Objective:To evaluate the sonodynamic effect of chlorin-e6, a sonosensitizing agent, on the proliferation of MDA-MB-231 cells. Methods:MDA-MB-231 and normal peripheral mononuclear cells (PMNCs) were treated with chlorin-e6 alone or combined with ultrasound. Cell proliferation was determined by MTT assay and cell morphology was observed under inversed fluorescence microscope. Results:Treatment with ultrasound (1.0 MHz, 1.0-2.0 W/cm~2,60 s) or chlorin-e6 (0.10-1.60 mg/mL) alone significantly inhibited the cell proliferation of both MDA-MB-231 and PMNC cells in a intensity-dependent and a dose-dependent manner, respectively. The 50% intensity of ultrasound for inhibiting the growth of MDA-MB-231 and PMNC cells was 1.23 W/cm~2 and 1.25 W/cm~2, respectively (P>0.05) and the IC_(50) of chlorin-e6 was 0.38 mg/mL and 0.77 mg/mL, respectively (P<0.05). Compared with single ultrasound (1.0 MHz,0.5 W/cm~2,60 s) or chlorin-e6 (0.05-0.20 mg/mL) treatment, the combination of ultrasound with chlorin-e6 significantly inhibited the proliferation of MDA-MB-231 cells (P<0.05), while had no synergistic inhibitory effects on the growth of PMNC(P>0.05). Compared with single ultrasound treatment (1.0 MHz, 0.5 W/cm~2,60 s) and single chlorin-e6 treatment (0.20 mg/mL), combination of ultrasound and chlorin-e6 significantly increased the death rate of MDA-MB-231 cells (P<0.05). Conclusion:Ultrasound combined with chlorin-e6 exerted specific inhibitory effect on the proliferation of MDA-MB-231 cells. Chlorin-e6 may be a promising sonosensitizing agent for the treatment of breast cancer.

10.3969/j.issn.2095-4344.2013.23.026

The solvent sublation technique was applied for the separation and enrichment of L-Arg using dodecylbenzene sulfonic (DBSA) as the surfactant, di (2-ethylhexyl) phosphoric acid (P204) as the extractant and n-heptane as the organic solution. The solvent sublation was compared with the floatation complexation extraction, foam floatation and solvent extraction. The experimental results showed that enrichment ratio of 16.2 and removal rate of 97.2% to L-Arg were obtained by the solvent sublation under the conditions of room temperature, 0.09 g/L L-Arg aqueous solution 250 mL, DBSA concentration 0.15 g/L, the initial pH 7.0, volume of n-heptane 10 mL, volume of P204 4.5 mL, gas flow rate of 200 mL/min. The study of the kinetics indicated that the solvent sublation process could be divided into three stages distinctly. The processes of the first stage and the second stage followed the first order kinetics equation; the process of the third stage followed the zero order kinetics equation. The separation mechanism of solvent sublation was also discussed.

Objective To investigate the activation state of and expressions of surface co-stimulatory molecules on peripheral CD4+ T cells from patients with pemphigus and healthy human controls.Methods Ninety patients with pemphigus including 24 patients with first-onset pemphigus,51 with quiescent pemphigus and 15 with recurrent pemphigus,as well as 30 healthy human controls were enrolled in this study.Peripheral blood samples were obtained from these subjects followed by lymphocyte isolation.Flow cytometry was performed to detect the expressions of CD69,intercellular adhesion molecule-1 (ICAM-1),inducible co-stimulatory molecule (ICOS),CD40 ligand (CD40L) and OX40 on CD4+ T cells.Statistical analysis was done by Mann-Whitney test using Graphpad 5.0 software.Results The expression rate of CD69 on peripheral CD4+ T cells from the healthy human controls was significantly lower than that from patients with pemphigus,patients with first-onset pemphigus,patients with quiescent pemphigus,and patients with recurrent pemphigus ((1.26 ± 0.19)％ vs.(2.46 ± 0.19)％,(2.77 ± 0.40)％,(2.15 ± 0.25)％ and (2.36 ± 0.35)％,all P ＜ 0.05).The patients with pemphigus also showed a significant increase in the expression rates of ICAM-1,CD40L and OX40 compared with the healthy human controls ((55.88 ± 1.67)％ vs.(47.75 ± 2.52)％,P＜ 0.05; (2.23 ± 0.22)％ vs.(0.73 ± 0.07)％,P＜ 0.01; (2.55 ± 0.29)％vs.(0.62 ± 0.17)％,P ＜ 0.01).No significant differences were observed between patients with different stages of pemphigus in the expression rates of CD69,ICAM-1,CD40L or OX40 (all P ＞ 0.05).The percentage of ICOS-expressing CD4+ T cells was significantly up-regulated in only patients with first-onset pemphigus as compared to the healthy controls ((3.73 ± 0.60)％ vs.(2.39 ± 0.16)％,P ＜ 0.05).Conclusions The peripheral blood CD4+ T cells from patients with pemphigus are in a relatively active state with up-regulated surface expressions of many costimulatory molecules,suggesting that CD4+ T cells are involved in the initiation and progression of pemphigus by interacting with B cells through co-stimulatory molecules.

Objective:To study the effects of obesity induced by high-fat diet on T lymphocyte subsets in the adipose tissue in mice.Methods:C57BL/6 male mice were randomly divided into 2 groups, the normal control group and high-fat diet group.After feeding 16 weeks, serum was separated and CHOL, TG, HDL, LDL and glucose levels were measured by automatic biochemical analyzer.The concentrations of TNF-αwere determined by ELISA kit.FACS was used to analyze the number of T cells and the percentage of subgroup in epididymal fat adipose tissue.Results:Compared with control group,body weight,weight gain,epididymal fat pad weight,perirenal fat weight,blood lipids,glucose and TNF-αwere significantly increased in high-fat diet group,but there were no difference in the thymus index and spleen index between the two groups.Compared with the control group,the mice fed a high-fat diet had increasing proportion of CD3+T cells,CD4+T cells and CD8+T cells in adipose tissue and there was a significant increase on the proportion of Th1 and Th17 sublineage in the HFD group.Conclusion:High-fat diet induced obesity can lead to the increasing proportion of CD3+T cells,CD4+T cells and CD8+T cells in epididymal fat pads and generate a progressive Th1 and Th17 bias.

Objective:To investigate the possible role of NKT cells in unexplained recurrent spontaneous abortion by measuring the NKT cell numbers,maturity and cytokine secretion of the placenta of mice with unexplained recurrent spontaneous abortion.Methods:Normal pregnancy model in hybrid by feeding CBA / J and BABL/C in a cage,and the model of unexplained recurrent spontaneous abortion was established by feeding CBA / J and DBA2/J in a cage.The number of NKT and CD3~+T cells was determined by flow-cytometry;Th1/Th2-relative cytokines were assayed by ELISA and T-bet expression was determinded by fluorescence quantitative PCR.Results:There was not significant change of CD3~+ T cells when compared between normal pregnancy and unexplained recurrent spontaneous abortion group (P＞0.05).In the course of normal pregnancy,the IFN-γ secreted by placenta lymphocytes decreased gradually,accompanied by the decline of NKT cell number and the proportion of mature cells;whereas in the course of unexplained recurrent spontaneous abortion,it was on the opposite.There was significant difference of T-bet mRNA expression between the two groups.T-bet mRNA expression was related to the proportion of mature NKT cells or placenta IFN-γ secretion by lymphocytes.Conclusion:Unexplained recurrent spontaneous abortion may be related to NKT cells disorders,NKT cells are of low-mature proportion and inadequate secretion of IFN-γ during early pregnancy,whereas are shown high-mature proportion and excessive secretion of IFN-γ during latter pregnancy;the anomaly of T-bet mRNA expression may be one of the factors leading to NKT cells disorder.

BACKGROUND AND OBJECTIVESonodynamic therapy (SDT) is a promising new approach for cancer therapy. The purpose of this study is to detect the effects of SDT on the cell proliferation of human lung adenocarcinoma cell SPCA-1, using Chlorin e6 as a sonosensitizing agent activated by ultrasound.

METHODSSPCA-1 and normal peripheral mononuclear cell (PMNC) were treated with ultrasound or Chlorin e6 alone and combined. Cell proliferation was determined by MTT assay, and cell morphology was studied by inverted microscope after 6 h treated.

RESULTS1.0 MHz ultrasound (1.0 W/ cm(2)-2.0 W/cm2 x 60 s) and Chlorin e6 (0.4 mg/mL-3.2 mg/mL) inhibited the cell proliferation of both SPCA-1 and PMNC cells in a intensity- and a dose-dependent manner respectively. Compared with the ultrasound (1.0 W/cm2 x 60 s) or Chlorin e6 (0.05 mg/mL-0.2 mg/mL) alone, the inhibitory effect on the cell proliferation was remarkably increased by the combination of ultrasound and chlorin e6 in SPCA-1 cells (P < 0.05), but no same effect was observed in PMNC cells (P > 0.05). Compared with the ultrasound (1.0 W/cm2 x 60 s) or chlorin e6 (0.2 mg/mL) alone, the combination treatment of ultrasound with Chlorin e6 induced more necrotic cells in SPCA-1 cells (P < 0.05).

CONCLUSIONThere was a significant selectively inhibitory effect of sonodynamic effect with Chlorin e6 on the SPCA-1 cell growth. Chlorin e6 may be a promising sonosensitizing agent for the treatment of non-small cell lung cancer.

Adenocarcinoma ; drug therapy ; therapy ; Cell Line, Tumor ; Humans ; Lung Neoplasms ; drug therapy ; therapy ; Porphyrins ; chemistry ; therapeutic use ; Radiation-Sensitizing Agents ; chemistry ; therapeutic use ; Ultrasonic Therapy ; methods

Objective To investigate the influence of limonin on the malignant biological behavior of non-small cell lung cancer (NSCLC) cells by regulating the protein tyrosine kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway. Methods CCK-8 method was applied to detect the survival rate of A549 cells treated with different concentrations of limonin (0, 5, 10, 25, 50, 75, 100 μmol/L). A549 cells were separated into normal culture (NC) group, low-dose limonin group (treatment with 10 μmol/L limonin for 24 h), medium-dose limonin group (treatment with 25 μmol/L limonin for 24 h), high-dose limonin group (treatment with 50 μmol/L limonin for 24 h), coumermycin A1 group (treatment with 10 μmol/L JAK2 activator coumermycin A1+50 μmol/L limonin for 24 h), and AG490 group (treatment with 10 μmol/L JAK2 inhibitor AG490+50 μmol/L limonin for 24 h). Clone formation assay was applied to detect the clones of each group of cells. Transwell assay was applied to detect cell migration and invasion, and flow cytometry was applied to detect apoptosis. Western blot analysis was applied to detect the protein expression levels of JAK2, p-JAK2, STAT3, p-STAT3, E-cadherin, N-cadherin, and vimentin in each group. Results The viability of A549 cells decreased significantly in a limonin concentration-dependent manner (P < 0.05), with IC₅₀ of 45.16±1.66 μmol/L. Concentrations of 10, 25, and 50 μmol/L were selected for subsequent experiments. The numbers of clones, migration, and invasion of A549 cells and the protein expression levels of IL-6, p-JAK2, p-STAT3, N-cadherin, and vimentin in the low-, medium-, and high-dose limonin groups significantly decreased, compared with those in the NC group, and the apoptosis rate and E-cadherin protein expression significantly increased (P < 0.05). The JAK2 activator coumermycin A1 attenuated the ability of limonin to inhibit the proliferation, migration, invasion, and other malignant biological behavior of A549 cells and attenuated the apoptosis ability. The JAK2 inhibitor AG490 enhanced the ability of limonin to inhibit the proliferation, migration, invasion, and other malignant biological behavior of A549 cells and enhanced the apoptosis ability. Conclusion Limonin can inhibit the malignant biological behavior of NSCLC cells, such as proliferation, migration, and invasion, by inhibiting the JAK2/STAT3 pathway.

This paper reviewed the mechanism and clinical application of Dahuang-Zhezhong Pill in Jin Gui Yao Lue. It was found that the anti-tumor effect of Dahuang-Zhezhong Pill was mainly achieved by inhibiting cell proliferation, inducing cell apoptosis, regulating immune function, inhibiting tumor angiogenesis, affecting tumor cell metastasis, and reversing cell resistance. It was used in the treatment of liver cancer, gastric cancer, esophageal cancer, lung cancer and other malignant tumors. Therefore, we should clarify the anti-tumor mechanism, the dominant dosage, the dominant tumor species of Dahuang-Zhezhong Pill, and standardize the dosage of treatment and adverse reactions.

OBJECTIVETo investigate the neonatal umbilical cord blood thyroid stimulating hormone (TSH) level in the universal iodized salt areas and put forward the cut-point, then analyze its application.

METHODSSeven provinces were selected where the pregnant women having satisfied urinary iodine levels, then the urinary samples of pregnant women and the neonates cord blood were collected for urine iodine and TSH tests, and the relative factors were also recorded.

RESULTSTotal 1 524 urine and cord blood samples were collected from pregnant women and their new borns respectively. The median urinary iodine of pregnant women was 246.0 micro g/L, and the median TSH was 3.58 mU/L. The TSH level among seven areas and the neonatal delivery type varied significantly.

CONCLUSIONSThe neonatal cord blood TSH was influenced by several factors and could not be controlled, thus not be suitable as a iodine deficiency disorders surveillance indicator.

Delivery, Obstetric ; Female ; Fetal Blood ; drug effects ; metabolism ; Humans ; Iodine ; pharmacology ; urine ; Pregnancy ; Thyrotropin ; blood