Objective To observe the effects of different dosages of granulocyte-macrophage colony-stimulating factor (GM-CSF) on generating the routine bone marrow dendritic cells, and supply suitable dosage of GM-CSF on preparation of dendritic cell vaccines used for different purpose. Methods Using low (5 ng/ml) and conventional (20 ng/ml) and high dosage( 50 ng/ml ) of GM-CSF combined interleukin-4 ( IL-4 ) to induce murine bone marrow dendritic cells were performed, The phenotypes (CD_(11c), CD_(80), CD_(86)) and functional properties of the DC were compared by FACS analysis and MLR. Results The DC induced by low dosage of GM-CSF were immature DC, expressing low CD_(11c), CD_(80), and CD_(86). DC induced by conventional dosage were functional mature, expressing higher CD_(11c), CD_(80), CD_(86), which could induce allogeneic T lymphocyte responses. DC induced by the high dosage GM-CSF were the most phetotypicaUy and functional mature cells, expressing the highest CD_(11c), CD_(80) CD_(86), which could induce the strongest allogeneic T lymphocyte responses. Conclusion The dosages of GM-CSF affect the maturation stage of dendritic cells. Low dosage of GM-CSF generated immature dendritic cells, but conventional dosage and high dosage generated mature dendritic cells. DC generated through high dosage of GM-CSF were the most mature in phenotype and function.

Aim To clone and express the 1.2kb cDNA fragment (1/753-2 934bp) of human integrin α 4 subunit. Methods The 1.2 kb cDNA fragment of human integrin α 4 subunit was amplified from HL-60 total RNA by RT-PCR, then it was subcloned into expression vector pGEX-3X and induced with IPTG. Results The 1.2 kb cDNA fragment of human integrin α 4 subunit was cloned. The sequencing indicated that there was only one missense mutation (Arg→ Gln) among the fragment, and this mutation won't affect antigenicity after analysed by GOLDKEY. Then the 1.2 kb cDNA was subcloned into expression vector pGEX-3X. The α 4 fragment was highexpressed in E.coli after induced with IPTG. Conclusion The 1.2kb cDNA fragment of α 4 subunit was obtained, and it was highexpressed in E.coli, it might be important for study on the function of α 4 integrins.

Objective To explore the effect of parvovirus B19 (VB19) infection on pediatric leukemia patients. Methods The pediatric leukemia patients were enrolled in the study in the Children's Hospital of Soochow University. Expression levels of VB19-DNA-PCR were detected using the polymerase chain reaction. Positive patients would be monitored and treated by conventional treatment as well until VB19 gene became negative. The data was compared according to the VB19 clearance time, clinical symptoms and blood counts to evaluate the effect. Results In the 3009 samples from 824 pediatric leukemia patients, there were 36 samples (1.2%) from 12 cases (1.5%) of pediatric leukemia paients with positive VB19 infection. Among the positive patients, 11 cases (1.9%) were from 582 with acute lymphoblastic leukemia (ALL) patients and 1 (0.45%) was from 212 with acute myeloid leukemia (AML). According to the treatment stage, 3 cases were in initially diagnosed period, 2 cases in early stage of consolidation chemotherapy, 4 cases in delayed enhanced chemotherapy period, and 3 cases in maintenance chemotherapy period. According to the treatment response, 4 cases were in continuous treatment, 2 cases were sensitive to treatment, and 3 cases were drug resistant. In the drug resistance group, 2 cases developed into the pure red cell aplastic anemia (PRCA). After treatment, one was recovered from PRCA with VB19 cleared, the other one remained PRCA with continuously positive VB19. Conclusions More VB19 virus infection in pediatric ALL happened in delayed enhanced chemotherapy period. The persistent presence of VB19 infection on pediatric leukemia patients is closely related with PRCA.

Objective: To investigate the mechanism of miR-503 modulates radio-resistance of esophageal squamous cell carcinoma (ESCC) by targeting excision-repair cross-complementing 1 (ERCC1). Methods: The expression level of miR-503 in radio-resistant ESCC tumor tissues and KYSE140 and KYSE140R cells was detected by qPCR. The miR-503 mimic, miR-503 inhibitor or si-ERCC1 was transfected into KYSE140 and KYSE140R cells.After radiation treatment, the colony formation assay and CCK-8 assay were used to detect the proliferation of KYSE140R cells. Flow cytometry was used to detect apoptosis of KYSE140R cells. WB was used to detect changes in protein expression of ERCC1. Dual luciferase reporter gene assay was used to validate the targeting relationship between miR-503 and ERCC1. Results: The expression level of miR-503 was down-regulated in radio-resistant tissues and ESCC cell lines (all P<0.01). Over-expression of miR-503 significantly inhibited cell proliferation and promoted apoptosis of KYSE140R cells (all P<0.01). Dual-luciferase reporter assay validated that ERCC1 was a target gene of miR-503, and miR-503 negatively regulated the expression of ERCC1. Over-expression of miR-503 significantly down-regulated the expression of ERCC1 in KYSE140 and KYSE140R cells (both P<0.01), inhibited cell proliferation (both P<0.01), but significantly increased apoptosis rate (all P<0.01); knockdown of ERCC1 exhibited a similar effect, while knockdown of both ERCC1 and miR-503 reversed the above effects. Conclusion: Over-expression of miR-503 up-regulated the radio-sensitivity of KYSE140R cells by targeting ERCC1.

Dizziness or vertigo is a common clinical symptom, and its underlying etiology is complex. Many clinicians are confused about its diagnosis and treatment. This article presents a case about chronic vestibular syndrome. And case appreciation and academic discussion are conducted by well-known domestic neurologists and otologists, so as to provide a good thinking model and basic ideas for the diagnosis and treatment of dizziness or vertigo, hoping to further improve the diagnosis and treatment level among clinicians.
Humans ; Dizziness/therapy* ; Vertigo/etiology* ; Vestibular Diseases/complications* ; Otolaryngologists

Objective To study the evolution of renal cell carcinoma treated in a single institution over the past 12 years.Methods A total of 1923 patients with renal cell carcinoma surgically treated from 1999 to 2010 were included in this study.Age at diagnosis,gender,symptoms,tumor size,TNM stage,histological subtype,Fuhrman grade and type of treatment were compared among 3 periods (Period 1:1999-2002,Period 2:2003-2006,Period 3:2007-2010).Results The number of patients grew fasty from 267 cases in the Period 1 to 1092 cases in the Period 3.The incidentally diagnosed cases increased significantly from 49.8％ to 73.6％ (P ＜0.01),while the mean age of patients at diagnosis,the male to female ratio and the proportion of young (＜40 years old) patients were not statistically different.The tumor size at diagnosis gradually decreased from 6.1 cm to 4.8 cm (P ＜ 0.01),and the proportion of small tumors less than 4 cm increased remarkably from 30.3％ to 54.4％ (P ＜ 0.01).Concerning the surgical approach,open surgery decreased from 98.5％ to 73.8％ (P ＜ 0.01),and laparoscopic surgery increased from 1.50％ to 26.2％ (P ＜ 0.01).Overall,the rate of nephron-sparing surgery increased from 5.2％ to 34.7％ (P ＜0.01) in all cases.The use of nephron-sparing surgery increased significantly from 17.9％ to 54.7％ in T1a subset and from 0.00％ to 14.8％ in T1b subset respectively (P ＜ 0.01).Furthermore,radiofrequency ablation was applied and the percent of this approach reached 2.47％ in Period 3.Conclusions The evolution of clinical-pathological characteristics and surgical treatment for renal cell carcinoma in a single institution is apparent over the last 12 years.With the increase of incidentally diagnosed cases and small renal tumors,nephron-sparing surgery has been widely performed in T1 subset instead of traditional radical nephrectomy.

ObjectiveTo analyze the incidence of benign lesions in patients undergoing surgery for presumed renal cell carcinoma (RCC) and investigate the correlation of tumor size and histopathological characteristics.MethodsFrom Jan 2003 to Sep 2010,1531 patients (1042 males,489 females with average age of 55.1 years (15 -89 years) underwent nephrectomy.There were 1123 radical nephrectomies and 408 partial nephrectomies for solitary renal cortical neoplasms presumed to be RCC in preoperative imaging study.The pathological tumor size,the percentage of benign lesions and histologic subtypes were analyzed retrospectively.The correlation of Fuhrman grading and tumor size in clear cell type RCC and papillary RCC was investigated as well.Results Pathological examinations revealed that there were 81 (5.3％)benign lesions of 1531 patients.The incidence of benign lesions was 7.8％ in renal masses smaller than 4.0cm,3.8％ in masses with 4.1 - 7.0 cm,and 1.1％ in masses larger than 7.0 cm ( P ＜ 0.01 ).Angiomyolipoma was the most predominant histologic subtype in benign renal lesions with a frequency of 69.5％,52.6％ and 33.3％ in ≤4.0 cm,4.1 - 7.0 cm and ＞ 7.0 cm subgroups,respectively.Oncocytoma was present in 13.6％,15.8％ and 33.3％ of the benign lesions in the above 3 subgroups (P =0.47).One thousand four hundred and fifty cases of malignancies (94.7％) were identified.The frequency of clear cell type RCC was 91.7％ in malignant tumors smaller than 4.0 cm,88.1％ in 4.1 - 7.0 cm malignancies,and 77.6％ in tumors ＞7.0 cm； the frequency of papillary RCC was 4.0％,4.2％ and 7.7％ in the above 3 subgroups,and the frequency of chromophobe cell RCC was 3.6％,5.2％ and 6.3％ accordingly ( P ＜0.01 ).One thousand three hundred and seventeen cases of clear cell type RCC and papillary RCC with definite Fuhrman grading were identified.The probability of high grade tumors increased with size,while the probability of low-grade lesions decreased (r =0.94,P ＜ 0.01 ),especially for tumors smaller than 11.0cm.ConclusionsPatients in the present study population show a low incidence of benign renal lesions.Benign lesions are less common in larger tumors than in smaller ones.The proportion of clear cell type RCC decreases in larger tumors,however papillary RCC and chromophobe cell RCC are increasing.The probability of high-grade tumors increases with size.The correlation between tumor size and histopathological characteristics may be helpful in patients counseling and decision-making.

Objective To construct recombinant lentiviral vector of microRNA-34a and observe the cell viability,cell cycle and apoptosis of hepatocellular carcinoma cells transfected with the vector system and treated with Doxorubicin.Methods Recombinant lentiviral vector containing microRNA-34a gene was constructed and transfected into 3 hepatocellular carcinoma cell lines,and cells were treated with Doxorubicin.The expression of microRNA-34a gene was detected by real-time PCR.The effect of microRNA-34a overexpression on hepatocellular carcinoma cells proliferation were quantified via MTT assay,cell cycle and apoptosis was evaluated by flow cytometry.Western blotting was used to evaluate the expression of cell cycle and apoptosis related protein.Results The successful construction of microRNA-34a recombinant lentiviral vector was confirmed by plasmid enzyme digestion and DNA sequencing.Compared with the control group,relative expression of microRNA-34a gene in hepatocellular carcinoma cells significantly increased ((HepG2:t=15.36,P＜0.01;Hep3B:t=36.75,P＜0.01;Bel-7402:t=24.17,P＜0.01)).Cells viability decreased (HepG2:t =7.12,P ＜ 0.01;Hep3B:t =8.89,P ＜ 0.01;Bel-7402:t =13.62,P ＜0.01),G1 phase cells increased significantly(HepG2:F =137.65,P ＜ 0.01;Hep3B:F =143.39,P ＜0.01;Bel-7402:F =1 306.47,P ＜ 0.01) and cell apoptosis increased(HepG2:F =386.14,P ＜ 0.01;Hep3B:F =881.94,P ＜ 0.01;Bel-7402:F =885.89,P ＜ 0.01).Conclusions MicroRNA-34a recombinant lentiviral vector (LV-hsa-mir-34a) transfected hepatocellular carcinoma cells overexpress microRNA-34a,reduce the malignant biological behavior.MicroRNA-34a recombinant lentiviral vector (LV-hsa-mir-34a) enhance the in vitro inhibitory effects of Doxorubicin on hepatocellular carcinoma cell lines.

The implementation of living donor liver transplantation not only extends the number of donors and effectively alleviates the shortage of organ donation, but also reduces the fatality of patients on the waiting list for liver transplantation. In recent years, with persistent development of laparoscopic techniques and instruments, laparoscopic donor liver hepatectomy has been widely applied in the field of living donor liver transplantation, which is also one of the research hotspots. It has multiple advantages of mild trauma, slight pain, accelerated postoperative recovery for the donors and slight psychological burden, etc. However, due to strict screening criteria for donors, complicated procedures and long learning curve, it has been carried out in a few transplantation centers nationwide. In this article, the development history and current status, specific surgical patterns, the problems and challenges of laparoscopic living donor liver procurement for liver transplantation were introduced, and the development direction of laparoscopic living donor liver procurement was predicted, aiming to provide reference for accelerating the development and enhancing clinical prognosis of living donor liver transplantation.