Objective To observe and analyze the clinical efficacy of docetaxel and S-1 in treatment of anthracycline-resistant triple-negtive breast cancer(TNBC).Methods 64 cases with TNBC admitted in People' Hospital of Shaoxing City from June 2009 to June 2011were selected as research object.The clinical data of these cases were analyzed retrospectively.Anthracycline had been used to treat the cases, but with no effect or recurrence.Then docetaxel combined with S-1 was applied to the cases.Clinical efficacy and adverse reactions of the chemotherapy were observed and analyzed. Results After treatment,the efficiency of 64 cases was 54.69%,and the disease control rate was 79.69%.During the treatment,the main adverse reactions were gastrointestinal reactions and bone marrow suppression,without death,and patients could tolerate the adverse event.Follow-up results showed that the median time of progression was 10.5 months.After two years,progression-free survival rate was 0.Conclusion Docetaxel combined with S-1 has good clinical efficacy for anthracycline-resistant triple-negtive breast cancer,with relatively mild side effects,which may be an ideal adjuvant chemotherapy method.

Objective To investigate the involvement of apoptosis in the lesions of patients with psoriasis. Methods The apoptosis was detected with terminal deoxynucleotidyl transferase mediated dUTP biotin nick end labeling (TUNEL), and the expression of p53, PCNA and apoptosis suppressing protein bcl 2 was assessed with immunoperoxidase technique in psoriatic lesions and normal skin. Results A large number of keratinocytes showing biochemical and morphologic features of cells undergoing apoptosis were observed in all the suprabasal layers of the psoriatic epidermis. The plaques from all patients analysed showed marked increase in the number of PCNA positive cells in the middle and basal keratinocytes, and a dramatic reduction in the number of bcl 2 positive cells in the basal cell layer. Conclusion The increased apoptosis of keratinocytes in the lesions of psoriasis might be a homeostatic mechanism to the hyperplasia of cells.

Objective To study the etiology of functional dyspepsia (FD) and compare the clincial responses of medical treatment and psychological intervention.Methods Psychological intervention (psychologic support),advices for daily life (eating habits) and medical treatment were administered to the 346 FD patients;and the therapeutic effects were compared.Results Complete remission:225 cases (65%);partial remission:52 cases (15%);no effect:69 cases (20%).The overall effective rate was 80%.Conclusion FD is a clinical syndrome focusing in upper abdomen,but without local or systematic evidence Psychological treatment is stressed.

Objective:To clone the single chain variable Fv of anti-idiotype monoclonal antibody against vibrio alginolyticus.Methods:Total RNA was extracted from hybridoma cell 2F4 secreting MAb against vibrio alginolyticus and cDNA was amplified by retropolymerase chain reaction(RT-PCR),the expression vector pTAT-AL1 was constructed for the recombinant V_H-V_L expression.The transformed E.coli BL21 cells were propagated and induced by IPTG.Results:The V_H gene contained 369 base pairs and encoded 123 amine acid residues;The V_L gene contained 339 base pairs and encoded 113 amine acid residues;There were four FRs three CDRs and two characteristic cysteine residues in the V_H and V_L gene,respectively.ELISA results showed the ScFv retained almost the same antigen affinity and specificity as its parent monoclonal anitbody.Conclusion:The single chain variable Fv of anti-idiotype monoclonal antibody against vibrio alginolyticus was constructed successfully and expression products was found in the periplasmic space and inclusion bodies.This ScFv might be a new generation of gene engineering vaccine of the anti-idiotype monoclonal antibody against vibrio alginolyticus in fishery.

Objective To construct the pCDNA3.1-ING4 eukaryotic expression vector and investigate its effect on the proliferation of melanoma cell line M14. Methods The targeted cDNA fragment encoding ING4 was cloned by reverse transcription-PCR with normal gastric mucosa from patients with gastric ulcer, and subcloned into eukaryotie expression vector pcDNA3.1. PCR and DNA sequencing were performed to identify the eukaryotic expression vector pCDNA3.1-ING4, which was then transfected into M14 cells with Lipofectamine 2000 reagent. The expression of ING4 was detected in untransfected, pCDNA3. 1-ING4-transfected and pcDNA3.1-transfected M14 cells by Western blot and immunocyto-chemistry, and cell proliferation by MTT assay. Results A fragment of expected size (750 bp) was amplified by PCR analysis, and DNA sequencing confirmed the correctness of the recombinant plasmid. As shown by immunocytochemistry, the percentage of cells positive for ING4 protein was significantly higher in pCD NA3. 1-ING4-transfected MI4 cells than in non-transfected M14 cells and pcDNA3.1-transfected M14 cells (71.80%±9.88% vs 4.20%±3.35%, P < 0.01). Western blot also revealed an increased expression of ING4 in pCDNA3.1-ING4-transfected cells. Decreased cell viability was observed in ING4-transfected cells compared with nontransfected cells and pCDNA3.1-transfected cells (both P < 0.01). Conclusions The pCDNA3. 1-ING4 eukaryotic expression vector has been constructed successfully, and M14 cells transfected by this recombinant plasmid could effectively express ING4 protein, which may inhibit the cell proliferation of M14 cells.

Objective:To detect genetic mutations in a case of severe epidermolysis bullosa simplex.Methods:Clinical data and peripheral blood samples were collected from the patient and her parents, and genomic DNA was extracted. Whole exome sequencing was performed to identify causative gene mutations in the patient, and then Sanger sequencing to verify the mutations among the family members.Results:A heterozygous mutation c.1429G>A at position 1429 in exon 7 of the KRT5 gene was identified in the patient, which led to the substitution of glutamic acid by lysine at amino acid position 477 (p.Glu477Lys) of keratin 5 encoded by the KRT5 gene. The mutation was not detected in her unaffected parents.Conclusion:A causative mutation c.1429G>A (p.Glu477Lys) in the KRT5 gene was identified in the patient with severe epidermolysis bullosa simplex, which was a de novo mutation.

Objective:To increase the solubility of the campher and oleum anisi stellati in the syrup of cough and reduce the precipitation of syrup.Methods: The technology of ? cyclodextrin inclusion was used in the experiment. Results: The experiment shows the method may meet the demand of production. Conclusions: The method is very good in the production of cough syrup.

Objective To discuss the clinical effects of treatment of femoral intertrochanteric fracture by DHS internal fixation. Methods 42 cases with femoral intertrochanteric fracture were treated by DHS internal fixation. According to AO classification, there were 16 cases of type A1,14 A2,and 12 A3. Results 36 patients were followed up for 6 to 18 months, with an average of 10 months. Their results were satisfactory. Conclusion Treatment of femoral intertrochanteric fractures with DHS internal fixation is an effective method.

Objective To develop an effective and broad immune protective vaccination strategy by using DNA and recombinant vaccinia-based H5N1 vaccines.Methods BALB/c mice were immunized with various prime-boost regimens by using different DNA ( pIRES-based or pVRC-based) and recombinant vaccinia (Tiantan strain, rTTV)-based H5N1 vaccines expressing multivalent antigens (HA, NA, M1 and M2).The differences of immunity induced by two DNA vaccines were compared between intradermal electro -poration (IDE) and intramuscular electroporation (IME) deliveries.Immune responses were analyzed by hemagglutination inhibition( HAI) assay, neuraminidase ( NA)-specific antibody measured by ELISA , mi-croneutralization assay and IFN-γELISPOT assay .Results High levels of humoral immunity and T cell re -sponses were induced in mice primed with DNA-based vaccine than those primed with rTTVb-ased vaccine . DNA priming by IDE resulted in higher levels of neutralizing antibody in mice than those by IME delivery . Higher levels of HAI and anti-NA antibodies as well as NA-specific T cell responses were induced by pVRC-based DNA prime than those by pIRES-based DNA prime .HA-specific T cell responses were also enhanced in mice primed with pVRC-based DNA than those primed with pIRES-based DNA by IME .Conclusion The prime-boost strategies by using DNA-based vaccine in combination with rTTV-based H5N1 vaccine could induce humoral and T cell responses in mice against multi-antigens .Immunities induced by vaccines in com-bination might be modulated by various prime regimes .The study provided references for the further develop-ment of novel H5N1 vaccine and the optimization of immunization programs of combined multi-antigen vac-cine candidates .

ObjectiveTo identify the presence of side population (SP) cells in human ovarian cancer cell line OVCAR-3 and to investigate whether SP cells have the characteristics of cancer stem cells.MethodsSP and non-SP (NSP) cells from OVCAR-3 were isolated by fluorescence-activated cell sorting after being stained by DNA-binding dye Hoechst 33342.Limiting dilution transplantation assay,realtime PCR,and drug sensitivity assay were performed to compare the tumorigenic ability,differentiation ability in vivo,the mRNA expressiou of stemness marker (Oct-4,Klf4,and Nanog) and ATP-binding cassette (ABC) transporter (ABCG2,ABCB1,and ABCC2),and response to multiple drugs (cisplatin,paclitaxel,doxorubicin,and mitoxantrone )between SP and NSP cells.ResultsA few of SP cells [ ( 1.13 ±0.39) ％ ] which were sensitive to reserpine were identified in OVCAR-3 cells.The injection of as few as 102 SP cells initiated tumors in two of five mice.Tumor latency was 52 -61 days.However,the NSP cells did not generate any tumors in mice until 104 NSP cells were injected (two of five mice).Tumor latency was 64 - 98 days.Tumorigenicity of SP cells was enhanced by at least 100-fold than that of NSP cells.The SP cells regenerated both SP [ ( 2.09 ± 0.73 ) ％ ] and NSP populations in vivo with a fraction size that was comparable to the original population.The mRNA expression ofstemness genes Oct-4,Klf4 and ABC transporters ABCG2,ABCC2 genes were elevated in SP cells compared to NSP cells,the fold changes were 1.95±0.41 (P＜0.05),4.26 ±0.63 (P＜0.01),3.22±0.36 (P＜0.01),and 1.76±0.26 (P＜0.01 ),respectively.The relative activity of SP and NSP cells were 0.757 ± 0.105 versus 0.474 ± 0.035 (P＜0.01),0.521 ±0.092 versus 0.384 ±0.073 (P＜0.05),0.742 ±0.051 versus 0.526 ±0.088 (P ＜0.01 ),and 0.690 ± 0.096 versus 0.466 ± 0.112 ( P ＜ 0.01 ) when they exposed to 0.25 μg/ml cisplatin, 0.01μmol/Lpaclitaxel, 0.25μmol/Ldoxorubicin, and0.05μg/mlmitoxantrone,respectively.ConclusionsSP cells from OVCAR-3 have enhanced self-renewal,differentiation,and tumorinitiating capacity compared to NSP cells.The mRNA expression of stemness genes and ABC transporters are markedly elevated in SP cells,which showed resistance to multiple chemotherapeutic drugs and have characteristics of cancer stem-like cells.Therefore,SP phenotype could be used as a marker to isolate the cancer stem-like cells in ovarian cancer.