Background and purpose:Osteosarcoma, a highly malignant bone tumor, develops rapidly. The current medicines for osteosarcoma present some limitations with serious side effects of long-term use. Isosteviol has the structure of tetracyclic diterpene which is the starting material of many anti-cancer drugs. However, its anti-tumor activity has been rarely reported. This study investigated the effect of isosteviol on proliferation of human osteosarcoma cell line U-2OS.Methods:The effect of isosteviol on U-2OS cell proliferation was assayed by MTT method. Cellular morphologic changes were observed under an inverted phase contrast microscope. The cell condition was observed with Hoechst 33342 and PI staining. Generation of reactive oxygen species and cell membrane potential were detected as well. The cell cycles were analyzed with lfow cytometry. The expressions of apoptosis-related proteins Bcl-2 and Bax were measured by Western blot assay.Results:The result indicates that isosteviol suppressed the growth of U-2OS cells in time- and concentration-dependent manner. Isosteviol could cause S phase cell cycle arrest at 24 h and apoptosis at 48 h. With the increased drug concentration, reactive oxygen species increased significantly, and the membrane potential gradually reduced. In addition, isosteviol treatment enhanced the expression of Bax but reduced that of Bcl-2.Conclusion:The inhibition of isosteviol on cell growth of U-2OS cells was possibly caused by promoting apoptosis through regulating the apoptosis-related protein expressions, such as the enhancement of Bax and reduction of Bcl-2 expression.

Objective To structure the model of acute carbon monoxid poisoning(ACOP)in rats. Evaluate the effectiveness of the poisoning on the pulmonary function and the significance of carbon monoxide hemoglobin(HbCO)and oxygenation index in diagnosis of acute lung injury(ALI)/acute respiratory distress syndrome(ARDS).Method Eighty healthy adult male Wistar rats were randomized into 4 groups.According to the concentration of CO,poisoning group was randomized into three groups(each group=20),group A,group B,group C.After poisoned,arterial blood was collected rapidly for arterial blood gas analysis.According to the pathological changes,the models were divided into ALI/ARDS group and non-ALI/ARDS group.Results Compared with control group,the incident rate of ALI/ARDS in group B(25%)and group C(55%)were significantly higher(P

Objective To explore the methods and curative effect of metallic self-expanding stent in inoperable malignant gas-troduodenal obstruction. Methods The data of 15 cases with gastroduodenal obstruction including 9 cases of carcinoma of head of pancreas and 6 cases of carcinoma of stomach were analyzed retrospectively. The operative procedures of the stent implanted and the tors accepted more radiation dose because the manipulation was under the fluoroscopy in a short distance and with a full field of view. sions, the postoperative eating habit and the development turnover of disease. The main death reasons were tumor transfer and sys-tem exhaustion. Conclusion To pay close attention to the details and main points of operative procedure is the key point to implant stent successfully for malignant gastroduodenal obstruction. The determinative factor to influence the curative effect is the develop-ment turnover of tumor.

BACKGROUND: Polyethylene glycol-polyethyleneimine/ferroso-ferric oxide (PEG-PEI/Fe_3O_4) was selected as drug carders in tumor treatment, which can increase drug loading capacity and targeting capacity.OBJECTIVE: To test the toxicity of PEG-PEI/Fe_3O_4 nano-magnetic fluid in vitro and in vivo. METHODS: When the prepared PEG-PEI/Fe_3O_4 nano-magnetic fluid reached nano level, 7702 and HpG2 cell lines were filtrated and diluted in 5-20 multiple, and detected by in vitro MTT toxicity test assay; in vivo hemolysis test and micronucleus test was used to test the toxicity and biocompatibility.RESULTS AND CONCLUSION: MTT assay results indicated that the toxicity grade of PEG-PEI/Fe_3O_4 nano-magnetic fluid to 7702 cell line was 0-1, which was harmless to natural hepatic cells; however, PEG-PEI/Fe304 nano-magnetic fluid had slight bystander restraining effect to HpG_2 cell line. Maximum hemolysis rate of the matedel was 0.372%, which was far less than 5%. The micronucieus test result indicated that PEG-PEI/Fe_3O_4 nano-magnetic fluid had no teratogenicity or mutagenicity.

In order to investigate the anti-proliferative effects of triptolide (TP) on 4T1 mice breast cancer cell line in vitro and in mouse model, as well as the possible mechanisms, we detected the effect of TP on cell proliferation by MTT assay or Crystal Violet Staining in our research. Flowcytometry combined with FITC-Annexin V/PI staining were used for detecting TP induced 4T1 cell apoptosis. The protein expression of ERalpha, p-ERalpha, ERbeta, p-ERbeta, ERK, p-ERK, p38, p-p38, SAPK/JNK, and p-SAPK/JNK was tested by western blotting. We also compare TP with chemotherapy drug doxorubicin in 4T1 tumor bearing BLAB/c mice model, the Xenogen bioluminescence imaging, H&E, and IHC result indicated that TP exhibits an anticancer proliferation activity. As a result, TP in 100, 10, 1, 0.1 micromol x L(-1), all inhibited the proliferation of 4T1 cells by MTT assay and Crystal Violet Staining. TP which concentrations is 10, 1, 0.1 micromol x L(-1) could induce the apoptosis of 4T1 cells and reduce the cell proliferation. TP in 200 microg x kg(-1) could inhibit the tumor growth in vivo. The anticancer proliferation of TP was involved in its effect on reducing expression of ERalpha, p-ERalpha, ERbeta, and p-ERbeta, but nothing to do with the activation of MAPK signaling pathway.
Animals ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; Diterpenes ; pharmacology ; therapeutic use ; Down-Regulation ; drug effects ; Epoxy Compounds ; pharmacology ; therapeutic use ; Female ; Lung Neoplasms ; secondary ; Mammary Neoplasms, Experimental ; drug therapy ; metabolism ; pathology ; Mice ; Mice, Inbred BALB C ; Phenanthrenes ; pharmacology ; therapeutic use ; Phosphorylation ; drug effects ; Receptors, Estrogen ; metabolism ; Tumor Burden ; drug effects

OBJECTIVETo explore the effects of the exogenous and endogenous reactive nitrogen metabolites (RNM) as NK cell inhibitors on NK cell-mediated killing of K562 cells and the influence of Tiopronin (TIP), glutamylcysteinylglycine (GSH) and histamine dihydrochloride (DHT) as RNM scavengers on reversing the suppressing effect of RNM.

METHODSThe exogenous ONOO(-) was administered in the NK+K562 culture system, then the RNM scavengers were added in the NK+K562+ONOO(-) culture system, respectively. The concentrations of RNM, TNF-beta and IFN-gamma, K562 cell inhibition rate (KIR) and the percentage of living NK cells were examined. IL-2+PHA were used as monocyte (MO) activators in the culture system of MO+NK+K562. Then TIP, GSH and DHT were administered and the parameters of NK cell activity were analyzed.

RESULTSAfter exogenous ONOO(-) was administered in NK+K562 culture system, the percentage of living NK cells was decreased from (93.17 +/- 2.57)% to (71.87 +/- 1.02)% (P < 0.01) and KIR was decreased from (67.47 +/- 2.64)% to (43.44 +/- 2.87)% (P < 0.01). When TIP, GSH and DHT were administered into the systems, the percentage of living NK cells was increased to (91.13 +/- 3.67)% (P < 0.05), (88.03 +/- 1.46)% (P < 0.05), (73.60 +/- 2.76)% (P > 0.05), respectively; KIR was increased to (61.58 +/- 1.89)% (P < 0.05), (60.68 +/- 2.07)% (P < 0.05) and (45.26 +/- 3.31)% (P > 0.05), respectively. When IL-2/PHA were administered in the NK+K562+MO culture system, RNM products was increased from (82.10 +/- 6.60) micromom/L to (193.65 +/- 5.95) micromom/L(P < 0.01);KIR was decreased from (90.64 +/- 3.06)% to (61.29 +/- 2.22)% (P < 0.01). When the TIP, GSH and DHT were administered in the systems, RNM products were decreased to (91.32 +/- 6.81) micromom/L (P < 0.05), (84.66 +/- 5.99) micromom/L (P < 0.05) and (188.92 +/- 5.00) micromom/L (P > 0.05), respectively; KIR was increased to (84.31 +/- 4.56)%(P < 0.05), (81.65 +/- 3.09)% (P < 0.05) and (72.20 +/- 4.10)% (P < 0.05), respectively.

CONCLUSIONNK Cell-mediated killing of K562 cells can be suppressed by exogenous and endogenous RNM administration. Both of TIP and GSH can protect NK cells by scavenging RNM and enhance the antineoplasmic activity of NK cells.

Cells, Cultured ; Coculture Techniques ; Glutathione ; pharmacology ; Histamine ; pharmacology ; Humans ; Interferon-gamma ; metabolism ; Interleukin-2 ; immunology ; pharmacology ; K562 Cells ; Killer Cells, Natural ; cytology ; immunology ; metabolism ; Lymphotoxin-alpha ; metabolism ; Monocytes ; cytology ; Peroxynitrous Acid ; pharmacology ; Reactive Nitrogen Species ; antagonists & inhibitors ; metabolism ; Tiopronin ; pharmacology

OBJECTIVETo observe the effect of composite factors, like long-term high-salt & fat diet and alcohol abuse on blood viscosity and blood pressure in rats, and compare with a model induced by high molecular dextran, in order to build a chronic hyperviscosity aminal model which is similar to human hyperviscosity in clinic and lay a foundation for efficacy evaluation on traditional Chinese medicines.

METHODMale SD rats were randomly divided into the normal group, the high molecular dextran (HMD) group and the high salt & fat and alcohol (HSFA) group. The HMD group was given normal diet and water for 23 day and then 10% HMD through tail vein for 5 days. The HSFA group was fed with high salt and high fat diets every day and alcohol for 20 h x d(-1) for 13 weeks. After the modeling, whole blood viscosity and plasma viscosity were measured in the 5th, 8th and 11th week. Blood pressure was measured in the 5d, 7h, and 10th week. Red cell count (RBC) and hematocrit (HCT) were measured in the 11th week. PAgT, Fb, ET-1, NO, PGI, TXA2 contents of the normal group and the HSFA group were measured in the 13th week, and IECa21 content was measured with flow cytometry. Result: After the modeling, the HMD group was in good conditions with glossy hairs and active behaviors. The HSFA group was depressed with withered hairs and less activities. During the 5th-11th weeks, the HMD group and the HSFA group showed higher values in high and low shear whole blood viscosity (WBV) than the normal control group. The plasma viscosity (PV) of HMD rats was significantly increased only in the 5th week, and that of HSFA rats significantly increased in the 8"' and 11th week, particularly in the 11'h week. In the 111h week, the HSFA group showed significant increases in RBC and HCT. After the modeling, the blood pressure of HMD rats showed no significant changes, but the blood pressure of HSFA rats significantly increased during 7' and 101h weeks, particularly in the 10"' week. In the 13th week, PAgT, IECa2+, Fb, ET-1 of HSFA rats significantly increased, but with decreases in NO and PGI2.

CONCLUSIONLong-term high salt & fat and alcohol diets can cause abnormal blood viscosity in rats. WBV significantly increased since the 5th week in rats, and PV increased since the 8th week. The mechanism for increasing BV may be: (1) increases in RBC, HCT, and IECa2+, (2) PAgT increase, (3) Fb content increase, or (4) TXA2/PGI2, ET-1/NO imbalance. Although the modeling time with the method is longer than that with the HMD method, the model is more stable and moderate, and could lead to abnormal increases in WBV and PV; Whereas the HMD method only induced transient increase in plasma viscosity and abnormal increase in SBP. The model is more similar to traditional Chinese medicine syndromes and pathogenesis, with higher value for studies on efficacy of traditional Chinese medicines.

Alcoholism ; blood ; metabolism ; Animals ; Blood Pressure ; Blood Viscosity ; Diet, High-Fat ; adverse effects ; Disease Models, Animal ; Ethanol ; adverse effects ; metabolism ; Humans ; Male ; Rats ; Rats, Sprague-Dawley ; Sodium Chloride, Dietary ; adverse effects ; metabolism

Electromagnetic Fields ; Magnetic Fields ; Microwaves ; adverse effects ; Television

Animals ; Endoplasmic Reticulum ; metabolism ; Ischemia ; metabolism ; Male ; Molecular Chaperones ; metabolism ; Pressure Ulcer ; metabolism ; Rats ; Rats, Sprague-Dawley ; Subcutaneous Tissue ; blood supply

Objective To quantitatively detect circulating DNA levels in the plasma of patients withcervical lesion and to determine the value for diagnosis of cervical lesion and cervical cancer . Methods Preoperative blood samples were collected from 53 cases of low-grade lesions, 49 cases of high-grade lesions, 44 cases of cervical invasive cancer and 70 cases of healthy women. Plasma DNA was extracted by magnetic bead method (BILATEST DNA kit). The quantity of plasma DNA was determined by duplex real-time quantitative PCR. Results Median plasma DNA level of invasive cervical cancer patients was 61. 59 mg/L (32. 06 - 162. 16 mg/L) , which was significantly higher than that of healthy women [16. 35 mg/L(11. 98 -22.71 mg/L), P < 0.01]. Among invasive cervical cancer patients, median plasma DNA level of squamous carcinoma patients was slightly higher than that of adenocarcinoma (50. 43 versus 47. 31 mg/L,P>0. 05). Median plasma DNA level of stage I patients was lower than that of stage Ⅱ- Ⅲ patients (46. 02 versus 71. 35 mg/L, P <0. 05). Conclusion Quantitatively detecting plasma circulating DNA may be with some application prospect in the diagnosis of cervical diseases.