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.

Objective To investigate the feasibility of treatment mode of end-to-end anastomosis of esophagus(EAS) af ter partial resection for early-stage cervical esophageal carcinoma(ECEA).Methods 7 patients were substantially confirmed as squsmous cell carcinoma of cervical esophagus by endoscopy,the nearest distance of the lesion from the incisors was 17cm,and the furthest was 20 cm,the maximum extent was 2.5 cm,and the minimum was 1 cm.None of them with longitudinal muscularis invasion.Confirmed by PET/CT or chest enhancement CT examination preoperatively,intrathoracic and cervical lymphatic metastasis was excluded,cT1 -2 N0 M0.Incisal margin length was not less than 1 cm,the maximum was 5 cm and the minimum was 3 cm.Meanwhile,the cervical lymph node should be dissected,and the average number was 6.43 per case.After surgery,all the patients were fixed by plaster slab to release the tension of anastomosis.Postoperative adjuvant radiotherapy or chemotherapy was received.Results None of the patients had severe postoperative complications,and the average hospital stay was 14.5 days.All the patients are alive,the longest follow-up lasts for 3 years and 4 months,all of them can take normal food,without anastomotic stenosis.Conclusion Treatment mode of EAS after partial resection for ECEA significantly decrease the operative damage,apparently improve the patient's quality of life(QOL),so that the patients can better receive adjuvant treatment subsequently; it is a feasible and effective method for cervical esophageal carcinoma at the early stage.

In recent years, with the gradual improvement of people's understanding of intestinal flora, exploring common complications after renal transplantation from the perspective of intestinal flora has become a research hotspot. Studies have demonstrated that the composition of the intestinal flora in renal transplant recipients has significant differences before and after surgery, and this difference is closely related to the occurrence and development of many complications after renal transplantation such as infection, rejection, diarrhea and so on, which could affect the prognosis of the recipients. This article reviewed the research progress on intestinal flora and renal transplantation in order to provide new treatment ideas and strategies for renal transplant recipients.

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.

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 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.

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.