Objective Effects of different factors on shoot tip culture in vitro were studied by using Dioscorea bulbifera as test material to find the media suitable to the shoot tip differentiation of D.bulbifera and the method of virus delection.Methods Plant tissue culture method was used in shoot tip culture and the symptom observation method,instruction plant method,and RT-PCR method were used in virus detection of virus-free plantlets.Results The best disinfection method for D.bulbifera shoot tips was firstly disinfected for 30 s with 70% alcohol and then disinfected for 12 min with 0.1% HgCl2;It is better for D.bulbifera to cut shoot tips in 0.5 mm length after 37 ℃ heat treatment for 7 d;The best proliferation medium of D.bulbifera shoot tips was MS+KT 2 mg/L+NAA 0.5 mg/L;The best rooting medium of regeneration buds from D.bulbifera shoot tips was 1/2 MS+NAA 0.5 mg/L;The best matrix of regeneration plantlets from D.bulbifera shoot tips was perlite-vermiculite (2∶1);RT-PCR Method was primary mean to detect potato virus Y (PVY) of regeneration plantlets from D.bulbifera shoot tips,and average rate of PVY-free was 86.5% by RT-PCR.Conclusion The virus-free culture system of D.bulbifera shoot tips is established for the first time,providing a technological basis for the rapid propagation and factory production of D.bulbifera virus-free plantlets.

Objective Effects of different factors on proliferation and rooting of stems with a bud were studied by using Dioscorea bulbifera as test material to optimize the rapid propagation system of D.bulbifera virus-free plantlets.Methods Plant tissue culture method was used in shoot tip culture and rapid propagation study,and RT-PCR method was used in virus detection of virus-free plantlets.Results The best proliferation medium of D.bulbifera stems with a bud was MS+KT 2 mg/L+6-BA 1 mg/L+NAA 0.5 mg/L;The best sucrose and agar concentration of D.bulbifera stems with a bud was 30 g/L and 0 g/L,respectively;The best rooting medium of D.bulbifera stems with a bud was 1/2 MS+IBA 0.1 mg/L+NAA 0.5 mg/L+PP_(333) 1 mg/L;The best transplanting matrix of regeneration plantlets from D. bulbifera stems with a bud was perlite-vermiculite(2:1);The best PP_(333) concentration of D.bulbifera regeneration plantlets for transplanting was 50 mg/L.Conclusion The rapid propagation system of D. bulbifera virus-free plantlets is established successfully for the first time,which provides a technological basis for factory production of D.bulbifera virus-free plantlets.

Objective To study the effects of several factors on bud proliferation and rooting of Dioscorea bulbifera stem with a bud.Methods Single factor test and plant tissue culture methods were applied.Results The combination of 6-BA or KT and NAA helped the proliferation of stem with a bud in D.bulbifera;high concentration sucrose led to callus growth of stem with a bud in D.bulbifera which didn′t help the proliferation of stem with a bud in D.bulbifera;Liquid culture also helped the proliferation of stem with a bud in D.bulbifera;In a certain range,the increase of NAA concentration helped the rooting of stem with a bud in D.bulbifera.But it will inhibit the rooting with concentration in a higher level.Conclusion The best proliferation culture medium of D.bulbifera,stem with a bud is MS + KT 2 mg/L +NAA 0.5 mg/L or MS + 6-BA 2 mg/L + NAA 0.5 mg/L;For proliferation of D.bulbifera stem with a bud,the best sucrose concentration is 30 g/L sucrose,the best culture mode is liquid culture;The best rooting culture medium of D.bulbifera stem with a bud is MS + NAA 2 mg/L.

Purpose To detect high mobility group protein A2 (HMGA2) expression in breast cancer, and to analyze its relationship with clinicopathological features and the levels of HMGA2 in different molecular subtypes of breast carcinomas. Methods An immu-nohistochemical study was undertaken for measuring the levels of HMGA2 in 58 breast carcinomas. Results ( 1 ) The expression of HMGA2 was 0, 62. 5%, 60. 0%, 100. 0% and 80. 0% in Lum A, Lum B, HER-2-OE, basal-like breast carcinoma (BLBC) and un-classification phenotype respectively ( triple negative breast cancer, 92. 3%) . High expression of HMGA2 was associated with the tri-ple-negative breast cancer (TNBC) subtypes (P<0. 01). (2) An association was identified between high expression of HMGA2, and high tumor grade, lymph node metastasis (P<0. 05), positive Ki-67, CK5/6 and EGFR (P<0. 05), negative ER and PR (P<0. 01), but no association was observed for tumor size and patients’age. Conclusion An association is identified between high ex-pression, and high tumor grade, lymph node metastasis, positive Ki-67, EGFR and CK5/6, negative ER and PR, that means a high expression of HMGA2 is associated with an adverse outcome in breast cancer. High expression of HMGA2 are associated with the TNBC subtypes. Thus recognizing HMGA2 as a rational target in TNBC. The results of the study have implications for therapeutic target iden-tification and the design of future clinical trials for TNBC.

Objective To identity whether there is muscle atrophy phenomenon in end-stage kidney disease patients and to detect the level of transcription factor Foxo1 and the activity of ubiquitin-proteasome system.Methods Twenty-two patients in chronic kidney disease (CKD) stage 5 were selected and their mean muscle cross sectional area was measured.mRNA and protein levels of Foxo1,Atrogin-1,MuRF1 in rectus abdominis biopsies obtained from consecutive patients were detected.Control biopsies were obtained from 8 healthy subjects during elective surgery for abdominal wall hernias and 6 subjects during elective surgery for adenomyosis.Results Compared with the control group,cross sectional area of muscle fibers decreased and the transcription and protein levels of Foxo1,Atrogin-1,MuRF1 were upregulated in CKD group(P＜0.05).Protein level of p-Foxo1 decreased in CKD group(P＜0.05).Conclusion There exist muscle atrophy phenomenon in CKD patients,which may associate with the upregulation of Foxo1 and activation of ubiquitin-proteasome system.

Objective To identify the imbalance of T cell-specific transcription factors T-bet/GATA-3,and to explore the modulation with dexamethasone and imiquimod in CD4+T cells from ovalbumin (OVA)sensitized mice.Methods CD4+T cells were obtained fromsingled-cell suspension of spleen(after lysis of RBC).ELISA assay was used to detect the concentrations of IL-4,IL-5 and IFN-γin superna tants and cell pellets,and the expression of T-bet and GATA-3 was detected by Western blot.Resuits In the control group,tIle low levels of IFN-γ were detected in the supernatants during 24 h.In OVA treatment group,the concentrations of IL-4,IL-5 were increased significantly,and the concentrations of IFN-γ were always low in the supernatants.In the dexamethasone treatment group,the concentrations of IFN-γ,IL-4 and IL-5 were all low in the supernatants during 24 h.In the imiquimod treatment group,the concentrations of IFN-γ were increased significantly,and the concentrations of IL-4 and IL-5 were decreased in the super natants.It worked at 6 h,and achieved the peak at 12 h,lasted over 24 h.In the control group,the expres sions of T-bet and GATA-3 were detected in CD4+T cells during 24 h.In OVA treatment group,the expressions of T-bet were decreased,and that of GATA-3 were increased rapidly in CD4+T cells.In dexam ethasone treatment group,the expressions of T-bet were always low in CD4+T cells,and that ofGATA-3 were no change during 24 h.In imiquimod treatment group,the expressions of T-bet were increased,andthat of GATA-3 were decreased in CD4+T cells.The protein expressions worked at 6 h.and achieved the peak at 12 h,lasted over 24 h.Conclusion The imbalance T cell-specific transcription factors T-bet/GA-TA-3 contributes to both high expression of GATA-3 and low expression of T-bet in CD4+T cells from OVA sensitized mice.Dexamethasone treatment inhibits the expression of T-bet in CD4+T cells and has no func tion in GATA-3.Imiquimod treatment modulates key master switches GATA-3 and T-bet that results in com mitting T helper cell to a TH 1 phenotype and imiquimod may play a key role in the regulation of TH2 cytokine responses in asthma.

Objective To study the transcriptional regulation of vp1667 by H-NS in Vibrio parahaemolyticus.Methods Total RNAs were extracted from Δhns and WT strains.Quantitative RT-PCR was carried out to calculate the transcriptional variation of vp1667 between Δhns and WT.Primer extension assay was also employed to detect the transcription start site and the promoter activity (i.e.the amount of primer extension products) of vp1667 in Δhns and that in WT.The promoter DNA region of vp1667 was amplified, purified, and cloned into the corresponding restriction endonuclease sites of pHRP309 that harbors a gentamicin resistance gene and a promoterless lacZ reporter gene.The recombinant pHRP309 plasmid was transformed into Δhns and WT, respectively, while β-galactosidase activity in cellular extracts was measured using a β-galactosidase enzyme assay system.The over-expressed His-H-NS was purified under native conditions with nickel loaded HiTrap Chelating Sepharose columns.The electrophoretic mobility shift assay (EMSA) and DNaseⅠ footprinting were then applied to analyze the DNA-binding activity of His-H-NS to vp1667 promoter region in vitro.Results and Conclusion The primer extension assay detected one transcription start site for vp1667, which was located at 28 bp upstream of vp1667, and its transcribed activity was under the negative control of the H-NS.The EMSA and DNaseⅠ footprinting assay results showed that His-H-NS was unable to bind to the promoter-proximal DNA region of vp1667, suggesting that H-NS indirectly inhibits the transcription of vp1667.

Purpose To study the levels and subcellular localization of β-catenin in 5 different molecular subtypes of breast carcino-mas. Methods An immunohistochemical study was undertaken for measuring the levels and subcellular localization ofβ-catenin in 58 breast carcinomas. Results ( 1 ) The cytoplasmic expression of β-catenin was 21. 1%, 50%, 60%, 100% and 60% ( TNBC 84. 6%) in Lumina A, Lumina B, HER-2-OE, basal-like breast carcinoma ( BLBC) and uncl phenotype respectively. High cytoplas-mic expression was associated with the BLBC and TNBC subtypes ( P<0. 05 ) . ( 2 ) An association was identified between high cyto-plasmic expression of β-catenin, and high tumor grade (P<0. 01), abnormal E-cadherin, positive Ki-67 and CK5/6 (P<0. 05), negative ER and PR (P<0. 01), but no association was observed for lymph node metastasis, tumor size and patients’age. Conclu-sion An association is identified between high cytoplasmic expression, and high tumor grade, positive Ki-67 and CK5/6, negative ER and PR, that means a high cytoplasmic expression ofβ-catenin is associated with an adverse outcome in breast cancer. High cytoplas-mic expression are associated with the BLBC and TNBC subtypes whcih recognizing Wnt signaling as a rational target in TNBC and BLBC. The results of the study have implications for therapeutic target identification and the design of future clinical trials for TNBC and BLBC.

Objective To compare the effect of video‐assisted thoracoscopic(VAT) surgery and conventional thoracotomy in emergency treatment of multiple rib fracture complicating pulmonary laceration to provide the reference for clinical treatment .Meth‐ods Forty‐seven cases of multiple rib fracture complicating pulmonary laceration in our hospital from April 2013 to April 2014 were selected and divided into the VAT group(n=32) and thoracotomy group(n=15) according to the willingness of patients .The two groups performed the thoracoscopic and traditional thoracotomy titanium nickel alloy rib plate treatment respectively .The sur‐gery situation ,complications and changes of perioperative blood gas levels were compared between the two groups .Results The op‐eration time ,intraoperative bleeding volume ,ICU hospitalization time ,total hospitalization time and postoperative analgesic in the VAT group were lower than those in the thoracotomy group ,the differences were statistically significant (P<0 .05);the VAS score on postoperative 1 d had statistical difference(P<0 .01) .Compared with before operation ,arterial PaO2 ,SaO2 and PaO2/FiO2 at postoperative 12 h in the two groups were increased ,while PaCO2 was decreased ,and the differences were statistically significant (P<0 .05);PaO2 ,SaO2 and PaO2/FiO2 at postoperative 12 h in the VAT group were higher than those in the thoracotomy group , while PaCO2 was lower than that in the thoracotomy group ,and the differences were statistically significant (P<0 .05) .The occur‐rence rate of complications had no statistical difference between the VAT group and thoracotomy group (3 .1% vs .6 .7% ,P>0 .05) .The excellent rate in the VAT group was 90 .6% ,which was higher than 66 .7% in the thoracotomy group ,and the differ‐ence was statistically significant (P<0 .05) .Conclusion Thoracoscopic internal fixation for the treatment of multiple rib fractures complicating laceration has the advantages of minimal trauma ,convenient operation and high safety ,could effectively alleviate the patient′s sufferring ,improve the living quality ,and be a better way of treatment .