Objective To study the effet of ET-1 and ECE on endothelial proliferation of autografted vein.Methods An animal model of the autogenous vein graft was established in 80 Wistar rats.The expression of ECE and EF-1 gene was determined by RT-PCR and immunohistological method to test the mRNA and protein expresion level respectively.Results PCNA positive smooth muscle cells appeared 6 hours after transplantation,increased with time,and reached a peak at 1 to 2 weeks.After 2 weeks,PCNA positive SMC in the tunica media began to decline and recovered to the 6 hour level at 8 weeks after the operation.mRNA of ECE increased with the time after the operation,reached a peak after 1-2 weeks,declined afterwards,and became stable at around 8 weeks.Expression change of ET1 was similar to the change of ECE,reached the peak after 1-2 weeks and was stable after 8 weeks(r=0.975).Conclusions The time and pattern of change of the pathologic process of intimal hyperplasia are in agreement with the expression of ET-1 and ECE,and suggests that ECE is involved in the process of intimal hyperplasia of vein graft.

Aquaporin1 (AQP1) is a member of a family of specific channel proteins which could mediate the trans-biofilm transportation of small molecules such as water.Recent studies have shown that AQP1 is highly expressed in cancer tissues.It also has an effect on the proliferation and migration of cancer cells, angiogenesis in cancer and so on.AQP1 is expected to be a marker of screening, diagnosis, treatment and prognosis at tumor early stage.

microRNAs (miRNAs) are short non-protein-coding RNAs,which play important roles in the cell proliferation,differentiation,apoptosis,as well as activation of oncogenic and antioncogenic signals.Researches show that the abnormal expressions of miRNAs are closely related to the tumorigenesis,histological type,diagnosis,treatment and prognosis of lung cancer.So miRNAs may be the most potential and promising therapeutic targets for lung cancer.

OBJECTIVE: To determine if greater efficacy could be achieved with the intranasal antihistamine azelastine and the intranasal corticosteroid fluticasone propionate used concurrently in the treatment of nasal obstruction of persistent non-allergic rhinitis. METHOD: A total of 162 persistent non-allergic rhinitis cases with moderate to severe nasal obstruction were randomized to treatment with the following: the combination therapy or nasal corticosteroids monotherapy. Efficacy was assessed by change from baseline in nasal obstruction score at week 2 and week 6 visits. The perceptions of global treatment satisfaction(convenience, side effects, cost and effectiveness) in both groups were analyzed. RESULT: In both groups, the nasal obstruction score assessment descended significantly at week 2 and week 6 visits versus at baseline (all P < 0.01). At week 2 and week 6 visits, the nasal obstruction score in the combination therapy groups were significantly improved than that in nasal corticosteroids monotherapy groups (all P < 0.01). The perceptions of global treatment satisfaction in the combination therapy groups were significantly better (P < 0.05). CONCLUSION Azelastine nasal spray and intranasal corticosteroid in combination may provide a substantial therapeutic benefit for patients with persistent non-allergic rhinitis, especially nasal obstruction. The combination therapy was well tolerated and safety.
Administration, Intranasal ; Adrenal Cortex Hormones ; therapeutic use ; Drug Therapy, Combination ; Histamine H1 Antagonists ; therapeutic use ; Humans ; Nasal Obstruction ; Phthalazines ; therapeutic use ; Rhinitis ; drug therapy

Objectives 99mTc-methoxyisobutylisonitrile (MIBI) SPECT imaging technology was used to observe the condition of tumor cell in-taking imaging agent in the C57BL/6J mice Lewis lung cancer model before and after using Ginsenoside Rg3 (short for Rg3).We aimd so as to discuss the feasibility of using this method to evaluate tumor multidrug resistance (MDR) status.Methods Mice Lewis lung cancer models were randomly divided into Rg3 group and the control group.After applying Rg3,semi-quantitative analysis was made on 99mTc-MIBI SPECT imaging and region of interest (ROI) to observe the multidrug resistance state of tumor and then the results were compared with the detection results of flow cytometry.Results The tumor intake ratio (T/N) difference between the control group and the Rg3 group in imaging,imaging before applying Rg3 and imaging after applying Rg3 were separately 15,60 and 120 min.The differences were statistical significant (P ＜ 0.01).The eliminate indexes (WR) of the control group and Rg3 group were positively related to P-gp protein expression positive cells detected by flow cytometry (P ＜ 0.05).Conclusions 99mTc-MIBI imaging is negatively related to P-glycoprotein (P-gp) expression in mice Lewis lung cancer cells,which can clearly show the multidrug resistance state of tumors and dynamically monitor the effect of Rg3 on multidrug resistance reversion of mice Lewis lung cancer.

Objective To study the clinical and prognostic value of CK19 mRNA-positive circulating tumor cells in early breast cancer patients. Methods We analyzed the peripheral blood in 50 patients with early breast cancer after surgery and before the initiation of any adjuvant treatment for the presence of CK19 mRNA-positive circulating tumor cells using a nest reverse polymerase chain reaction assay. All patients were followed up. Results CK19 mRNA-positive cells were detected in 40.0 %(20/50) of patients with early breast cancer, 12.5 %(3/24) of patients with breast benign lesions, but 5 %(1/20) in healthy individuals (P =0.017,P =0.004); 11 to 20 of them relapsed during the follow-up period (P =0.002). There was no significant association between the detection of CK19 mRNA-positive cell and the patients' menstrual status, tumor stage, tumor size, etc (P ＞0.05). Detection of peripheral-blood CK19 mRNA-positive cells was associated with reduced median relapse-free interval in early breast cancer patients (P =0.007). Conclusion CK19 mRNA is one of the molecular markers for the detection of circulating tumor cells in early breast cancer. Detection of peripheral blood CK19 mRNA-positive cells might be an important predictive value as a marker of relapse in early breast cancer patients.

Objective To explore the influence of Rapamycin (RAPA) on apoptosis of acute lymphoblastic leukemia EU-4 cells induced by Methotrexate (MTX).Methods EU-4 cells were pretreated 0.5 h with 5 pμg/L,10 μg/L,25 μg/L,50 μg/L and 100 μg/L RAPA.RAPA untreated group was set up as control group.The cells were collected and the expression of LC-3 was assayed by using Western blot.The apoptosis of EU-4 cells was detected by using flow cytometry (FCM).The effects of different concentrations of RAPA pretreatment on autophagy and apoptosis of EU-4 cells were observed,and the pretreatment concentration of RAPA was determined.EU-4 cells were divided into RAPA pretreatment group and untreated group.The cells were treated with 0.05 μmol/L,0.10 pμmol/L and 0.20 μmol/L MTX for 24 h,respectively.The apoptotic rate was detected by using FCM.Western blot was performed to test the expression of LC-3 protein,while the absorbance(A562) was measured by using microplate reader,and the protein concentration in the sample was calculated according to the standard curve.Results The apoptotic rates of EU-4 cells in the control group and the different concentrations of PAPA pretreatment group showed that the control group,5 μg/L,10 μg/L,25 μg/L,50 μg/L and 100 μg/L RAPA were (7.51 ±0.32)％,(7.33 ±0.41)％,(7.71 ± 0.51) ％,(7.63 ± 0.38) ％,(7.80 ± 0.43) ％ and (16.66 ± 0.87) ％,respectively.The apoptotic rates of EU-4 cells in the 5 μg/L,10 μg/L,25 μg/L and 50 μg/L PAPA pretreatment group had no significant difference from those in the control group(t =0.427,-0.417,-0.297,-3.561,all P ＞ 0.05).The apoptotic rate of EU-4 cells in 100 μg/L PAPA pretreatment group was significantly higher than that in control group,and the difference was significant (t =-28.815,P ＜ 0.01).Combined with the results of Western blot and FCM,50 μg/L was used as the pretreatment of PAPA.The apoptosis rates of EU-4 cells in PAPA pretreatment group were (50.23 ± 2.11) ％,(66.88 ± 2.89) ％ and (73.11 ± 2.67) ％ after treated with 0.05 μmol/L,0.10 μmol/L and 0.20 μmol/L MTX for 24 h,respectively.The apoptotic rates of EU-4 cells in PAPA unpretreatment group were (22.53 ± 1.67) ％,(42.82 ± 2.26) ％ and (53.22 ± 1.93)％ after treated with 0.05 μmol/L,0.10 μmol/L and 0.20 μmol/L MTX for 24 h,respectively.The apoptotic rates of EU-4 cells in RAPA pretreatment group were significantly higher than those in untreated group in the same concentration of MTX treatment after 24 h,and the differences were significant(t =12.693,66.148,14.429;all P ＜ 0.01).Conclusion With RAPA pretreatment,relative low dose MTX can induce a great deal of acute lymphoblastic leukemia EU-4 cells apoptosis.

AIM: To study the survival, transfer and distribution of bone marrow CD34+/CD45+ cells from transgenic GFP mouse after transplanted into the completed transversional spinal cord rat model. METHODS: The bone marrow cells isolated from transgenic GFP mice were cultured in vitro. The cultured cells were identified by anti-CD34 and anti-CD45 monoclonal antibodies, and were transferred into the end of transection spinal cord. Paraformaldehyde was infused into the left ventricle of the rat model at the 24 h, 48 h, 1 week, 2 weeks, 4 weeks and 8 weeks after cell transplantation. Through sank and frozen, the spinal cord was sectioned at 10 ?m thickness. The green fluorescence positive cells were observed under the fluorescence microscope. CD34+/CD45+ cells were identified by immunohistochemistry staining. RESULTS: Green fluorescence positive cells were found at the head and the end of the completed transection part of spinal cord. Most of the green fluorescence positive cells were distributed in the gray substance of spinal cord. CD34+/CD45+ cells were found by immunohistochemistry staining. CONCLUSION: CD34+/CD45+ cells survived in spinal cord of SD rat, and migrated to the head of the transection part. The distance of migration was extended by the time.

BACKGROUND AND OBJECTIVEThe mouse lung cancer orthotopic model includes spontaneous lung cancer model and endotracheal transplanted model, and etc. The spontaneous lung cancer needs longer time and does not ensure the rate of the generation of the tumor; as for endotracheal transplanted model, the position and size of the tumor are instable. In this study, the 3LL cell line was orthotopically transplanted into the lung of the C57BL/6 mice, compare to the heterotopic model, to discuss their stability and transfer-characteristics. And this study was also to optimize the method of establishing lung cancer orthotopic animal model.

METHODSDifferent quantity of 3LL cells were inoculated into the left oxter of C57BL/6 mice to establish the heterotopic model; or suspended with Matrigel then inoculated into the left lung of C57BL/6 mice to establish orthotopic model. The survival-time of the mice was examined. The tissue was collected for the subsequent histology assay after euthanizing the mice. Microvessels density (MVD) was observed and counted by immunohistological chemistry. CD44v was detected by flow cytometry.

RESULTSTTumor-form-rate of the heterotopic group were 100%, 66.7%, 16.7%, respectively, and had no macroscopic transfer. Tumor-form-rate of the orthotopic group were 100%, 100%, 83.3%, respectively, and had widespread transfer in contralateral chest and the lung. The median survival time of the orthotopic group (38, 35, 23 days) were less than the heterotopic group (82, 72, 50 days). MVD of the orthotopic group (120.2 +/- 9.73) was higher than the heterotopic group (92.6 +/- 7.12). The expression of CD44v of orthotopic (26.46 +/- 1.56)% was higher than the heterotopic group (23.13 +/- 1.02)%.

CONCLUSIONThe lung cancer orthotopic model which established by 3LL cells transplanted into the lung of the mice is simple, dependable, repeatable and has stronger transfer characteristics than the heterotopic model.

Animals ; Carcinoma, Lewis Lung ; Cell Line, Tumor ; Disease Models, Animal ; Female ; Lung Neoplasms ; Male ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Random Allocation

OBJECTIVETo explore the advantages, feasibility and limitations of hepatic arterial infusion under temporary hepatic circulation occlusion.

METHODSTwelve rabbits were randomly divided into two groups: hepatic artery infusion group (HAI group) and hepatic artery infusion under temporary hepatic circulation occlusion group (HAI-THCO). Microcatheters were separately inserted into the proper hepatic artery and right hepatic vein. For the HAI group, 5-Fu (10 mg/ml and 100 mg/kg) was infused into the common hepatic artery with a high pressure injector for 10 minutes. For the HAI-THCO group, the common hepatic artery and hepatic portal vein were temporarily occluded for 15 minutes using artery clamp when 5-Fu was being infused. For the two groups, at 2, 5, 10, 15, 20 and 30 min after the start of infusion, blood samples of the hepatic flow were collected from the right hepatic vein and of the systemic blood flow from the inferior vena cava, 1 ml at each time point. The blood drug concentration of these blood samples was determined by high performance liquid chromatography (HPLC).

RESULTSExcept that at 20 and 30 min after infusion, in the HAI group, the blood drug concentration of hepatic circulation was significantly higher than that of systemic circulation (P < 0.05). But in the HAI-THCO group, the blood drug concentration of hepatic circulation was significantly higher than that of systemic circulation at all the time points (P < 0.05). The hepatic circulation blood drug level of the HAI-THCO group was always significantly higher than that of the HAI group (P < 0.05), but the systemic circulation blood drug concentration of the HAI-THCO group was always lower (P < 0.05). The hepatic circulation maximum concentration (Cmax) of blood drug concentration of the HAI-THCO and HAI groups was (23.057±3.270) µg/ml and (4.408±1.092) µg/ml, respectively, and the Cmax of HAI-THCO group was significantly higher (P < 0.001), being 5.23 times of that of HAI group. The systemic circulation Cmax of the two groups was (1.456±0.217) µg/ml and (2.335±0.669) µg/ml, respectively, and the Cmax of HAI group was 1.60 times higher than that of the HAI-THCO group (P = 0.022). The hepatic circulation AUC of HAI-THCO and HAI groups was (368.927±52.416) µg·min·ml(-1) and (65.630±14.928) µg·min·ml(-1), respectively. The AUC of HAI-THCO group was 5.62 times higher than that of the HAI group (P < 0.001). The systemic circulation AUC of the two groups was (27.193±3.948) µg·min·ml(-1) and (45.301±12.275) µg·min·ml(-1), respectively. The AUC of HAI group was 1.67 times higher than that of the HAI-THCO group (P = 0.014).

CONCLUSIONSHAI-THCO is a simple and effective regional hepatic infusion chemotherapy technique. It can be performed through occluding the common hepatic artery and the hepatic portal vein by balloon catheter. HAI-THCO can not only increase the blood drug concentration in the hepatic circulation, but also decrease the blood drug concentration in the systemic circulation, therefore, distinctly lowering the systemic toxicity.

Animals ; Coronary Occlusion ; Fluorouracil ; Hemodynamics ; Hepatic Artery ; Hepatic Veins ; Infusions, Intra-Arterial ; methods ; Liver ; Liver Circulation ; Portal Vein ; Rabbits