Compared to the conventional two-dimensional (2D) culture,in vitro three-dimensional cell culture (3DCC) can mimic the microenvironment more exactly.Compared to in vivo animal models,3DCC is less time-consuming and can be real-time recorded.Thus,3DCC fills the gap between 2D culture and animal models,which is used for oncologic research quickly.In 3DCC,the characteristics of cancer cells,such as morphology,the capability of invasion and migration and the resistance to chemo-radiotherapy,are quite different from those in 2D culture.The development of 3DCC technology provides a platform for oncology,organization engineering and gene or drug screening.

Objective To explore the mechanism of Suzijiangqi decoction to passage recombination of asthma model by studying the effect of Suzijiangqi decoction on passage recombination and the relation between it and the content of TNF and ET of asthma model. Methods Asthma model was established by OVA atomized and inhaled, the effect of Suzijiangqi decoction on the lung histomorphometric and the content of TNF and ET in serum or plasma/BALF of asthma model were observed. Results Suzijiangqi decoction obviously improved the lung histomorphometric and significantly inhibited the content of TNF and ET. Conclusion Suzijiangqi decoction can obviously control passage recombination. It is relate to inhibiting the content of TNF and ET in serum or plasma/BALF of asthma model.

Objective To investigate the roles of cystatin C (CysC) and serum creatinine (Scr) in acute renal injury of patients with shock. Method A total of 71 patients with shock, 42 male and 29 female, were enrolled from February 2006 to June 2007. Patients with kidney disease or renal insufficiency were excluded. All of patients were assigned to 4 groups as per the duration of shock. The blood samples were taken from patients for measurements of CysC and Scr during the periods of 1 hr,2 hr,and 4 hr of shock and 72 hr and 7 days after correction of shock. The corrected GFR (cGFR) and decreased GFR (dGFR) were calculated. The levels of Scr and dGFR could be used to classify the acute renal injury into stages according to the Acute Kidney Injury Diagnosis Criteria. The positive detection rates of different methods were compared. The levels of CysC, Scr and cGFR were statistically analyzed. Data were studied by using Pearson's correlation analysis, Results The elevation of CysC appeared sooner than that of Scr in all shock patients. Contrarily, the high level of CysC lowered to normal level much slower than that of SCR after correction of shock. The CysC increased 1 hour after shock. The GFR was negatively correlated with CysC and Scr, especially in the early stage of shock. Conclusions The renal dysfunction appears in the early stage of shock. The CysC assayed is more sensitive in the stage 1 of renal injury than Scr.

The ultrastructural characteristics of the mouse mammary adenocarcinoma cell line (MA782/5S-8102) have been studied by transmission electron microscope. The results show that the cells were irregular in the shape, with abundant microvilli on cell surface. The nucleus was abnormal in shape with the nuclear membrane sunk or swollen, the nucleus/cytoplasmic ratio (N/C) was high. There were many free ribosomes, abundant mitochondrias, dilated rough endoplasmic reticulum in the cytoplasm. Golgi apparatus, lysosomes and lipid droplets were present. Moreover, many cytoplasmic type-A particles and a few immatural type-B particles at the surface of the cell were observed.

Objective To investigate the effets of three mood stabilizers on ouabain?induced ERK1/2 phosphorylation in astrocytes. Methods As?trocytes were treated with different agents and divided into different groups accordingly,namely,the control group with saline,the group with oua?bain,the group with mood stabilizers(lithium carbonate,carbamazepine,sodium valproate)and the group with ouabain+mood stabilizers. The phosphorylation of ERK1/2 in each group was analyzed by Western blot. Results Compared with saline and mood stabilizer groups,the phosphoryla?tion of ERK1/2 was increased in the ouabain group,with statistical significance(P<0.05). There was no significant difference in ERK1/2 phosphoryla?tion between the group with ouabain+mood stabilizers and the control or mood stabilizer group. Conclusion The three kinds of mood stabilizers can inhibit ouabain?induced ERK1/2 phosphorylation in astrocytes.

Objective To evaluate the safety and efficacy of transcatheter arterial embolization (TAE) of massive polycystic liver disease (PLD).Methods A total of 21 patients with symptomatic PLD were enrolled.The patients consisted of seventeen women and four men (aged 36-64 years,mean age,49 years).Transcatheter superselective embolization was performed with the mixture of N-butylcyanoacrylate (NBCA) and iodized oil.All patients underwent contrast enhanced computed tomography (CT) of the liver before TAE and at every 3 months for the first half year after TAE,and at 6-monthly intervals thereafter.Laboratory data,including routine blood tests and liver enzymes,were collected.T test was used for statistics.Results All procedures were successful without serious complications.There was no obvious improvement during the first three months.At follow-up of 6-12 months,symptoms notably improved in 18 of 21 patients,and these patients experienced further relief of the symptoms in the follow-up period.TAE failed to benefit in 3 patients,but there were no complaints of worsening of the symptoms.At follow-up CT,the total liver volume and total intra-hepatic cyst volume decreased significantly (t =6.75,7.73,P ＜0.01)compared with pre-TAE in 18 patients at 12 months after TAE.The total liver volume decreased from (8270 ± 3016) cm3 to (6120 ± 2680) cm3 and the total intra-hepatic cyst volume decreased from (7120 ±3070) cm3 to (4560±2488) cm3.Mild elevation of the liver enzymes returned to the normal range within 1 month in all patients.Conclusions It is suggested that transcatheter super selective embolization with the mixture of NBCA and iodized oil is a safe and effective treatment for PLD patients.This technique is a supplemental option for traditional therapy.

Objective To investigate the effect of histone deacetylase inhibitor LBH589 on proliferation,apoptosis and drug resistance of chemoresistant acute myeloid leukemia cells HL60/ADM.Methods HL60/ADM cells were treated with LBH589.Proliferation,apoptosis and adriamycin IC50 were evaluated by MTT assay and AnnexinV-FITC/PI stain.The change in MRP1 expression and intercellular adriamycin accumulatiom were analyzed by flow cytometry.Results Effective proliferative inhibition and apoptotic induction in HL60/ADM cells were observed after treatment with 10-80 nmol/L LBH589 with maximal effect detected after treatment with 70 nmol/L LBH589 for 60 hours.However,inhibition ratio remain unchanged with the further increase of drug dose and incubation time (P ＞ 0.05).Downregulation of MRP1 [(93.90±4.20) ％ vs (76.19±6.53) ％],upregulation of adriamycin accumulation [(8.53±0.68) ％ vs (25.67±1.34) ％] and decrease in adriamycin IC50 [(6.833±0.319) μg/ml vs (1.382±0.104) μg/ml] were induced by the treatment with 20 nmol/L LBH589 (P ＜ 0.01),whose reversal fold was 4.9.The expression of acetylated histone 3 after treatment with LBH589 was higher than that before treatment (P ＜ 0.01).However,relative p-Akt levels after treatment for 24 h and 48 h were 1.07±0.09 and 0.59±0.01,respectively,which were lower than that before treatment (2.03±0.12) (P ＜ 0.01).Meanwhile,expression levels of p53 were 0.57±0.04 and 1.31±0.09,respectively,which were higher than that before treatment (0.21 ±0.02) (P ＜ 0.01).Conclusion Treatment with LBH589 has the capability of inhibiting proliferation and inducing apoptosis,as well as increasing intercellular adriamycin accumulation and sensitivity through downregulation of MRP1 expression and inhibition of PI3K-Akt signaling pathway in HL60/ADM cells.

Objective:To investigate the cytotoxic effects of CTL cell induced by DCs loaded with exosomes derived from hepatoma Huh-7 cells(T-exo).Methods: Exosomes derived from hepatoma Huh-7 cells were isolated and purified by combination of ultrafiltration centrifugation and sucrose density gradient centrifugation.Morphology of exosomes was observed under transmission electron microscopy and the expression of CD 9,CD63,HSP70 and AFP was detected by Western blot.DCs were induced with peripheral blood monocytes isolated from healthy donors.Flow cytometry was used to analysis surface markers of the DCs loaded with T-exo.WST-1 light absorption measurement was adopted to evaluate the T cell proliferation ability.Annexin-V/PI Flow cytometry were respectively used to examined cytotoxicity against the tumor cells.Results:Exosomes isolated and extracted from culture supernatant of Huh-7 cells presented as circular or elliptical vesicle with bilayer membrane , unequal in size , and with diameter of 50 to 100 nm.Western blot showed that the T-exo expressed CD9,CD63,HSP70 and AFP molecules.DCs loaded with T-exo caused significantly higher T cell pro-liferation and cytotoxic effect against AFP positive Huh-7 cells as compare to gainst AFP negative SMMC 7721 cells and un-loaded control group ( P<0.05 ).Conclusion: T-exosome loaded-DC can promote proliferation and induce significant cytotoxic effect of CTL against Huh-7 cells.

BACKGROUND:The main component of cartilage, type Ⅱ col agen gene expression in chondrocyte is positively correlated with SOX9 concentration in a dose-dependent manner.
OBJECTIVE:To observe the variation of SOX9 and type Ⅱ col agen mRNA content at different periods in the differentiation process (osteogenic, chondrogenic, adipogenic induction) of mesenchymal stem cel s, and to explore the correlation of SOX9 expression and type Ⅱ col agen.
METHODS:Bone marrow mesenchymal stem cel s were isolated from 4-week-old Kunming mice, and cultured in vitro to passage 3. The cel phenotype was identified with flow cytometry. Cel s were divided into three groups and subjected to three kinds of induction conditions favorable for adipogenic, chondrogenic and osteogenic differentiation, and each group was observed at three time points. In addition, the non-induced cel s were used as a control group. The total RNA of cel s was extracted at 3, 7, 14 days after induction, and SOX9 and type Ⅱ col agen mRNA was quantified with reverse transcription-polymerase chain reaction. The induced cel s were stained by immunofluorescence to observe the differentiation and perform statistical analysis.
RESULTS AND CONCLUSION:Passage 3 bone marrow mesenchymal stem cel s grew wel , and cel phenotype was confirmed as stem cel s by flow cytometry. The staining results showed that, the cel s differentiated into chondrocytes, adipocytes and osteoblasts. The SOX9 mRNA levels in the induced cel s were the highest in chondrogenic differentiation group, then in osteogenic differentiation group, and the lowest in adipogenic differentiation group. Type Ⅱ col agen mRNA levels in the induced cel s were the highest in chondrogenic differentiation group, then in adipogenic differentiation group, and the lowest in osteogenic differentiation group. SOX9 expression in chondrogenic differentiation group increased at 3 and 7 days, and then decreased at 14 days. While type Ⅱ col agen expression increased at 3, 7, 14 days. SOX9 mRNA levels increased as the osteogenic differentiation, while type Ⅱ col agen expression gradual y decreased. There was no significant difference in the SOX9 mRNA expression between adipogenic differentiation group and control group (P>0.05), while type Ⅱ col agen expression was not regularly changed. Experimental findings suggest that, critical effect of SOX9 in chondrogenic differentiation is better than that in osteogenic and adipogenic differentiation. SOX9 is associated with type Ⅱcol agen, which may alter along with the SOX9 in the early chondrogenic differentiation;SOX9 may play a fine-tuning role in the process of chondrogenic and osteogenic differentiation.

Objective:To explore the role of NKG2D ligand MHC-I related molecule A (MICA) in chemotherapy combined with NK cell immunotherapy in patients with advanced esophageal cancer after surgery. Methods:A total of 90 patients with esophageal cancer from Fujian Provincial Tumor Hospital were divided into three groups after surgery:40 patients of chemotherapy alone, 25 patients of chemotherapy combined with NK cell therapy with negative expression of MICA (MICA-group), and 25 patients of chemotherapy combined with NK cells therapy with positive expression of MICA (MICA+group). The efficacy was then compared. Results:Compared with the chemotherapy alone and MICA-groups, the positive rates of CD3+, CD4+T cells, NK cells, and the CD4+/CD8+ratio in peripheral blood from MICA+group were higher than those before treatment (64.2%± 6.4%vs. 51.3%± 5.6%, 39.8%± 8.2%vs. 29.5%± 3.2%, 25.3%± 2.1%vs. 16.4%±4.3%, 1.4%± 0.5%vs. 1.1%± 0.7%;P<0.05). Meanwhile, the levels of T-reg cells were lower than those before treatment (6.3%± 4.5%vs. 17.3%± 2.4%, P<0.05). No significant difference was observed between the disease control rate and response rate. Chemotherapy-induced neutropenia and peripheral neurotoxicity symptoms were significantly improved, and time to progression (TTP) and overall survival (OS) were significantly prolonged (P<0.05). No statistically significant difference was observed between the chemotherapy alone group and MICA-group (P>0.05). Conclusion:Treatment with chemotherapy and autologous NK cells on patients with advanced esophageal carcinoma and MICA positive expression can be safely transfused with only minor side effects and can effectively improve a patient's immune system, quality of life, and survival.