Objective:To investigate the existence of side population cells with the potency of stem cells in human gallbladder carcinoma cell line GBC-SD and the differences in ABCG2,Oct-4 and CD34 expression among SP cells,non-SP cells and GBC-SD cells.Methods:SP and non-SP cells were sorted from GBC-SD cells by fluorescence-activated cell sorting(FACS).The expression of ABCG2,Oct-4 and CD34 in SP cells,non-SP cells,and GBC-SD cells was detected by reverse transcription-polymerase chain reaction(RT-PCR),Western blot,flow cytometry(FCM)and immunofluorescence chemistry.Results:SP cells with stem cell potency were isolated from GBC-SD cells with a proportion of(0.64±0.08)%.The metastatic ability of SP cells was obviously higher than that of non-SP cells and GBC-SD cells(P<0.05).The expression of ABCG2 was significantly higher in SP cells than in non-SP cells and GBC-SD cells[(89.56±3.86)%vs.(1.32±0.49)%and(12.37±1.61)%,P=0.001].The expression of Oct-4 in these cells was(94.87±1.40)%,(88.16±2.34)%and(90.17±1.61)%,respectively(P>0.05).CD34 was neady absent in these cells on protein level[(1.78±0.51)%vs.(0.63±0.21)%and(0.96±0.381)%,P>0.05)],but it was highly expressed in non-SP cells and GBC-SD cells and absent in SP cells off mRNA leve;.Conclusion:SP calls which hava the potency of stem cells,exist in human gallbladder carcinoma GBC-SD cell line and have the phenotype of ABCG2+Oct-4+CD34-.

[Summary] Diabetic nephropathy is one of the major chronic microvascular complications of diabetes ,which is the leading cause of end‐stage renal disease ,as well as the main cause of death in diabetic patients. Glomerular endothelial cell is an important component of the glomerular filtration barrier ,which is directly related to the materials of circulation ,and it can be easily damaged by glucose ,lipid and inflammatory factors. Under the hyperglycemia ,the PKC pathway ,the polyol pathway and oxidative stress were activated ,producing an excess of advanced glycation end products and reactive oxygen species ,which damage the endothelial nitric oxide synthase ,reduce the generation of nitric oxide ,while produce a large number of Ang Ⅱ. Ang Ⅱ damage the endothelial cell. In addition ,there are crosstalk between glomerular endothelial cells and endothelial cells ,which also cause endothelial cell injury. Here ,we reviewed the role of endothelial cell injury in the pathogenesis of diabetic nephropathy.

ObjectiveTo explore the diagnosis and the surgical operation of concealed penis, and raise the therapeutic efficacy.Methods16 cases of concealed penis were reviewed retrospectively.ResultsAll 16 cases were treated by Devine surgery and got the satisfactory effect, the penis recovered to its good external appearance. All 16 cases had been followed up for four months to two years. The revelation of penis and its external appearance were all satisfied. No complications such as hydrophallus, skin ischemia, haematoma and erection ache occurred.ConclusionThe real concealed penis should be strictly distinguished from buried penis, microphallus and redundant prepuce, and suitable operation pattern can obtain satisfactory effect.

Objective To investigate the value of the bedside index for severity in acute pancreatitis (BISAP),Ranson's,APACHE Ⅱ and computed tomography severity index (CTSI) scoring system in evaluating the severity of acute pancreatitis.Methods The clinical data of 385 patients with acute pancreatitis who were admitted to the Zhongnan Hospital of Wuhan University from 2005 to 2011 were retrospectively analyzed.The values of 4 scoring systems including BISAP,Ranson's,APACHE Ⅱ and CTSI in predicting the incidences of severe acute pancreatitis,local complications and death were investigated by Chi-square test and receiver operating characteristic curv e.Odds ratio (OR) was calculated.The differences of areas under the curves (AUC) were analyzed using the Z test.Results The incidences of severe acute pancreatitis,local complications and mortality of patients with BISAP score ≥ 3 were 64.4％ (56/87),16.1％ (14/87) and 8.0％ (7/87),which were significantly higher than 13.4％ (40/298),6.4％ (19/298) and 0.3 ％ (1/298) of patients with BISAP score ≤ 2 (x2 =93.4,8.1,19.7,P ＜ 0.05).The incidences of severe acute pancreatitis,local complications and mortality of patients with Ranson's score≥3 were 52.7％ (48/91),22.0％ (20/91) and 7.7％ (7/91),which were significantly higher than 16.3％ (48/294),4.4％ (13/294) and 0.3％ (1/294) of patients with Ranson's score ≤2 (x2 =49.2,27.3,18.5,P ＜0.05).The incidences of severe acute pancreatitis,local complications and mortality of patients with APACHE Ⅱ score ≥ 8 were 46.6％ (27/58),20.7％ (12/58) and 8.6％ (5/58),which were significantly higher than 21.1％ (69/327),6.4％ (21/327) and 0.9％ (3/327) of patients with APACHE Ⅱ score≤7 (x2 =17.0,12.8,14.4,P ＜0.05).The incidences of severe acute pancreatitis,local complications and mortality of patients with CTSI score ≥4 were 51.4％ (19/37),51.4％ (19/37),16.2％ (6/37),which were significantly higher than 22.2％ (77/347),4.0％ (14/347),0.6％ (2/347) of patients with CTSI score≤3 (x2 =15.1,95.3,40.1,P ＜ 0.05).The sensitivity,specificity,positive and negative predictive values of BISAP were 58％,89％,64％,86％,respectively,and the AUC was 0.848,which were significantly higher than the other 3 systems (Z =2.02,4.22,4.78,P ＜ 0.05).The sensitivity,specificity,positive and negative predictive values of CTSI were 58％,95％,51％ and 96％,respectively,and the AUC was 0.926,which was significantly higher than the other 3 systems (Z =3.99,3.24,4.06,P ＜ 0.05).The sensitivity,specificity,positive and negative predictive values of BISAP were 88％,79％,8％ and 100％,respectively,and the AUC was 0.855,with no significant difference compared with the other 3 systems (Z =0.81,0.03,0.14,P ＞ 0.05).Conclusions The accurate rate of BISAP in predicting the severe acute pancreatitis is higher than Ranson's,APACHE Ⅱ and CTSI.The accurate rate of CTSI in predicting the incidence of local complications is higher than the other 3 systems.There is no significant difference of the 4 systems in predicting the mortality.The BISAP scoring system is helpful in early diagnosis of severe acute pancreatitis,and making the individualized treatment plan,thus improving the prognosis of patients.

Aim Toinvestigatewhetherepidermal growth factor receptor ( EGFR )-containing exosomes could induce tumor-specific regulatory T cells, and the effects of those T cells on tumor protein-specific CD8+Tcells.Methods TheexosomeswithEGFRwerepu-rified from NSCLC tumor, which modulated tolerogenic property of DCs. Then the induced TolDCs generated tumor-specific Tregs, with which the tumor protein-specific CD8 + T cells were suppressed. Results 80%exosomeswereEGFRpositivefromLCpatients while less than 2% exosomes were EGFR positive from control lung tissue. After exposed to the exosomes in the culture for 7 days, the IDO+ DCs proportion was much higher than that in control group ( 80. 8% ± 3. 2% vs 65. 6% ± 6. 4%, P <0. 05 ) . The induced Tregs was also higher ( 24. 1% ± 5. 2% vs 4. 2% ± 2. 3%,P<0. 01 ) , which suppressed the proliferation of CD8+ T cells(5. 4% ± 0. 2% vs 86. 7% ± 9. 3%, P<0.01).Conclusion Thepurifiedexosomesin-duce tolerogenic DCs. Coculture of the tolerogenic DCs and Th0 cells generates the tumor antigen-specific reg-ulatory T cells. The Tregs could suppress the tumor an-tigen specific CD8+ T cells.

Human cytomegalovirus (HCMV) is a DNA virus and serious opportunistic pathogen for both newborn and immunocompromised individuals.To research technique for gene silence and antiviral agents, ribozyme M1GS-T6 was constructed from external guide sequences(EGSs)that consist of a sequence complementary to HCMV UL54 gene RNA and M1 RNA, the catalytic RNA subunit of RNase P from Escherichia coli. The results showed that M1GS can efficiently cleave the mRNA sequence encoding UL54 protein in vitro.

Objective To develop a real-time polymerase chain reaction(PCR)system to determine transcriptional level of MexAB-OprM multidrug efflux pump gene and to investigate the impact of androgra- pholide on MexAB-OprM gene transcription in Pseudomonas aeruginosa.Methods The fragments of mexB gene of mexAB-oprM operon and 30S rRNA gene rpsL were amplified and cloned into two plas- mids respectively.These plasmids were used as external standards for real-time PCR.Real-time PCR was applied to measure the mRNA transcripition of mexB and rpsL gene in Pseudomonas aeruginosa growing in medium with different concentrations of andrographolide.Results The plasmids for standard curve were constructed successfully.The relative mexB mRNA expressions in 50,100,150 and 200?g/mL andrographolide were 0.04?0.03,0.06?0.07,0.09?0.03 and 0.04?0.03 respectively, which were significantly lower than that in the control(0.24?0.04,P0.05).Conclusion Andrographolide can reduce the transcriptional level of MexAB-OprM,which may he one mechanism for its anti-infection effect.

Objective To investigate the clinicopathological features,diagnosis,treatment,prognosis and causative agent of a case of eumycetoma on the submaxilla.Methods A case of eumycetoma diagnosed in our department was assessed for its clinical and pathological features as well as mycologic and molecular identification.Related literature was reviewed.Results The patient was primarily characterized by swelling of the submaxilla,with multiple sinuses draining many black granules.Pathologic examination revealed a pyogenic granulomatous inflammation,and a number of lotus rhizome node-like hypha were observed in tissue samples through PAS staining.Sequence analysis of multiple loci of the isolates,including ITS 1,ITS2 and D1/D2,showed that it was mostly similar to Madurella mycetomatis with a homology of 97%.Conclusion This is a case ofeumycetoma on the submaxilla induced by a novel species of Madurella.

Objective To explore the relationship between fluconazole resistance and expression of multidrug resistant protein genes,including CDR1,CDR2,MDRI.Methods The total RNA was extracted from fluconazole-resistant and -susceptible Candida albicans isolates,and cDNA was synthesized.The expression of CDR1,CDR2 and MDRl genes was then detected by quantitative real-time PCR.The?CT (threshold cycle) value was obtained by subtracting the CT value of 18S rRNA from that of the targeted gene.Results The?CT value of CDR2 was significantly lower in fluconazole-resistant isolates than in fluconazole- susceptible isolates (7.52?2.53 vs.9.28?3.15,t=2.367,P

Objective To explore the influence of fluorine on mRNA and protein expression of the insulin-like growth factor-1 (IGF-1) and its receptor of rat osteoblasts.Methods Osteoblasts were isolated from rat bone by enzyme digestion.Different fluorine concentration [0 (control),10-7,10-6,10-5,10-4,10-3 mol/L] was add to the second generation osteoblasts.The IGF-1 in the culture medium was determined by radioimmunoassay (RIA) at different fluorine concentration and different time (24,48 h).The expression of IGF-1 receptor was measured by the method of fluorescent quantitation PCR and the expression of protein IGF-1 receptor was measured by Western blotting.Results ①With increased dose of fluoride exposure,IGF-1 concentration in the osteoblastic culture medium increased first and then decreased at 24,48 h,respectively.Compared to the control group [(38.83 ± 3.48)ng/L],IGF-1 concentration of the 24 h 10-6 mol/L group[(65.45 ± 4.84)ng/L] was higher,and the difference was statistically significant(P ＜ 0.05).The same result was also shown in the 48 h 10-5 mol/L group [(59.14 ± 1.53)ng/L] to its corresponding control group [(33.79 ± 1.84)ng/L,P ＜ 0.05].②The mRNA expression of IGF-1 receptor of the 24,48 h 10-5 mol/L groups (0.0055 ± 0.0004,0.0262 ± 0.0040) was significantly higher than their corresponding control groups (0.0022 ± 0.0001,0.0073 ± 0.0008,all P ＜ 0.05).③With increased dose of fluoride exposure,the protein expression of IGF-1 receptor increased first and then decreased ;the expression of 24 h 10-5 mol/L group (1.39 ± 0.16) was compared with the corresponding control group (0.86 ±0.12),and the difference was statistically significant (P ＜ 0.05) ; the expression of 48 h every fluorine group was also compared with the corresponding control group,and the difference was not statistically significant(all P＞ 0.05).Conclusions Fluorine can affect the mRNA and protein expression of osteoblastic IGF-1 and its receptor.It indicates that IGFS signal transduction pathways play an important role in fluorine regulation of bone metabolism.