Approximately 70％ advanced breast cancer will develop bone metastases.Bone metastases can cause a series of complications such as skeletal-related events,seriously affecting the quality of life of the patients.Therefore,it is very important to choose the reasonable and effective treatments.Bisphosphonates are the standard treatments of bone metastatic breast cancer,denosumab may also be a reasonable alternative to bisphosphonates in the near future,while chemotherapy,endocrinotherapy and targeted therapy are the basic treatments of advanced breast cancer,combined use of various treatments will get the best efficacy.

Objective To explore the cytotoxicity of the cytotoxic T lymphocyte (CTL) induced by SW480 sonicate sensitized dendritic cells (DC) on the colon cancer cell line SW480. Methods PBMC were separated from the HLA-A*0201 donor and DC were cultured with rhGM-CSF, rhIL-4 and rhTNF-α. The same donor's primary CTL were stimulated by DC loaded with SW480 sonicate. The cytotoxicity of CTL on SW480 (HLA-A*0201 positive) and K562 (HLA-A*0201 negative) was determined by the MTT method. Results The cytotoxicity of the CTL on SW480 was stronger than that on K562 (P ＜0.05). Conclusion The DC vaccine can stimulate specific CTL which can trigger cytotoxic activity on the target cells and this cytotoxicity is related to MHC restriction.

Objective To investigate the effects of DRD1 rs265981,rs4532,rs686 and rs265976 polymorphisms on response to clozapine in resistant schizophrenic patients.Methods DRD1 genotype was determined by SNaPshot SNP technique for 154 patients with resistant schizophrenia.Clinical symptoms were evaluated by Positive and Negative Syndrome Scale (PANSS),and the responder was defined as a reduction of 50％ on PANSS score from baseline after the patients were administered orally clozapine for 8 weeks.Results The frequencies of rs265981 genotypes,alleles and rs265976 genotypes had significant differences between clozapine responder group (88 cases) and nonresponder group(66 cases) in total clinical efficacy ( x2 =10.215,P =0.004 ; x2 =4.082,P =0.041 ; x2 =14.083,P =0.007 ).The frequencies of rs265976 genotypes had significant differences between clozapine response (58 cases) and nonresponse (96 cases) to negative symptom ( x2 =9.805,P =0.046).Conclusion The polymorphisms of DRD1 gene rs265981 and rs265976 may relate with clinical response to clozapine in resistant schizophrenias.rs265981 T/T,allele T and rs265976 genotype A/A are likely to be predictive factors to the improvement of total clozapine therapeutic effects.rs265976 genotype A/A are likely to be predictive factors of negative symptom with treatment of clozapine.

Objective To quantify the expression levels of calpain 1 and caspase-3 in lichen planus (LP) lesions and their relationship with the apoptosis in keratinocytes.Methods Biopsy samples were obtained from the lesions of 20 patients with LP and normal skin of 10 healthy controls,and embedded in paraffin.Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) was used to evaluate the apoptosis in keratinocytes,and immunohistochemical staining to detect the expressions of calpain 1 and caspase-3,in these tissue specimens.Data were processed by SPSS 13.0 software.Comparison analysis was carried out by t test for apoptosis index,and by rank sum test for the expressions of calpain 1 and caspase-3.Spearman correlation analysis was conducted to evaluate the relationship between these parameters.Results The apoptosis index of keratinocytes was higher in LP lesions than in the normal skin (67.59 ± 13.50 vs.28.26 ± 7.56,t =8.52,P ＜ 0.01).Significantly increased expressions of calpain 1 and caspase-3 were observed in the epidermis of LP lesions compared with the normal skin (T =78.00 and 77.00,respectively,both P ＜ 0.01).The expressions of both calpain 1 and caspase-3 were positively correlated with apoptosis index of keratinocytes (r =0.71 and 0.74,respectively,both P ＜ 0.01).Conclusions The expressions of calpain 1 and caspase-3 are upregulated in LP lesions,which may be closely associated with the accelerated apoptosis in keratinocytes.

Objective To analyse the problems presented in external quality assessment (EQA)for blood cytomorphology and propose measures for improvement.Methods Statistical analysis was performed on EQA results for blood cytomorphology from 2011 to 2013 in Shaanxi Province,including general information of participant laboratories,coincident rate,and reported incorrect results.EQA for blood cytomorphology was held two times every year in Shaanxi province,with ten pictures inclu-ding bone marrow and peripheral blood smear each time.The compact disc including twenty pictures was given to partici-pants by EMS.Participants reported two EQA results in April and September each year.The center statisticed the EQA re-sults and provided the EQA reports to every participants in June and November.Results Participating laboratories increased from 76 in 2011 to 163 in 2013.The ratio of laboratories with the coincidence rate≥80% was 80%,47%,44%,55%,77%and 96% respectively.The number for single cell with the coincidence rate≥80% was 38.The coincidence rate of peripheral blood cells was higher than that of the bone marrow on the whole.Causes of incorrect results included cell lines’misclassifi-cation,growth stage’s misclassification,insufficient identification of abnormal cytomorphology,and so on.Conclusion The identification of blood cytomorphology was unbalanced in different leveled hospitals in Shaanxi Province.To develop EQA of blood cytomorphology definitely has a positive role in improving the experimenters’skill of identifying cytomorphology.

Objective To investigate the risk factors of intrauterine hepatitis B virus (HBV)infection and its preventive measures.Methods 254 cases of pregnant women with positive HBsAg and their neonatuses from Shaanxi People’s Hospi-tal were selected as the research object.The serum of pregnant women were tested HBV markers and HBV-DNA,their neo-natal umbilical cord blood were only detected HBV markers.Results In 254 samples,the positive rates of HBsAg and HBeAg within their neonatal umbilical cord blood were 5.12% and 13.78% respectively;the positive rate of HBV infection in neonatal umbilical cord blood among maternal HBeAg-positive group was 53.33%,far higher than 3.61% in HBeAg-neg-ative group (χ2 =99.44,P <0.001);the positive rates of HBsAg and HBeAg in neonatal umbilical cord blood were elevated along with the increase of maternal HBV-DNA copy (Hc= 13.50,66.70;P < 0.001).The positive rate of HBV-DNA in pregnant women with HBeAg-positive group was 100%,far more than 19.59% in HBeAg-negative group (P <0.001).Con-clusion Both maternal positive HBeAg and HBV-DNA are risk factors for neonatal intrauterine HBV infection.Prenatal HBeAg screening is predominant,especially in some hospitals without real-time quantitative assay.An effective measure to reduce intrauterine HBV infection is to be pregnant when HBV DNA and HBeAg are turned into negative after HBV inter-vention and treatment for women in reproductive age.

BACKGROUND:Studies have shown that the number of osteoblasts is often decreased after osteoporosis, and osteoblast replacement therapy becomes a new target for the treatment of osteoporosis. OBJECTIVE:To observe the osteogenic differentiation of human bone marrow mesenchymal stem cels cultured in dexamethasone, vitamin C and beta-glycerophosphate. METHODS:Mesenchymal stem cels were isolated and purified from adult bone marrow using human lymphocyte separation medium. The expression of cel surface markers was detected by flow cytometry. Cel ultrastructure was observed by transmission electron microscope. Then, the bone marrow mesenchymal stem cels were cultured in osteogenic induction medium containing dexamethasone, vitamin C andβ-glycerophosphate, and RT-PCR was used to detect the bone morphogenetic protein-2 mRNA expression after osteogenic induction. RESULTS AND CONCLUSION:A large number of adherent cels were visible as fibrous growth at 2 weeks after culture and strongly expressed CD44, CD29, but did not express CD34, CD45. These cels could be induced to differentiate into osteoblasts, and express bone morphogenetic protein-2 mRNA. Alizarin red staining and alkaline phosphatase staining were positive for the cels. These findings suggest that human bone marrow mesenchymal stem cels cultured in dexamethasone, vitamin C and beta-glycerophosphate can differentiate into osteoblasts, and has a potential for the treatment of osteoporosis.

Objective:To study the curative efficacy of budesonide combined with huaiqihuang granules in the treatment of children with asthma and its effects on the serum leukotriene D4(LTD4),nerve growth factor (NGF),tissue inhibitor of metalloproteinase-1 (TIMP-1) levels.Methods:90 cases of children with asthma who were treated from March 2014 to March 2015 in our hospital were selected and divided into the observation group (n=45) and the control group (n=45) according to the random number table.Both groups of children were given bronchial antispasmodic agent,oxygen,antibiotics,glucocorticoid and other conventional treatment after admission,the control group was given budesonide atomization inhalation treatment on the basis of conventional treatment,0.5 mg budesonide were add in 3ml saline for continuous inhalation,twice a day,while the observation group was treated with Huaiqihuang Granules on the basis of the control group,twice a day.After 12 weeks of treatment,the curative effect,disappearance time of wheezing,cough relief,pulmonary rales,wheezing and other symptoms,forced vital capacity (FVC),serum LTD4,NGF,TIMP-1 levels,CD3+,CD4+,CD8+ of both groups were compared.Results:After treatment,the total effective rate of observation group was significantly higher than that of the control group [93.33％(42/45) vs 77.77％(35/45)](P＜0.05);disappearance time of breath,cough,moist rales and wheezes were significantly shorter than those of the control group[(4.32± 1.03)d vs (6.08± 1.24)d,(5.60± 1.12)d vs (7.21± 1.30)d,(3.19± 0.98)d vs (4.98± 1.02)d,(3.25± 1.03)d vs (5.89± 1.35)d](P＜0.05);the FEV1,FEV1/FVC were significantly higher than those of the control group [(92.63± 10.01)L/s vs (78.36± 9.19)L/s,(95.37± 11.72)％ vs (80.19± 10.23)％](P＜0.05);the serum LTD4,NGF,TIMP-1 levels were significantly lower than those of the control group [(7.24± 0.86)ng/mL vs (12.68± 1.01vng/mL,(68.18± 9.01)pg/mL vs (80.78± 10.24)pg/mL,(34.16 ± 5.06)ng/mL vs (49.76 ± 5.47)ng/mL] (P ＜0.05);the CD3+,CD4+ were significantly higher than those of the control group [(66.15± 7.20)％ vs (62.03± 6.85)％,(45.13± 7.90)％ vs (37.42± 7.06)％],CD8+ was significantly lower than that of the control group[(34.16± 5.06)％ vs (49.76± 5.47)％] (P＜0.05).Conclusion:Budesonide combined with Huaiqihuang granules was effective for children with asthma,which could enhance the immune function,improve the lung function,reduce the serum levels of LTD4,NGF,TIMP-1.

Objective To explore the role of miRNA-148a in bladder tumorous development and progression.Methods Ex-pression of miRNA-148a was assessed in 35 bladder carcinoma tissues and 16 non-carcinoma tissues by fluorescence quanti-tative real time PCR,and correlation with clinical features was evaluated.Target genes and transcription factors of miRNA-148a were predicted using bioinformatic analysis,then TF-miRNA-148a-target genes network diagram was built and the tar-get genes was analyzed of gene ontology enrichment and KEGG pathway.Results Expression of miRNA-148a was lower in bladder carcinoma tissues than in non-carcinoma tissues(0.000 8±0.000 2 vs 0.002 1±0.000 5)(t=2.46,P <0.05),but its expression was no statistical significance in different groups of gender,age,pathological classification,clinical stage, lymph node metastasis (P >0.05).268 target genes of miRNA-148a were predicted by three softwares at the same time,60 transcription factors were predicted and the binding sites with combination scroes above 80 was 657.The target genes of miRNA-148a was enriched in many biological processes,such as neuron differentiation,generation of neurons,neuron projec-tion development,cytoplasmic mRNA processing body,cytoplasm(P <0.001).They also participated in p53 signaling path-way,proteoglycans in cancer-homo sapiens,pathways in cancer,prostate cancer,protein processing in endoplasmic reticulum, focal adhesion and so on(P <0.05).According to TF-miRNA-148a-target genes network diagram,miRNA-148a was regula-ted by SP1,ESR1,AP1,MYC and BRCA1,genes of IGF1,P27kip1 ,NCOA1,PTEN,SERPINE1 might be regulated by miR-NA-148a.Conclusion miRNA-148a which was significantly down-regulation in bladder carcinoma tissues may be participate in bladder tumorous development and progression,bioinformatics analysis provides some ideas for further research.

Objective To investigate the influence of Herba Epimedium, a kidney-strengthening Chinese herb, on the regenerate ossification of nano-hydroxyapatite/collagen/ polylactic acid (nHAC/PLA) in the repair of rabbit mandibular defect. Methods Sixteen rabbit models with 10 mm × 8 mm subperiosteal mandibular defect were established. The rabbits were randomly divided into 2 groups, namely Herba Epimedium group and control group. In both groups, the left mandibular defect region (short for left side) was not filled with any material, while the right mandibular defect region ( short for right side) was filled with nHAC/PLA. On week 2, 4, 8, 12 after modeling, rabbits in each group were killed, and then the macro-view, imageology and histology of the bilateral mandibles were observed. The data of the regenerate bone areas were calculated and analyzed with analysis of variance (ANOVA) . Results The imageological results showed bone mineral density in the right side of Herba Epimedium group and the control group, and in the left side of Herba Epimedium group were higher than those in the left side of the control group on week 2, 4, 8, 12 after modeling. The histological results showed that the regenerate ossification in the right side of Herba Epimedium group and the control group was superior to that in the left side of the corresponding groups, with more regenerative bone and vessels. In the right side of Herba Epimedium group and the control group, amount of bone-like tissues were grown into nHAC/PLA, and the maturity degree of bone tissue was increased with the prolongation of time. And the changes were more obvious in the right side of Herba Epimedium group, showing as the remaining of nHAC/PLA having been surrounded and replaced by regenerative bone tissue. Regenerative bone area was increased in both sides of Herba Epimedium group, which differed from the same side of the control group (P<0.05) . The differences of the regenerative bone area were also significant between the left side and right side of the same group (P<0.05) . Conclusion Herba Epimedium can effectively accelerate the repair of mandibular defect, and nHAC/PLA is of good biocompatibility, osteoinduction and bone conduction, and can be expected for bone defect repairing in clinic.