organs.Upon activation,vagal efferents release acetylcholine,which influences organ function.In addition,vagal afferents convey information regarding the mechanical and chemical environment of the organ to the central nervous system (CNS).This bidirectional communication provides a mechanism for reflex regulation of the biological function of the organ.In the lung,the vagus nerve modulates airway tone,perfusion and secretion,in addition to its effects on breathing pattern.Recently,the vagus nerve has been recognized to play a role in the pathogenesis of lung disease via neuro-immune interactions.The vagus nerve has significant influences in pulmonary diseases,such as asthma,chronic obstructive pulmonary disease (COPD),acute respiratory distress syndrome (ARDS),pulmonary fibrosis and lung cancer.In disease,the nerve is activated by cytokines,chemokines,and other mediators from many cell types to convey immunologic information to the CNS,which may alter disease outcome.Activation of the vagus nerve also releases neuropeptides to modulate immune cell behavior and can evoke the cholinergic anti-inflammatory pathway that regulates lung inflammation.Understanding the role of the vagus nerve in neuro-immune interaction may contribute significantly to the clinical management of pulmonary diseases.

Objective To evaluate the association of gastric hyperplastic polyps with colorectal neo-plasia.Methods Data of consecutive patients who underwent esophagogastroduodenoscopy (EGD)and colonoscopy between January 2011 and December 2013 were reviewed retrospectively.They were compared with patients without gastric polyps who also underwent EGD in the same period.The relationship between gastric polyps and colorectal neoplasia was analyzed.Results The incidence of colorectal neoplasia in gas-tric hyperplastic polyps group was higher than that of the control group [24.0% (46 /192)VS 10.4%(40 /384),P =0.000].An increased incidence of colorectal adenomas in gastric hyperplastic polyps group was found,but there was no difference in the incidence of colorectal cancer in gastric hyperplastic polyps group and control group[2.1%(4 /192)VS 2.3%(9 /384),P =1.000].Stratification analysis suggested that the incidence of colorectal neoplasia in gastric hyperplastic polyps group who aged over 50 was signifi-cantly higher than that in the control group[28.5%(43 /151)VS 10.6%(29 /274),P =0.017],regard-less of different genders and the number of gastric hyperplastic polyps.Conclusion The incidence of color-ectal neoplasia in gastric hyperplastic polyps has significantly increased,especially in those aged over 50 years,regardless of different genders and the number of gastric hyperplastic polyps.Such patients should undergo colonoscopy to detect colorectal neoplasia.

Objectives To investigate the prevalence of heterogeneous vancomycin intermediate Staphylococcus aureus(hVISA) and the sensitivity of hVISA to novel antibiotics,and to explore the risk factors and infection attributable mortality associated with hVISA infection.Methods A total of 456 methicillin resistant Staphylococcus aureus (MRSA) isolates were isolated in Zhongshan Hospital from January,2008 to November,2010.All MRSA isolates were investigated for hVISA by two agar screening methods BHIA5T (brain-heart infusion containing teicoplanin 5 mg/L)or BHIA6V (brain-heart infusion containing vancomycin 6 mg/L),as well as macroEtest method(MET).Possible hVISA isolates were tested by modified population analysis profile-area under the curve (PAP-AUC).The minimal inhibitory concentrations(MICs) of vancomycin,teicoplanin and linezolid were determined by microbroth dilution as recommended by Clinical Laboratory Standards Institute(CLSI).The contribution difference between hVISA and vancomycin susceptible Staphylococcus aureus (VSSA) in different MIC range was compared.A retrospective case-control study of the patients with hVISA infection or VSSA infection was carried out and statistical analysis was performed using t test,Mann-Whitney test,x2 test and Fisher exact test.Results A total of 105 isolates of hVISA were screened by BHIA5T and BHIA6V (23.0％) with other 23 isolates by MET(5.0％) and 21 by PAP-AUC(4.6％).All isolates were 100％ sensitive to vancomycin,teicoplanin and linezolid.The vancomycin MIC [(1.76 ±-0.16) mg/L] in hVISA group was significantly higher than that in VSSA group[(1.09 ± 0.07)mg/L,P ＜ 0.01],which was a potential risk factor for hVISA infection.The retrospective study showed chronic obstructive pulmonary disease (COPD) was also a risk factor for hVISA infection of the lower respiratory tract.No significant difference in infection attributable mortality was showed between the hVISA group and the VSSA group.Conclusions The overall prevalence of hVISA in Zhongshan Hospital is estimated as 4.6％,while the prevalence of hVISA isolated from blood is as high as 12.5％.All isolates are 100％ sensitive to vancomycin and linezolid.COPD is a risk factor for hVISA infection of the lower respiratory tract.

Objective To investigate the effects of berberine on tumor-associated macrophages (TAM)and the expression of cyclooxygenase-2 (COX-2)of intestinal polyps in Apc(Min/+) mice.Methods A total of 20 Apc(Min/+) mice,four weeks old,were equally divided into the control group and the berberine group,10 in each group.The mice of the control group drank plain water,while the mice of berberine group drank water with 0.1 % berberine.After 12 weeks,all the mice were sacrificed.The intestine and colon were isolated,and the numbers of polyps were counted.The expression of F4/80,inducible nitric oxide synthase-2 (iNOS),macrophage mannose receptor (MR)and COX-2 was detected by immunohisto-chemistry method.The relative expression of COX-2 at protein level was measured by Western blotting. The t test was performed for comparison between two independent groups.Results The total number of intestinal polyps,the number of small intestinal polyps and the number of colon polyps of the berberine group (11 .50±2.05 ,10.50±1 .77 and 1 .00±0.46,respectively)were all less than those of the control group (30.63±1 .69,28.00±2.00 and 2.63±0.74,respectively),and the differences were statistically significant (t=16.727,16.952 and 3.162,P =0.001 ,0.001 and 0.010,respectively).The percentage of F4/80 positive cells in the stroma of polyps of the berberine group ((17.40 ±4.23 )%)was less than that of the control group ((31 .24±6.34)%),and the difference was statistically significant (t =5 .327, P =0.043).The percentage of iNOS positive cells in the stroma of polyps of the berberine group ((7.43± 1 .78 )%) was higher than that of the control group ((2.72±0.68)%), and the difference was statistically significant (t=7.335 ,P =0.004).The percentage of MR positive cells in stroma of polyps of the berberine group ((19.52±1 .54)%)was less than that of the control group ((12.63±0.68)%),and the difference was statistically significant (t=5 .634,P =0.016).The percentage of COX-2 positive cells in stroma of polyps of berberine group ((3.38 ± 0.51 )%)was less than that of the control group ((7.60±0.57 )%),and the difference was statistically significant (t = 7.234,P = 0.001 ).The relative expression of COX-2 at protein level of polyps of the berberine group was lower than that of the control group. Conclusion Berberine may take the role in inhibiting the growth of intestinal polyps in Apc(Min/+) mice through interfering the differentiation of TAM in polyps and suppression the expression of COX-2.

Objective To evaluate the efficacy of Lianhua Dingchuan Tablets in bronchial asthma.Methods Fifty BALB/C mice were randomly and equally divided into control (Con) group,ovalbumin (OVA) group,dexamethasone (DEX) group,high-dose Lianhua group,low-dose Lianhua group.The mice were sensitized and challenged with OVA plus aluminium hydroxide to establish asthmatic model and were pre-treated 30 minutes before challenge.Specific airway resistance (sRaw) was used to evaluate airway hyperresponsiveness,and airway inflammatory changes were measured.ELISA and Magnetic Luminex(R) were used to quantified the levels of IL-4,IL-13 and INF-γ.Results Airway resistance significantly decreased in DEX group and High-dose Lianhua group (P＜0.05).Levels of inflammatory cells and IL-13 in BALF evidently reduced in DEX group,high-dose Lianhua group and low-dose Lianhua group (P ＜ 0.05),while IL-13 level in serum only decreased in DEX group.There was no significant changes in the levels of IL 4 and INF γ among those groups.Conclusions Lianhua Dingchuan Tablets might relieve the symptoms of asthma by reducing IL-13 level and inhibiting the airway inflammation.

Objective To explore the miRNA regulating the potential cancer-promoting gene CCL18 in cutaneous malignant melanoma.Methods Bioinformatics analysis was conducted by using online software miRanda and TargetScan,so as to predict the miRNA targeting CCL18 gene.Three kinds of C CL18 3'UTR dual-luciferase reporter vectors,including mutant 3'UTR vector (mutant 3'UTR group),wildtype 3'UTR vector (wild-type 3'UTR group) and empty vector (blank control group),as well as miRNA vectors carring selected miRNAs were constructed according to human gene sequence analysis,and then were used to co-transfect 293T cells.After 48-hour treatment,the cells were lysed for detection of luciferase activity.Real-time fluorescence-based quantitative PCR was performed to measure the expression of CCL 18 and selected miRNA in 14 fresh malignant melanoma tissue specimens and 14 paracancerous normal skin tissue specimens (control tissues),and their correlations were analyzed.Results Online software analysis showed that some miRNAs were identified to target the 3'UTR of CCL18 gene,including miR-183,miR-128 and miR-33a.Luciferase reporter vectors and miRNA vectors were constructed successfully.As luciferase activity assay showed,when miR-183 and miR-128 were bound to the CCL18 3'UTR,the luciferase activities were significantly higher in their mutant 3'UTR groups (11.63 ± 0.42;8.80 ± 0.49) than in their wild-type 3'UTR groups (4.86 ± 0.39;5.01 ± 0.54;both P ＜ 0.05) and blank control groups (2.41 ± 0.13;2.39 ± 0.05;both P ＜ 0.01),while there were no significant differences between miR-33a-hinding mutant 3'UTR group (6.41 ± 0.47) and miR-33a-binding wild-type 3'UTR group (6.16 ± 0.22,P ＞ 0.05).Real-time fluorescence-based quantitative PCR revealed higher mRNA expression of the CCL18 gene (3.52 ± 1.68),but lower expression of miR-183 (0.49 ± 0.32),miR-128 (0.30 ± 0.20) and miR-33a (0.46 ± 0.40) in the malignant melanoma tissues compared with the control tissues.The mRNA expression of the CCL18 gene was negatively correlated with the expression of miR-128 (rs =-0548,P ＜ 0.05),but showed no significant correlations with the expression of miR-183 and miR-33a (both P ＞ 0.05).Conclusion miR-128 may play a role in regulating the potential malignant melanoma-promoting gene CCL18.

Objective To investigate the expression of regulators of G protein signaling(RGS), including RGS2, RGS3 and RGS4 in OLETF rats, as well as the effects of metformin on these expressions. Methods LETO rats were used as control group. Eight-week-old male OLETF rats were assigned to two guoups randomly:model and trial(metfomin dose during 8~(th) to 22~(nd) weeks:300mg kg~(-1)·d~(-1);during 23rd to 28th weeks:400 mg·kg~(-1) ·d~(-1))groups. Expressions of RGS mRNA in aorta and heart werequantified by real-time PCR. Results RGS2, RGS3 and RGS4 mRNA of the thoracic aorta and left ventricle were significantly higher in model group than in control group (P<0.01). Compared with model group, metformin significantly reduced their mRNA in trial group (P<0.01). Conclusions Upregulation of RGS2, RGS3 and RGS4 mRNA expression in the thoracic aorta and left ventricle of OLETF rats is in correlation with cardiovascular lesions; while downregulation of their expression is in correlation with the action of metformin.

Objective To screen microRNAs(miRNAs)related to early mycosis fungoides(MF). Methods A high?throughput miRNA PCR array was used to determine miRNA expression profiles in skin lesions of 6 patients with early MF (early MF group) and 6 patients with lichen planus (control group), followed by screening of differentially expressed miRNAs between the two groups. Then, real?time fluorescence?based quantitative PCR(RT?qPCR)was performed to verify the differentially expressed miRNAs in lesional specimens from 13 patients with early MF and 13 patients with eczema or lichen planus, as well as in Myla cells and normal human T?lymphocytes. Results The high?throughput miRNA PCR array showed that the expressions of hsa?miR?378a?5p, hsa?miR?107 and hsa?miR?302c?3p were significantly higher in the early MF group than in the control group(all P<0.05). For skin lesions, the results from RT?qPCR were similar to those from the miRNA array assay. Compared with normal human peripheral blood T?lymphocytes, Myla cells showed significantly increased expressions of hsa?miR?378a?5p and hsa?miR?107, which was consistent with the results from the miRNA array assay. However, no significant difference was observed in the expression of hsa?miR?302c?3p between the two kinds of cells. Conclusion MiRNA expression profiles in early MF are different from those in inflammatory skin diseases.

Objective To measure histidine triad nucleotide?binding protein 1(HINT1)protein expression and gene promoter methylation, and to analyze the relationship between HINT1 gene promoter methylation and clinical pathological features of melanoma. Methods Fifty?six patients with melanoma and 51 patients with nevus were enrolled as subjects and controls, respectively. Methylation?specific PCR (MSP) was performed to measure the methylation of HINT1 gene promoter in lesional and paratumoral tissue specimens from the patients with melanoma, as well as in lesional specimens from the patients with nevus. Immunohistochemistry was carried out to quantify the expression of HINT1 protein in these tissue specimens. Results MSP showed that the methylation rate of HINT1 gene promoter was significantly higher in melanoma tissues than in paratumoral and nevus tissues(76.8%[43/56]vs. 33.9%[19/56]and 35.3%[18/51], χ2 = 20.810 and 18.749, respectively, both P < 0.05), but was insignificantly different between paratumoral and nevus tissues(χ2=0.022, P>0.05). Immunohistochemistry revealed that the expression rate of HINT1 was 21.4%(12/56)in melanoma tissues, compared to 82.4%(42/51)in nevus tissues(χ2 = 39.633, P <0.01). There was a significant difference in the methylation rate of HINT1 promoter between HINT1?positive and ?negative melanoma tissues(6/12 vs. 37/44[84.1%], P<0.05), and between Clark levelⅠ-ⅡandⅢ-Ⅴmelanoma tissues(59.1%[13/22]vs. 88.2%[30/34],χ2=6.365,P=0.012). Conclusions HINT1 protein is lowly expressed in melanoma, which may be associated with high methylation of its gene promoter. Moreover, the high methylation ofHINT1 gene promoter may be involved in the initiation and progression of melanoma.