Objective To detect the expression of Ezrin in giant-cell tumor of bone,and to investigate its cilincal significance. Methods 60 cases of biopsy which had been confirmed as bone giant-cell tumors in our hospital from January 2008 to December 2013 were set as observation group;tumor tissues from 8 cases of reactive new bone in nonmalignant bone diseases,12 cases of osteoid osteoma and 11 cases of osteoblastoma in the corresponding period were set as control group. Protein and gene levels of Ezrin were tested with Western blotting method and real-time PCR detection,simultaneously proceeded the corresponding analysis combined with the clinical data of patients;60 cases of bone giant-cell tumor patients accepted tumor resection and pros-thesis replacement,2 courses of preoperative chemotherapy;mitochondria morphological changes of tumor tissue and Ezrin protein and genetic changes were observed before and after chemotherapy. Results In the giant-cell tumors of bone,the Ezrin protein mainly located in the cytoplasm,and its expression positive rate was much higher than that in reactive new bone of nonmalignant bone diseases(19. 7% ),osteoid osteoma(21. 2% )and osteoblastoma(20. 7% );the difference was statistically significant(χ2 = 4. 18,P = 0. 024),but no statistical difference in the Ezrin expression among the groups of osteosarcoma,osteoid osteoma and osteblastoma(χ2 =6. 18,P = 0. 087). In the giant-cell tumors of bone tissue after chemotherapy,mitochondria pyknosis and the phenomenon of liquid cavitation was less than that before the treatment,and Ezrin protein expression decreased and gene levels reduced[(23. 99 ± 1. 49)vs(20. 11 ± 1. 11),t = 5. 03,P = 0. 018)]. Conclusion The expression of Ezrin in giant-cell tumor of bone is much higher than other benign bone tumor,and it could be a biological marker for differentiating benign and malignant bone tumor. Early intervention in Ezrin may be helpful for reatment of giant-cell tumor of bone.

AIM: To explore the influence of CD47 molecules on the maturation and function of cultured dendritic cells (DCs). METHODS: Monocyte cell-derived DCs were propagated in granulocyte-macrophage colony-stimulating factor (GM-CSF) and lipopolysaccharide (LPS) plus interleukin (IL)-4, in the presence or absence of anti CD47 monoclonal antibodies (anti-CD47 mAbs). Flow cytometry was used to detect the cell surface phenotype. The concentration of IL-12P70 in supernatant was measured by ELISA technique. The antigen-presenting functions of DCs were determined in one-way mixed leukocyte reaction by Brdu-ELISA. Electrophoretic mobility shift assays (EMSA) was used to examine NF-?B activity. RESULTS: The anti-CD47 mAbs markedly suppressed the expression of CD80,CD86,CD83,CD1a,HLA-DR on the surface of DCs (P

AIM: To investigate the effects of transforming growth factor ?1 (TGF-?1) on murine-derived dendritic cells (DC). METHODS: Murine bone marrow cells were cultured with GM-CSF and TGF-?1 to develop TGF ?-DC. Then they were stimulated by lipopolysaccharide (LPS). Their phenotypes were assessed by flow cytometry (FCM). The allogeneic stimulating capacity of DC was measured by mixed lymphocyte reaction (MLR) using BrdU ELISA method. IL-12 p70 protein was detected by ELISA and the expressions of Toll like receptor 4 (TLR4) on DCs were measured by semi-quantitative RT-PCR and FCM. RESULTS: Compared to immature DC (imDC) cultured with GM-CSF alone, the expressions of CD_80, CD_86, I-A~b and CD_40 in TGF ?-DC were lower. The TGF ?-DC was resistant to maturation by LPS. Maturation resistance was evident from a failure to up-regulate CMs, to stimulate larger T cell proliferation and to increase secretion of IL-12 p70. Down-regulation of TLR4 expression on TGF ?-DC was also found. CONCLUSION: TGF-?1 inhibits the expression of co-stimulatory molecules on DC. It is resistant to maturation stimulus (LPS) and might be linked with TLR4 down-regulation.

Objective To investigate the carcinogeic role of miR-365 in cuntanerous squamous cell carcinoma (cSCC).Methods Normal HaCaT cells were divided into control and irradiation groups,the later was exposed by UVB irradiation (50 J/m2).MicroRNA expression profiles of the two groups were analyzed by microRNA array.The expression variations of miR-365 in HaCaT,A431,Tca8113 and HSC-1 cells were validated by qRT-PCR analysis.The colony-forming and invasion capacities were dectected by colony forming assay and Transwell migration assay in vitro,respectively.HaCaTpre-miR365-2 highly expressing miR-365 was constructed by retroviral vector infection.Tumorigenicity evaluation was carried out by subcutaneously inject of the cells at the right back flank of nude mice.Results There were 30 microRNAs differentially expressed in HaCaT cells after UVB irradiation and miR-365 was one of the most sensitive miRNAs(as high 6.7 times as control).Expression of miR-365 in all the cSCC cell lines A431,Tca8113 and HSC-1 were significantly higher than that in HaCaT cell,in which the maximum was A431 (15.67 ±1.12 times,P ＜ 0.01),and the minimum was TcaS113 (4.72 ± 0.85 times,P ＜ 0.05).Knockdown of miR-365 in cSCC cell lines significantly inhibited the colony forming ability (t =13.68,P ＜ 0.05) and cell migration (t =19.98,P ＜ 0.05) in vitro.HaCaT cells overexpressing miR-365 by transient transfection significantly increased the ability of colony formation (t =7.11,P ＜ 0.05) and cell migration (t =22.03,P ＜0.05) in vitro.In addition,HaCaTpre-miR-365-2 cell line stably expressing miR-365 could successfully establish tumors in nude mice.Conclusions MiR-365 is an oncogene for cutaneous squamous cell carcinoma.

Objective To discuss the surgical method of resection of petroclival tumors used temporal base transtentorial approach.Methods Analyzed 26 cases of petroclival tumors.All of them were surgically treated under intraoperative neurophysiological monitoring by temporal base transtentorial approach.Firstly the supratentorial part of tumors were resected by pieces,and then the tentorium was cut open,in order to resect residue of the subtentorial part of tumors.This surgical maneuver was applied to reduce the retraction to surrounding vessels,nerves and brain stem,as well to protect important structures and to finally achieve radical removal of the tumors.The efficacy and complications were observed.Results Among 26 cases,19 cases (73.1％,19/26) achieved total removal,5 cases (19.2 ％,5/26) achieved subtotal removal and 2 cases (7.7％,2/26) achieved great partial removal.There was no surgery-related death.Eleven cases suffered from partial neurological deficit.All cases were followed up for 3 months to 4 years,3 cases underwent complete recovery,4 cases underwent partial recovery,and 4 cases underwent permanent deficit.Conclusions Using temporal base transtentorial approach to resect petroclival tumors is convenient,applicable,safe and with minimal injury and with high proportionality of total resection.Under intraoperative neurophysiological monitoring,this approach may be an ideal choice for surgical treatment of these tumors.

Objective To evaluate the effect of early continuous renal replacement therapy(CRRT)on patients with acute kidney injury(AKI)induced by severe sepsis. Methods A prospective study was conducted. 160 patients with AKI induced by severe sepsis admitted to intensive care unit(ICU)of Qingyuan People's Hospital between October 2009 and June 2013 were enrolled. According to the starting time of CRRT,the patients were randomly divided into two groups:an earl y treatment group and a regular treatment group(each,80 cases). All patients were treated in accordance with the principle of the cluster of severe sepsis. In early treatment group,the patents began to receive CRRT when the amount of urine output was<0.5 mL?kg-1?h-1 and had taken continuous conventional treatment over 6 hours,but the curative effect was not ideal. In the regular treatment group,the patients began to apply CRRT when the amount of urine output was<0.5 mL?kg-1?h-1 and had taken continuous conventional treatment persistently over 24 hours or difficultly corrected acidosis or heart failure developed. The changes in urine output,creatinine,saturation of blood oxygen,and albumin,and 28-day survival rate,length of ICU stay,time of organ support were measured and compared between the two groups to evaluate the therapeutic significance of the early CRRT. Results The mean length of ICU stay(day:6.5±1.7 vs. 8.2±1.6),length of organ support(day:5.3±1.8 vs. 6.0±1.4),the recovery time of urine amount(day:3.4±0.9 vs. 4.8±0.5)in early treatment group were significantly shorter than those in regular treatment group(all P<0.01);recovery ratio of urine in 1 week in early treatment group was significantly higher than that in regular treatment group(46.2%vs. 28.8%,P<0.05). The creatinine recovery ratio in 1 week in early treatment group was lower than that in regular treatment group(55.0%vs 78.8%),but there was no statistically significant difference(P>0.05). Early treatment group and regular treatment groups of creatinine recovery time were similar(day:5.1±1.3 vs. 5.2±1.2). 28-day survival rate in early treatment group was higher than that in regular treatment group(41.2%vs. 35.0%),but there was no statistically significant difference(P>0.05). Conclusion It is found that there is no evidence to suggest that early CRRT may improve the prognosis of the patients with AKI induced by severe sepsis,but it may be in favor of shortening the time of urine recovery,length of stay in ICU,length of organ support and length of dialysis.

Objective:To compare the efficacy and safety of gemcitabine and epirubicin curing for patients with non-muscle invasive bladder cancer (NMIBC).Methods:From October 2014 to October 2017, 86 patients with NMIBC diagnosed by transurethral resection of bladder tumor (TURBT) in Wenling Hospital Affiliated to Wenzhou Medical University were analyzed retrospectively. Among them, 42 were treated with gemcitabine (gemcitabine group) and the other 44 with epirubicin (epirubicin group). In gemcitabine group, there were 37 males and 5 females. The average age was 63.9 (48-81) years old. 30 cases were single tumor while 12 cases were multiple. 35 cases with tumor diameter less than 3cm and 7 cases with tumor diameter greater than 3cm. There were 28 cases in T a stage and 14 cases in T 1 stage. 13 patients’ tumor were high grade and 29 patients’ tumor were low grade. In the epirubicin group, there were 36 males and 8 females. The average age was 65.4 (48-88) years. 31 cases were single tumor while 13 cases were multiple. 36 cases with tumor diameter less than 3cm and 8 cases with tumor diameter greater than 3cm. There were 30 cases in T a stage and 14 cases in T 1 stage. 15 patients’ tumor were high grade and 29 patients’ tumor were low grade. There was no significant difference in the above general information between the two groups ( P > 0.05). The two groups were treated with epirubicin or gemcitabine within 24 hours after operation, and bladder perfusion once a week was performed continuously after the first week of operation, a total of 8 times, and after that once a month till one year after operation. The clinical efficacy and adverse reactions were compared between the two groups. Kaplan-Meier was used to compare the recurrence free survival time of tumor after operation. The prognostic factors were analyzed by Cox proportional hazards model. Results:The adverse reactions of the two groups were mainly bladder irritation, gross hematuria, fever, nausea and vomiting. The incidence of bladder irritation and gross hematuria in epirubicin group was 25.0% (11 / 44) and 18.2% (8 / 44), which were significantly higher than those in gemcitabine group [7.1% (3/42) and 2.4% (1/42)], and the difference was statistically significant ( P<0.05). There was no significant difference in other adverse reactions between the two groups ( P> 0.05). Kaplan-Meier survival analysis showed that the median tumor recurrence free survival time of gemcitabine group was 29.7 (6.3-58.8) months, and the 1-year and 2-year tumor recurrence free survival rates were 71.4% (30/42) and 45.2% (19/42), respectively; the median tumor recurrence free survival time of epirubicin group was 28.8 (4.5-57.8) months, and the 1-year and 2-year tumor recurrence free survival rates were 70.5% (31/44) and 47.7% (21/44), respectively. There was no difference in the tumor recurrence free survival rates between the two groups( P>0.05). Cox analysis showed that age ( HR=1.1, 95% CI 1.034-1.113) and tumor grade ( HR=12.2, 95% CI 5.776-25.680) were independent risk factors for prognosis. Conclusions:The efficacy of bladder perfusion chemotherapy with gemcitabine and epirubicin in patients with NMIBC was not significantly different, but the incidence of adverse reactions with gemcitabine was lower, which was safe and reliable. The risk factors affecting postoperative survival rates of NMIBC were patient's age and tumor grade.

Objective To explore the guiding role of serum procalcitonin (PCT) in antibiotic therapy during acute exacerbation of chronic obstructive pulmonary disease (AECOPD). Methods Totally 72 patients with AECOPD were randomly divided into PCT group (n=40) and conventional therapy group(n=32).For patients in PCT group,the use of antibiotics was based on PCT serum levels,antibiotics were stopped when PCT＜0.25 μg/L,while in conventional treatment group,the use of antibiotics was based on clinical symptoms of patients.The main observation indexes included ratio of antibiotic usage,time of antibiotics use,hospital stay,clinical efficacy,aggravating cases and death cases. Results There were no significant difference in clinical efficacy between the two groups(82.5％ vs.75.8％,x2 =0.217,P=0.641),however,the ratio of antibiotics usage in PCT patients was significantly lower than conventional therapy group (47.5％ vs.71.9％,x2 =4.346,P=0.037),average time of antibiotics use and days of hospital stay were shorter in PCT treatment group than conventional therapy group [(6.84±3.27) d vs.(10.22±3.67)d,x2 =3.116,P=0.003; (11.7±5.2) d vs.(20.3±8.7) d,x2 =5.202,P=0.000].There were no difference in double infection incidence (2.5％ vs.18.8％,x2 =3.657,P=0.056),aggravating cases (3 cases vs.4 cases,x2 =0.097,P=0.756) and mortality (2.5％ vs.6.3％,x2 =0.039,P=0.843) between the two groups.Conclusions Serum PCT level may be an appropriate indicator to guide antibiotic therapy in patients with AECOPD in view of its effective decreases of excessive use of antibiotics,double infection opportunities and hospitalization time.

Objective To study on the characterization of MICA/B genetic polymorphism in a northern Chinese Hunan Han population.Methods Ninty-five unrelated individuals were involved in this study and MICA/B genotypes were determined by two methods:PCR-sequence-specific primers (PCR-SSP) and PCR-sequence-based typing (PCR-SBT).Results In northern Hunan Han population,eleven MICA alleles were found,among which MICA * 010 (28.95％),MICA * 008 ∶ 01 (20.53％) and MICA * 002 ∶ 01 (15.79％) were the common alleles.Five MICA-STR(short tandem repeat) alleles were found,among which MICA * A5 (37.89％) and MICA * A5.1 (21.05％) predominated.In this population,ten MICB alleles were found.The common alleles were MICB * 005 ∶ 02/* 010 (58.42％),MICB * 002 ∶ 01 (10.00％),and MICB * 008 (7.89％).Two kinds of MICA-MICB haplotypes were MICA * 004-MICB * 004 ∶ 01 and MICA * 010-MICB * 005 ∶ 02/010 in significant linkage disequilibrium.This study also showed MICA/B gene with high polymorphism in different populations.Conclusion MICA/B alleles distribution in northern Hunan Han population with its unique characteristics.

Objective To investigate the way that nasopharyngeal carcinoma (NPC) and NPC stem cells develops resistance to cisplatin through anti-reactive oxygen species mechanism. Methods Using CCK-8 cell counting kit, we measured the half inhibitory concentration of cisplatin against NPC cellsCNE-2and NPC stem cellsCNE-2S, and compared their resistant index. We examined the differences in the reactive oxygen species (ROS) levels, total glutathi?one (GSH) levels, and total superoxide dismutase (SOD) levels between CNE-2 and CNE-2S at different concentrations of cisplatin administration (0.1,0.5 and 1.0μmol·L-1). Using q-PCR, we determined the mRNA expression level of GSS, GCLC, GCLM, SOD1 and SOD2 after 48 hours administration of cisplatin at 1 μmol · L-1. Protein expression level of SOD2 was also tested using Western Blot after 48 hours administration of cisplatin at 1μmol · L-1. Upon silencing the SOD2 in NPC cell through siRNA, Trypan blue was used to analyze cell survival after cisplatin was administrated at 1μmol · L-1. Results The inhibition concentration of cisplatin against CNE-2 was higher than that against CNE-2S (μmol · L-1:9.8 ± 1.1 vs 2.4 ± 0.6,P<0.05). ROS levels in CNE-2 and CNE-2S both rise with cisplatin administration, but ROS levels of CNE-2 before and after cisplatin treatment were both higher than those in CNE-2S (P<0.05). The total gluta?thione levels in CNE-2 and CNE-2S were both increased after 1μmol·L-1 cisplatin treatment but there is no significant dif?ference in levels of glutathione between these two cell lines. After treated with cisplatin, SOD level were increased in both CNE-2S and CNE-2, but it is higher in CNE-2S than that in CNE-2 (P<0.05). The mRNA levels of GSS, GCLC, GCLM, and SOD1 were not different significantly between in CNE-2 and in CNE-2S with or without cisplatin treatment. However, SOD2 in CNE-2S were higher than that in CNE-2 on both mRNA and protein levels (P<0.05). Silenced SOD2 disrupted the resistance of cisplatin in CNE-2S. Conclusion These data suggest that NPC stem cells (CNE-2S) enhance its drug re?sistance to cisplatin through highly expression of SOD2 which posed anti-ROS capacity.