Objective To investigate the inhibitory effects and mechanism of ginsenoside Rg3 on vasculogenic mimicry of human breast cancer MCF-7 cell line. Methods The MCF-7 cells at logarithmic growth phase were obtained, and were cultured with different concentrations (0, 20, 50, 100, 150 and 300 mg/L) of ginsenoside Rg3. Cells cultured without Rg3 were served as controls. The IC50 were determined by CCK8 assay and anti-angiogenic effects were performed for testing the potential of tube-like structure (TLSs) formation. The expression levels of VEGF-A, MMP 9 and HIF-1αwere detected by Western blotting and real-time PCR. The secreted contents of VEGF-A and MMP9 were detected by enzyme linked immunosorbent assay (ELISA). Results The ginsenoside Rg3 suppressed the proliferation of MCF-7 cells in a dose-dependent manner, in which IC50 was (115.34±8.50) mg/L. The formation numbers of TLSs in MCF-7 cells were significantly inhibited by Rg3 in concentration dependent manner in 50 mg/L, 100 mg/L and 150 mg/L for (19.0 ± 1.0), (15.0 ± 1.5), and (10.0±1.7) vs. controls (22.0±1.8, F=150.805, P<0.05). The mRNA and protein expression levels of VEGF-A, MMP9 and HIF-1αprotein were inhibited by 50 mg/L,100 mg/L and 150 mg/L Rg3 vs. controls (P<0.05). Meanwhile, the contents of VEGF-A in MCF-7 cell supernatant was down-regulated by 50 mg/L,100 mg/L and 150 mg/L Rg3 vs. controls (P<0.05). The contents of MMP-9 in MCF-7 cell supernatant was down-regulated by 100 mg/L and 150 mg/L Rg3 vs. controls (P<0.05). There was no significant difference in MMP-9 expression between 50 mg/L group and control group. Conclusion The ginsenoside Rg3 is able to inhibit the vasculogenic mimicry of MCF-7 cells, which may be related with the down-
regulation of VEGF-A, MMP9 and HIF-1α.

Objective To detect mitochondrial gene 3243 A to 1507 Chinese and analyze their clinical characteristics.Methods Classical PCR－ RFLP was used to detect the mutation and statistical analysis was performed on the data after stratification in accordance with presence or absence of diabetes.Results (1)In this study,prevalence of mitochondrial gene 3243 A to G mutation was 1.16～ 1.20％ .(2)Compared with MDM(mitochondrial diabetes),M－ NGT(normal glucose to lerance with mutation)showed no significant difference in body fat parameters,blood lipid and C－ peptide except for early onset and hearing loss(P=0.0028).(3)Compared with type 2 DM,MDM presented characteristics of early onset,weight loss,hearing loss,islet cell function decrease and maternal hereditary.Conclusion Follow－ up should be given to the carrying the mitochondrial gene 3243 A to G mutation but presenting generally normal clinical manifestation in order to give the patient in－ time diagnosis and treatment.

Objective To compare the clinical outcomes between two different femur rotating osteotomy methods for kneeling ability recovery after total knee arthroplasty (TKA).Methods From January 2012 to December 2014,88 patients underwent TKA were selected for a retrospective study and were divided into two groups based on the methods to determine femoral rotation.Forty-eight patients were in measured resection group,while 40 patients in gap balancing group.The patients in both groups underwent fixed-bearing tibia prosthesis.There were no statistical significance between the two groups in gender,age,BMI and knee varus angle (P＞0.05).The knee varus angle,ROM,Oxford knee score (OKS) and American Knee Society (KSS) knee score were collected to assess malformation correction,kneeling ability and functions at pre-operation,one and two years postoperatively.Results The operation duration and blood loss in measured resection group were 80±19 min and 348±121 ml,while these data in gap balancing group were 82±23 min and 315 ± 100 ml respectively (P＞0.05).Patients in measured resection group were followed up 24-59 months (mean 43± 11 months),while the followed-up duration in gap balancing group was 25-58 months (mean 47±10 months).No major complications such as infection loosen and instability were occurred.Varus angles in measured resection group at postoperative 1 year and 2 year postoperative were 1.2°±0.4° and 1.0°±0.2° respectively,while those in gap balancing group were 0.9°±0.2° and 0.8°±0.3° (P＞0.05).The scores of the seventh item of OKS in measured resection group at 1 year and 2year follow-ups were 2.79±1.02 and 2.75± 1.03 respectively,while those in the gap balancing group were 1.90±0.85 and 1.80±0.83 (P＜0.01).ROM in the measured resection group at 1 year and 2 year postoperative were 102.08°± 15.60° and 102.08°±15.60° respectively,while those in the gap balancing group were 112.50°±18.32° and 113.00°±18.09° (P＜0.05).KSS in measured resection group at postoperative 1 year and 2 years were 154.63±31.12 and 154.63±31.26 respectively,while those in the gap balancing group were 170.55±22.67 and 173.45±22.52 (P＜0.05).Conclusion The method of measured resection and gap balancing to confirm femoral rotation during TKA can both achieve favorable kneeling ability and clinical outcomes,while gap balancing show superiority on kneeling ability recovery,ROM and clinical outcomes at 2-year postoperative improvement.

Objective:To search for the optimum formula for the self emulsification of evening primrose oil. Methods: Through solubility experiment, orthogonal screen and drawing phase diagram, taking the degree of emulsifying, the volume of the rest oil and emulsion particle size as parameters,the oil, non ionic surfactant and flux for the optimum formulation were screened. Results: In the formulation of evening primrose oil, taking Tween 85 as the non ionic surfactant and propylene glycol as the co surfactant can get the best effect of emulsifying. Conclusion: The optimum formula for self emulsification of evening primrose oil is evening primrose oil, Tween 85 and propylene glycol,with their proportion being 5∶7∶3.

Objective To estimate the effect of microRNA (miRNA) let-7 expression on human esophageal squamous cell carcinoma(ESCC) and the relationship between let-7 level and clinicopathological parameters. Methods ESCC cell line (Eca109) was transfected with let-7 or its inhibitor by RNAi and cell transfection techniques. Normal cultured Eca109 cell was served as negative control. The proliferation of Eca109 cell was detected by MTT. The expression of let-7 in Eca109 cells and 45 paired ESCC tissues and corresponding para-cancerous tissues were measured using real-time quantitative polymerase chain reaction (qRT-PCR). The relationship between let-7 level and clinicopathological parameters in patients with ESCC was analyzed. Results The A value of let-7 in Eca109 cells transfected with let-7 was lower than negative control (P=0.005), while it was higher in Eca109 cells transfected inhibitor than that in negative control 72 hours after transfection. In comparison with negative control, the expression of let-7 in Eca109 cells transfected with let-7 was increased 33% (1.33 vs 1.00,P=0. 039) and it was decreased 50% in Eca109 cells transfected with inhibitor (0.50 vs 1.00,P=0. 014). The ratio of let-7 expression in ESCC tissue and para-cancerous tissue was 0.66 ± 0.47 with significant differece (P= 0.001). Moreover, The level of let-7 expression in Han patients with ESCC was lower than Kazakh patients with ESCC (0.48±0.43 vs 0. 88±0.51,P=0. 019). The level of let-7 expression in poorly differentiated ESCC tissue was lower than well differentiated ESCC tissue (0.42±0.30 vs 0.84±0.38,P=0. 015). The level of let-7 expression in patients with lymph node metastasis was lower than those without lymph node metastasis (0.50±0.35vs 0. 80±0.52,P=0. 032) . Conclusion It is demonstrated that let-7 can inhibit the carcinogenesis and development of ESCC. The level of let-7 expression is associated with cell differentiation,lymph node metastasis and nationalities.

Objective To investigate the protection effects and possible mechanism of apoptotic effect of compound Xiancao granules on renal ischemia reperfusion injury ( IRI ) in rats. Methods Renal IRI rat model was established by clipping bilateral renal artery.The rats were divided into model control group (n=10), compound Xiancao granules group (n=10) and sham operation group (n=10).All Serum creatinine (Cr) and blood urea nitrogen (BUN) were determined to evaluate kidney function after IRI.Superoxide dismutase (SOD), glutathione peroxidase (GSH-PX) and malondialdehyde (MDA) activity in kidney were measured by colorimetric method. Expression of apoptotic regulatory genes Bcl-2 and Bax in renal tissue were detected by Western blotting.Renal tissue sections were stained by hematoxylin and eosin (HE) and examined under a light microscope. Results SOD and GSH-PX levels of the compound Xiancao granules group (278.1±16.2),(155.96±20.58) U?mg-1 were significantly higher than those of the model control group (196.3±12.1),(109.34±17.81) U?mg-1 (P<0.05). MDA, BUN and Cr (12.49±1.07) nmol?mL-1,(8.9±2.7) mmol?L-1,(149.7±8.5) μmol?L-1 were significantly reduced in the compound Xiancao granules group as compared with those of the model control group (17.32±1.26) nmol?mL-1,(14.6± 3.3) mmol?L-1,(206.1±11.2) μmol?L-1(P<0.05).Bax and Bcl-2 mRNA expression levels of sham operation group were significantly lower, and the Bax and Bcl-2 mRNA expression levels of model control group were significantly enhanced ( Bcl-2:P<0. 05, Bax: P<0. 01 ) . Bax mRNA expression level of compound Xiancao granules group was significantly decreased as compared with that of model control group ( P<0.05) , but Bcl-2 mRNA expression level of compound Xiancao granules group was significantly enhanced as compared with that of model control group (P<0.01). Conclusion These results suggested that compound Xiancao granules has protection effect on renal IRI in rats. The mechanisms may be related to its antioxidant activity and expression of the apoptotic genes such as Bcl-2 and Bax.

Abstract] Objective To analyze the molecular characteristics of the full-length genome of a new-ly isolated genotypeⅤJapanese encephalitis virus (JEV) strain (XZ0934) in China and the first genotypeⅤJEV strain ( Muar) isolated in Malaya 60 years ago.Methods Several softwares including ClustalX 2.0.9, DNAStar 7.1, Bioedit 7.2.5 and MEGA6.06 were used to conduct sequence alignments and phylo-genetic analysis.Results The full-length genomes of XZ0934 strain (isolated in Tibet, China in 2009) and Muar strain (isolated in Malaya in 1952) were composed of 10 983 and 10 988 nucleotides, respective-ly.The XZ0934 strain was highly similar with the Muar strain showing the homology of 90.6%in nucleotide (nt) and 98.3%in amino acid (aa).The open reading frame (ORF) of the two genotype Ⅴ JEV strains encoded 3433 aa while the ORF of other four genotypes (Ⅰ-Ⅳ) (10 299 nt) encoded 3432 aa.Compared with JEV strains of other genotypes, a serine were inserted into the NS4A gene of JEV strains genotype Ⅴand 10-14 nucleotides were inserted into the downstream of the ORF stop codon in 3′-untranslated region. Phylogenetic analysis of E sequences of all JEV strains genotypedⅠ-Ⅴrevealed that in the cluster of geno-typeⅤ, XZ0934 and 10-1827 ( isolated from mosquitoes in South Korea, 2010) stains formed a branch and were divergent from that of Muar strain indicating that there were molecular genetic differences among geno-typeⅤJEV strains after a 60 years hiatus.Conclusion The two genotypeⅤJEV strains showed high lev-els of identity in nucleotide sequences and amino acid sequences with serine insertion in the NS4A gene. However, there were molecular genetic differences between genotypeⅤJEV strains isolated after a 60 years hiatus.

Objective To investigate the combined effect and mechanism of metformin (Met) and 2-deoxy-D-glucose (2DG) on cell proliferation and apoptosis in liver cancer cells HepG2 and Hep3B.Methods Wst-1 reagent was used to determine the anti-proliferation effects after treatments with Met and 2DG alone or combined in HepG2 and Hep3B cells.Microscopy was used to observe cell morphological changes after treatments with Met and 2DG alone or combined in HepG2 and Hep3B cells.Cell apoptosis was observed by flow cytometry after treatment of different kinds of drugs.Western blotting was used to analyze the protein expressions of Caspase-3,PARP,Mcl-1 of HepG2.Results The survival rate of HepG2 cells in the combination group was (22.48 ± 0.51)％,and compared with the control group (100.00 ± 5.05)％,Met group (80.68 ±5.10)％ and 2DG group (72.56 ±4.34)％,the differences were statistically significant (P ＜ 0.001;P ＜ 0.001;P =0.001).The survival rate of Hep3B cells in the combination group was (29.16 ± 1.34) ％,and compared with the control group (100.00 ± 1.23) ％,Met group (59.58 ± 1.92) ％ and 2DG group (33.87 ± 1.95) ％,the differences were statistically significant (P ＜ 0.001;P ＜ 0.001;P =0.001).Microscopy observation showed that combined treatment of Met and 2DG caused less viable adherent cells of HepG2,but more floating dead cells.While the combination group also caused a decrease in the density of Hep3B cells,but did not significantly increase the shedding of cells.The apoptosis of HepG2 cells in the combination group was (39.63 ± 0.21) ％,and compared with the control group (7.12 ± 0.14) ％,Met group (12.56 ± 0.35) ％ and 2DG group (15.16 ± 1.93) ％,the differences were statistically significant (P ＜0.001;P ＜ 0.001;P =0.001).The apoptosis of Hep3B cells in the combination group was (12.58 ± 1.03) ％,and compared with the control group (2.82 ± 0.51) ％ and Met group (8.98 ± 0.86) ％,the differences were statistically significant (P ＜ 0.001;P =0.007),but compared with the 2DG group (12.40 ± 1.78) ％,the difference was not statistically significant (P =1.000).Furthermore,Western blotting demonstrated that the combined treatment induced evident Caspase-3 activation and poly ADP-ribose polymerase (PARP) cleavages,and decreased expression of Mcl-1.Conclusion The combination of Met and 2DG can effectively inhibit cell proliferation of HepG2 and Hep3B,and induce apoptosis of HepG2 cells.The mechanism may be involved with Caspase-3 activation,cutting PARP substrate and decreasing Mcl-1 protein.

Chemokines and chemokine receptors involve in biological activity and pathological process widely.It has been reported that many tumor cells overexpress functional chemokines.In lung cancer,chemo-kines involve in its proliferation,apoptosis,invasion and metastasis.Chemokines and chemokine receptor over-expressed in lung cancer can be used as specific target for pertinent anti-tumor treatment.

Objective To investigate the expression of the cyclin D1 in pubertal mammary hypertrophy and micromastia. Methods The expression of cyclin D1 was detected by SP immunohisto-chemistry in 18 cases of pubertal mammary hypertrophy and 12 cases of micromastia. Results The expression rate of cyclin D1 within breast tissue was 83. 33 % and 16. 67 % in pubertal mammary hypertrophy and micromastia, respectively. There was significant difference between the two groups (P<0. 01). Conclusion The expression level of the cyclin D1 within breast tissue in pubertal mammary hypertrophy is higher than that in micromastia.