CXCR4, a receptor for stromal cell-derived factor-1 (SDF-1, also know as CXCL12) and a member of the chemokine receptor family, has been shown to play an important role in the proliferation, migration, invasion and metastasis of breast cancer cells. CXCR4 expression is an early event in breast cancer development and may serve as a biomarker for early detection and diagnosis of breast cancer. Expression of CXCR4 has been demonstrated to correlate with metastasis and poor prognosis in breast cancer. Inhibition of CXCR4 expression may be a potential target of breast cancer treatment.

Objective To establish a mathematic model of correlativity of human cytomegalovirus(HCMV) infection between mothers and their newborns.Method The plasma HCMV-IgM content,HCMV-DNA levels in plasm and peripheral blood mononuclear cells (PBMCs) were assayed with ELISA and PCR technique.Results There was a evident correlation between the two generation,and there was largest correlativity between HCMV-DNA level of mother's plasma and HCMV congenital infection rate,its spearman rank correlative coefficient was 0 7018(P

The essential oil of curcuma phaeocaulis valeto n can be extracted by supercritical fluid CO2 modified with ethanol. The recov ery of active compound, curdione, could be improved when supercritical fluid ext raction with trapping by silica gel column was used and then eluted the analytes with hexane, ether and ethanol. The relative recovery of curdione increased fro m 10.7% to 34.7%.

Objective To compare the clinical outcomes between proximal femoral nail antirotation (PFNA) combined with salmon calcitonin and PFNA only for the treatment of unstable intertrochanteric fractures in elderly patients.Methods From January 2009 to December 2011,120 elderly patients with intertrochanteric fracture were randomly divided into two groups:calcitonin group and control group.Patients in calcitonin group were treated with PFNA combined with salmon calcitonin,while patients in control group were treated with PFNA only.According to Evans-Jensen classification,60 patients in calcitonin group (28 males and 32 females,with an average age of 75.1 years) were divided into 20 cases of type Ⅱ A,32 cases of type Ⅱ B and 8 type Ⅲ.Sixty patients in control group (27 males and 33 females,with an average age of 74.9 years) were divided into 22 cases of type Ⅱ A,32 cases of type Ⅱ B and 6 type Ⅲ.Bone healing was assessed with X-ray and bone mineral density (BMD) was measured by dual energy X-ray absorptiometry.Harris hip and SF-12 score,complications,adverse effect of salmon calcitonin and subsequent fragility fractures were evaluated postoperatively.Results One-hundred and thirteen patients were followed up for at least 2 years.In 6 months after surgery,there were 4 cases of delayed healing in control group.However,all fractures were healed in 12 months after surgery.No significant difference was found between the two groups in BMD preoperatively.The changes in BMD were significantly different between the two groups in 6 months,1 year and 2 years after surgery.No significant difference was found between the two groups in Harris hip and 1-year SF-12 score while the 2-year SF-12 score was significantly different between the two groups.There was 1 patient in calcitonin group who suffered from subsequent fragility fracture in 3 months after surgery,while there was 6 patients in control group during 13 to 23 months postoperatively.Conclusion PFNA combined with salmon calcitonin achieves good effect for the treatment of unstable intertrochanteric fractures in elderly patients.

The gene encoding thermostable lactate dehydrogenase (Tm-LDH) was cloned into the plasmid pHsh from Thermotoga maritima, and expressed in Escherichia coli JM 109. The recombinant protein was purified to homogeneity by a simple step, heat treatment. The recombinant enzyme had a molecular mass of 33 kDa. The optimal temperature and pH of Tm-LDH were observed 95 degrees C and 7.0. The purified enzyme had a half-life of 2 h at 90 degrees C, and exhibited better stability over a pH range from 5.5 to 8.0. The K(m) and V(max) values were 1.7 mmol/L, 3.8 x 10(4) U/mg of protein for pyruvate, and 7.2 mmol/L and 1.1 x 10(5) U/mg for NADH, respectively. The expression of Tm-LDH in T7 system could not obtain high efficiency, but it has been soluble over-expression in pHsh system and reached 340 mg/L. The superior stability and productivity of Tm-LDH will lay the foundation of its industrial-scale fermentation and application in the NAD regeneration.
Cloning, Molecular ; Enzyme Stability ; Escherichia coli ; metabolism ; L-Lactate Dehydrogenase ; biosynthesis ; Molecular Weight ; Recombinant Proteins ; biosynthesis ; Temperature ; Thermotoga maritima ; enzymology

[ABSTRACT]AIM:Toinvestigatetheeffectofsilencingisocitratedehydrogenase2(IDH-2)genebysmallinter-fering RNA (siRNA) on the biological characteristics of human small cell lung cancer cell line NCI -H446.METHODS:IDH-2 expression was knocked down in human small cell lung cancer cell line NCI -H446 by siRNA-IDH-2.The expression level of IDH-2 was determined by real-time PCR and Western blotting .The cell proliferation was measured by CCK-8 as-say , the protein expression of MAPK p 42 was detected by Western blotting , and the cell cycle was analyzed by flow cytome-try.The migration was observed using Transwell cell migration system .BALB/c nude mice were subcutaneously injected on the back with NCI-H446 cells transfected with siRNA-IDH-2/negative control siRNA or non-transfected cells to study the tumor growth .RESULTS:siRNA-IDH-2 remarkably down-regulated the expression of IDH-2 and MAPK p42 in the NCI-H446 cells.siRNA-IDH-2 inhibited both the proliferation and migration abilities of NCI-H446 cells, and the cell cycle was arrested in S phase as compared with negative control group .Additionally, the volume of xenograft tumors in siRNA-IDH-2 group was significantly decreased as compared with control group .CONCLUSION:siRNA-IDH-2 down-regulates the expres-sion of IDH-2 in NCI-H446 cells, reduces the cell migration efficiency and inhibits the tumor growth in vitro and in vivo.

Objective To investigate whether hydrogen can regulate the expressions of inflammatory factors by ultraviolet B (UVB)-induced human HaCaT keratinocytes through the autophagy pathway.Methods Cultured HaCaT keratinocytes were divided into several groups:blank control group receiving no treatment,hydrogen group cultured in hydrogen-rich medium,three UVB groups irradiated with UVB at 1,10,50 mJ/cm2 respectively,three UVB + hydrogen groups irradiated with UVB at 1,10,50 mJ/cm2 respectively followed by culture in hydrogen-rich medium,UVB + 3MA group pretreated with the autophagy inhibitor 3MA for 1 hour followed by UVB radiation at 50 mJ/cm2,UVB + rapamycin group pretreated with the autophagy activator rapamycin for 1 hour followed by UVB radiation at 50 mJ/cm2,UVB + 3MA +hydrogen group pretreated with 3MA for 1 hour followed by UVB radiation at 50 mJ/cm2 and culture in hydrogen-rich medium,UVB + rapamycin + hydrogen group pretreated with rapamycin for 1 hour followed by UVB radiation at 50 mJ/cm2 and culture in hydrogen-rich medium.After additional culture with or without hydrogen for 12 hours,methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate cellular proliferative activity,Western-blot analysis to measure the expressions of autophagy-associated protein 1 light chain 3 (LC3) and Beclin 1,and enzyme-linked immunosorbent assay (ELISA) to measure the supernatant levels of inflammatory factors including tumor necrosis factor (TNF)-α,interleukin (IL)-1β,IL-6 and high mobility group protein B1 (HMGB1),and a test kit was used to determine the level of lactate dehydrogenase (LDH).Results Compared with the blank control group,the 10-and 50-mJ/cm2 UVB groups showed significantly increased release of LDH,expressions of LC3 and Beclin1 and supernatant levels of TNF-α,IL-1 β,IL-6 and HMGB 1,but decreased cellular proliferative activity (all P ＜ 0.05).Hydrogen significantly attenuated the release of LDH,down-regulated the supernatant levels of TNF-α,IL-1β,IL-6 and HMGB1,but up-regulated cellular proliferative activity as well as LC3 and Beclin1 expressions in the 10-and 50-mJ/cm2 UVB + hydrogen groups compared with the 10-and 50-mJ/cm2 UVB groups respectively (all P ＜ 0.05).In addition,the levels of TNF-α,IL-1β,II-6 and HMGB1 were significantly higher in the 50-mJ/cm2 UVB + 3MA group than in the 50-mJ/cm2 UVB group,and higher in the 50-mJ/cm2 UVB + 3MA + hydrogen group than in the 50-mJ/cm2 UVB + hydrogen group,but lower in the 50-mJ/cm2 UVB + rapamycin group than in the 50-mJ/cm2 UVB group (all P＜ 0.05).Conclusion UVB radiation can increase the expressions ofautophagy-associated proteins,and hydrogen-rich medium can down-regulate the expressions of inflammatory factors by UVB-induced HaCaT cells through the autophagy pathway.

AIM: To evaluate the genotype, muscle histopathology and ultrastructure in dko mice. METHODS: Dystrophin/Utrophin-deficient double knockouts (dko) mice were obtained from university of Oxford, UK. Genotype of filial generation of heterozygote was evaluated by PCR-SSP. HE staining and fluorescent immunohistochemistry by SABC-Cy3 were used to detect striated muscle of dko mouse, and the muscle ultrastructure was observed by transmission electron microscope(TEM). RESULTS: In 112 filial generation mice, there were 28 mdx (25.0%), 26 dko (23.2%) and 58 heterozygote (51.8%), which coincided with the law of Mendelian inheritance. HE staining showed that the myocytes were not very uniform, there were phenomenon of round outline, centrally nucleated fibers, widening interspace, inflammatory cell infiltration and connective tissue proliferation in dko mice. There were no any immunofluorescent expression of dystrophin and utrophin in sarcolemma in dko mice. TEM showed sarcolemma breakage, separation and edema, and loose myofibril texture, inflammatory cell infiltration and connective tissue proliferation in dko mice. CONCLUSION: PCR-SSP is a very quick and accurate way for genotype evaluation of filial generation. The pathophysiology of dko mouse was very similar to Duchenne muscular dystrophy (DMD), and dko mouse is an ideal animal model for study of DMD clinical therapy.

Objective To investigate the diagnosis and treatment of the accessory breast cancer with breast cancer.Methods Five patients with accessory breast cancer were admitted,among of them,1 case with breast cancer,and their documents were retrospectively analyzed.Results Of the 5 cases,1 case underwent modified radical mastectomy,the other 4 cases underwent accessory breast enlarged resection and axillary lymph node dissection.They all received chemotherapy and radiotherapy and 1 case received tamoxifen therapy.Four cases were followed up for 2 years,there was no recurrence and metastasis,1 case was received CAP sequenced chemotherapy.Conclusions Accessory breast cancer is rare but aggressive.The diagnosis is mainly depended on imaging results and postoperative pathology.Combined therapy principle dominated by surgery should be followed.The operation of accessory breast enlarged resection and axillary lymph node dissection should be indicated.

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.