Objective To explore the regulatory effect of miR-6838-5p on DDR1 gene expression and proliferation of breast cancer cells.Methods The expression levels of miR-6838-5p in normal breast epithelial cells and breast cancer cells were detected using qRT-PCR,and the potential target genes of miR-6838-5p was predicted using TargetscanV 8.0.Double luciferase reporter gene experiment was performed to verify the binding between miR-6838-5p and DDR1.Breast cancer MCF-7 cells were transfected via liposome,miR-6838-5p mimic,miR-6838-5p inhibitor,DDR1 siRNA,DDR1-overexpresisng vector,or both miR-6838-5p mimic and DDR1-overexpressing vector,and the changes in cell proliferation were examined with CCK-8 and EdU assays;Western blotting was used to detect the expression of DDR1.The mediating role of DDR1 in miR-6838-5p overexpression-induced inhibition of MCF-7 cell proliferation was verified in a nude mouse model bearing MCF-7 cell xenografts.Results The expression of miR-6838-5p was significantly lower in breast cancer cells than in normal breast epithelial cells.In MCF-7 cells,miR-6838-5p overexpression induced significant inhibition of cell proliferation.Dual luciferase reporter gene experiment demonstrated a binding relationship between miR-6838-5p and DDR1(P<0.01).Western blotting showed that miR-6838-5p overexpression significantly lowered DDR1 expression in MCF-7 cells,and DDR1 overexpression promoted proliferation of the cells;co-transfection of the cells with DDR1-overexpressing vector significantly attenuated the inhibitory effect of miR-6838-5p mimic on cell proliferation.In the tumor-bearing nude mice,the xenografts overexpressing miR-6838-5p showed a significantly smaller volum with obviously the expression of DDR1.Conclusion Overexpression of miR-6838-5p inhibits breast cancer cell proliferation by regulating DDR1 expression.

AIM:To investigate the molecular mechanism of a novel bromobenzene substituted trifluoromethylbenzo Cyclopentanone WW02 inhib-iting the viability and proliferation of human lung cancer A549 and H1299 cells.METHODS:The ef-fect of different concentrations of WW02(6.25,12.5,25,50 μg/mL)on cell viability and prolifera-tion of A549 and H1299 were measured using CCK-8 and EdU methods.After 24 hours of stimulation of A549 and H1299 cells with different concentra-tions of WW02,the changes in Akt and mTOR phos-phorylation levels under different concentrations of WW02 were detected through Western blot as-say.Macromolecular docking was carried out be-tween WW02,AKT and mTOR through MOE Dock.RESULTS:After treating A549 and H1299 cells with WW02 using different concentrations(6.25,12.5,25,50 μg/mL),the activity of A549 and H1299 cells decreased in a concentration dependent manner compared with the DMSO control group(P<0.05).The proliferation of cells showed a concentration dependent decrease compared to the DMSO con-trol group(P<0.05).Compared with the DMSO con-trol group,after 24 hours of WW02 stimulation,the phosphorylation levels of Akt and mTOR in A549 cells decreased under the concentration of WW02(12.5,25,50 μg/mL,P<0.05).Compared with the DMSO control group,the phosphorylation levels of Akt and mTOR in H1299 cells decreased af-ter 24 hours of WW02 stimulation(25,50 μg/mL,P<0.05).Based on pattern analysis,it was found that WW02 had a strong binding with Akt and mTOR,with the highest score of-8.3 kcal/mol for WW02 and mTOR,while the highest score for WW02 and Akt was-7.3 kcal/mol.CONCLUSION:WW02 inhib-its the activity and proliferation of lung cancer A549 and H1299 cells,and its mechanism of action may be achieved by directly binding to Akt and mTOR proteins to inhibit Akt and mTOR phosphory-lation.

Objective To explore the regulatory effect of miR-6838-5p on DDR1 gene expression and proliferation of breast cancer cells.Methods The expression levels of miR-6838-5p in normal breast epithelial cells and breast cancer cells were detected using qRT-PCR,and the potential target genes of miR-6838-5p was predicted using TargetscanV 8.0.Double luciferase reporter gene experiment was performed to verify the binding between miR-6838-5p and DDR1.Breast cancer MCF-7 cells were transfected via liposome,miR-6838-5p mimic,miR-6838-5p inhibitor,DDR1 siRNA,DDR1-overexpresisng vector,or both miR-6838-5p mimic and DDR1-overexpressing vector,and the changes in cell proliferation were examined with CCK-8 and EdU assays;Western blotting was used to detect the expression of DDR1.The mediating role of DDR1 in miR-6838-5p overexpression-induced inhibition of MCF-7 cell proliferation was verified in a nude mouse model bearing MCF-7 cell xenografts.Results The expression of miR-6838-5p was significantly lower in breast cancer cells than in normal breast epithelial cells.In MCF-7 cells,miR-6838-5p overexpression induced significant inhibition of cell proliferation.Dual luciferase reporter gene experiment demonstrated a binding relationship between miR-6838-5p and DDR1(P<0.01).Western blotting showed that miR-6838-5p overexpression significantly lowered DDR1 expression in MCF-7 cells,and DDR1 overexpression promoted proliferation of the cells;co-transfection of the cells with DDR1-overexpressing vector significantly attenuated the inhibitory effect of miR-6838-5p mimic on cell proliferation.In the tumor-bearing nude mice,the xenografts overexpressing miR-6838-5p showed a significantly smaller volum with obviously the expression of DDR1.Conclusion Overexpression of miR-6838-5p inhibits breast cancer cell proliferation by regulating DDR1 expression.

OBJECTIVE: To observe the effects of acupuncture combined with bloodletting on the expression of inflammatory factors in serum, the morphology of sensitized skin tissue and the mast cell degranulation in urticaria rats, and to explore the potential mechanism of this therapy for urticaria. METHODS: Among 42 SD rats of SPF grade, 6 rats were randomly collected for the preparation of sensitized antiserum; and the rest 36 rats were randomized into a blank group, a model group, a positive drug group, an acupuncture group, a bloodletting group and a combined treatment group (acupuncture + bloodletting), 6 rats in each one. The rat model of urticaria was established by passive cutaneous anaphylaxis. In the positive drug group, loratadine (1 mg•kg^-1) by gavage was administered once a day. In the acupuncture group, 1 h after gavage with 0.9% NaCl (1 mL), acupuncture was delivered at "Baihui" (GV 20), "Zhongwan" (CV 12), and bilateral "Quchi" (LI 11) and "Xuehai" (SP 10) for 15 min, once daily . In the bloodletting group, 1 h after gavage with 0.9% NaCl (1 mL), bloodletting was operated at "Dazhui" (GV 14) and bilateral "Geshu" (BL 17), around 0.1 mL of bleeding volume at each point, once daily. In the combined treatment group, 1 h after gavage with 0.9% NaCl (1 mL), the interventions as the acupuncture group and the bloodletting group were adopted, once daily. All the interventions started on day 6 of modeling, lasting 2 weeks. After intervention completion, antigenic stimulation was performed in the rats of each group. Using ELISA, the levels of serum immunoglobulin E (IgE), tryptase (TPS), interleukin-4 (IL-4), interleukin-5 (IL-5), tumor necrosis factor-α (TNF-α) were detected. The diameter of the blue spots of the sensitized skin on the back was measured with ruler in each rat. The morphology of sensitized skin tissue was observed using HE staining, and the degranulation of mast cells was observed using Toluidine blue staining. RESULTS: Compared with the blank group, in the model group, the levels of serum IgE, TPS, IL-4, IL-5 and TNF-α increased (P<0.01), the diameter of blue spot on the sensitized part of the rat back was larger (P<0.01), the degranulation rate of mast cells was elevated (P<0.01), and there were obvious inflammatory cell infiltration and edema in the dermis of sensitized skin tissue on the rat back. Compared with the model group, the serum levels of IgE, TPS, IL-4, IL-5 and TNF-α were reduced in the positive drug group, the acupuncture group, the bloodletting group and the combined treatment group (P<0.01, P<0.05); skin blue spot diameter was smaller in the positive drug group and the combined treatment group (P<0.05); the degranulation rate of mast cells decreased in the positive drug group, the acupuncture group, the bloodletting group and the combined treatment group (P<0.01); and the dermal edema, inflammatory infiltration were attenuated in the positive drug group, the acupuncture group, the bloodletting group and the combined treatment group. Compared with the acupuncture group and the bloodletting group, the serum levels of IgE, TPS, IL-4, IL-5 and TNF-α, as well as the degranulation rate of mast cells in the sensitized tissue were lower in the positive drug group and the combined treatment group (P<0.05). CONCLUSION Acupuncture combined with bloodletting effectively suppress mast cell degranulation in the sensitized skin tissue on the back of urticaria rats, and ameliorate the histopathological morphology. Its effect mechanism may be related to inhibiting the differentiation and proliferation of helper T cells 2 and regulating the humoral immune response.
Animals ; Rats ; Mast Cells/immunology* ; Acupuncture Therapy ; Rats, Sprague-Dawley ; Male ; Cell Degranulation ; Humans ; Bloodletting ; Urticaria/immunology* ; Female ; Acupuncture Points ; Tumor Necrosis Factor-alpha/blood* ; Interleukin-4/blood* ; Interleukin-5/blood* ; Combined Modality Therapy ; Immunoglobulin E/blood* ; Disease Models, Animal

Objective To evaluate the influencing factors of compliance and the compliance changes before and after intervention on chronic hepatitis B patients who oral nucleos(t)ide analogue (NA), by using the continuing care service items provided by WeChat platform. Methods A total of 67 patients with chronic hepatitis B were selected from January 2013 to June 2015. WeChat health care teams and WeChat groups were established to conduct continuing care intervention for 6 months by providing services, such as authoritative scientific diagnosis and treatment information, online consulting service, expert outpatient appointment, etc. Results The score of Morisky Medication Adherence Scale (MMAS-8) in patients of different culture levels of illiteracy (4.75 ± 2.20) points, primary school or junior high school graduate (5.73±1.87) points,high school or vocational school graduate (6.31±1.45) points,university diploma and above (7.24 ± 1.23) points were significant difference (χ2=22.113, P < 0.01). The score in patients previous used of nucleoside drugs (5.76±1.89) points was lower than that in patients not previous used of nucleoside drugs (6.62±1.60) points, and there was a significant difference between the two groups(Z=-2.636, P<0.01). Significant difference (χ2=10.844, P<0.01) was shown between groups of≥3 kinds drugs combined with the score 7.02 ± 1.08, 1-2 kinds drugs combined with the score 7.02 ± 1.08 and no combined medication with score 6.04±1.95. Different medical insurance types of free of charge (7.60±0.48) points,residents medical insurance (6.54 ± 1.52) points,new rural cooperative (5.17 ± 2.34) points and self-paying medical service (5.93±1.91) points showed significant difference(χ2=15.630, P<0.01). The scores in other diseases showed no significant difference (Z=0.401, P > 0.05) both in combination or non-combination. Compared to 6.45 ± 1.69 of pre-intervention, the total compliance score 6.86 ± 1.53 showed significant increased after intervention (Z=-2.551, P < 0.05), and the number of people with high compliance after the intervention increased while the number of people with middle and low compliance after the intervention decreased, and significant differencewas observed in two groups (χ2=9.521, P<0.01). Conclusions After the intervention of continuing care based on WeChat platform, the patients' compliance with nucleoside antiviral drugs were significant improvement than those of pre-intervention.