Purpose:To investigate the relationship between cell apoptosis and dephosphorylated RB protein in human breast cancer. Methods:In our work,human breast cell lines (MCF-7/S,the chemosensitive cell line and MCF-7/ADR,the chemoresistent cell line)were evaluated. Chemosensitivity of two cell lines was evaluated by the MTT colorimetric assay;the expressive levels of dephosphorylated RB protein were detected with immunocytochemistry. Apoptosis rates were determined by flow cytometry(FCM). Results:ADR inhibited proliferation of chemosensitive cell line MCF-7/S ,the 50% inhibition concentration (IC 50 ) was 0.128 ?g/ml;And IC 50 of MCF-7/ADR was 10.89 ?g/ml. The chemotherapeutic sensitivity of MCF-7/S was more than that of MCF-7/ADR by 86 times . Before treatment with ADR,phosphorylated RB protein was positive in two cell lines,but dephosphorylated RB protein was negative;After treatment of different concentration ADR,when the concentration of ADR was increased,expression of dephosphorylated RB protein elevated accordingly in MCF-7/S,but no significant change in MCF-7/ADR. Apoptosis and cell cycle was detected by FCM assays shows ADR induced apoptosis of MCF-7/S more than MCF-7/ADR(P0.05).

Objective To develop the methods for using 0.015 cc pinpoint chambers, 0.007 cc miniature chambers and diode detector to measure Multi-leaf collimator (MLC) small field in IMRT.Methods MAX4000 and Unidos electrometers were connected with different types of small chambers and diode detectors.MLC shaped fields of10 cm×10 cm, 6 cm×6 cm, 4 cm×4 cm, 3 cm×3 cm, 2 cm× 2 cm were defined at 100 cm SSD.The field sizes for the Varian accelerator were defined by the tertiary MLC, while the secondary jaws were kept at 10 cm × 10 cm field, with the monitor units of 250 MU.Each field was measured three times to obtain the average value.The readings of all small fields were normalized to 10 cm × 10 cm field values for comparison of measured and published output factors.Results The relative deviations of the MLC small field output factors from the published outputs are 1.0％ , 1.7％ , 1.5％ and 2.4％, respectively, for Unidos electrometer connected with 0.015 cc pinpoint chamber;0.2％, 0.8％, 0.8％ and 1.4％, respectively, for Unidos electrometer connected with 0.007 cc miniature chamber;and 0.1％, 0.5％, 0.5％ and 0.9％, respectively, for MAX4000 electrometer connected with 0.007 cc miniature chamber.Conclusions The 0.015 cc chamber-measured MLC output factors for 3 cm × 3 cm and 2 cm × 2 cm fields are excellent.As required by IAEA, the relative deviations of the measured output factor from the published output factor are within ± 2％ for 2 cm × 2 cm fields and ± 3％ for larger fields.The results measured using 0.007 cc chamber are better than those measured using 0.015 cc chamber.The measured results using the diode detector, normalized to the 10 cm × 10 cm field, are consistent with the minimum requirements and excellent when being normalized to the 4 cm × 4 cm field.For dosimetric consideration, MLC small field output factor should be measured using small chamber and diode detector.The method is accurate and reliable, therefore, all measured output factors for MLC small fields should be input into radiation treatment plan system.

Objective To investigate the relationship of matrix metalloproteinases(MMP?3 and MMP?9)and the tissue inhibitor TIMP?1 with left ventricular hypertrophy(LVH)in patients with primary hypertension. Methods Totally 140 patients with primary hypertension and 132 healthy controls were included. Matrix metalloproteinase?3(MMP?3),matrix metalloproteinase?9(MMP?9)and tissue inhibitor metalloproteinase?1 (TIMP?1)were measured. All subjects were taken echocardiography examination ,then left ventricular mass index(LVMI)was calculated. Re?sults MMP?3,MMP?9,TIMP?1(488.32±100.32 vs 314.59±99.78;340.56±43.21 vs 290.15±33.98;389.16±57.53 vs 243.45±62.31;P<0.001) and LVMI(113.7±9.9 vs 88.3±10.4,P<0.001)in patients with primary hypertension were significantly higher than those in controls. In a multiple stepwise regression analysis with LVMI as the variable,it was found that age,SBP,MMP?3,MMP?9 and TIMP?1 were main determinants for LVMI (r2=0.78,P<0.001). 3. Patients with primary hypertension were divided into two subgroups according to LVMI,i.e.,hypertension with LVH (group A)and hypertension without LVH(group B). SBP,MMP?3,MMP?9 and TIMP?1(178±31 vs 166±25;490.14±99.13 vs 405.56±53.12;340.56±43.21 vs 290.15±33.98;393.45±47.69 vs 301.58±39.57;P<0.05)of group A were significantly higher than those of group B. Conclusion MMP?3,MMP?9 and TIMP?1 are influencing factors for LVH in patients with primary hypertension.

To explore the antibacterial activity and mechanism of total alkaloids and berberine from Coptidis Rhizoma on Aeromonas hydrophila, and determine the effect of total alkaloids and berberine from Coptidis Rhizoma on minimum inhibitory concentrations, permeability and fluidity of cell membrane, conformation of membrane proteins and virulence factors of A. hydrophila. The results showed that both total alkaloids and berberine from Coptidis Rhizoma had antibacterial activities on A. hydrophila, with minimum inhibitory concentrations of 62.5 and 125 mg · L(-1), respectively. Total alkaloids and berberine from Coptidis Rhizoma could increase the fluidity of membrane, change the conformation of membrane porteins and increase the permeability of bacteria membrane by 24.52% and 19.66%, respectively. Besides, total alkaloids and berberine from Coptidis Rhizoma significantly decreased the hemolysis of exotoxin and the mRNA expressions of aerA and hlyA (P < 0.05, P < 0.01), the secretion of endotoxin and the mRNA expression of LpxC (P < 0.05, P < 0.01). The results suggested that the antibacterial activity of total alkaloids and berberine from Coptidis Rhizoma on A. hydrophila may be related to the bacteria membrane injury. They inhibited the bacterial growth by increasing membrane lipid fluidity and changing conformation of membrane proteins, and reduced the secretion of virulence factors of A. hydrophila to weaken the pathogenicity.
Aeromonas hydrophila ; drug effects ; genetics ; metabolism ; Alkaloids ; pharmacology ; Anti-Bacterial Agents ; pharmacology ; Bacterial Proteins ; genetics ; metabolism ; Bacterial Toxins ; biosynthesis ; Berberine ; pharmacology ; Cell Membrane ; drug effects ; genetics ; metabolism ; Coptis ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Membrane Fluidity ; drug effects ; Rhizome ; chemistry

Objective To determine the expression of cathepsin D (CD), cathepsin H (CH) and cathepsin L (CL) in human primary hepatocellular carcinoma (HCC), and to investigate their mechanisms. Methods The protein expression of CD, CH and CL in hepatic specimens consisted of control (n = 17), HCC (n = 37) and paracancerous (n = 28) tissues were detected by immunohistochemistry. The relative values of CD, CH and CL protein expression were examined with absorbent density analysis. Data were analyzed using SPSS11. 5 software. The univariate analysis was used to compare the difference among groups. Results The mean absorbance of CD, CH and CL proteins in HCC tissues (1.21± 0.33, 0. 89 ± 0.22 and 1.16± 0. 25, respectively) were significantly higher in comparison with those in control tissues (0. 19 ± 0. 07, 0. 24 ± 0. 12 and 0. 28 ± 0. 14,respectively) and in paracancerous tissues (0.27±0.13,0. 31± 0.14 and 0. 36±0.15)(all P values =0.0001). Whereas there was no difference between control and paracancerous tissues with respect to CD, CH and CL proteins (P ＞0. 05). Three proteins immunohistochemically appeared as a lot of diffused spots and stripes staining in cytoplasma of HCC tissues,but only a few scatted spots staining was found in control and paracancerous tissues. Conclusion The high expression of CD, CH and CL protein in primary HCC may be important markers for carcinogenesis and malignant progress.

ObjectiveTo investigate the expression of programmed death receptor ligand 1 ( PD-L1 ) of peripheral blood mononuclear cells (PBMCs) in patients with hepatic cystic echincccccosis (HCE) and its relation with interferon-γ.MethodsThe clinical data of 63 patients with HCE who were admitted to the First Affiliated Hospital of Xinjiang Medical University from June 2010 to February 2011 were retrospectively analyzed.All patients were divided into HCE active group (38 patients) and HCE non-active group (25 patients) according to the system established by the World Health Organization's Informal Working Group on Echinocoecosis.Twenty patients with hepatic hemangioma or healthy individuals were recruited in normal control group.The positive rate of PD-L1 expression was detected by flow cytometry and immunocytochemistry.The expression of interferon-γ was detected by enzyme-linked immtmosorbent assay (ELISA).All data were analyzed by the t test,one-way analysis of variance,LSD test and chi-square test.The relationship between the expression of interferon-γ and positive rate of PD-L1 expression was analyzed by the Pearson test.ResultsThe results of flow cytometry showed that the positive rates of PD-L1 expression in the HCE active group,HCE non-active group and normal control group were 12.1％±3.8％,10.9％ ± 2.5％ and 9.1％ ±2.5％,respectively.There was a significant difference in the positive rate of PD-L1 expression between the HCE active group and normal control group (t =3.327,P ＜ 0.05 ).The results of immunohistochemistry showed that the positive rates of PD-LI expression in the HCE active group,HCE non-active group and normal control group were 11.9％ ± 3.4％,i0.6％ ± 2.9％ and 9.5％ ± 3.6％,respectively.There was a significant difference in the positive rate of PD-L1 expression between the HCE active group and normal control group (t =2.470,P ＜ 0.05 ).The expressions of intefferon-γ in the HCE active group,HCE non-active group and normal control group were ( 141 ± 38 ) μμg/L,( 124 ± 32 ) μg/L and ( 105 ± 42 ) μg/L.There wasasignificant difference in the expression of interferon-γ between the HCE active group and normal control group ( t =3.280,P ＜ 0.05).The results of flow cytometry and immunohistochemistry revealed that the positive rate of PD-L1 expression was positively correlated with the expression of interferon-γ( r =0.59,0.61,P ＜ 0.05 ).Conclusion With the help of interferon-γ,PD-L1 may play an important role in promoting the immune.evasion of echinococcus.

To study the effects of alkaloids from Coptidis Rhizoma on low-density lipoprotein receptor (LDLR) mRNA expression and antihyperlipedemic levels. The LDLR mRNA expression were detected by real time fluorescence quantitative PCR, and the levels of total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL-c) and high-density lipoprotein cholesterol (HDL-c) in serum were measured at the first and last examination. The results show that, after the drug treatment, compared with the model group, each drug group showed a lipid-lowering effect. Especially, coptisine, palmatine, jatrorrhinze were significantly reduced TC, TG, LDL-c (P < 0.05, P < 0.01), and increased HDL-c (P < 0.01). In addition, they also increased mRNA expression of the LDLR in liver and HepG2 cells. The results showed that alkaloids from Coptidis Rhizoma can regulate lipid metabolism disorder, and coptisine have the best lipid-lowering effect.
Alkaloids ; administration & dosage ; Animals ; Cholesterol ; metabolism ; Cricetinae ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Hyperlipidemias ; drug therapy ; genetics ; metabolism ; Hypoglycemic Agents ; administration & dosage ; Lipid Metabolism ; drug effects ; Lipids ; blood ; Lipoproteins, LDL ; metabolism ; Mesocricetus ; Receptors, Lipoprotein ; genetics ; metabolism ; Triglycerides ; metabolism

Objective To compare the imaging quality of articular cartilage of the knee with 3D-sampling perfection with applica-tion optimized contrast using different flip angle evolutions (3D-SPACE),3D-true fast imaging with steady-state precession (3D-True FISP)and 2D-fast-spin-echo-proto-density(2D-FSE-PD)sequences.Methods 40 healthy volunteers and 20 patients of knee joints were examined with 3D-SPACE,3D-True FISP and 2D-FSE-PD sequences at 1.5T MRI.Signal-noise ratio (SNR),contrast-to-noise ratio (CNR)and lesion visualization of articular cartilage were compared.Results 3D-SPACE showed the highest SNR of cartilage and CNR of fluid/cartilage among the three sequences (P <0.05).3D-SPACE had the better capability for showing the lev-el I 、level Ⅱcartilage injury comparing with 3D-True FISP,but no significant difference between the cartilage injury at level Ⅲ and level Ⅳ.For all levels of cartilage injury,3D sequence was better than the 2D sequence.Conclusion Compared with the 3D-True FISP sequence and 2D-FSE-PD sequence,3D-SPACE sequence can show the structure of knee and knee cartilage injury better.

Objective To investigate the expression and significance of mRNAand exon mutationof NT5C2 gene in acute lymphoblastic leukemia (ALL) bone marrow.Methods Case control study design was used in this study.Bone marrow samples were collected from ALL patients in Anhui Provincial Cancer Hospital in recent 4 years.The patientswere divided into the initial diagnosis group , the complete remission group and the recurrence group.And they could specifically be divided into 36 patients initially diagnosed, 36 patients who achievedcomplete remission and 16 patients who relapsed with children B -ALL,15 patients initially diagnosed,15 patients who achievedcomplete remission and 9 patients who relapsed with children T -ALL, 18 patients initially diagnosed,18 patients who achievedcomplete remission and 12 patients who relapsed with adult B-ALL, and 11 patients initially diagnosed,11 patients who achievedcomplete remission and 6 patients who relapsed with adult B -ALL.The initial diagnosis,complete remission and recurrence samples were matched.8 children and 8 adults without hematologic malignanciewere used as controls .Real-time PCR was performed to detect the level of NT5C2 mRNAin ALL patients.The exons of NT5C2 gene were cloned and sequenced for the common mutations in all cases .The results of NT5C2 mRNA levels in different groups were performed using non -parametric test by SPSS16.0 analytics software, and then non-parametric test together with correlation analysis was analyzed between NT 5C2 mRNA levels of different initial diagnosis groups and gender, age, leukocyte level and risk classification .Results (1)The expression of NT5C2 mRNA levels of recurrence group were higher than that of initial diagnosis group ,complete remission group and controls in children and adult B -ALL respectively(P <0.01).(2)NT5C2 mRNA expression in children and adult T-ALL showed no difference in initial diagnosis ,complete remission, recurrence group and controls (P >0.05).(3)NT5C2 mRNA expression of initial diagnosis group in children and adult B -ALL and T-ALL was not correlated with risk classification (P >0.05).(4)A newheterozygousmutation p.P414A of NT5C2 was discovered in a recurrencesample.Conclusions (1) High expression ofNT5C2 mRNA is associated with recurrence inchildren and adult B-ALL, and it may be an indicator of monitoring recurrence .(2)The incidence of exons mutation of NT5C2 gene in ALL is low in China.

Locoweeds are presently defined as those species of the genera Oxytropis and Astragalus (family Leguminosae) that specifically contain the key toxic constituent,swainsonine.After ingesting locoweeds,livestock can develop poisoning disease characterized by chronic dysfunction of the nervous system,which causes severe economic losses to the pastoral areas.In addition,swainsonine has attracted a great attention from toxicology and medicine fields,due to its dual role of toxicity and pharmacological activity.This review not only summarizes the latest research progress of toxicity and its poisoning mechanism,pharmacological activity,source,and biosynthesis pathway of swainsonine,but also speculates the possible regulatory enzymes involved in the synthesis pathway.Moreover,the future research on swainsonine is also looked ahead,which provide references for the prevention and treatment of locoism.