Objective To investigate the micrometastatic lesion of tumor stem cells in the axillary swollen lymph nodes of breast cancer patients and the influence of miR30a on its invasive ability ,and to explore the feasibility of miRNAs anti‐breast cancer treatment .Methods The tumor stem cell‐like breast cancer cells were separated from the axillary swollen lymph nodes in breast cancer patients and cultured .miR30a oligonucleotide fragment was synthesized and transfected into human primary generation breast cancer cells by using adenovirus ,meanwhile the breast cancer cell line MDA‐MB‐231 was taken as the experimental control and the transfection efficiency was assessed by the fluorescence microscopy .The changes of tumor cell proliferation and invasiveness before and after transfection were detected by the Transwell chamber in vitro invasion assay .Western blot was used to detect the ALDH1 ,Vimentin and N‐Cadherin protein expression .Results The Transwell chamber in vitro invasion assay showed that the pri‐mary generation breast cancer cell had more strong invasive ability than MDA‐MB‐231 cell line ,their invasion indexes were (75 .3 ± 3 .2)% and (58 .4 ± 2 .8)% respectively ,the difference was statistically significant (P<0 .05) ,and after transfecting miR30a ,the in vitro invasiveness ability in these two kinds of cells were significantly weakened and their invasion indexes were (21 .4 ± 1 .9)% and (28 .2 ± 2 .3)% respectively ,the difference compared with the control group showed the statistical significance (P<0 .05) .Conclu‐sion The ALDH expression in partial axillary hyperplasia and swollen lymph nodes in the patients with breast cancer is increased , and the tumor micrometastasis may exist ,which should be completely cleaned in operation .miR30a inhibits the expression and inva‐sive ability of tumor stem gene .

Objective:To investigate the effects of two SNP sites of delta-like ligand protein-1 (DLL1) gene rs2738822 (C>T) and rs9459988 (T>G) and gene expression on bone marrow suppression after neoadjuvant chemotherapy for breast cancer.Methods:Breast cancer patients who received neoadjuvant chemotherapy were selected as study subjects, including 90 patients with severe bone marrow suppression and 72 patients with mild bone marrow suppression. Patient’s demographic characteristics and laboratory test indicators were collected. Two SNP sites of DLL1, rs2738822 and rs9459988, were genotyped by capillary electrophoresis and section analysis (SNaPshot) . The relative mRNA expression of DLL1 gene was detected by quantitative reverse polymerase chain reaction (QRT-PCR) method.Results:For The rs2738822 of DLL1 gene, the genotype distribution difference between severe and mild bone marrow suppression groups was statistically significant ( χ2=8.622, P=0.013) . Compared with CC genotype, CT and TT genotype carriers had a higher risk of severe bone marrow suppression, with an OR value of 2.746 (1.335-6.882) and 3.054 (1.282-8.143) , respectively. The dominant model results showed that TT OR CT carriers had a significantly higher risk of severe bone marrow suppression than THOSE with CC genotype [ OR=2.976 (1.231-4.963) ]. For rs9459988, there was no significant difference in genotype distribution between severe bone marrow suppression group and mild bone marrow suppression group ( χ2=2.149, P=0.342) . Results of the dominant model showed that TG or GG carriers had a significantly higher risk of severe bone marrow suppression than TT carriers, with an OR value of 2.046 (1.053-5.611) . The relative mRNA expression level of DLL1 gene was 1.15±0.23 in patients with severe bone marrow suppression, which was significantly lower than that in patients with mild bone marrow suppression (2.64±0.51) ( t=6.381, P<0.001) . For rs2738822, with the increase of T allele, the relative mRNA expression level of DLL1 gene decreased gradually ( P<0.05) . For rs9459988, the relative mRNA expression level of DLL1 gene in patients with mutant allele G was also significantly lower than that in wild-type CC carriers ( P<0.05) . Conclusion:Mutations of DLL1 genes rs2738822 and rs9459988 are related to the occurrence of severe bone marrow suppression after neoadjuvant chemotherapy for breast cancer, and can be used as a genetic marker to predict the degree of bone marrow suppression after neoadjuvant chemotherapy for breast cancer patients.

Objective:To investigate the effect of c-maf gene on the MM cells' proliferation and invasion activity. Methods:Lipo-fectin Reagent was used to transfect c-maf siRNA into multiple myeloma cell of RPMI8226. The mRNA expression level of c-maf was detected by RT-PCR.Cell growth curve was measured by MTT assay. Transwell chamber test was used to measure MM cells' in vitro in-vasion activity. The cell cycle distribution were assessed by flow cytomentry. The protein expression levels of survivin,MMP-2, MMP-9, ARK5 and cyclin B1 were detected by Western blot. We also detected the activity of Caspase-3/7. Results:The c-maf siRNA was effectively transfected into cells and the mRNA expression of the c-maf gene was inhibited.MTT test and Transwell chamber test showed that the proliferation and in vitro invasion activity of transfected cells were significantly lower than those of other two groups (P<0.05). Cell cycle of c-maf siRNA transfected group cells was arrested in G2/M phase. The expression levels of survivin,MMP-2, MMP-9,ARK5,cyclin B1 and the activity of Caspase-3/7 between c-maf siRNA transfected group and the other two groups were sta-tistically different (P<0.05). Conclusion:c-maf gene by c-maf siRNA can remarkably inhibit proliferation and invasion of multiple my-eloma cell lines of RPMI8226. C-maf gene may be used as the target for multiple myeloma gene therapy.

Objective:To investigate regulatory immune response induced by DNA vaccination encoding T cell receptor V?5.2 or V?2.1 chain predominantly displayed on ovalbumin (OVA)-specific T cell clone.Methods:BALB/C mice were vaccinated with pcDNA3.1 encoding T cell receptor (TCR) V?5.2 or V?2.1 chain respectively.Using RT-PCR,transcription of the recombined plasmids was analysed. Cell proliferation was measured by 3H-TdR incorporation.CTL response was assayed by JAM test.Immuno-fluorescent assay was used to examine the anti-TCR antibody level and the number V?2 +T cells.Results:RT-PCR analysis showed that the recombined plasmids can be transcripted in vivo and in vitro.DNA vaccine with TCR variable chain effectively induced TCR-specific humoral and cellular immune response,V?2 +T cells was not depleted by V?2.1 TCR-DNA vaccination,but rather was anergy.Conclusion:Regulatory immune response can be induced by DNA vaccination encoding TCR V? or V? region in normal mice.

ObjectiveTo investigate the effects and molecular mechanism of endoplasmic reticalam stress (ERS) on albumin-induced apoptosis in renal proximal tubular cells (HKCs). MethodsWestern blot was performed to detect the relationship of the expression of glucose-regulated protein 78 (GRF78) and CCAAT/enhancer-binding protein-homologous protein (CHOP) with the action time and concentration of haman serum albumin (HSA). Expression levels of CHPO mRNA and protein in HKCs after CHOP siRNA transfection were examined by real-time fluorescence quantitative PCR and Western blot respectively. Annexin-V-FITC and PI doable staining cytometry was used to detect the apoptosis of HKCs induced by HSA and influenced by CHOP siRNA. Results(1)After HKCs were stimulatde by 0, 5, 10, 20 g/L albumin for 24 hours respectively, the expression of GRP78, CHOP and HKCs apoptosis were increased with the albumin concentration (P<0.01). After HKCs were stimulated by 20 g/L albumin for 0, 6, 12, 24, 36 hours respectively, the expression of GRP78 was up-regulated at 6-hour, while CHOP and HKCs apoptosis were increased at 12-hour, and significant differences were found among groups (P<0.01). (2) CHOP siRNA significantly inhibited albumin-induced HKC CHOP mRNA and protein expression, as well as HKC apoptosis (P<0.01). ConclusionsRenal tubular cells exposed to high protein load result in EBS. ERS may subsequently lead to tubular damage by activation of pro-apoptosis factor CHOP.

Objective To investigate the predictive value of coagulation state on the occurrence of no-reflow phenomenon after primary percutaneous coronary intervention (PCI) in patients with acute myocardial infarction (AMI). Methods A total of 187 consecutive patients with the first AMI underwent PCI within 12h post-onset of symptom. The clinical features and angiographic findings were collected. According to the thrombolysis in myocardial infarction (TIMI) flow grade with related artery and myocardial blush grade(MBG), the patients were divided into the no-reflow group (TIMI ≤ 2, or MBG ≤ 1) and the normal reflow group. Blood samples were taken immediately on admission before coronary angiography. The levels of plasma von Willebrand factor(vWF), P-selectin(Ps) and Tissue factor(TF) were measured by enzyme-linked immunosorbent assay. Results 23.5%patients of 187 patients developed the no-reflow phenomenon. The plasma level of vWF and Ps and TF were (4 574 ± 1 677) U/L and (16.8 ± 5.1) ng/mL and (283 ± 81) ng/L in the no-reflow group, and (4 074 ± 1 063) U/L and (14.8 ± 4.2) ng/mL and (254 ± 54) ng/L in the normal group, with significant differences (P = 0.020, 0.010 and 0.007, respectively). The hypercoagulation patients in the no-reflow group were much more than patients in the normal reflow group (P = 0.003). Multivariate stepwise logistic regression analysis revealed that hypercoagulation was independent predictor of no-reflow phenomenon ( OR = 2 . 361 , 95%CI 1 . 083 ~ 5 . 148 , P = 0.031). Conclusion The high levels of plasma vWF, Ps and TF present the evidences of hypercoagulation, which might imply the development of no-reflow after PCI.

Objective To investigate the methylation status of PCDH8 gene in pancreatic carcinoma.Methods Methylation of PCDH8 gene in 2 samples of normal pancreatic tissues and 6 pancreatic carcinoma cell lines (PANC1, ASPC1, BxPC3, CFPAC, PaTu8988 and SW1990) was detected by the methylationspecific PCR (MSP) method. The expression of PCDH8 mRNA was detected with 5-Aza-2-deoxycytidine (5-Aza-dC) treatment, a kind of DNA methyltransferase (DNMT) inhibitor in 6 pancreatic carcinoma cell lines by real-time-PCR. Results The methylation of PCDH8 gene was not detected in normal tissues, while it was partially methylated in PANC1, BxPC3, CFPAC and it was totally methylated in PaTu8988, ASPC1, SW1990.PCDH8 mRNA was expressed in PANC1, SW1990, PaTu8988 and the relative quantities of mRNA expression (RQ) were 1.576 ± 0.648, 0.013 ± 0.008, 0.002 ± 0.001; PCDH8 mRNA was not expressed in BxPC3,CFPAC, ASPC1. After 5-Aza-dC treatment, PCDH8 mRNA was expressed in PANC1, ASPC1, BxPC3,CFPAC, PaTu8988, SW1990 and the relative quantities of mRNA expression all significantly increased, and they were 7. 463 ± 2.628, 10. 696 ± 1.539, 7.852 ± 2.762,421.815 ± 1.493, 118.595 ± 4.089, 6.690 ±1.884. Conclusions The methylation of PCDH8 gene may be the major mechanism of down-regulated expression of PCDH8 gene in pancreatic carcinoma.

Objective To investigate the RADIL mRNA expression in pancreatic carcinoma and to evaluate its clinical significance.Methods Fluoesecent quantitative PCR (FQ-PCR) was used to detect the RADIL mRNA expression in 40 patients with pancreatic carcinoma and adjacent tissue and in 5 healthy adult with normal pancreatic tissue and to observe its relationship with clinicopathologic parameters.Results RADIL mRNA was expressed in pancreatic carcinoma and adjacent tissue, as well as normal pancreatic tissue, and the relative expression was 2.263 ± 3.826, 5.425 ± 8.858 and 8.559 ± 4.214, respectively.There was statistically significant difference among the three groups (P ＜0.05 ).RADIL mRNA expression was closely related with the metastasis and differentiation grade ( r = -0.312 and -0.294, P ＜ 0.05 ), however, it was not significantly related to tumor site, tumor size, CA19-9, TNM staging, sex and age.Conclusions RADIL gene may have an inhibitory effect on the pancreatic cancer.

Objective To review our single institutional 10-year experience in complex chest wall reconstruction and identify a working algorithm based on our retrospective analysis.Methods A retrospective analysis of 87 patients who underwent chest wallreconstruction in our department from January 2005 to December 2015.Fifty female patients and 37 male patients who underwent the above procedure were reviewed retrospectively.The median age of the patients is 52.3 years (24-75years).Histologic diagnosis including squamous-cell carcinoma (n =10),soft tissue sarcoma(n =22),chondrosarcomas(n =13) and metastasis from breast cancer(n =42).Type of skeletal defect including partial ribs/sternum defects in 19 cases,soft tissue defects alone in 33 cases,complicated composite chest wall defects involving multiple layers(soft tissue,ribs/sternum,and intrathoracic organs) in 35 cases.Sole methylmethacrylate/polypropylene mesh was used for small sized rib defects in 26cases.Methylmethacrylate/polypropylene mesh sandwich prostheses was used in 28 cases with extensive skeletal reconstruction after partial sternectomy and rib resection.The chest wall defects were repaired with pedicled internal mammary artery perforator flap(3 cases),pedicled deep superior epigastric artery perforator flap(4 cases),pedicled pectoralis major flap(8 cases),free anterolateral thigh perforator flap(9 cases),free deep inferior epigastric artery perforator flap(17 cases),pedicled lateral thoracic flap(5 cases),pedicled latissimus dorsi flap(17 cases),pedicled rectus abdominis flap(15 cases),free deep inferior epigastric artery perforator flap combined with pedicled rectus abdominis flap (4 cases),pedicled bipaddled latissimus dorsi flap(5 cases).11 cases with extensive full-thickness defects of the chest wall,the skeletal reconstruction was achieved with prosthetic sandwich and then covered with the omental flap,further free flaps were harvested for skin and soft tissue repairing.Results 1 case with pedicled rectus abdominis flap partial necrosis was noted,free anterolateral thigh flap was used for repairmen after further revision.1 case with edicled bipaddled latissimus dorsi flap,necrosis of the distal 1/4 part of one paddle was noted,healed with dressing therapy,no secondary skin grafting was required.Postoperative venous congestion occurred in 2 cases with deep inferior epigastric artery flap transplantation,in which both skin flaps exhibited venous crisis within 24 h after surgery.The reexploration procedures were successful in both cases and the flaps survived totally.All other flaps survived.The mean follow-up was 31 months,ranged from 9 to 72 months.No tumour extirpation was noted,functional and appearance results were satisfied.Conclusion According to the size and location of chest wall defect,different pedicled and free flaps should be chosen to achieve optimal outcome.Free flaps are efficient for large complex chest defects reconstruction.

Objective To study the incidence and duration of residual dizziness after successful repositioning treatment in patients with benign paroxysmal positional vertigo(BPPV) and the clinical factors associated with the residual dizziness.Methods A total of 202 cases of confirmed BPPV patients,61 males and 141 females with the average age of 54.78± 13.71 years old,were followed up for 2 months after successful particle repositioning.The incidence and duration of residual dizziness were analyzed.The risk factors for residual dizziness were analyzed by logistic regression.Results A total of 202 cases of confirmed BPPV were included in this study,and 113 cases complained of residual dizziness.Over the time,residual dizziness disappeared gradually.The differences of the age,the duration of vertigo before treatment,recurrent,and underlying diseases between the two group were significant (P ＜0.05),while the side,the gender,the incubation period of BPPV,the duration time of BPPV,and the types of canals were not associated with the residual dizziness(P＞0.05).The logistic regression analysis showed that the duration of vertigo before treatment and the age were the risk factors for residual dizziness.Conclusion More than half of the patients included in this study complained of residual dizziness after particle repositioning,and the symptoms disappeared naturally.The duration of vertigo before treatment and the age were the risk factors for residual dizziness.